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Biomedical subjects

H Ohtake

Publications and source records attributed to H Ohtake.

At least 109 records · Page 6Linked to original sources

Surgical treatment of patients with Wolff-Parkinson-White syndrome and associated Ebstein's anomaly.

Ebstein"s anomaly is the most common congenital heart disease associated with the Wolf-Parkinson-White syndrome. Between November 1973 and March 1993, we surgically treated 42 patients with Wolff-Parkinson-White syndrome and Ebstein's anomaly. The patient's ages ranged from 5 months to 59 years (mean 35.3 +/- 14.0 years). There were a total of 52 accessory pathways, 48 of which were located in the right (65%) or posteroseptal (29%) area. A left-sided accessory pathway was seen in only two patients (3.8%). Division of all right-sided accessory pathways was done during normothermic cardiopulmonary bypass with the heart beating; cryocoagulation was applied together with scalpel dissection of the atrioventricular groove. Division of the left-sided accessory pathways was done with the use cold potassium cardioplegic arrest. Thirty-five of these patients underwent tricuspid valve operation for Ebstein's anomaly and 11 of them underwent tricuspid valve replacement with a bioprosthesis. All 52 accessory pathways were successfully divided, although two patients required reoperation because of tachycardia caused by accessory pathways in different positions. Three hospital deaths (7.1%) occurred. There were no late deaths during the follow-up period (mean 94.3 +/- 52.4 months), but two patients required repeat tricuspid operation because of progression of the tricuspid regurgitation. Because no repeat operations were required during long-term follow-up patients who underwent valve repair or valve replacement, correction should be indicated in some patients.

Adult↗

Different growth control of the two human thyroid cell lines of adenomatous goiter and papillary carcinoma.

To study the growth control of human thyroid cells in different stages of differentiation, we established two human thyroid cell lines of adenomatous goiter and papillary carcinoma. A 59-year-old female patient with adenomatous goiter was operated in September 1991, and a 27-year-old female patient with papillary carcinoma in May 1990. The thyroid cell lines were established by successive passage without cellular or genetic manipulations such as fusing other cell lines or oncogenic viral infection. These cell lines, human adenomatous goiter cells (hAG) and human papillary thyroid carcinoma cells (hPTC), exhibited a flattened polygonal shape and proliferated as a monolayer in cell culture. The doubling time of the hAG cells was 60 h in Ham's F12 medium supplemented with 10% fetal bovine serum, and that of the hPTC cells, 18 h in the same medium. Both cell lines expressed mRNA for TSH receptor and secreted cAMP into the medium during incubation with thyrotropin (TSH) at concentrations as low as 0.01 mU/mL. The effects of activators of protein kinase A (PKA), protein kinase C (PKC), tyrosine kinase (TK), and estradiol (E2) on proliferation of the hAG cells and the hPTC cells were assessed by measuring cellular DNA content in 24-well plates with diaminobenzoic acid. TSH stimulated proliferation of the hAG cells, but it inhibited proliferation of the hPTC cells. Since TSH activates two signaling pathways, the adenyl cyclase-PKA system and phospholipase C-PKC system, we tested effects of dibutylyl cAMP (dBC) and phorbol myristate 13-acetate (PMA), separately. dBC stimulated proliferation of the hAG cells, but it inhibited that of the hPTC cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenoma↗

Effect of perinatal hypothyroidism on expression of cytochrome c oxidase subunit I gene, which is cloned by differential plaque screening from the cerebellum of newborn rat.

Early development of the central nervous system is influenced by several hormones including thyroid hormone. This study was designed to clone the gene whose expression is changed in association with perinatal hypothyroidism in the rat cerebellum. Rats were sacrificed at 15 day-old postnatal age (P15) and their cerebella were removed. Poly (A)+ RNA was extracted to construct a cDNA library using lambda gt 10 cloning vector. Differential plaque screening was then performed using 32P-labeled antisense cDNA synthesized from poly (A)+ RNA of the methimazole-treated (hypothyroid) P15 rat cerebellum (hypothyroid probe), and of the euthyroid P15 rat cerebellum (euthyroid probe). The clones, which hybridized strongly to the euthyroid probe and weakly or not at all to the hypothyroid probe, were isolated. Sequence analysis of these clones revealed that all isolated clones encode cytochrome c oxidase subunit I (COX I), which is located in the mitochondrial DNA. The decrease in COX I gene expression was not seen in the animals, which received methimazole treatment and daily replacement of thyroid hormone. In situ hybridization detection showed not only overall decrease in COX I gene expression but also change in distribution of hybridization signal in the cerebellar cortex of hypothyroid rat. Such change was not observed in the T4-replaced animals. Based on the evidence that thyroid hormone greatly influences brain development, the results of the present study indicate that the terminal enzyme of mitochondrial respiratory chain, COX I is one of the important target molecules regulated by thyroid hormone in the newborn rat cerebellum.

Animals↗

Molecular cloning and characterization of a chemotactic transducer gene in Pseudomonas aeruginosa.

A Pseudomonas aeruginosa mutant, defective in taxis toward L-serine but responsive to peptone, was selected by the swarm plate method after N-methyl-N'-nitrosoguanidine mutagenesis. The mutant, designated PCT1, was fully motile but failed to show chemotactic responses to glycine, L-serine, L-threonine, and L-valine. PCT1 also showed weaker responses to some other commonly occurring L-amino acids than did the wild-type strain PAO1. A chemotactic transducer gene, denoted pctA (Pseudomonas chemotactic transducer A), was cloned by phenotypic complementation of PCT1. Nucleotide sequence analysis showed that the pctA gene encodes a putative polypeptide of 629 amino acids with a calculated mass of 68,042. A hydropathy plot of the predicted polypeptide suggested that PctA may be an integral membrane protein with two potential membrane-spanning regions. The C-terminal domain of PctA showed high homology with the enteric methyl-accepting chemotaxis proteins (MCPs). The most significant amino acid sequence similarity was found in the region of MCPs referred to as the highly conserved domain. The pctA gene was inactivated by insertion of a kanamycin resistance gene cassette into the wild-type gene, resulting in the same observed deficiency in taxis toward L-amino acids as PCT1. In vivo methyl labeling experiments with L-[methyl-3H]methionine showed that this knockout mutant lacked an MCP with a molecular weight of approximately 68,000.

Amino Acid Sequence↗

Isolation and characterization of chemotaxis mutants and genes of Pseudomonas aeruginosa.

Two chemotaxis-defective mutants of Pseudomonas aeruginosa, designated PC1 and PC2, were selected by the swarm plate method after N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis. These mutants were fully motile but incapable of swarming, suggesting that they had a defect in the intracellular signalling pathway. Computer-assisted capillary assays confirmed that they failed to show behavioral responses to chemical stimuli, including peptone, methyl thiocyanate, and phosphate. Two chemotaxis genes were cloned by phenotypic complementation of PC1 and PC2. From nucleotide sequence analysis, one gene was found to encode a putative polypeptide that was homologous to the enteric CheZ protein, while the other gene was cheY, which had been previously reported (M. N. Starnbach and S. Lory, Mol. Microbiol. 6:459-469, 1992). Deletion and complementation analysis showed that PC1 was a cheY mutant, whereas PC2 had a double mutation in the cheY and cheZ genes. A chromosomal cheZ mutant, constructed by inserting a kanamycin resistance gene cassette into the wild-type gene, changed its swimming direction much more frequently than did wild-type strain PAO1. In contrast, cheY mutants were found to rarely reverse their swimming directions.

Amino Acid Sequence↗

Expression of prolactin gene in human decidua during pregnancy studied by in situ hybridization histochemistry.

The prolactin (PRL) gene is known to be expressed not only in the anterior pituitary but also in the decidualized human endometrium. This study was designed to detect the site of synthesis of PRL during pregnancy by in situ hybridization histochemistry. Decidual and trophoblast tissues from early pregnancy were obtained from patients undergoing therapeutic abortion at 8-10 weeks of gestation. Term placentae were obtained from patients with uncomplicated deliveries at 38-40 weeks. Sections of these tissues were hybridized with 35S-labeled RNA probe complementary to human PRL mRNA. Specific hybridization signals were distributed over the decidual cells in early and term pregnancy. In the decidua capsularis of early pregnancy, labeled cells were concentrated close to the amniotic cavity, although decidual cells were distributed evenly. In the decidua parietalis, almost all decidual cells were labeled, but no specific labeling was seen in the endometrial glands or capillary endothelium. In the decidua basalis, greater signals were always detected over the decidual cells in early pregnancy than in term pregnancy, when sections, which were hybridized with the same probe and exposed simultaneously, were compared. No specific hybridization was detected in the trophoblast cells. These results not only confirm that PRL is specifically synthesized in the decidual cells but also indicate that there are regional and periodical differences in PRL gene expression in the decidual cells during pregnancy.

Blotting, Northern↗

[A construction of a PACS and reporting system linked with the hospital information system in Gunma University Hospital].

A PACS (Picture Archiving and Communication System) and reporting system were introduced into Gunma University Hospital. These systems were linked with Hospital Information System (HIS), enabling us to refer to images, such as those of CR, CT, MRI and scintigraphy, at wards and conference rooms within a reasonable times. Bone scintigraphy and a report on it are illustrated as an example. A standardized protocol for the interface between PACS and the imaging format must be established without delay for the progress of PACS.

Computer Communication Networks↗

Development of a new intraoperative radiofrequency ablation technique using a needle electrode.

We studied a new intraoperative technique for radiofrequency ablation using a needle electrode. The ventricles of 12 mongrel dogs were ablated. The needle electrode was inserted to a depth of 3 mm through the epicardium. Macroscopically, tissue defect was found only at the needle insertion site. The degenerated area had a well-demarcated bullet shape with microscopical coagulation necrosis. This characteristic shape allows the ablation of deeper areas of the myocardium by deeper insertion of the ablation needle. Compared with conventional epicardial surface ablation, this new method could be used to achieve deeper coagulation. For any given energy level and conductance, the ratio between the volume and the depth of the lesion with this new method is smaller than that of lesions made by the conventional radiofrequency ablation method. With this new method, the energy spreads vertically, a characteristic suitable for deep ablation. This may result from the shape of the needle electrode itself. For ablation of deep targets, this new method is superior to the conventional approach because less myocardium is coagulated and the target can be ablated with greater efficiency, requiring the destruction of less tissue. In conclusion, the needle electrode procedure can achieve a narrower ablation for deeper targets than can the conventional technique.

Animals↗

Surgical treatment of Wolff-Parkinson-White syndrome in infants and children.

Electrophysiologic features and surgical results were examined in 55 pediatric patients who underwent surgical accessory pathway division for Wolff-Parkinson-White syndrome. There were 31 male and 24 female patients ranging in age from 4 months to 15 years (mean age, 9.8 +/- 4.2 years; 25 patients were less than 10 years old; 4 patients were less than 12 months). Eleven of these patients had associated congenital heart disease and underwent concomitant surgical procedures to treat those conditions. Preoperative effective refractory period of antegrade accessory pathways, the right atrium, atrioventricular node, and cycle length during reentrant tachycardia were shorter in pediatric patients than in adult patients. Antegrade accessory pathways showed right predominance more frequently in the pediatric group than in the adult group. Surgical techniques included an endocardial approach (an epicardial approach was used in 1 patient) and concomitant operation for combined heart disease. The early mortality rate was 3.6%, whereas no late deaths occurred during the follow-up period of 96.8 +/- 54.9 months (maximum follow-up, 205 months). The absolute cure rate was 92%. There were no significant differences in early and late mortality between pediatric and adult patients. Surgical treatment of the Wolff-Parkinson-White syndrome in pediatric patients is as safe and effective as in adults. Considering the potential complications from prolonged fluoroscopic exposure during catheter ablation, surgical division of accessory pathways in children is a promising modality for the treatment of Wolff-Parkinson-White syndrome in selected cases.

Adolescent↗

Surgical treatment of arrhythmogenic right ventricular dysplasia: long-term outcome.

Eight male patients ranging from 15 to 51 years old (mean age, 36.3 years) underwent surgical treatment of ventricular tachycardia (VT) associated with arrhythmogenic right ventricular dysplasia. One patient had an associated left ventricular aneurysm. The earliest activation site was detected for 15 lesions, and delayed potentials were recorded during sinus rhythm in all patients. On the basis of the epicardial mapping, the origins of the VT foci in the right ventricle were resected. Cryoablation on the surrounding myocardium was performed. There were no surgical deaths or postoperative fatal complications. During long-term follow-up, there has been no recurrence of VT and no congestive heart failure in the 6 patients without left ventricular involvement. The 2 patients with LV involvement died late of either congestive heart failure or development of VT originating from the left ventricle. In conclusion, a surgical approach consisting of myocardial excision and cryocoagulation offers a curative treatment of VT associated with arrhythmogenic right ventricular dysplasia and yields excellent long-term results when the VT origin is well identified in the right ventricle.

Adolescent↗

In situ hybridization detection of TSH beta subunit gene expression in the serum-free primary culture of the adult rat pituitary.

This study was designed to construct the primary culture system to detect the change in TSH beta subunit (TSH beta) gene expression in individual cells. Adult, male Wistar rats were sacrificed by transcardial perfusion of 0.25% trypsin solution under pentobarbital anesthesia (50 mg/kg body weight). Their anterior pituitaries were removed, dispersed and cultured for 1, 2, 3, or 6 days with or without 1 nM triiodothyronine (T3) under the serum-free condition. In some cultures, TRH was added to a final concentration of 1 microM on 6, 12 or 24 h before fixation. Then the culture media were removed to measure TSH concentration. Cells were fixed with paraformaldehyde and hybridized with 35S-labeled RNA probe complementary to TSH beta mRNA. Emulsion autoradiography was subsequently performed. T3 treatment markedly suppressed relative cellular levels of TSH beta mRNA on 2, 3 and 6 days after the onset of culture (day 2, 3 and 6) and suppressed TSH secretion on day 3 and 6. TRH treatment increased TSH beta mRNA on 12 and 24 h after the treatment on day 2 and 3 but did not increase TSH beta mRNA on day 6. TSH concentration in the culture medium was increased by TRH treatment on 6, 12 and 24 h after the treatment on day 2, on 12 h and 24 h on day 3, and 24 h on day 6. On day 2 and 3, although T3 treatment suppressed basal level of TSH beta mRNA, TRH-induced increase in TSH beta mRNA was not suppressed by T3 treatment. These results show that the thyroid hormone and TRH regulate TSH beta gene expression independently. Our culture system may provide a useful model to examine the action of individual substances on a specific subpopulation of the anterior pituitary cells.

Animals↗

Myocardial coagulation by intraoperative Nd:YAG laser ablation and its dependence on blood perfusion.

To investigate whether the efficacy of intraoperative laser ablation therapy is affected by myocardial blood perfusion, we irradiated 11 canine hearts through the epicardium with a Nd:YAG laser (10, 20, and 30 J/mm2) using air as the medium. Compared coagulated myocardial depth, width, and volume obtained in the red beating heart was compared with those in the white nonbeating heart infused with 0 degrees C saline (cardioplegic model) via the coronary artery. Histologically, the macro- and microscopical findings were very similar. At each level of energy, the width and volume of red myocardium coagulated were significantly larger than those of white myocardium (P < 0.01). At 30 J/mm2, the depth of coagulation of red myocardium was significantly larger than that of white myocardium (P < 0.01). The coagulated volume of the white myocardium was about 60% of that of the red myocardium. Nd:YAG laser energy was absorbed by blood (red color = hemoglobin), and more energy was transferred in the higher temperature myocardium. During intraoperative Nd:YAG laser ablation, the presence of blood perfusion in cardiac tissue is thought to be an important factor affecting safe irradiation.

Animals↗

Production and release of polyphosphate by a genetically engineered strain of Escherichia coli.

A recombinant strain of Escherichia coli MV1184, which contains plasmid-borne genes encoding the phosphate-specific transport (Pst) system and polyphosphate (polyP) kinase, accumulated high levels of Pi and released polyP into the medium. PolyP could be separated from the culture supernatant by DEAE-Toyopearl 650M chromatography and identified by high-resolution 31P nuclear magnetic resonance spectroscopy. Once E. coli recombinants accumulated high levels of polyP, they released polyP concomitantly with Pi uptake. PolyP release did not accompany the decrease in the cell density, indicating that it is not simply a result of cell lysis. PolyP release ceased when Pi became depleted in the medium and resumed upon addition of Pi to the medium. When Pi uptake was inhibited by 0.1 mM carbonyl cyanide m-chlorophenylhydrazone (CCCP), no polyP release was observed. Furthermore, neither Pi uptake nor polyP release occurred when cells were incubated at 4 degrees C. These findings suggest that the occurrence of polyP release is a possible mechanism that limits a further increase in the cellular polyP concentration in E. coli recombinants. High-resolution 31P nuclear magnetic resonance spectroscopy also detected a surface pool of polyP in intact cells of the E. coli recombinant. The polyP resonance increased when cells were treated with EDTA and broadened upon the addition of a shift reagent, praseodymium. Although the mechanism of surface polyP accumulation is unclear, surface polyP seems to serve as the source for polyP release.

Biological Transport, Active↗

Cloning and characterization of a Pseudomonas aeruginosa gene involved in the negative regulation of phosphate taxis.

Pseudomonas aeruginosa PAO1 exhibited a positive chemotactic response to P(i). The chemotactic response was induced by P(i) limitation. An alkaline phosphatase (AP) constitutive mutant showed a chemotactic response to P(i), regardless of whether the cells were starved for P(i). Sequence analysis and complementation studies showed that the P. aeruginosa phoU gene was involved both in the regulation of AP expression and in the induction of P(i) taxis. However, unlike AP expression, P(i) taxis was not regulated by the phoB gene product.

Alkaline Phosphatase↗

[Expression of prolactin gene in human decidua].

UNLABELLED: This study was designed to detect the localization of prolactin gene expressing cells in the human utero-placental unit during pregnancy. METHODS: Human endometrium was obtained at hysterectomy from normally cycling women who underwent surgery for myoma uteri. Decidual and trophoblast tissues from early pregnancy were obtained by curettage from patients undergoing therapeutic abortion at 8-10 weeks of gestation. Term placenta was obtained from patients with uncomplicated deliveries at 38-40 weeks. Total RNAs of these tissues were extracted to perform Northern blot hybridization (NBH) with the radiolabelled human pituitary prolactin cDNA probe. Frozen sections of these tissues were cut and processed for in situ hybridization (ISH) with the radiolabelled RNA probe complementary for human prolactin mRNA. RESULTS: 1. By NBH, approximately a 1.3 kilobase (kb) size band was detected in the total RNA obtained from late secretory endometrium (day 26), decidua of early pregnancy and placenta of term pregnancy, and the decidua had the most significant signal. No hybridization signal was detected in the chorionic villi in early pregnancy. 2. By ISH, a significant hybridization signal was detected in the cytoplasm of the decidual cells in early and term pregnancy. No hybridization signal was detected in the endometrial grandsand trophoblast cells. CONCLUSION: These results indicate that prolactin is synthesized in the decidual cells in the human uterus during pregnancy.

Animals↗