[Current topics of receptor proteins for heat-stable enterotoxin produced by enterotoxigenic Escherichia coli].
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Adenosine Triphosphate↗
Biomedical subjects
A Wada
Publications and source records attributed to A Wada.
[Affinity of estrogen binding in the cultured spinal ligament cells: an in vitro study using cells from spinal ligament ossification patients].
This immunological study investigated the effects of estrogen as a potential causative factor for spinal ligament ossification (such as ossification of the posterior longitudinal ligament, OPLL; ossification of the yellow ligament, OYL; and ossification of the anterior longitudinal ligament, OALL). The serum total estrogen (estron + estradiol + estriol) level in the OPLL patients and controls was measured by radioimmunoassay. To determine any difference in the affinity of the estrogen, 3,17 beta-estradiol receptors of cultured spinal ligament cells obtained from OPLL patient were detected by receptor binding assay, and compared with cells from controls. Additionally, to evaluate the responses of cultured spinal ligament cells to stimulation by 3,17 beta-estradiol, examined the production of bone Gla protein (BGP) in medium, the rate of tritiated thymidine (3H-TdR) uptake, and change in affinity of transforming growth factor-beta 1 (TGF-beta 1) receptor on those cells with 3,17 beta-estradiol added (E2+ group), and compared the results with those when 3,17 beta-estradiol was not added (E2- group). The serum total estrogen level was significantly higher in OPLL patients than in controls, and the level increased with increasing extent of ligament ossification. Cultured cells obtained from OPLL patients had receptors with a higher affinity for 3,17 beta-estradiol than did cells from controls. Cells obtained from OPLL patients responded to the stimulation by 3,17 beta-estradiol, accelerated BGP production, and elevated the 3H-TdR uptake. However, cells from controls showed no change in the stimulation by 3,17 beta-estradiol.(ABSTRACT TRUNCATED AT 250 WORDS)
Adult↗
[Prostate and prostatic carcinoma: comparison of gadolinium-enhanced MR images and histopathologic findings].
The purpose of this study was to determine the patterns of enhancement in normal prostate, and to compare them with the enhancement patterns and histopathologic findings in patients with prostatic carcinoma. Seventeen patients who had no urogenital disease and 27 who underwent radical prostatectomy or prostatic biopsy were included in this study. All patients were evaluated with T1-, T2-weighted and gadolinium-enhanced T1-weighted images. With gadolinium enhancement the inner gland showed more enhancement than the peripheral zone in the normal prostate. The carcinoma tissue was enhanced more than the peripheral zone in 13 patients, about the same as the peripheral zone in 11 patients, and less than the peripheral zone in three patients. The three patients whose cancer tissue demonstrated no enhancement had poorly differentiated adenocarcinoma of the solid type. In conclusion, gadolinium-enhanced T1-weighted imaging is not reliable for routine use in MR imaging of the prostate, but may be useful to evaluate histologic types in patients with prostatic carcinoma.
Adolescent↗
Disruption of cell-cell adhesion in an inbred strain of hereditary cardiomyopathic hamster (Bio 14.6).
OBJECTIVE: Disarrangement of cardiomyocytes is a pathological characteristic of dilated cardiomyopathy. Hereditary cardiomyopathic hamster Bio 14.6, a model of dilated cardiomyopathy, displays disorder of cardiomyocyte arrangement. The aim of this study was to analyse the disturbance of cell alignment from the point of view of the cell-cell adhesion system in Bio 14.6. METHOD: Cardiomyopathic hamster Bio 14.6 was used as a model of dilated cardiomyopathy. Histological study was performed by light and electron microscopy. Disorder of the adherens junction-specific cell-adhesion molecule (A-CAM) was analysed by immunofluorescent microscopy and immunoblotting with anti-A-CAM antibody. RESULTS: Hematoxylin-eosin staining revealed that intercalated disks were identifiable less clearly in cardiomyopathy than in a normal cardiac muscle. It was disclosed by electron microscopy that cardiomyocytes adhered to each other with reduction in subsarcolemmal electron density at intercalated disks in Bio 14.6 compared with normal hamsters. We examined the localization of the A-CAM molecule in heart by immunofluorescent microscopy. In contrast to normal cardiac samples, fluorescence was weak in intensity and unclearly demarcated in the Bio 14.6 hamsters. We measured the content of A-CAM in the heart. In Bio 14.6 hamsters, the content of A-CAM was 60 +/- 11% of that measured in normal adult hamsters. A-CAM was reduced to a lesser extent (81 +/- 12%) in the newborn hamsters. CONCLUSIONS: In Bio 14.6 hamster, structural disturbance of the intercalated disks was found on histological examination of the heart. Biochemically, A-CAM, which plays a role in intercellular adhesion in intercalated disk areas, decreased significantly. These results suggest that cardiomyopathy may be accompanied by structural disruption of cell-cell adhesion in intercalated disk regions, which may lead to the pathological feature of disarranged cardiomyocytes.
Animals↗
Batchwise purification of specific tRNAs by a solid-phase DNA probe.
A simple and efficient method for purifying a specific tRNA in a single microcentrifuge tube was developed. Oligodeoxyribonucleotides (about 30 mer) with sequences complementary to the 3' side of target tRNAs were synthesized with an aminohexyl linker at the 5' end, immobilized on a silica gel at a high concentration, and used as a solid-phase probe. A mixture of tRNAs was added to a suspension of the solid-phase probe in 2.4 M tetraethylammonium chloride and incubated for 10-30 min. Only a target tRNA hybridized with the immobilized probe at appropriate temperatures and was eluted out by heating. The solid-phase probe showed a large hybridization capacity (up to 17 A260 units/g dry gel) and specific and quantitative recovery of the target tRNA. The intactness of recovered tRNAs was ascertained by both Donis-Keller sequencing and aminoacylation experiments. These features show the usefulness of the solid-phase probe method as a reliable tool for purifying tRNAs whose gene sequences are known.
Base Sequence↗
Preoperative Discrimination of Fibroadenoma Which Is Clinically and Cytologically Indistinguishable from Breast Carcinoma through Clonal Analysis of Fine Needle Aspirate of Tumor: Report of a Case.
Recently, it has been demonstrated that breast carcimoma is monoclonal and fibroadenoma is polyclonal in origin. In the present case report, this observation was successfully applied to a preoperative differential diagnosis of a fibroadenoma clinically and cytologically indistinguishable from carcimoma. Case report: A45-year old female presented for clinical examination with a history of breast lump. A firm tumor measuring 2 x 2 cm was palpable in the upper-outer quadrant of @the left breast. The margin of the tumor was partially ill-defined and its mobility was restricted. A tumor shadow with a partially ill-defined margin was revealed by mammography. Ultrasonographic examination showed and irregularly-shaped, hypo-echoic tumor accompanied by an acoustic shadow. Fine needle aspiration biopsy yielded positive cytology. Based on these results, the tumor was diagnosed as breast carcinoma. However, clonal analysis of fine needle aspirates showed a polyclonal pattern, strongly indicating that the tumor was not a carcinoma but benign disease; most probably fibroadenoma. Thus, an open surgical biopsy was performed. Histoglogical examination revealed that the tumor was indeed a fibroadenoma with epithelial hyperplasia. This case suggests the usefulness of clonal analysis of fine needle aspirates in differentiating fibroadenoma from carcinoma of the breast.
Journal Article↗
Ca(2+)-dependent cosecretion of adrenomedullin and catecholamines mediated by nicotinic receptors in bovine cultured adrenal medullary cells.
Bovine cultured adrenal medullary cells (4 x 10(6)) contained 4266.5 +/- 370.0 fmol of immunoreactive adrenomedullin and 373.4 +/- 32.6 nmol of catecholamines. Nicotinic (but not muscarinic) receptors mediated the Ca(2+)-dependent co-secretion of adrenomedullin and catecholamines, with the molar ratio of adrenomedullin/catecholamines secreted into the medium being equal to the ratio stored in the cells. The concentration-response curve of carbachol for adrenomedullin secretion (EC50 42 microM) was similar to that for catecholamine secretion (EC50 63 microM). Reverse phase HPLC analysis showed that immunoreactive adrenomedullins in the cells and secreted into the medium were both eluted exclusively at the position almost identical to synthetic human adrenomedullin[1-52]NH2.
Adrenal Medulla↗
Congenital malformations in the offspring of male mice treated with ethylnitrosourea at the embryonic stage.
Ethylnitrosourea (ENU) was i.p. injected into ICR female mice at 25 or 50 mg/kg on day 10 of gestation, and male newborns treated with ENU at the embryonic stage were obtained. The treated males were aged for 10 weeks and then mated to untreated females of the same strain. The male germ cells used in the copulation correspond to primordial germ cells (PGC) at the time of ENU treatment. The F1 offspring were sampled as fetuses on day 18 of gestation and inspected for external and skeletal malformations. Evidence of F1 teratogenesis due to the mutagenized PGC was obtained when the fetuses from the males treated with 25 mg ENU/kg were inspected the frequencies of skeletally malformed fetuses in the ENU-treated series and the control being, respectively, 1.0% (14/1,360) and 0.3% (3/1,017). The fetuses from 50 mg/kg-treated males did not show a significant increase in malformations, most probably reflecting a high vulnerability of PGC to the killing effect of ENU. The findings in this study suggest that germ cell stage at PGC is at risk for the induction of congenital malformations by environmental chemicals.
Abnormalities, Drug-Induced↗
Isoflurane inhibits nicotinic acetylcholine receptor-mediated 22Na+ influx and muscarinic receptor-evoked cyclic GMP production in cultured bovine adrenal medullary cells.
The effects of isoflurane on 22Na+ influx, 45Ca2+ influx, catecholamine secretion and cyclic GMP production induced by three kinds of secretagogue (nicotinic agonists, veratridine and a high concentration of K+) have been investigated using cultured bovine adrenal medullary cells. (1) Isoflurane (1-6%) inhibited catecholamine secretion stimulated by carbachol, nicotine and dimethyl-4-phenylpiperazinium in a concentration-dependent manner. Isoflurane suppressed carbachol-evoked 22Na+ influx and 45Ca2+ influx at concentrations similar to those which suppressed catecholamine secretion. The inhibition of catecholamine secretion by isoflurane was not overcome by increasing the concentration of carbachol. (2) The inhibitory effects of isoflurane on veratridine-induced 22Na+ influx, 45Ca2+ influx and catecholamine secretion became evident when the concentration of isoflurane was raised to 4-6%, i.e. 2-3 fold higher than the concentrations (1-2%) employed clinically. (3) High K(+)-evoked 45Ca2+ influx and catecholamine secretion were not affected by isoflurane (1-6%). (4) Isoflurane (1-6%) attenuated the production of cyclic GMP caused by muscarine, but not that caused by atrial natriuretic peptide or by sodium nitroprusside. These results suggest that isoflurane, at clinical anesthetic concentrations, inhibits nicotinic acetylcholine receptor-mediated cell responses as well as muscarinic receptor-mediated cyclic GMP production in adrenal medullary cells.
Adrenal Medulla↗
Functional relation between nitric oxide and noradrenaline for the modulation of vascular tone in rat mesenteric vasculature.
As previously reported, N omega-nitro-L-arginine (L-NNA), an inhibitor of nitric oxide (NO) synthesis, decreased transmural field stimulation (TFS)-induced noradrenaline overflow from the isolated perfused rat mesenteric vasculature attached to the intestine. The decrease was attenuated by L-arginine. This suggests that NO may increase noradrenaline release (Yamamoto et al. 1993). The present experiments with this preparation were done in order to monitor changes in vascular perfusion pressure caused by TFS or by noradrenaline infusion in parallel with those in the noradrenaline outflow caused by TFS in the presence of atropine (0.1 mumol/l) (to block acetylcholine-induced release of endothelial NO) and of indomethacin (3 mumol/l) (to inhibit L-NNA-induced production of vasoconstrictor prostanoids). (1) TFS (2-10 Hz) caused a frequency-dependent increase in noradrenaline overflow and perfusion pressure. (2) L-NNA (10 and 30 mumol/l) caused a concentration-dependent inhibition of TFS-induced noradrenaline overflow, whereas the TFS-induced pressure increase was augmented by L-NNA in a concentration-dependent manner. At any given concentration of L-NNA, the potentiation of vasoconstriction by L-NNA became greater in magnitude as the frequency of the TFS was raised. (3) Infusion of noradrenaline (0.38-6 nmol) caused a dose-dependent increase in perfusion pressure up to a value comparable with that caused by TFS. The pressure increase in response to noradrenaline infusion was also enhanced by L-NNA, relatively, to a greater extent than the enhancement, by L-NNA, of the pressure response to TFS.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Characterization of [3H]brevetoxin binding to voltage-dependent sodium channels in adrenal medullary cells.
We have previously reported that in bovine adrenal chromaffin cells Ptychodiscus brevis toxin-3 (PbTx-3) does not alter the veratridine-induced 22Na influx when given alone, but increases the influx of 22Na when co-applied with either alpha- or beta-scorpion venom (Wada et al. 1992). In the present study, we characterized [3H]PbTx-3 binding in bovine adrenal chromaffin cells. [3H]PbTx-3 binding was saturable, reversible and of high-affinity with an equilibrium dissociation constant (Kd) of 32.0 +/- 4.9 nmol/l and a maximum binding capacity (Bmax) of 6.2 +/- 1.2 pmol/4 x 10(6) cells (4.5 +/- 0.9 pmol/mg cell protein). A Hill plot revealed the lack of cooperative interaction among the binding sites. Unlabelled PbTx-3 inhibited [3H]PbTx-3 binding with an IC50 of 31 nmol/l. However, tetrodotoxin, veratridine, alpha- and beta-scorpion venom, or veratridine in combination with either alpha- or beta-scorpion venom did not alter [3H]PbTx-3 binding. All these results suggest that PbTx-3 binds to a site (site 5) distinct from the previously known four toxin binding sites, which does not gate voltage-dependent Na channels by itself, but is specifically involved in the allosteric modulation of Na channels in adrenal medullary cells.
Adrenal Medulla↗
Absence of hemodynamic tolerance to nicorandil in patients with severe congestive heart failure.
To evaluate whether hemodynamic tolerance develops to nicorandil, a nitrate and potassium channel opener, 14 patients with chronic heart failure (CHF) were treated with nicorandil and 11 were treated with nitroglycerin (GTN). Doses of GTN or nicorandil were titrated to achieve a > or = 20% reduction in pulmonary capillary wedge pressure (PCWP) within 1 hour, and the infusion was maintained at a constant rate for 24 hours. Both groups of patients had comparable hemodynamic parameters before drug infusions were started. The fall in PCWP was identical after 1 hour infusion of either GTN or nicorandil. In the GTN group, PCWP was not significantly different from the baseline value at 12 hours; however, in the nicorandil group, PCWP remained significantly lower than the preinfusion value for 24 hours. During the study period changes in plasma atrial natriuretic peptide (ANP) concentrations paralleled and correlated with changes in PCWP (r = 0.84, p < 0.001). These findings indicate that CHF patients develop hemodynamic tolerance to GTN within 12 hours of continuous infusion, but not to nicorandil, which remained hemodynamically effective during the 24-hour period of infusion. Furthermore, plasma ANP concentration may be a useful noninvasive index of hemodynamic tolerance during GTN or nicorandil therapy in patients with CHF.
Adult↗
Dexamethasone inhibits production of interleukin-6 by cultured cardiac myxoma cells.
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Aged↗
Fundamental significance of DNA mass-sequencing factory for biological sciences in future.
We are now entering an era of the mass-analysis of genetic information, which will signal the beginning of the study of living organisms on the basis of their most detailed plan: the DNA base-sequence. The researcher's essential requirement in reading and deciphering the DNA base-sequence is the precision, speed, reliability, and low cost of such operation. I have long stressed that these requirements can only be satisfied by establishing a centralized organization, a DNA sequencing factory, where a large-scale and international operation of the sequencing can be carried out with the assistance of high technologies and under an administration which equitably represents the research interests of researchers worldwide. In this context, the basic concept and aims of the Japanese project which was begun in 1981 are described, detailing the advantages of such a factory with a mega-base daily output. Scientists and engineers are devising a high speed and automated DNA sequencing system by coordinating available DNA-processing technologies, particularly in the field of computer/robotic engineering. Typical of the achievements of this enterprise is the recent progress made by RIKEN in the development of a DNA mass-sequencing line which aims the production of raw base-sequence data of 0.1 mega-base per day (machine throughput). Finally, the elemental units in the current DNA sequencing procedures and the means of DNA processing are displayed on a family tree network to illustrate their functional interrelation in the current scheme of DNA analysis. It is hoped that this will provide a clue to finding new dimensions in DNA processing.
Base Sequence↗
Relation between endothelin-1 spillover in the lungs and pulmonary vascular resistance in patients with chronic heart failure.
OBJECTIVES: The aim of this study was to clarify the origin of plasma endothelin-1 and to determine the relation between pulmonary vascular resistance and endothelin-1 secretion in the pulmonary circulation in patients with chronic congestive heart failure. BACKGROUND: Plasma levels of endothelin-1, a potent endothelium-derived vasoconstrictor peptide, are increased in congestive heart failure, but the source has not been clarified. Recent studies have indicated a relation between endothelin-1 and pulmonary hypertension. We therefore evaluated the contribution of endothelin-1 secretion in the pulmonary circulation to the regulation of pulmonary vascular resistance in patients with chronic heart failure. METHODS: A comparison was made of the plasma levels of endothelin-1 between the main pulmonary artery and the pulmonary capillary wedge region, as well as between the femoral artery and the femoral vein in 62 patients with chronic heart failure. Stepwise multivariate regression analysis was used to detect independent predictors of pulmonary vascular resistance among the various vasoconstrictor hormones in these patients. RESULTS: There was no significant difference in plasma endothelin-1 levels between the femoral artery and vein. In contrast, plasma endothelin-1 increased significantly from the main pulmonary artery to the pulmonary capillary wedge region ([mean +/- SEM] 3.1 +/- 0.23 vs. 4.6 +/- 0.36 pg/ml, p < 0.01), and the increase was related to the severity of heart failure. Among the various vasoconstrictor factors, such as plasma active renin concentration, plasma angiotensin II, plasma norepinephrine, femoral venous plasma endothelin-1 and pulmonary endothelin-1 spillover, only endothelin-1 spillover in the lungs showed an independent and significant correlation with pulmonary vascular resistance (r = 0.82, p < 0.001). CONCLUSIONS: The main source of circulating endothelin-1 is not the peripheral vascular bed but the pulmonary vascular bed in patients with chronic heart failure. In addition, endothelin-1 secretion in the lungs may regulate the pulmonary vascular resistance in patients with chronic heart failure. These findings are consistent with a significant role for endogenous endothelin-1 in the pathophysiology of heart failure, especially in the pulmonary circulation.
Adult↗
Cardiac myxoma with Gamna-Gandy bodies: case report with MR imaging.
We describe a patient with a left atrial tumor that showed very unusual magnetic resonance (MR) imaging features. Postsurgical histologic examination showed characteristics of myxoma, and Gamna-Gandy bodies were found in the tumor. We believe that the unusual MR appearance was caused by the hemosiderin in the Gamna-Gandy bodies.
Aged↗
Pig intestinal membrane-bound receptor (guanylyl cyclase) for heat-stable enterotoxin: cDNA cloning, functional expression, and characterization.
A cDNA encoding the receptor protein for a heat-stable enterotoxin (STa) produced by enterotoxigenic Escherichia coli was cloned from intestinal epithelial cells of a 10-week-old pig. The cDNA had an open reading frame of 3,219 base pairs and coded for a protein with 1,073 amino acid residues. The mature protein consisted of 1,050 amino acid residues with a molecular mass of ca. 121 kDa and was 87% and 82% identical with the human and rat protein, respectively. The CHO cell line overexpressing the pig recombinant STa receptor specifically bound to a photoaffinity-labeled analog of STa and showed marked elevation of the cellular content of cGMP in response to STa.
Amino Acid Sequence↗
Regulation of the Escherichia coli hfq gene encoding the host factor for phage Q beta.
The host factor (HF-I) for phage Q beta RNA replication is a small protein of 102 amino acid residues encoded by the hfq gene at 94.8 min on the Escherichia coli chromosome. The synthesis rate of HF-I at the exponential-growth phase is higher than at the stationary phase, and it increases concomitantly with the increase in cell growth rate. The intracellular level of HF-I is about 30,000 to 60,000 molecules per cell, the majority being associated with ribosomes as one of the salt wash proteins. Taken together, we suggest that HF-I is one of the growth-related proteins.
Allolevivirus↗