[Diagnosis of anemia--analysis of erythrocyte ferritin].
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Anemia↗
Biomedical subjects
T Miyazaki
Publications and source records attributed to T Miyazaki.
Signal transduction mediated by the reconstituted IL-2 receptor. Evidence for a cell type-specific function of IL-2 receptor beta-chain.
The binding of IL-2 to its specific receptor (IL-2R) triggers various cellular events including the induction of nuclear proto-oncogenes (c-fos, c-jun and c-myc genes) and the proliferation of hemopoietic cells. In the present study, we have established NIH 3T3 fibroblasts in which the three IL-2R subunits, the alpha-chain (IL-2R alpha), the beta-chain (IL-2R beta), and the gamma-chain (IL-2R gamma), are constitutively expressed. The resulting cell lines express high affinity IL-2R on their cell surface at levels comparable with those of IL-2-responsive lymphoid cells. We observed that the high affinity IL-2R in NIH 3T3 fibroblasts can mediate the IL-2-stimulated tyrosine phosphorylation of p42/p44 (mitogen-activated protein kinase) and the induction of the c-fos, c-jun and c-myc genes. In NIH 3T3 fibroblasts the high affinity IL-2R bearing a deletion of a region rich in acidic amino acids (the "acidic" region) in the IL-2R beta-chain failed to induce the tyrosine phosphorylation of MAP kinase as well as the expression of the all three nuclear proto-oncogenes. On the other hand, our previous studies had demonstrated that the high affinity IL-2R bearing the same mutant IL-2R beta-chain retained the ability to induce c-myc gene in response to IL-2 in a murine IL-3-dependent pro-B cell line, BAF/B03. Hence, these results reveal the underlying complexity of signal transduction among different cell types. The inability of the reconstituted high affinity receptor to mediate the IL-2-induced proliferation of NIH 3T3 fibroblasts suggests that induction of the three nuclear proto-oncogenes and the tyrosine phosphorylation of mitogen-activated protein kinase in NIH 3T3 fibroblasts are not sufficient to induce cellular proliferation.
3T3 Cells↗
p53 gene mutation in the chronic phase was not detected in the myeloid crisis of a chronic myelocytic leukemia case.
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Adult↗
Electronic structure of hole-doped Sr1+xLa1-xFeO4 studied by UPS and XAS.
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Journal Article↗
Hepatocyte growth factor specifically binds to sulfoglycolipids.
Hepatocyte growth factor (HGF) is a heparin-binding pleiotropic factor that acts on a variety of epithelial cells. The interaction of human HGF with glycolipids was studied by overlaying them with 125I-HGF on thin layer chromatograms and by a solid-phase assay using lipids adsorbed on microtiter plates. Among various glycolipids tested, HGF was found to bind to sulfoglycolipids, including galactosylceramide sulfate (SM4), lactosylceramide sulfate (SM3), and gangliotriaosylceramide bis-sulfate. In contrast, HGF failed to bind to gangliosides or neutral glycolipids. HGF binding to SM4 was strongly inhibited by dextran sulfate, heparin, and fucoidan, whereas neither keratan sulfate nor hyaluronic acid had any inhibitory activity. When glycolipids from a renal cancer cell line, SMKT-R3, which overexpresses sulfoglycolipids, were developed on a thin layer chromatogram, SM4 and SM3 were the only glycolipids that bound HGF. We further examined the effect of the incorporation of glycolipids into SMKT-R3 cells on HGF binding to the cells. The incorporation of SM4 into the cells enhanced HGF binding to SMKT-R3 cells, while that of galactosylceramide, a precursor of SM4, had no effect. These observations indicated that SM4 exogenously incorporated into the cell membranes could react with HGF and suggested that endogenous sulfoglycolipids on SMKT-R3 cells might function as reservoirs for HGF.
Carcinoma, Renal Cell↗
Cell-surface sulfoglycolipids are involved in the attachment of renal-cancer cells to laminin.
We investigated the role of sulfoglycolipids on human renal-cell carcinoma cells (SMKT-R3) in the attachment to a substrate adhesive protein, laminin. SMKT-R3 cells over-express sulfoglycolipids, including SM2, SM3 and SM4. When acidic glycolipid fractions, were extracted from SMKT-R3 cells, separated by HPTLC, and then overlaid with laminin, laminin bound specifically to SM3 and SM4. A monoclonal antibody, Sulph-1, reacting with SM3 and SM4 inhibited attachment of the cells to laminin but not to fibronectin, in a dose-dependent manner. In addition, when exogenous SM4 was incorporated into the cells, their attachment to laminin, but not to fibronectin, was enhanced. On the other hand, the incorporation of GalCer, which is a precursor of SM4, had no effect on adherence of the cells to laminin or to fibronectin. We also assayed haptotaxis, tumor-cell migration along a gradient of substratum-bound laminin. The incorporation of SM4 into the cells caused an approximately 3-fold increase of the haptotactic response to laminin compared with non- or GalCer-incorporation. These results taken together suggest that sulfoglycolipids on renal-cancer cells are involved in attachment to laminin and that they can modulate the metastatic potential of renal-cell carcinoma cells.
Carcinoma, Renal Cell↗
Hepatocyte growth factor elevates the activity levels of glycolipid sulfotransferases in renal cell carcinoma cells.
Accumulation of sulfoglycolipids associated with markedly elevated activity levels of glycolipid sulfotransferases has previously been demonstrated in the human renal cell carcinoma cell line, SMKT-R3. To elucidate the regulatory mechanisms of sulfoglycolipid synthesis in SMKT-R3 cells, the effects of various growth factors on the metabolic enzymes of sulfoglycolipids were investigated. Hepatocyte growth factor (HGF) significantly increased the activity levels of the sulfotransferases in a dose-dependent manner, but did not change that of arylsulfatase A, which hydrolyzes sulfoglycolipids. Scatchard analysis of 125I-HGF binding to SMKT-R3 cells indicated that the cells expressed high-affinity receptors for HGF with a Kd of 36 pM and 750 sites/cell. Furthermore, metabolic labeling with [35S]sulfate revealed that the addition of HGF to the culture medium of the cells resulted in an increment of sulfoglycolipid synthesis. Therefore, these observations suggest that HGF can function as a regulatory factor in sulfoglycolipid synthesis through the modulation of the sulfotransferase activity levels in renal cell carcinoma cells. In addition, HGF stimulated the proliferation and motility of SMKT-R3 cells, suggesting that HGF has multiple biological activities in renal cell carcinoma cells.
Animals↗
Characteristics of Na+ current in Schwann cells cultured from frog sciatic nerve.
Characteristics of voltage-dependent currents in cultured frog Schwann cells were investigated by the whole-cell clamp technique. An inward current was detectable at a membrane potential level more positive than -50 mV and reached a maximum value at about -10 mV, while no rectifying channel was present. The inward current was carried by Na+ ions, because the extrapolated reversal potential of the current agreed with the calculated ENa, and the current was sensitive to tetrodotoxin. The membrane potential for half-maximal inactivation was -82 mV. The inactivation curve indicated that more than 90% of the Na+ channels were inactivated at the resting membrane potential, suggesting that the cultured frog Schwann cells could not generate an action potential under physiological conditions. The time constant for the inactivation at a maximum current was 5.3 ms (-10 mV, 13 degrees C). The electrophysiological characteristics of the Na+ current in the cultured frog Schwann cells were compared with those in other tissues. This Na+ current was quantitatively different from that observed in the amphibian node of Ranvier but was similar to that in the mammalian Schwann or glial cells, especially in the more hyperpolarized half-maximal inactivation potential and in the slower inactivation time course.
Animals↗
Alteration of aldolase isozymes in serum and tissues of patients with cancer and other diseases.
We studied the alteration of aldolase isozymes in the serum and tissues of patients with cancer and other diseases using radioimmunoassays specific for aldolase A, B, and C subunits. Aldolase B was predominantly found in adult liver, where aldolase A and C were distinctly low. Aldolase A and B showed almost the same concentration in fetal liver, while in neonatal liver aldolase B protein concentrations were much higher than aldolase A. In contrast, aldolase A was the predominant isozyme found in hepatoma and gastric cancer tissues, whereas aldolase B was distinctly low in hepatoma tissues, and extremely low in gastric cancer tissues. These results suggest that the aldolase A is a more fetal type of liver isozyme than the aldolase B and C, and aldolase B is a more differentiated type of liver isozyme than aldolase A and C. Serum FDP aldolase activities were elevated in half of patients with liver diseases, all patients with muscle diseases and a few patients with cancer. Serum aldolase A levels were elevated in patients with muscle diseases and cancer, but not elevated in patients with liver diseases. In contrast, serum aldolase B levels were elevated in patients with liver disease, but not elevated in patients with muscle diseases and other diseases without liver injury. Serum aldolase B levels showed a trend to decrease in cancer patients with normal GPT levels. Serum aldolase A/B ratios were significantly increased in cancer patients with normal GPT levels, whereas they showed the decreased levels in patients with liver diseases.(ABSTRACT TRUNCATED AT 250 WORDS)
Fructose-Bisphosphate Aldolase↗
Detection of Candida enolase antibody in patients with candidiasis.
The serodiagnostic value of candida enolase antibody was evaluated in 27 patients with systemic candidiasis. The glycolytic enzyme enolase was prepared from Candida albicans IFM 40009 and used as the antigen for immunoblot assays. IgG class antibody was found in the sera of 17 of 27 patients (62.9%) obtained during the early course of the infection. Serial sampling of sera increased the number of patients with positive Candida enolase antibody, which was detected in 25 of 27 patients (92.5%). None of the patients with localized candidiasis showed a positive IgG titer more than 1:100. The specificity of Candida enolase IgG antibody was 95%. This antibody was observed in various candida species: Candida albicans (6/7, 85.7%); C. parapsilosis (7/9, 77.9%); C. tropicalis (5/5, 100%); C. guilliermondii (4/4, 100%); and C. glabrata (1/3, 33.3%).
Adult↗
Electron spin resonance study of radiosterilization of antibiotics: ceftazidime.
Fundamental information on radiosterilization of ceftazidime was obtained by electron spin resonance (ESR) measurement of free radicals produced in gamma-ray-irradiated ceftazidime at 4.2-295 K. Three types of free radicals are produced by gamma-irradiation. The first one shows septet lines in an ESR spectrum and decays at 230 K: it is assigned as a .C(CH3)2COOH radical. The second one shows triplet lines and decays at 293 K: it is assigned as iminoxyl radicals (> C = N-O.). The third one shows a broad singlet line and survives even at 295 K. The plausible assignment of this spectrum is discussed. It is concluded that an O-C bond of the gamma-irradiated ceftazidime is ruptured, producing the iminoxyl radicals and .C(CH3)2COOH radicals. The yields of free radicals increase linearly with the increasing dose of gamma-irradiation up to 10 kGy at 295 K as well as 77 K. Half of the radicals that show a broad singlet line survive at 295 K upon storage of the irradiated ceftazidime for 159 days. It is concluded that the ESR measurement of free radicals gives a good method for discrimination of irradiated antibiotics from non-irradiated ones and an estimation of an irradiation dose.
Ceftazidime↗
Estimation of irradiation dose of radiosterilized antibiotics by electron spin resonance: ampicillin.
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Ampicillin↗
Expression of adhesion molecules on human hematopoietic progenitor cells at different maturational stages.
In this report we examined the expression of several adhesion molecules on human hematopoietic progenitor cells at different maturational stages. Human hematopoietic progenitor cell-enriched fractions were prepared from bone marrow cells by depleting lymphocytes and monocytes (CD2+, CD14+ and CD19+ cells). These cells were separated into adhesion molecule-positive and -negative cell populations by immunomagnetic separation methods and then assessed for their ability to form various colony forming cells (CFC). CD44 and CD49d were expressed on multipotent hematopoietic progenitor cells, or mixed colony forming units (CFU-Mix), erythroid burst forming units (BFU-E), granulocyte-macrophage CFU (CFU-GM) and erythroid CFU (CFU-E). Leu8 was expressed on CFU-Mix, BFU-E and some populations of CFU-GM, but not CFU-E. CD11a was expressed on some populations of CFU-Mix, CFU-GM and BFU-E. CD54 was expressed only on some populations of CFU-GM. These results suggest that Leu8, CD44, CD49d and CD11a appear to play important roles in the differentiation and proliferation of human hematopoietic progenitor cells at different maturational stages in the bone marrow microenvironment.
Antigens, CD↗
Blockade of ATP-sensitive potassium channels by 5-hydroxydecanoate suppresses monophasic action potential shortening during regional myocardial ischemia.
We tested 5-hydroxydecanoate (5-HD), a specific blocker of ATP-sensitive potassium channels (IK.ATP), to determine if mitigates electrophysiologic changes produced by regional myocardial ischemia in vivo. A sequence of 5-minute occlusion of the distal LAD and 30-minute reperfusion was repeated while recording the monophasic action potential (MAP) and bipolar electrogram (EG) from the epicardial center of the ischemic myocardium in anesthetized dogs. 5-HD (30 mg/kg, i.v.) or glibenclamide (0.15 or 0.3 mg/kg, i.v.) was administered before the third occlusion, and the data were compared to the second occlusion data. 5-HD did not affect baseline MAP duration at 90% and 50% repolarization (APD90, APD50) before LAD occlusion but suppressed occlusion-induced shortening of APD90 (16 +/- 2% during the second occlusion vs. 5 +/- 3% during the third occlusion, n = 8, p < 0.01) and APD50 (16 +/- 3% vs. 10 +/- 3%, n = 8, p < 0.05). Pretreatment with glibenclamide also suppressed occlusion-induced MAP shortening and eliminated an additional effect of 5-HD (n = 3). 5-HD did not affect the occlusion-induced increase in duration and activation time of EG. 5-HD, as well as glibenclamide, suppressed regional ischemia-induced MAP shortening, probably by blocking activation of IK.ATP, without affecting conduction delay. These differential effects of 5-HD on repolarization and conduction during the early phase of regional ischemia might have the potential to suppress reentrant ventricular arrhythmias.
Action Potentials↗
The Eiken Latex test for detection of a cryptococcal antigen in cryptococcosis. Comparison with a monoclonal antibody-based latex agglutination test, Pastorex Cryptococcus.
A latex agglutination test for cryptococcal antigen, the Eiken Latex test (Eiken, Tokyo, Japan), was compared with a monoclonal antibody-based agglutination assay, Pastorex Cryptococcus (Diagnostics Pasteur, Marneur-la-Coquette, France). In a murine model of disseminated cryptococcosis, the kinetics of the antigen titers by the Eiken Latex were similar to those by the Pastorex Cryptococcus, but sensitivity was much higher. In HIV-negative patients with pulmonary cryptococcosis, a cryptococcal antigen was detected in 6 of 10 patients by the Eiken Latex test and in only 3 of those patients by the Pastorex Cryptococcus. The results indicate that the Eiken Latex is more sensitive for the detection of the cryptococcal antigen, even in non-disseminated cryptococcosis. The sensitivity and specificity of the Eiken Latex were examined using 195 sera from 25 patients with pulmonary cryptococcosis and 170 patients with non-cryptococcosis. The cutoff value of > or = 1:8 showed a sensitivity of 76% (19/25) and a specificity of 98.9% (168/170).
Agglutination Tests↗
In vitro and in vivo antifungal activities of liposomal amphotericin B, and amphotericin B lipid complex.
The in vitro and in vivo antifungal activities of liposomal amphotericin B (L-AMPH) and amphotericin B lipid complex (ABLC), which is composed of amphotericin B and the phospholipids dimyristoyl phosphatidylcholine and dimyristoyl phosphatidylglycerol, were compared with those of conventional amphotericin B (Fungizone, AMPH). The acute intravenous toxicity was markedly lower in BALB/c mice; 50% lethal doses (LD50s) were 2.75 mg/kg in AMPH, 32.9 mg/kg in L-AMPH and > 75 mg/kg in ABLC. In vitro antifungal activities against Candida albicans, C. parapsilosis, C. tropicalis, C. glabrata, and C. krusei were evaluated by the agar plate dilution method. The activities were unchanged against C. albicans, but MICs increased more than four fold in 18 of the 20 strains other than C. albicans in L-AMPH and in 9 of the 20 in ABLC. L-AMPH and ABLC were as efficacious as AMPH in the treatment of mice infected with C. albicans, and at a dose of 0.5 and 1.0 mg/kg of body weight, ABLC was more efficacious on survival A ten-times larger dose (10 mg/kg) of L-AMPH and ABLC was administered to mice with 100% survival, suggesting improved tolerability as compared to amphotericin B.
Amphotericin B↗
A rapid method for detecting barbiturates in serum using EI-SIM.
A simple and rapid method for analysis of barbiturates in serum has been developed. In order to extract and clean barbiturates in serum, a separation column packed with Extrelut and Florisil was used, and the eluate was directly analyzed by means of electron impact selected ion monitoring (EI-SIM). Selected ions used were base peak ions of 10 barbituartes, and the internal standard used was allobarbital or secobarbital. The calibration curves were linear over the range 0.5-5 ng. Extraction of replicate serum samples containing 20 micrograms/1.5 ml and 5 micrograms/1.5 ml resulted in a recovery of 87.2-105.2% and 81.6-104.6%, respectively, with the exception of phenobarbital, which was 151.9% and 172.1%, respectively. Secobarbital was also analyzed in the serum of 13 patients who had been given secobarbital intravenously. In 3 out of 10 cases, secobarbital levels greater than 1 micrograms/ml were detected more than 72 h after administration. This method seems to have possibilities for clinical use.
Adult↗
Evaluation of mixed chimerism by two-step polymerase chain reaction amplification of hypervariable region MCT118 after allogeneic bone marrow transplantation.
We assessed the origin of peripheral blood cells and bone marrow cells of 92 samples obtained from 19 patients after allo BMT by two-step polymerase chain reaction (PCR) amplification of MCT118, one of the variable number of tandem repeat regions (VNTR) which has a different length in 19 of 32 sibling pairs examined, that can detect the DNA pattern of a minor cell population of only 1% without using radioisotopes. Mixed chimerism (MC) was detected in the hematopoietic cells of four patients. Two patients who showed progressive MC developed relapse of leukemia 3 months and 4 months after the detection of MC, and two patients died with bone marrow hypoplasia 61 days and 7 months after BMT. These data suggest the clinical importance of analyzing the correlation of MC and clinical complications after BMT by this method, which can be used to monitor MC in about two thirds of allo BMT patients with an adequate sensitivity.
Base Sequence↗