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Biomedical subjects

S Takehara

Publications and source records attributed to S Takehara.

At least 19 recordsLinked to original sources

Mice containing a human chromosome 21 model behavioral impairment and cardiac anomalies of Down's syndrome.

Trisomy 21 (Ts21) is the most common live-born human aneuploidy; it results in a constellation of features known as Down's syndrome (DS). Ts21 is the most frequent cause of congenital heart defects and the leading genetic cause of mental retardation. To investigate the gene dosage effects of an extra copy of human chromosome 21 (Chr 21) on various phenotypes, we used microcell-mediated chromosome transfer to create embryonic stem (ES) cells containing Chr 21. ES cell lines retaining Chr 21 as an independent chromosome were used to produce chimeric mice with a substantial contribution from Chr 21-containing cells. Fluorescence in situ hybridization and PCR-based DNA analysis revealed that Chr 21 was substationally intact but had sustained a small deletion. The freely segregating Chr 21 was lost during development in some tissues, resulting in a panel of chimeric mice with various mosaicism as regards retention of the Chr 21. These chimeric mice showed a high correlation between retention of Chr 21 in the brain and impairment in learning or emotional behavior by open-field, contextual fear conditioning and forced swim tests. Hypoplastic thymus and cardiac defects, i.e. double outlet right ventricle and riding aorta, were observed in a considerable number of chimeric mouse fetuses with a high contribution of Chr 21. These chimeric mice mimic a wide variety of phenotypic traits of DS, revealing the utility of mice containing Chr 21 as unique models for DS and for the identification of genes responsible for DS.

Animals↗

Construction of 700 human/mouse A9 monochromosomal hybrids and analysis of imprinted genes on human chromosome 6.

As an in vitro assay system for the identification of human imprinted genes, a library of human/mouse A9 monochromosomal hybrids containing a single, intact bsr-tagged human chromosome of known parental origin, derived from normal human fibroblasts, has been previously generated by microcell-mediated chromosome transfer (MMCT). To supplement this assay system, we constructed additional 700 A9 monochromosomal hybrids, using a pSTneo or pPGKneo selection marker. To validate the A9 hybrids, we screened them with chromosome-specific polymorphic markers, and identified the hybrids containing either human chromosome 6, 7, 14, 18, or 21 of known parental origin. Matching paternal and maternal chromosome pairs of A9 hybrids were identified for chromosomes 6, 7, 14, and 18. The paternal-specific expression of ZAC (zinc finger protein, which regulates apoptosis and cell cycle arrest) and HYMAI (hydatidiform mole-associated and imprinted transcript), and the maternal-specific methylation of a CpG island within an imprinted domain on human chromosome 6q24, were maintained in A9 hybrids. For an example, we profiled the expression of expressed sequence tags (ESTs) and the methylation of CpG islands in the 300-kb imprinted domain around 6q24, which may be associated with cancers and transient neonatal diabetes mellitus (TNDM). Thus, the 700 A9 hybrids should be useful for various aspects of imprinting studies.

Animals↗

Specific impairment of cardiogenesis in mouse ES cells containing a human chromosome 21.

Down syndrome (DS) leads to cardiac defects which are common and significant in babies with DS. We recently generated chimeric mice carrying a human chromosome (hChr) 21. The contribution ratio of embryonic stem (ES) cells containing a hChr 21 was specifically low in the heart, compared to other organs, and cardiovascular malformations were observed, suggesting that an additional copy of hChr 21 also disrupts the normal development of heart in mice. Here we describe that the presence of hChr 21 in ES cells delays the appearance of beating cardiomyocyte during differentiation, whereas differentiation into other cell types is not disrupted. Furthermore, the defect in cardiogenesis was restored following the deletion of a specific region of hChr 21. Therefore, we conclude that the imbalance of specific gene(s) on hChr 21 may lead to the disturbance of cardiogenesis and that this may be a useful system to model and investigate the cardiac defects of human DS.

Animals↗

Stability of transferred human chromosome fragments in cultured cells and in mice.

Chromosome fragments represent feasible gene delivery vectors with the use of microcell-mediated chromosome transfer. To test a prerequisite for a gene delivery vector, we examined the stability of human chromosome fragments (hCFs) in cultured cells and in trans-chromosomic (Tc) mice. Fragments of human chromosomes 2 (hCF(2-W23)), 11 (hCF-11) and 14 (hCF(SC20)) tagged with neo were introduced into the TT2F mouse ES cells, and retention of the hCFs was examined by FISH during long-term culture without selection. In contrast to the gradual loss of hCF(2-W23) and hCF-11, hCF(SC20) remained stable over 70 population doublings in the ES cells. The hCF(SC20) was also stable in cultured human tumor cells and chicken DT40 cells. We have previously generated chimeric mice using the ES cells harboring the hCF(2-W23) or hCF(SC20), followed by production of Tc mice. Although both the hCF(2-W23) and hCF(SC20) persisted in cells of Tc mice as an additional chromosome and were transmitted to offspring, the hCF(SC20) was more stable than the hCF(2-W23) in F1 and F2 mice. The present study shows that the stability of hCFs in Tc mice differs with tissue types and with genetic background used for successive breedings. Thus, the hCF(SC20), which was relatively stable in both mouse and human cells, may be a promising candidate for development as a gene delivery vector.

Animals↗

Telomere maintenance in telomerase-deficient mouse embryonic stem cells: characterization of an amplified telomeric DNA.

Telomere dynamics, chromosomal instability, and cellular viability were studied in serial passages of mouse embryonic stem (ES) cells in which the telomerase RNA (mTER) gene was deleted. These cells lack detectable telomerase activity, and their growth rate was reduced after more than 300 divisions and almost zero after 450 cell divisions. After this growth crisis, survivor cells with a rapid growth rate did emerge. Such survivors were found to maintain functional telomeres in a telomerase-independent fashion. Although telomerase-independent telomere maintenance has been reported for some immortalized mammalian cells, its molecular mechanism has not been elucidated. Characterization of the telomeric structures in one of the survivor mTER(-/-) cell lines showed amplification of the same tandem arrays of telomeric and nontelomeric sequences at most of the chromosome ends. This evidence implicates cis/trans amplification as one mechanism for the telomerase-independent maintenance of telomeres in mammalian cells.

Animals↗

[Prolactinoma in a child showing high MIB-1 labeling index: a case report].

We report a very rare case of a prolactin secreting pituitary tumor (prolactinoma) which occurred in a 12-year-old boy. The tumor showed an extremely high MIB-1 index. The clinical implication in the postoperative management of childhood prolactinoma is discussed. The patient showed right third nerve palsy, and MRI revealed a pituitary tumor invading the right cavernous sinus. Preoperative hormonal evaluation showed a very high prolactin level (2800 ng/ml). The patient underwent transsphenoidal surgery, and the third nerve palsy disappeared just after the procedure. MIB-1 index obtained by using immunostaining was 18.9%. Postoperative prolactin level remained high (2200 ng/ml), and the patient was treated with 10 mg/day of bromocriptine. Prolactinomas in children with high MIB-1 index show resistance to treatment with bromocriptine. In the postoperative management of a childhood prolactinoma, it should be considered how to control sufficiently high serum prolactin level to expect sexual development while preserving other normal residual pituitary functions. If control with bromocriptine, fails radiation treatment should be adopted with careful observation of the increase in height and the progress of sexual development of the patient.

Bromocriptine↗

[Endovascular treatment of ruptured intracranial aneurysms using platinum coils in patients over 80 years of age].

Endovascular treatment of intracranial aneurysms using plantinum coils was performed in 10 patients over 80 years of age. The patients ranged in age from 81 to 96 years. Preoperative Hunt and Kosnik grading revealed one patient in grade I, 4 in grade II, 3 in grade III, and 2 in grade IV. The aneurysms were located in the internal carotid artery in 6 patients, in the anterior communicating artery in 1, and in the middle cerebral artery in 3 respectively. Intra-aneurysmal occlusion was accomplished in 8 patients with total or subtotal occlusion in 6 and partial occlusion in 2. In one patient with a peripheral middle cerebral artery aneurysm, the parent artery was occluded just proximal to the aneurysm. In the remaining one patient in grade III, an aneurysm associated with wide neck could not be occluded because of coil protrusion into the parent artery. In 9 patients who were successfully treated, all five patients in grade I or II and one in grade III resulted in good outcome, but the other three patients in grade III and IV showed poor outcome. No evidence of recurrent hemorrhage was noted during the follow-up period ranging from 2 months to 4 years. Endovascular coil embolization is a useful therapeutic alternative for treating ruptured intracranial aneurysms in patients over 80 years of age, especially those in Grade I or II.

Aged↗

Concanavalin A-immobilized polystyrene nanospheres capture HIV-1 virions and gp120: potential approach towards prevention of viral transmission.

To establish an effective tool for the prevention of HIV-1 transmission, lectin-immobilized polystyrene nanospheres were synthesized and examined for their HIV-1 capture activity. When concanavalin A (Con A) was immobilized on the surface of polystyrene nanospheres (400 nm in diameter) with poly(methacrylic acid) branches and incubated with HIV-1 suspension at room temperature for 60 min, the nanospheres (Con A-NSs) achieved a >3.3 log and a 2.2 log reduction of viral infectivity in HIV-1 (IIIB strain) suspension at a concentration of 2 and 0.5 mg/ml, respectively. Meanwhile, Con A-free nanospheres, which were not immobilized with Con A, achieved only a 0.29 log reduction at 0.5 mg/ ml. Con A-NSs (2 mg/ml) could also reduce the gp120 level of III(B) and HE strains to <7.1% and 5.5% of each control, respectively. The combination of Con A-NS treatment followed by filtration with a microporous membrane efficiently removed virion-free gp120 as well as infectious viral particles from HIV-1 suspension. Electron microscopic examination demonstrated that HIV-1 virions were trapped on the surface of Con A-NSs. Thus, Con A-NSs can capture HIV-1 virions and gp120 with high affinity, and may have potential as an effective tool for the prevention of HIV-1 transmission.

Concanavalin A↗

Examination of prognostic factors after resection of pulmonary metastasis of osteosarcoma by multivariate analysis.

Prognostic factors for the case of pulmonary metastasis of osteosarcoma was examined using the multivariate analysis. 1) The 3-year survival rate after operation of primary focus in 23 cases was 34.8% and the 5-year survival rate was 30.4%. The 3-year survival rate after the first operation of pulmonary metastasis was 30.4% and the 5-year survival rate was 12.2%. No deaths attributed to operation were found. 2) Gender, age, histological type, solitary or multiple pulmonary metastasis, laterality, number of pulmonary metastatic tumors, TDT and DFI were examined, and the laterality, number of pulmonary metastatic tumors and DFI were considered to be useful prognostic factors. DFI among them was most closely associated with postoperative survival days. 3) A significant difference in the number of excised nodules was found between the DFI less than 90 days group and the DFI not less than 90 days group, and the number in the former group was significantly larger. 4) It was suggested that postoperative observation and close examination for pulmonary metastasis at about 3 months intervals after of primary focus are important, and also, positive or negative pulmonary metastasis as the time of examination, particularly, the number metastatic nodules of less than 5 or not less than 5, are reflected in the prognosis. 5) Expected survival time after the first operation pulmonary metastasis appeared possible to estimate as 3.0 x DFI days.

Adolescent↗

A case of pulmonary metastases of synovial sarcoma surviving for a long period after active surgery.

The patient was a 24-year-old woman. She underwent marginal excision of synovial sarcoma originating in the right femoral soft tissue at the age of 18, followed by local radiation therapy and chemotherapy. At the age of 20, abnormal shadow was indicated in bilateral lungs on plain roentgenography of the chest, and she underwent open-chest surgery for excision of bilateral pulmonary metastatic tumors. She was subjected to bilateral thoracotony repeatedly thereafter for a total of 5 times, and all of the metastatic foci detected in each operation were resected. Consequently, she has been surviving for a long period of about 7 years. The therapeutic outcome of tumor of bone soft tissue is greatly affected by its pulmonary metastasis. Although the therapeutic outcome has recently been improved by large dose chemotherapy and active excision of metastatic foci in the lung, survival time in the case of synovial sarcoma, giant cell sarcoma and Ewing's sarcoma is not very long yet. In the present study, we observed a patient who had lung metastasis of synovial sarcoma of which prognosis is generally considered poor has been surviving for a long time of 7 years owing to active excision of pulmonary metastatic foci. The relation of the present prognostic factors to those in the past were compared.

Adult↗

[Basilar artery aneurysm associated with agenesis of unilateral internal carotid artery: two case reports].

Two patients presented with a basilar artery aneurysm associated with agenesis of the right internal carotid artery. In both cases, the right middle cerebral artery was supplied by the basilar artery via the enlarged right posterior communicating artery, and the right anterior cerebral artery was supplied by the left internal carotid artery via the anterior communicating artery. The first patient was a 65-year-old woman who suffered from a subarachnoid hemorrhage due to rupture of a basilar bifurcation aneurysm, which was demonstrated as a "de novo" aneurysm 13 years after successful clipping of the anterior communicating artery aneurysm. Delayed surgery was planned, but she died due to recurrent hemorrhage. The second patient was a 67-year-old woman who had had a history of progressive left sided weakness over the previous few years. Conventional angiography and 3 dimensional CT angiography showed a large aneurysm of the basilar artery trunk with a wide neck. Endovascular embolization was performed with mechanically detachable coils, and the aneurysm was incompletely occluded. The patient regained complete strength in the left arm after the treatment, but the follow-up angiography at 5 months disclosed recanalization of the treated aneurysm, associated with shift of the packed coils. Hemodynamic stress resulting from unique collateral circulation with agenesis of the internal carotid artery may cause a predisposition to "de novo" aneurysm formation or recanalization of an occluded aneurysm with coils.

Aged↗

[Case of benign metastaxsizing leiomyoma of the lungs].

A 51-year-old woman with bilateral multiple pulmonary metastases of uterine leiomyoma was reported. The patient had undergone a hysterectomy for uterine myoma in 1976. In 1981, routine chest roentgenograms showed three nodular lesions in the right lung with well-defined margins. The patient received no treatment because she was asymptomatic. In January 1993, she consulted a gynecologist because of acute lower abdominal pain and an abdominal mass, and bilateral pulmonary tumors were found. The retroperitoneal solid tumor was extirpated in the first operation, and histological findings showed that it was a benign leiomyoma. The pulmonary tumors, extirpated in the second operation, were benign leiomyoma, also. The clinical course and histological findings suggested that this was a case of benign metastasizing leiomyoma. This disease is rare, 21 case have been reported in Japan.

Female↗

[Prognostic factors of resected pulmonary metastases of osteogenic sarcoma].

From December 1979 to December 1994, 22 patients with osteogenic sarcoma underwent a total of 34 thoracotomies for resection of pulmonary metastases in our department. We did a retrospective study to identify prognostic factors. The tumor doubling time (TDT) calculated for 21 patients before the first (if multiple) resection was not correlated with days of survival after resection of the primary tumor, nor was it correlated with days of survival after the first (if multiple) resection of the pulmonary metastatic tumors. Of the nine patients who underwent at least two thoracotomies, the TDT calculated from X-ray films taken before the second (or later) thoracotomy was significantly shorter than the TDT calculated before the preceding thoracotomy (p < 0.05). The disease-free interval (DFI) after resection of the primary tumor was correlated with the days of survival after that resection (p < 0.001) and also with survival after resection of the pulmonary metastatic tumors (p < 0.01). This DFI of patients who survived at least 2 years from the date of the resection of the primary tumor was significantly longer than in patients who survived less than 2 years from that date (p < 0.05). The most significant prognostic factor was this DFI. The cumulative survival rate was 42.4% at three years and 33.9% at five years, with an operative mortality of 0%.

Adolescent↗

[Multiple dural arteriovenous fistulas following extensive sinus thrombosis: a case report].

A 43-year-old man suffered generalized convulsions following severe headache. Initial angiography showed a dural arteriovenous fistula (DAVF) involving the right transverse-sigmoid sinus, and no contrast filling of the superior sagittal sinus, bilateral transverse sinuses, the left sigmoid sinus and the straight sinus. The diagnosis was extensive sinus thrombosis associated with DAVF. Transarterial embolization was performed for the DAVF. The follow-up angiography at 7 days postembolization showed no contrast filling of the DAVF. But the follow-up angiography at 6 months disclosed the appearance of multiple DAVF's involving the superior sagittal sinus, the left transverse sinus and the straight sinus. Each DAVF was supplied by different dural arteries. Coagulation tests revealed deficiency of plasma protein S. The pathogenesis of DAVF is controversial. But our case suggests that multiple isolated lesions indicated the presence of a congenital dural vascular abnormality, which was transformed to multiple DAVF's by sinus thrombosis.

Adult↗

Optic nerve injury demonstrated by MRI with STIR sequences.

We studied nine patients with optic nerve injury associated with closed head trauma by magnetic resonance imaging (MRI) with short inversion time inversion recovery (STIR) sequences on 11 occasions from 4 days to 14 years after the injury: three studies were within 17 days and eight over 4 months to 14 years. MRI revealed abnormal high signal in 10 of the 11 injured nerves. MRI 4 days after the injury showed no abnormality.

Adult↗

The pharmacological properties of Y-23684, a benzodiazepine receptor partial agonist.

1. The pharmacological properties of a benzodiazepine receptor (BZR) partial agonist, Y-23684 were investigated in comparison with those of diazepam, a conventional BZR full agonist. 2. Y-23684 and diazepam showed high and selective affinity for the BZR with Ki values of 41 and 5.8 nM, respectively. 3. In contrast to diazepam, variability was noted in the anticonvulsive potency of Y-23684 depending on convulsants (bicuculline, pentylenetetrazol and maximal electrical shock). Y-23684 produced the most potent protective effect against bicuculline in rats and mice with ED50S of 1.3 and 1.2 mg kg-1, respectively. 4. In rat conflict models (Geller-Seifter and water-lick tests), Y-23684 produced an antipunishment action at doses 2-4 times lower than diazepam. In contrast to diazepam, Y-23684 did not affect unpunished responding up to 50 mg kg-1 in the Geller-Seifter test. 5. In other rat models of anxiety (social interaction and elevated plus-maze tests), Y-23684 was as efficacious as and ten fold more potent than diazepam. In a mouse model of anxiety (exploration (light/dark box) test), Y-23684 was as efficacious and two fold less potent as diazepam. In these paradigms, Y-23684 showed a selective anxiolytic profile over a wide dose-range without loss of efficacy and sedative action. 6. The impairment of motor coordination (rotarod) and potentiation of CNS depressants (ethanol and hexobarbitone) by Y-23684 was much weaker than that of diazepam. 7. These results suggest that Y-23684 would be a potent and selective anxiolytic agent in man with less side-effects than conventional BZ-anxiolytics.

Animals↗

Synthesis and structure-activity relationships of 2,3-dihydrobenzofuran-7-carboxamide derivatives as potent serotonin-3 (5-HT3) receptor antagonists.

A series of N-(azabicyclo-3-yl)-2,3-dihydrobenzofuran-7-carboxamide derivatives were synthesized and evaluated for serotonin-3 (5-HT3) receptor antagonistic activities assessed by 5-HT3 receptor binding (in vitro) and by the ability to antagonize the von Bezold-Jarisch reflex in rats (in vivo). In these compounds, 1-azabicyclo[2.2.2]oct-3-yl derivatives were more potent than 8-methyl-8-azabicyclo[3.2.1]oct-3-yl derivatives for 5-HT3 receptor antagonistic activities. The introduction of methyl groups at position 2 of the dihydrobenzofuran ring increased the pharmacological activities (dimethyl > monomethyl > dihydro). Furthermore, the stereoisomers of dimethyl-, monomethyl-, and dihydrobenzofuran derivatives were prepared to evaluate the stereoselectivity of their 5-HT3 receptor binding affinities. Concerning the basic part, the compounds bearing (S)-1-azabicyclo[2.2.2]octan-3-yl moiety were more potent than their counterparts. With respect to the methyl substituent at position 2 of the dihydrobenzofuran ring, the rank order of the potency was dimethyl > or = (2S)-methyl > (2R)-methyl > dihydro. These results suggest that the (2S)-methyl group of the dihydrobenzofuran part contributes to the enhancement of the pharmacological activity. Among these compounds, (S)-N-(1-azabicyclo[2.2.2]oct-3-yl)-5-chloro-2,3-dihydro-2,2- dimethylbenzofuran-7-carboxamide hydrochloride (24) showed the highest affinity for 5-HT3 receptors (Ki = 0.055 nM), and the most potent antagonistic activity on the von Bezold-Jarisch reflex (ED50 = 0.18 microgram/kg i.v.).

Animals↗