[A case of nephrotic syndrome with a peculiar glomerular lesion].
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Adult↗
Biomedical subjects
A Murayama
Publications and source records attributed to A Murayama.
[Analysis of the glucocorticoid receptor of bovine tracheal smooth muscle].
Though the therapeutic efficacy of glucocorticoids on bronchial asthma has been attributed mainly to their anti-allergic and anti-inflammatory effects, the possibility of intervention of glucocorticoids in bronchial asthma might be an important parallel process. However, little is known about the mechanism of glucocorticoid action on tracheal muscle and there is insufficient basic information about the glucocorticoid receptor of tracheal muscle. Accordingly we set out to analyze the glucocorticoid receptor of bovine tracheal muscle. Bovine tracheal muscle was used because it is plentiful enough for biochemical analysis. Our results showed that a small amount (0.08 pmol/mg.protein) of glucocorticoid binder of high affinity (kd = 4.6 nM) was present in the cytosol of bovine bronchial muscle. The binder did not show any affinity for estradiol, dihydrotestosterone or andronsterone, but showed a low affinity for progesterone. Furthermore, the binder underwent salt-dependent variation of its molecular form (9.2S under low salt condition and 4.6S under high salt condition) typical of steroid receptors. These results indicated the presence of a glucocorticoid receptor in the tracheal muscle, suggesting that glucocorticoid acts on bronchial muscle in therapeutic practice on bronchial asthma.
Animals↗
Expression of soybean glycinin subunit precursor cDNAs in Escherichia coli.
As the cDNAs encoding A1aB1b and A2B1a subunit precursors of the glycinin A2 subfamily contain a unique NcoI site sequence, (A)CCATGG, occurring at their translation initiation sites, plasmids were constructed to direct the synthesis of those precursor proteins by inserting NcoI/PstI fragments derived from those cDNA clones into the NcoI/PstI-pKK233-2 expression vector in Escherichia coli MV1190, respectively. The resultant plasmids directed the expression of 57-kDa protein components that have molecular masses in agreement with those of the in vitro translation products directed by glycinin A2 subfamily mRNAs, by the addition of isopropyl beta-D-thiogalactoside. These proteins, which comprised as much as 1% of the total bacterial protein, are immunoprecipitable with rabbit antibodies specific for glycinin subunits. This procedure makes glycinin subunits available as a model for studying structure-function relationships in seed proteins using site-directed mutagenesis. This is the first expression of glycinin-like storage protein in E. coli.
DNA↗
Effect of molybdate on the molecular organization of the estrogen receptor system of porcine uterus.
Molybdate was shown to have complex effects in modulating the molecular organization of the constituents of the estrogen receptor (ER) system of porcine uterus. We showed previously the presence of one basic ER molecule (vero-ER) (sedimentation coefficient, 4.5S; Stokes radius, 44 A) and ER-binding factors (ERBFs) ["8S" ER-forming factor ("8S" ER-FF), (component A) X (component B)6; "6S" ER-FF, (component B)6; "5S" ER-FF, component A] in the porcine uterus [Fukai, F. & Murayama, A. (1981) J. Biochem. 95, 1697-1704]. Molybdate regulates the specific interaction of vero-ER with ERBFs in a complex way. The apparent Kd value (6.7 X 10(-10) M) of vero-ER with "8S" ER-FF in the presence of molybdate (30 mM) was decreased remarkably as compared with that (2.7 X 10(-9) M) in the absence of molybdate. In contrast, the apparent Kd value (3.7 X 10(-9) M) of vero-ER with "5S" ER-FF observed in the presence of molybdate (30 mM) was increased over ten-fold as compared with that in the absence of molybdate. Meanwhile, the affinity (Kd, 5 X 10(-9) M) of vero-ER for "6S" ER-FF was scarcely influenced by molybdate. These results reveal the mechanism by which molybdate selectively stabilizes "8S" ER. Molybdate further affected the molecular constitution of ERBFs. The dissociation of "8S" ER-FF into component A and component B, which takes place under hypertonic (0.4 M KCl) conditions at higher temperature (25 degrees C), was suppressed almost completely by molybdate (30 mM).(ABSTRACT TRUNCATED AT 250 WORDS)
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Capacitive effects on canine coronary artery diastolic pressure-flow relationships.
The extent of capacitive effects on coronary diastolic pressure-flow relationships was investigated by proposing a theoretical model and by actually measuring the time constants of the coronary and systemic circulation. The dynamic pressure-flow relationship obtained by an aortic declining pressure perfusion and the static pressure-flow relationship obtained by constant pressure reservoir perfusion were compared to support the theoretical model, which revealed that the capacitive effects could be quantified by the alpha value, i.e. the ratio of the time constants of the coronary and systemic circulations. In 10 anesthetized dogs, the two independent methods disclosed that capacitive effects are limited and diastolic outflow pressure in the coronary circulation is the static zero-flow pressure intercept determined by the vascular tonus and intramyocardial pressure.
Animals↗
Association and dissociation of estrogen receptor with estrogen receptor-binding factors is regulated by Mg2+.
It was recently shown that uterine estrogen receptor (ER) is translocated from the cytoplasm into the nucleus in the form of vero-ER X E (basic ER molecule bound with estradiol), and the translocation is inhibited by the specific binding of vero-ER X E with the cytoplasmic protein factors designated as ER-binding factors (ERBFs) ["5S" ER-forming factor ("5S" ER-FF), "6S" ER-FF, "8S" ER-FF] [Murayama, A. & Fukai, F. (1983) J. Biochem. 94, 511]. It was found that the specific interaction of vero-ER X E with the ERBFs is regulated by Mg2+ at physiological concentrations. The apparent Kd values of vero-ER X E for the ERBFs ["5S" ER-FF, 2.7 X 10(-9) M; "6S" ER-FF, 6.0 X 10(-8) M; "8S" ER-FF, 3.5 X 10(-8) M] observed in the presence of 1 mM Mg2+ were all increased over 10 times as compared with in the absence of Mg2+. The inhibitory effects of the ERBFs on the nuclear translocation of vero-ER X E were reduced by Mg2+.
Animals↗
Action of calpain on the basic estrogen receptor molecule of porcine uterus.
Basic estrogen receptor (ER) molecule (vero-ER) of the cytosol of porcine uterus was purified 1,200-fold after successive chromatographies on phenyl-Sepharose, hydroxylapatite, and DEAE-cellulose, followed by Sephadex G-150 gel filtration. The purified vero-ER was completely free from endogenous protease and ER-binding factor. The action of Ca2+-dependent cysteine proteinase (calpain) on vero-ER was studied by utilizing the purified receptor and calpains from porcine uterus (endogenous calpain), porcine kidney, and human erythrocytes. Proteolysis of vero-ER was followed by monitoring the disappearance of the binding capability of vero-ER with "8S" ER-forming factor. Vero-ER was proteolyzed by both the endogenous and the exogenous calpains in the presence of Ca2+. The calpains did not attack vero-ER in the absence of Ca2+. The results indicated the absolute requirement by calpain for Ca2+ for the limited hydrolysis of vero-ER. Uterine cytosol was shown to contain, in parallel with calpain, a protease which does not require Ca2+ for the limited proteolysis of vero-ER. The strongly hydrophobic domain of vero-ER, recently shown to be indispensable for the nuclear translocation of vero-ER (Murayama, A. & Fukai, F. (1983) FEBS Lett. 158, 255), was preferentially destroyed by both the Ca2+-requiring and -nonrequiring enzymes. It was assumed that calpain might intervene in the estrogen action by diminishing irreversibly the amount of the cytoplasmic ER capable of translocating into the nucleus.
Animals↗
[Vasodilator therapy in ischemic heart disease].
A comparative study of venodilating and veno-arterial dilating drugs was conducted for clinical patients and for experimental animals. The subjects were 21 patients with ischemic heart disease associated with left ventricular failure. Fourteen patients received venodilators alone (Group I), and seven were administrated venoarterial dilators alone (Group II). At the time of the maximum effect, the two groups showed similar improvement. In Group I, the left ventricular filling pressure (LVFP) was reduced from 24 to 16 mmHg and the cardiac index (CI) was increased from 2.5 to 2.8 l/min/m2. In Group II, the LVFP was similarily reduced from 27 to 16 and the CI increased from 2.4 to 2.9. The effects of each drug on the total vascular resistance/LVEP were similar in both groups. In animals with partial occlusion of the left anterior descending artery, intravenous administration of nitroglycerin (TNG) or nitroprusside (NPS) lowered the aortic pressure stepwise at intervals of 10 mmHg from a mean of 130 mmHg. As a result, segmental shortening was improved either by TNG or NPS. However, when the systolic aortic pressure was decreased below 100 mmHg, segmental shortening of the ischemic area was aggravated in dogs receiving either drug. The end-diastolic length showed no difference between the dogs administrated TNG and NPS. When the effects of TNG and nifedipine (NFP) were compared to those in dogs with the old infarcted myocardium, both drugs produced improvement in hypokinetic segments. TNG induced a decrease in the end-diastolic length, whereas NFP revealed no changes, reflecting different modes of improvement.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
The strong hydrophobic domain of the activated estrogen receptor of porcine uterus.
Basic estrogen receptor (ER) molecule (vero-ER) of porcine uterus, which was previously shown to be the activated ER necessary to translocate from the cytoplasm into the nucleus, possesses a strongly hydrophobic nature. The strong hydrophobicity of vero-ER was concealed through binding with ER-binding factors (ERBFs). Vero-ER lost its strong hydrophobicity and its capability to bind with ERBFs after limited proteolysis by endogenous protease. The strong hydrophobic domain of vero-ER, indispensable for the nuclear translocation, was assumed to be located near the binding site with ERBFs.
Animals↗
Studies on the molecular mechanism of the translocation of estrogen receptor from the cytoplasm into the nucleus.
A detailed study of the molecular mechanism of the translocation of estrogen receptor (ER) from the cytoplasm into the nucleus was undertaken in an in vitro system of porcine uterus. The capabilities of vero-ER . E (basic ER molecular bound with estradiol) (sedimentation coefficient 4.5S; Stokes radius 44 A) and the complexes ["5S" ER . E, (vero-ER . E) . (component A); "6S" ER . E, (vero-ER . E) . (component B)6; "8S" ER . E, (vero-ER . E) . (component B)6 . (component A)] with ER-binding factors (ERBFs) to translocate into the isolated nuclei were estimated by subtracting the amounts of ER adsorbed by the nuclear envelopes from those of ER bound to the whole nuclei. The results strongly supported our previous assumption that vero-ER . E translocates into the nuclei, and the complexes with ERBFs do not. The results suggested also that the binding site of vero-ER to ERBFs is required to be unoccupied in the process of the translocation of ER from the cytoplasm into the nucleus. The presence of a cytoplasmic factor (component C) which binds specifically with "5S" ER . E under low salt conditions was indicated. The complex, ("5S" ER . E) . (component C), was shown to possess relatively high affinity towards nuclear envelopes, but not to translocate into the nuclei.(ABSTRACT TRUNCATED AT 250 WORDS)
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[Multiserial ECG-gated computed tomographic method: Evaluation of left ventricular volumes, wall thickness and wall motion].
The purpose of this study was to investigate the potential usefulness of multiserial ECG-gated computed tomography (CT) (GE CT/T 8800) for a quantitative assessment of left ventricular volume, wall thickness and wall motion in 15 patients with coronary heart disease. In order to obtain ideal left ventricular short-axis images, the gantry was tilted to the cephalad 15 degree with patients on the right lateral position. Five ECG-gated scans (5.6 sec for one scan) were performed in each patient with intravenous injection of contrast material (Urografin 76). Left ventricular volumes were computed by a modified Simpson's rule and segmental wall motion was analyzed at three levels (base, papillary muscle and apex) by tracing the endocardial outline at end-diastole and end-systole. Left ventricular systolic and diastolic volumes ( LVSV , LVDV ) had a good correlation with those measured by left ventriculography (LVG) (r = 0.75 and 0.66). Wall thickness of the interventricular septum and left ventricular posterior wall obtained by CT was correlated with those by M-mode echocardiography (r = 0.82 and 0.71). Comparative analysis of segmental wall motion by CT, two-dimensional (2 D) echocardiography and left ventriculography was done. The 2 D echocardiographic segments with normal and abnormal wall motion were accurately identified by CT, best at the papillary muscle level (91%), and less at the base (71%) and the apex (73%). Similarly, 85% of left ventriculographic segments could be detected by CT. In conclusion, ECG-gated computed tomography proved to be a useful method for evaluating left ventricular function in coronary heart disease.
Adult↗
Inhibition of the translocation of estrogen receptor from the cytoplasm into the nucleus by estrogen receptor-binding factors.
The molecular mechanism of the translocation of estrogen receptor (ER) from the cytoplasm into the nucleus was studied in an in vitro system taking into account the specific interactions of the basic constituents of the uterine ER-system which had been reported previously (Murayama, A. et al. (1980) J. Biochem. 88, 1457). It was found that ER is translocated from the cytoplasm into the nucleus in the form of vero-ER . estradiol (basic ER-unit bound with estradiol; sedimentation coefficient, 4.5S; Stokes radius, 44 A; Kd for estradiol, approximately 10(-10) M) dissociated from coexisting estrogen receptor-binding factors (ERBFs). ERBFs inhibited the translocation of vero-ER into nuclei through binding with it. The apparent equilibrium dissociation constants (Kd) of ERBFs for vero-ER were estimated by Scatchard analysis to be 2.6 X 10(-10) M ["5S" ER-forming factor ("5S" ER-FF)], 3.0 X 10(-9) M ("6S" ER-FF), and 2.5 X 10(-9) M ("8S" ER-FF), respectively. The inhibitory effects of ERBFs on the binding of vero-ER to the nucleus paralleled their association constants.
Animals↗
[Echocardiographic findings of a patient with cardiac amyloidosis and left ventricular outflow obstruction].
This report described 56-year-old male with cardiac amyloidosis, whose echocardiographic and hemodynamic findings suggested hypertrophic obstructive cardiomyopathy. M-mode echocardiography demonstrated asymmetric septal hypertrophy (2.1 cm/1.6 cm), systolic anterior motion of the mitral valve, and pericardial effusion. Two-dimensional echocardiography revealed marked hypertrophy of the septum and papillary muscles with characteristic fine grannular sparkling appearance of the myocardium. Bilateral heart catheterization showed normal hemodynamics except a pressure gradient of 35 mmHg within the left ventricular outflow tract. Moreover, no hemodynamic evidence of restrictive cardiomyopathy was observed. Myocardial biopsy was performed at the time of pericardiectomy, and diffuse amyloid deposits were identified between myocardial fibers. It is of particular interest that cardiac amyloidosis presented the form of ventricular hypertrophy associated with left ventricular outflow obstruction, probably due to uneven deposition of amyloid.
Amyloidosis↗
[Evaluation of mitral valve, subvalvular structures and valvular flexibility in mitral stenosis by two-dimensional echocardiography].
UNLABELLED: In order to identify the determinants for surgical procedures in mitral stenosis, we evaluated two-dimensional echocardiographic findings of the mitral valve and subvalvular structures in 35 patients undergoing open mitral commissurotomy (OMC) or valve replacement (MVR). As indices of a degree of subvalvular shortening and valvular flexibility, the distance between the mitral ring and the tip of the anterior mitral leaflet was measured by the LV long-axis view by in both midsystole (S) and early diastole (D). As a possible major determinant for MVR, a degree of valvular calcification (C) was semi-quantatively scored according to the extent of abnormally strong echo density. In nine of 11 patients undergoing MVR, a main reason for selecting MVR was a marked thickening or shortening of subvalvular structures. In patients in whom OMC was feasible, the degree of improvement of the mitral valve area (delta MVA) was assessed by the pre- and post-operative mitral valve areas (MVA) measured on the LV short-axis view, which were averaged 0.15 and 1.38 cm2, respectively. RESULTS: The index C was significantly higher in cases with MVR than those with OMC (9.2 +/- 2.6 vs 4.7 +/- 2.3 points, p less than 0.001), although there was a significant overlap between these two groups and index C did not correlate with delta MVA in the OMC patients. Similarly, the value S was significantly smaller in patients undergoing MVR than those undergoing OMC (1.2 +/- 0.4 vs 0.7 +/- 0.2 cm, p less than 0.001), though S did not correlate with delta MVA. On the other hand, the index of valve flexibility D--S was smaller in patients undergoing MVR (0.5 +/- 0.3 vs 0.8 +/- 0.3 cm, p less than 0.05) and correlated well with delta MVA (delta MVA = 0.699 x (D--S)+0.007, R = 0.678, p less than 0.02) in patients undergoing OMC. Furthermore, in all patients undergoing OMC with D--S greater than or equal to 0.8 cm, delta MVA was above 0.5 cm2, contrasting with delta MVA of 0.5 cm2 or less in 6 of 7 patients with D--S less than 0.7 cm. Using these indices, surgical procedures were successfully predicted in another 7 prospectively studied patients and predicted delta MVA in 4 patients was quite comparable with actual delta MVA. It was concluded that measurements of S and D by two-dimensional echocardiography are useful, 1) to predict patients requiring MVR and 2) to predict patients with inadequate delta MVA in whom OMC is surgically feasible.
Adult↗
Protection of steroid hormone receptors by protease inhibitors.
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Animals↗
Cytoplasmic estrogen receptor system of gilt uterus. Partial purification of component B labeled with [14C]iodoacetic acid.
Analysis of the cytoplasmic estrogen receptor (ER) system of gilt uterus by protecting ER from proteolysis showed that the ER system of gilt uterus is similar to that of cow uterus described previously. There was only one native unit molecule [native "4S" ER (sedimentation coefficient, 4.5S; Stokes radius, 45 A; molecular weight, 82,000)] with specific affinity towards estradiol. This molecule was further designated as vero-ER. "8S" ER-forming factor [("8S"ER)-FF] (Stokes radius, 51 A) was separated from the cytosol under hypotonic (low salt) conditions, and this was further dissociated into component A ["5S" ER-forming factor, ("5S"ER)-FF] (Stokes radius, 37 A) and component B (Stokes radius, 18.5 A) in the presence of sodium thiocyanate. Vero-ER was proteolyzed in the absence of Ca2+ ion by a cytoplasmic protease into modified "4S" ER (sedimentation coefficient, 4.5S; Stokes radius, 35 A; molecular weight, 65,000) which was further designated as secto-ER. The constituents of the cytoplasmic ER system of cow and gilt uteri cross-reacted with each other to undergo similar molecular assembly as in their own systems. When analyzed for various uterine specimens, fluctuation of ("8S"ER)-FF-level (1-50 unit/g tissue) was more remarkable than that of ER-level (3-6 x (10(-12) mol/g tissue). Component B labeled with [14C]iodoacetic acid was purified over 1,500-fold. Mixture of vero-ER and component B labeled with [14C]iodoacetic acid formed "6S' ER with 14C-activities under hypotonic conditions. This clearly excluded the possibility that component B is a catalyzer of self-association (dimerization) of vero-ER.
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Characterization of estrogen receptor, estrogen receptor binding factors and a protease in the nuclei of gilt uterus.
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Animals↗
Pulmonary embolism and hemolytic anemia in a patient with cold agglutinin disease.
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Anemia, Hemolytic, Autoimmune↗