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Biomedical subjects

T Kameda

Publications and source records attributed to T Kameda.

At least 19 recordsLinked to original sources
PubMed2006

SWI/SNF complex is essential for NRSF-mediated suppression of neuronal genes in human nonsmall cell lung carcinoma cell lines.

Mammalian chromatin remodeling factor, SWI/SNF complex contains a single molecule of either Brm or BRG1 as the ATPase catalytic subunit. Here, we show that the SWI/SNF complex forms a larger complex with neuron-restrictive silencer factor (NRSF) and its corepressors, mSin3A and CoREST, in human nonsmall cell lung carcinoma cell lines. We also demonstrate that the strong transcriptional suppression of such neuron-specific genes as synaptophysin and SCG10 by NRSF in these non-neural cells requires the functional SWI/SNF complex; these neuronal genes were elevated in cell lines deficient in both Brm and BRG1, whereas retrovirus vectors expressing siRNAs targeting integral components of SWI/SNF complex (Brm/BRG1 or Ini1) induced expression of these neuronal genes in SWI/SNF-competent cell lines. In cell lines deficient in both Brm and BRG1, exogenous Brm or BRG1 suppressed expression of these neuronal genes in an ATP-dependent manner and induced efficient and specific deacetylation of histone H4 around the NRSF binding site present in the synaptophysin gene by a large complex containing the recruited functional SWI/SNF complex. Patients with Brm/BRG1-deficient lung carcinoma have been reported to carry poor prognosis; derepression of NRSF-regulated genes including these neuron-specific genes could contribute to enhance tumorigenicity and also would provide selective markers for Brm/BRG1-deficient tumors.

Base Sequence↗
PubMed2001

Structure of Bombyx mori silk fibroin before spinning in solid state studied with wide angle x-ray scattering and (13)C cross-polarization/magic angle spinning NMR.

The structure of a crystalline form of Bombyx mori silk fibroin, commonly found before the spinning process (known as silk I), has been proposed as a repeated beta-turn type II-like structure by combining data obtained from solid-state two dimensional spin-diffusion nuclear magnetic resonance and rotational-echo double-resonance (T. Asakura et al., J Mol Biol, in press). In this paper, the WAXS pattern of alanine-glycine alternating copolypeptide, (Ala-Gly)(15) with silk I form which was used for a silk I model of B. mori silk fibroin was observed. The pattern calculated with the silk I model proposed by us is well reproduced the observed one, indicating the validity of the proposed silk I model. In addition, two peptides of the other repeated sequences which contain Tyr or Val residues in the silk fibroin,23 were synthesized; (Ala-Gly-Tyr-Gly-Ala-Gly)(5) and (X-Gly)(15) where X is Tyr for the 7th, 15th and 23th residues, and Val for the 11th residue and Ala for other residues. There are no sharp peaks in the WAXS patterns, and therefore both samples are in the non-crystalline state. This is in agreement with the (13)C CP/MAS NMR result, where the conformation is mainly random coil.

Amino Acid Sequence↗
PubMed2001

A repeated beta-turn structure in poly(Ala-Gly) as a model for silk I of Bombyx mori silk fibroin studied with two-dimensional spin-diffusion NMR under off magic angle spinning and rotational echo double resonance.

The structure of a crystalline form of Bombyx mori silk fibroin, commonly found before the spinning process (known as silk I), was proposed by combining data obtained from two-dimensional spin-diffusion nuclear magnetic resonance under off magic angle spinning, rotational-echo double-resonance (REDOR), previously reported X-ray diffraction analyses and 13C NMR chemical shifts. Instead of B. mori silk fibroin with silk I structure, we used the sequential model peptide (Ala-Gly)15. The structure of the sequential model peptide is characterized as silk I after dissolving the peptide in 9 M LiBr and then dialyzing against water. Moreover, 13C or 15N-labeled sites may be introduced easily at any position in (Ala-Gly)(15) by the solid phase synthesis method for these NMR experiments. The torsional angles of (Ala-Gly)15 with silk I structure were determined as (-60(+/-5) degrees, 130(+/-5) degrees ) and (70(+/-5) degrees, 30(+/-5) degrees ) for Ala and Gly residues, respectively. The formation of the intra-molecular hydrogen bonding along the chain was confirmed from REDOR NMR by determination of the inter-atomic distance between the nitrogen and carbon atoms comprising the intra-molecular hydrogen bonding. The structure is named a repeated beta-turn type II-like structure.

Alanine↗
PubMed2001

Genetic analysis of a variant luteinizing hormone in an infertile woman.

In a woman with infertility and an endometrial polyp, we analyzed the nucleotide sequence of the beta-subunit of her luteinizing hormone which showed anomalous immunogenecity in that it was recognized by time resolved-fluoroimmunoassay but not by immunoradiometric assay. The sequence analysis showed two substitutional mutations of beta-subunit Trp (TGG) to Arg (CGG) and Ile (ATC) to Thr (ACC). The variant luteinizing hormone might have had some relation to the infertility and the endometrial polyp. been described [2, 4, 13-15]. Although the role of the variant LH is still unclear, it has different biologicl activity as compared to normal LH and may play a role in causing infertility, menstrual disorder, endometriosis and spontaneous miscarriage [1, 3, 4, 9, 10, 17]. We now report a study of the DNA sequence of beta-subunit of anomalous LH in an infertile patient with an endometrial polyp.

Adult↗
PubMed2001

An evaluation of the intraoperative transit time measurements of coronary bypass flow.

OBJECTIVE: The intraoperative measurement of the coronary bypass flow enables the identification of technical errors while the sternum is still open. The transit-time flow method is able to effectively measure the internal thoracic artery graft flow. The aim of the present study was to analyze the factors which affected the bypass flow rate. METHODS: We measured the blood flow of 291 in situ internal thoracic artery (ITA) and 190 saphenous vein (SV) grafts constructed in 171 patients undergoing coronary artery bypass grafting from December 1996 to March 2000 using this method during the surgery. All patients underwent postoperative coronary angiography before the patients were discharged. The blood flow rate of all bypass grafts constructed was assessed after the patients were weaned from cardiopulmonary bypass. RESULTS: The mean flow rate of all ITA grafts was 65.1+/-36.7 ml/min and that of all SV grafts was 56.4+/-29.9 ml/min. According to analyses using correlation tests, the graft flow was found to significantly correlate with the grafted perfusion areas and the diameter of the bypassed coronary arteries. However, no significant difference was observed between the flow rates of the ITA grafts with and without stenosis or string phenomenon, but significant (P<0.0001) correlation was observed between the occurrence of a string sign and the degree of proximal stenosis of the recipient coronary artery. Regarding SV grafts, the mean flow rate of occluded grafts (29.2+/-20.5 ml/min) was significantly (P<0.0001) less than non-occluded grafts (56.4+/-29.9 ml/min). CONCLUSIONS: The bypass flow was affected by such a large number of factors that only measuring the bypass flow rate could not sufficiently predict either stenosed or narrowed grafts. However, ITA grafts bypassed to the coronary arteries with less stenosis were shown to more easily become narrowed.

Adult↗
PubMed2001

Acceleration of the formation of cultured epithelium using the sonic hedgehog expressing feeder cells.

Sonic hedgehog (Shh) regulates the principal possesses in many developmental stages, including the epithelial-mesenchymal interaction. The extraordinary acceleration of signaling by Shh is responsible for the development of human basal cell carcinomas and trichoepitheliomas; they might originate from the very immature keratinocytes, including the stem cells. We tried to utilize the mitogenic effect of Shh to accelerate the formation of cultured epithelium, which is already used in the medical field practically. To this end, we transfected shh cDNA into a Swiss-3T3 cell line, widely used as a feeder for keratinocytes, and established a Shh expressing cell line. The lethally irradiated Shh expressing feeder cells remarkably accelerated the growth of keratinocyte colonies obtained from the human neonatal foreskin, and the formation of well-stratified cultured epithelium, which is rich in immature small keratinocytes, expressing cytokeratin 14. This acceleration was suppressed by the addition of cyclopamine, a specific inhibitor of Shh signaling. These data indicate that the Shh is a promising mitogen to improve the technology for cultured epithelium formation.

3T3 Cells↗
PubMed2001

Electron paramagnetic resonance study on free radical scavenging and/or generating activity of dopamine-4-O-sulfate.

The free radical scavenging and/or generating activity of dopamine-4-O-sulfate was examined and compared with that of dopamine. In humans, dopamine mostly exists in two isomeric forms of sulfate ester conjugates as metabolites; i.e., dopamine-3-O-sulfate and dopamine-4-O-sulfate in the circulation. Dopamine is generally believed to be oxidized by molecular oxygen or another reactive oxygen species under physiological conditions, to form oxidized dopamine derivatives that are cytotoxic. However, it is not known whether dopamine conjugates are generated on interaction with reactive oxygen species or not. In the present study, we measured the susceptibility to oxidization of dopamine-4-O-sulfate by using electron paramagnetic resonance (EPR) spectroscopy and optical absorption spectrometry. Dopamine was easily oxidized and dopamine-derived radicals appeared, whereas dopamine-4-O-sulfate was not oxidized under physiological conditions. Furthermore, dopamine-4-O-sulfate did not react with a strong oxidizing agent, sodium periodate. These results suggest that dopamine-4-O-sulfate has resistance against autoxidation, and seems to be a stable metabolite of dopamine.

Dopamine↗
PubMed2001

Coronary artery bypass grafting within 30 days of an acute myocardial infarction.

UNLABELLED: Early surgical intervention is now often considered for symptomatic patients after an acute myocardial infarction. Conversely coronary artery bypass grafting soon after an acute myocardial infarction poses substantial risks. The present study was performed to evaluate the results of Coronary artery bypass grafting soon after an acute myocardial infarction. METHODS: From November 1991 to November 1999, 478 consecutive patients underwent coronary artery bypass grafting and 68 of these underwent an operation within 30 days of AMI. The data of these patients were analyzed retrospectively. Univariate and multivariate analyses of many variables were performed regarding operative mortality. RESULTS: Operative mortality (7.4%) was significantly higher in the patients with an acute myocardial infarction than in the patients without it (0.8%) during the same period as the subjects of this study. Coronary artery bypass grafting without arterial grafts was solely determined to be the predictor of survival. The survival curve demonstrated better long-term results in patients undergoing bypass grafting with arterial grafts than in patients undergoing bypass grafting with venous grafts alone. CONCLUSIONS: If hemodynamic conditions can not be stabilized, then coronary artery bypass grafting using arterial grafts, when indicated, should be performed even early after AMI.

Acute Disease↗
PubMed2000

Cloning and function of rabbit peroxisome proliferator-activated receptor delta/beta in mature osteoclasts.

Osteoclasts modulate bone resorption under physiological and pathological conditions. Previously, we showed that both estrogens and retinoids regulated osteoclastic bone resorption and postulated that such regulation was directly mediated through their cognate receptors expressed in mature osteoclasts. In this study, we searched for expression of other members of the nuclear hormone receptor superfamily in osteoclasts. Using the low stringency homologous hybridization method, we isolated the peroxisome proliferator-activated receptor delta/beta (PPARdelta/beta) cDNA from mature rabbit osteoclasts. Northern blot analysis showed that PPARdelta/beta mRNA was highly expressed in highly enriched rabbit osteoclasts. Carbaprostacyclin, a prostacyclin analogue known to be a ligand for PPARdelta/beta, significantly induced both bone-resorbing activities of isolated mature rabbit osteoclasts and mRNA expression of the cathepsin K, carbonic anhydrase type II, and tartrate-resistant acid phosphatase genes in these cells. Moreover, the carbaprostacyclin-induced bone resorption was completely blocked by an antisense phosphothiorate oligodeoxynucleotide of PPARdelta/beta but not by the sense phosphothiorate oligodeoxynucleotide of the same DNA sequence. Our results suggest that PPARdelta/beta may be involved in direct modulation of osteoclastic bone resorption.

Acid Phosphatase↗
PubMed2000

A study of conformational stability of polyglycine and poly(L-alanine), and polyglycine/poly(L-alanine) blends in the solid state by (13)C cross-polarization/magic angle spinning NMR.

13C Cross-Polarization/Magic Angle Spinning nmr and T(1rhoH) experiments of polyglycine (PG), poly(L-alanine) (PLA), and PG/PLA blends prepared from dichloroacetic acid solution have been carried out, in order to elucidate the conformational stability of these polypeptides in the solid state. From these experimental results, it was clarified that the conformations of PG and PLA in their blends are strongly influenced by intermolecular hydrogen-bonding interactions that cause their miscibility at the molecular level.

Carbon Isotopes↗
PubMed2000

Posterior ventricular septal perforation in an 80-year-old man.

Ventricular septal perforation is a complication of acute myocardial infarction that is fatal unless treated surgically. Posterior ventricular septal perforation remains particularly difficult to repair successfully. We report an 80-year-old man with postinfarction posterior ventricular septal perforation who successfully underwent surgical repair using a technique similar to that described by Daggett et al. and was discharged in good health without postoperative septal shunt.

Aged↗
PubMed2000

Stroke after coronary artery bypass grafting in patients with cerebrovascular disease.

BACKGROUND: Stroke has been associated with a significantly increased mortality from coronary artery bypass grafting (CABG). To determine the predictors of stroke in patients undergoing CABG, we collected data on 472 consecutive patients. METHODS: From March 1991 to March 1999, all patients undergoing CABG at our institution underwent routine duplex scanning of the extracranial carotid and vertebral arteries. Seven patients with symptomatic carotid stenosis were treated by carotid endarterectomy (CEA) before CABG. RESULTS: There was a 10-fold increase in mortality (12.5%) associated with postoperative stroke. Many variables were analyzed by a multivariate technique and the severity of extracranial carotid artery stenosis was determined to be the only independent predictor of postoperative stroke (p < 0.01). None of the patients with carotid artery occlusion and none of the patients who underwent CEA before CABG experienced a stroke. CONCLUSIONS: To reduce the stroke rate, the indications for prophylactic CEA may be extended for asymptomatic patients with carotid artery stenosis greater than 75%.

Aged↗
PubMed2000

Aortic arch repair using hypothermic circulatory arrest technique associated with pharmacological brain protection.

OBJECTIVE: Hypothermic circulatory arrest is a standard procedure for the treatment of aortic arch. However, there is a time limit for this procedure. There is now an urgent need to develop prophylactic measures to extend the time limit. We have used a pharmacological mixture of thiopental, nicardipine and mannitol for all patients undergoing circulatory arrest since 1991 to extend the safe limit. The purpose of this study was to analyze the neurological complications demonstrated by these patients and to evaluate the brain-protective effects of our measure. METHODS: The clinical records of 75 consecutive patients undergoing an aortic arch repair using a hypothermic circulatory arrest technique during the past 8 years were retrospectively reviewed. Systemic cooling was continued until a total disappearance of EEG activity. Prior to circulatory arrest, 15 or 30 mg/kg of thiopental, 20 mg of nicardipine and 300 ml of mannitol were infused into the venous reservoir of a cardiopulmonary bypass circuit. Graft replacement was performed in all patients and the extent of replacement was a total aortic arch in 43 patients, a distal aortic arch in 17, a hemiarch in 13 and a distal aortic arch and a total descending aorta in two. RESULTS: The duration of circulatory arrest ranged from 16 to 80 min (mean 41.5 min), and it exceeded 45 min in 37 patients. Operative mortality was 10.7% and two patients died of stroke. Three patients had permanent and three other patients had transient neural deficits. The incidence of stroke was 8.0% as a whole, and no correlation between the incidence of neurological complications and the duration of circulatory arrest was found. A multivariate analysis showed that the duration of circulatory arrest was determined as a predictor of neither operative mortality nor postoperative stroke. CONCLUSIONS: The findings of the present study suggest that our pharmacological brain protection appears to be effective for safely extending hypothermic circulatory arrest.

Adult↗
PubMed2000

Application of efficient and specific gene transfer systems and organ culture techniques for the elucidation of mechanisms of epithelial-mesenchymal interaction in the developing gut.

Epithelial-mesenchymal interactions are very important in the development of the vertebrate gut. In the avian embryonic stomach (proventriculus), expression of embryonic chick pepsinogen (ECPg) gene, which is specific to developing glandular cells in stomach epithelium, is regulated by mesenchymal influence. Molecular mechanisms of tissue-specific transcriptional regulation of the ECPg gene and the molecular nature of the mesenchymal signals were analyzed using a combination of the classic organ culture system and gene transfer strategies. In the present review, three methods for the introduction of DNA into tissues are described: lipofection, electroporation and retroviral infection, and characteristics of each system are discussed.

Animals↗
PubMed2000

Analysis of cartilage maturation using micromass cultures of primary chondrocytes.

A micromass culture (MM-C) system of primary immature chondrocytes for functional analysis of soluble factors involved in the maturation step of cartilage was previously developed. Ectopically expressed BMP-2 was shown to induce the expression of the Ihh and Noggin genes. Here it is demonstrated that, upon longer culture, secreted bone morphogenetic protein-2 (BMP-2) further promotes the maturation step as judged by the induction of type X collagen and BMP-6 expression, which are known to be detectable in the later phase of cartilage maturation. Induction of all of these genes by secreted BMP-2 was not inhibited by ectopic expression of parathyroid hormone-related peptide (PTHrP) induced by retrovirus vector infection, although the same virus vector showed strong inhibitory effects on the expression of type X collagen gene or alkaline phosphatase activity in mature chondrocytes. These results suggest that the maturation-promoting activity exhibited by BMP-2 is dominant over the suppressive effect of PTHrP in immature chondrocytes. When the BMP-6 gene was introduced into the same virus vector as that used for BMP-2, it induced the same sets of genes (Ihh, Noggin, type X collagen and endogenous BMP-6) as BMP-2 did. These results also suggest that BMP-6 would autonomously maintain and/or promote a later stage of chondrocytic maturation.

Animals↗
PubMed2000

Results of a total aortic arch replacement for an acute aortic arch dissection.

OBJECTIVES: Recently, the immediate results of a surgical repair for an acute aortic arch dissection have dramatically improved. However, a total aortic arch replacement is recommended in a limited number of patients with an intimal tear located in the aortic arch. We have performed a total aortic arch replacement for all such patients with an acute aortic arch dissection since September 1995. METHODS: During the past 4 years, 27 consecutive patients who had an aortic arch dissection underwent a total aortic arch replacement. Twenty-five patients underwent an emergency operation. In 5 patients the intimal tear was located in the aortic arch, but in the rest of the patients, it was located in the ascending aorta or the proximal descending aorta. To obliterate any false channels, gelatin-resorcin-formol glue was used. RESULTS: The hospital mortality was 11%, and no cerebral complications were observed. Postoperative aortography and computed tomography showed no evidence of any persisting false channels in 15 patients (65%). During the follow-up period (ranging from 5 months to 4 years), two patients underwent a reoperation because of the recurrence of a dissection at the sinus of Valsalva. All patients, except for one who died after a reoperation, are still alive and free from any serious events at this writing. CONCLUSIONS: Resecting both the ascending and transverse aorta, irrespective of whether the intimal tear is located in the aortic arch, may be an acceptable alternative at experienced centers because of its low mortality and good midterm results.

Adult↗
PubMed2000

BMPs are necessary for stomach gland formation in the chicken embryo: a study using virally induced BMP-2 and Noggin expression.

Epithelial-mesenchymal interactions are necessary for the normal development of various digestive organs. In chicken proventriculus (glandular stomach), morphogenesis and differentiation of the epithelium depend upon the inductive signals coming from underlying mesenchyme. However, the nature of such signals is still unclear despite extensive analyses carried out using experimental tissue recombinations. In this study we have examined the possible involvement of bone morphogenetic proteins (BMPs) in the formation of stomach glands in the chicken embryo. Analysis of the expression patterns of BMP-2, -4 and -7 showed that these BMPs were present in the proventricular mesenchyme prior to the initiation of the proventricular gland formation. BMP-2 expression, in particular, was restricted to the proventriculus among anterior digestive organs. Virus-mediated BMP-2 overexpression resulted in an increase in the number of glands formed. Moreover, ectopic expression of Noggin, which antagonizes the effect of BMPs, in the proventricular mesenchyme or epithelium, led to the complete inhibition of gland formation, indicating that BMP signals are necessary for the proventricular gland formation. These findings suggest that BMPs are of prime importance as mesenchymal signals for inducing proventricular glands.

Animals↗