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Biomedical subjects

T Terao

Publications and source records attributed to T Terao.

At least 343 records · Page 19Linked to original sources
PubMed1991

The effect of staurosporine on Ca2+ signals in rat basophilic leukemia (RBL-2H3) cells.

We examined the effect of staurosporine on cytosolic calcium response in rat basophilic leukemia (RBL-2H3) cells using fura-2 as a fluorescent indicator of calcium ion. Staurosporine at a dose of 30 nM inhibited antigen-stimulated Ca2(+)-influx into the cells from the extracellular environment. In contrast, the drug at this concentration inhibited neither the mobilization of Ca2+ from intracellular stores nor inositol 1,4,5-trisphosphate (IP3) formation. At a high concentration (300 nM), however, staurosporine completely inhibited the cytosolic calcium responses as well as IP3 formation. These results indicate that staurosporine, if used at an appropriate concentration, can be used to discriminate Ca2(+)-influx from extracellular environment from mobilization of the ion from intracellular stores. These results also suggest that protein kinases, possibly protein kinase C, are involved in the calcium influx of RBL-2H3 cells from the extracellular environment. Serotonin release was strongly inhibited by the drug at 30 nM staurosporine. Since the inhibition of serotonin release and suppression of cytosolic calcium increase in response to the antigen were in parallel, we concluded that the inhibition of serotonin release from RBL-2H3 cells caused by the drug was elicited by the suppression of Ca2(+)-influx into the cells.

Alkaloids↗
PubMed1991

Reduction of 2,3,5-trimethyl-6-(3-pyridylmethyl)-1,4-benzoquinone by PB-3c cells and biological activity of its hydroquinone.

2,3,5-Trimethyl-6-(3-pyridylmethyl)-1,4-benzoquinone (CV-6504) has inhibitory activities on both thromboxane A2 synthase and 5-lipoxygenase as well as scavenging activity against active oxygen species. The latter two activities are closely related to the quinone moiety, which is reduced to a hydroquinone in the living body. Comparison of these two activities for both the quinone and hydroquinone showed that the hydroquinone form had superior activities. Concerning the reduction mechanism by PB-3c cells we can see that superoxide dismutase (SOD) has no influence on the rate of reduction, but dicumarol almost completely inhibits the reduction at a concentration greater than 1 x 10(-6) M. Therefore, it can be concluded that CV-6504 is reduced mainly by two electron donating enzymes without the intermediary of a semiquinone radical and that the resulting hydroquinone inhibits lipid peroxidation as well as 5-lipoxygenase activity.

Animals↗
PubMed1991

The relationship between granulocyte elastase-like activity of cervical mucus and cervical maturation.

Granulocyte elastase (elastase)-like activity in 119 samples of cervical mucus from pregnant women was measured. Though elastase-like activity in normal pregnant women was low in 2nd and 3rd trimesters (less than 20 units), it increased during labor (1st stage of labor: 70 +/- 50 U/l; 2nd stage: 72 +/- 56). Elastase-like activity in pregnant women with imminent premature delivery (24-30 gestational weeks) was also high (78 +/- 57). Elastase-like activity of cervical mucus correlated with Bishop score in both normal term and preterm labor. Elastase-containing granulocytes were identified by immunohistochemistry in ripened cervix in normal and preterm labor. Some granulocytes degranulate and release elastase into the cervical stroma. Others migrate through the basement membrane of the endocervix to reach the cervical canal. Elastase could not be demonstrated in immature cervix as assessed by Bishop score. The staining intensity of elastase in the cervix correlated closely with elastase-like activity in cervical mucus. Thus elastase-like activity in cervical mucus reflects the degree of cervical maturation. These results lead one to speculate that elastase is one of the neutral proteases which depolymerize and degrade collagen in the ripening cervix.

Cervix Mucus↗
PubMed1991

[A questionnaire survey on micturition problems among institutionalized elderly].

Prevalence of micturition problems among 1,023 institutionalized elderly was surveyed by a questionnaire. The reply was obtained from 821 elderly (80.3%) including 276 males and 545 females with ages averaged 77 years (range 61 to 96 years). They had neither highly impaired performance status nor severe dementia. Micturition problems were complained by 38% of male responders, where micturition difficulty was the most common. On the other hand, 23% of female responders answered having micturition problems and urinary frequency was the most common. One hundred and twenty-four of total responders (15%) replied to have urinary incontinence; 8% in men and 19% in women. Over half of patients with marked objective incontinence denied its presence or refused to answer. Seventy-one per cent of the incontinent elderly had no intention to receive medical care. From the above facts, it seems that many elderly have micturition problems and the incidence of urinary incontinence is latently higher than complained at survey.

Aged↗
PubMed1991

[Circulating serum STN antigen as a prognostic marker in epithelial ovarian cancer].

Circulating serum sialyl Tn (STN) antigen levels were measured in 89 patients with epithelial ovarian cancer. Survival at 5 years for patients with STN-negative (serum STN levels less than 50 U/ml) versus STN-positive tumors (serum STN levels greater than or equal to 50 U/ml) was 76.9% versus 10.8%, respectively (p less than 0.05). The overall survival probability was worse in patients with STN-positive sera. Percent progression-free survival at 5 years for patients with STN-negative versus STN-positive tumors was 51.9% versus 5.4%, respectively (p less than 0.05). The overall progression-free survival was shorter in patients with STN-positive sera. Multivariate regression analysis revealed that positive STN antigen level in sera is an independent predictor of poor prognosis in epithelial ovarian cancer.

Adult↗
PubMed1991

Isolation and characterization of zinc coproporphyrin I: a major fluorescent component in meconium.

We isolated a substance from an organic extract of meconium (in neutral pH) that exhibited a porphyrin-like fluorescence peak at 580 nm on excitation at 405 nm and deduced its structure. Spectrometric data suggest that the substance is not free coproporphyrin I or free coproporphyrin III, but is a chelate of coproporphyrin I with Zn2+. We also detected a chelate of coproporphyrin III with Zn2+ by HPLC. These substances may be useful as new indicators of the presence of meconium.

Amniotic Fluid↗
PubMed1991

[The relation between granulocyte elastase activity in cervical mucus and gestational cervical polyp].

Granulocyte elastase activity in cervical mucus (elastase activity) was measured in pregnant women (10-20 gestational weeks) with cervical polyp and controls (gravida with no cervical polyp). Elastase activity in the cervical polyp group was 86 +/- 44 U/l, and that in the control group was 22 +/- 13 (p less than 0.01). Elastase activity after polypectomy was 44 +/- 24. It decreased significantly compared with prepolypectomy (p less than 0.05). A high level of elastase activity continued in cases without polypectomy. Immunohistochemical staining of elastase for cervical polyps was also performed. In the low elastase staining group there was low elastase activity (46 +/- 26 U/l). Elastase activity in the moderate staining group was 89 +/- 31, and that in the intense staining group was 114 +/- 31. As for the prognosis of patients with cervical polyps, the occurrence rate of chorioamnionitis is 9% in the control group, 14% in the polyp with polypectomy group and 40% in the polyp without polypectomy group. There was a significant difference between the polyp without polypectomy group and other groups (p less than 0.01). These data suggest that cervical polyp is a focus of inflammation and that it may cause chorioamnionitis. We conclude that cervical polyp during pregnancy should be removed. If polypectomy could not be performed, local anti-inflammatory and anti-infectious therapy would be needed.

Cervix Mucus↗
PubMed1990

A single cell observation of staurosporine effect on the Ca2+ signals in rat basophilic leukemia cells.

A digital imaging fluorescence microscope was used to study the effect of a protein kinase inhibitor staurosporine on the antigen-dependent calcium signals in an individual rat basophilic leukemia cell (RBL-2H3). Although dose dependency of staurosporine was different from one cell to another, staurosporine inhibited, at low concentration, the calcium influx from the external medium into RBL-2H3 cells. At high concentration, however, it inhibited both the removal of calcium ion from internal stores and the calcium influx from the external medium. These results indicated that staurosporine is necessary for the inhibition of the calcium influx from the external medium and that a protein kinase (possibly protein kinase C) is involved in the calcium influx from the external medium into the cytoplasm.

Alkaloids↗
PubMed1990

Human growth hormone-stimulated growth of human cultured lymphocytes (IM-9) and its inhibition by phorbol diesters through down-regulation of the hormone receptors. Possible involvement of phosphorylation of a 55,000 molecular weight protein associated with the receptor in the down-regulation.

We report in this paper that human growth hormone (hGH) stimulates the growth of human cultured IM-9 lymphocytes in a low concentration (3%) of serum. The hormone-stimulated growth was inhibited with the phorbol diesters phorbol 12-myristate 13-acetate and phorbol 12,13-dibutylate (PDBu). The binding experiments of 125I-hGH to the phorbol diester-treated cells and to their detergent-solubilized receptors revealed that the phorbol diesters caused internalization of the hGH receptors from the cell surfaces but did not significantly affect their affinity (Ka = 8.5 x 10(9) M-1). About half of the receptors (1.4 x 10(3)/cell) were internalized in 30 min at 37 degrees C, and the half-effective doses of phorbol 12-myristate 13-acetate and PDBu were 5 and 35 nM, respectively. When culture was continued after washing with the culture medium, the phorbol diester-treated cells recovered their hGH-responsive growth, and the number of the surface hGH receptors was restored. The down-regulation of the hormone receptor was also induced with another phorbol diester, phorbol 12,13-didecanoate, but not with the phorbol or phorbol monoesters phorbol 12-myristate and phorbol 13-acetate. The synthetic activators of protein kinase C 1-oleoyl-2-acetyl-glycerol and N-(6-phenyl-hexyl)-5-chloro-1-naphthalenesulfonamide had an effect similar to that of the phorbol diesters. Staurosporine and sphingosine, inhibitors of protein kinase C, inhibited the phorbol diester-caused down-regulation with a half-inhibitory dose (IC50) of 8 nM and 130 microM, respectively. This suggests that protein kinase C was involved in the reaction. When 32Pi-loaded IM-9 cells were stimulated with PDBu at 37 degrees C, the phosphorylation of Mr 55,000, 88,000, and 114,000 proteins increased rapidly. The PDBu-stimulated phosphorylation of 55,000 protein was also inhibited by staurosporine at 10 nM, which was a comparable concentration to inhibit the phorbol diester-induced down-regulation of hGH receptors. Furthermore, among these proteins, the 55,000 protein was specifically coisolated with the hGH receptors by three different experiments: 1) immunoprecipitation by anti-hGH antibody; 2) immunoisolation using protein A-cellulose columns; and 3) affinity purification by hGH-fixed agarose gel. These results suggest that phorbol diesters reduce the hGH-stimulated growth of cultured IM-9 lymphocytes by the down-regulation of hGH receptors and that the receptor-associated 55,000 protein may be involved in this regulation through phosphorylation by protein kinase C.

Alkaloids↗
PubMed1990

Characterization of a new type of variant of rat basophilic leukemia 2H3 cells presenting a different pattern of calcium signal.

We selected a new type of variant (designated 3C7) derived spontaneously from parental RBL-2H3 cells. 3C7 cells showed lower contact inhibition, anchorage dependency, and serotonin release activity than those of RBL-2H3 cells. We conclude that 3C7 cells are a transformant of RBL-2H3 cell with greater malignancy. The production of inositol bisphosphate and the release of Ca2+ from intracellular stores induced by IgE-antigen stimulation were enhanced in 3C7. Oscillation of [Ca2+]i in individual 3C7 cells was observed by a digital imaging microscopic technique. We propose that 3C7 cells are a useful model system for studies on the mechanisms of stimulus-secretion coupling and the relationships between malignant alterations and disorders of signal transduction.

Animals↗
PubMed1990

Antigenic determinants on rat metallothionein: fine epitope mapping for a murine monoclonal antibody and rabbit polyclonal antisera.

In order to determine the epitope of metallothionein (MT) to a murine monoclonal antibody (MT 189-14-7) which had been produced by immunization with rat MT 2 (Kikuchi et al. (1988) Mol. Immunol. 25, 1033-1036), various lengths of synthetic oligopeptides were tested for their inhibitory activities in competitive radioimmunoassay (RIA). The amino-terminal acetylated pentapeptide, AcMDPNC, exhibited an inhibitory activity comparable to that of native MTs, whereas the acetylated tetrapeptide, AcMDPN, and the deacetylated heptapeptide, MDPNCSC, were much less inhibitory. The results suggest that the major part of the epitope structure of MT to the MT 189-14-7 monoclonal antibody is located within the amino-terminal acetylated pentapeptide, AcMDPNC. The specificities of polyclonal rabbit anti-MT antisera raised against the same immunogen were also determined by using various animal MTs and synthetic peptides as inhibitors in the RIA. Among three antisera tested, two reacted with several amino-terminal oligopeptides similarly to the MT 189-14-7 antibody. The major epitope structures to these polyclonal antibodies were shown to be located within the acetylated tetrapeptide, AcMDPN. Another antiserum contained at least two different populations of antibodies: one consisted of antibodies reactive with the amino-terminal synthetic peptides, while the other was not reactive with them. These results suggest that, in the rabbit also, the amino-terminal region common to various animal MTs can be an epitope to antibodies raised against rat MT, as shown in the mouse. Moreover, the results indicate that the synthetic amino-terminal peptides are useful for determination of the specificity of polyclonal rabbit anti-MT antibodies, which have been widely used for the quantification of MTs.

Amino Acid Sequence↗
PubMed1990

Inactivation of human tumor cell pro-urokinase by granulocyte elastase.

Supernatant obtained from granulocytes stimulated in the presence of cytochalasin B by the chemotactic peptide N-formyl-norleucyl-leucyl-phenylalanyl-norleucyl-tyrosyl- lysine displayed an inhibitory effect on the plasmin-dependent conversion of tumor urokinase-type plasminogen activator proenzyme (pro-uPA) to the active form of uPA. Moreover, the supernatant was also found to inhibit the fibrinolytic activity of human vulva (A431) and breast (MCF7) carcinoma cell lines, which contain large amounts of pro-uPA, by 87% and 96%, respectively. By using eglin C (elastase inhibitor) and a monoclonal antibody to elastase (proteolytic activity blocker of the enzyme), elastase was identified as the key enzyme of the supernatant in these phenomena. Purified elastase converted pro-uPA to an enzymatically inactive molecule composed of two polypeptide chains of Mr = 33,000 and 22,000 linked to each other by a disulfide bond. Elastase-containing granulocytes were identified by immunohistochemistry techniques in the tissues of squamous cell carcinoma and adenocarcinoma of uterus. The cells were found close to the tumor cells and in the stroma surrounding the tumor nests. By immunohistochemical staining, uPA was also found in the tumor cells. Evidently, elastase released by chemotactically activated granulocytes, which are attracted into tumor tissues, may inhibit the conversion of pro-uPA to enzymatically active uPA in the tumor cells.

Breast Neoplasms↗
PubMed1990

The third nationwide survey in Japan of vitamin K deficiency in infancy.

The occurrence of hemorrhagic disease due to vitamin K (VK) deficiency beyond the neonatal period has come under investigation in Japan. In 1980 the 1st nationwide survey was conducted in Japan by Nakayama and others, and was followed by the 2nd nationwide survey in 1985 by Hanawa. The present survey was designed to further monitor the incidence of this disease in Japan during the 3-year period from July 1985 to June 1988. Questionnaires were sent to 1,315 hospitals having more than 200 beds, located throughout Japan. Responses were received from 775 hospitals, for an answer rate of 58.9%. The total number of reported cases was 175, including 129 idiopathic type, 28 secondary type and 18 near-miss type. In this survey it was revealed that the incidence rate of the idiopathic type of vitamin K deficiency in infancy (VKDI) has decreased remarkably, to about one-fourth that reported in the first survey. The declining incidence rate of VK deficiency in Japan is considered to be the result of ever more widespread prophylactic administration of VK during the neonatal period, as most occurrences of VK deficiency in infancy are preventable by prophylactic administration of VK from the neonatal period. However, in 16 cases of the idiopathic type of VK deficiency found in the present survey, VK had been administered at least once during or after the neonatal period. This shows the heterogeneity of this condition.

Female↗
PubMed1990

Radioimmunoassay of nivalenol in barley.

Antibodies against nivalenol (NIV) tetraacetate (Tetra-Ac-NIV) were prepared by immunizing rabbits with a hemisuccinate derivative of 8-hydroxy-3,4,7,15-tetraacetyl-12, 13-epoxytrichothece-9-en conjugated to bovine serum albumin. A radioimmunoassay system with one of these sera was developed to measure NIV contamination in barley. The detection limit for Tetra-Ac-NIV was about 0.5 ng/ml. The relative cross-reactivities of the antiserum with Tetra-Ac-NIV, acetyl T-2 toxin, and scirpenol triacetate, which were determined by the competitive radioimmunoassay, were 1, 0.78, and 0.56, respectively. Other derivatives showed no cross-reactivity. For the determination of NIV in a barley sample, NIV was extracted from the sample with acetonitrile-water (7:3), defatted with hexane, and then acetylated with acetic anhydride to form Tetra-Ac-NIV. The reaction mixture was loaded onto a C18 cartridge to remove excess reagents and impurities. Tetra-Ac-NIV was eluted from the cartridge with 50% methanol in water, and the eluate was subjected to radioimmunoassay. Analysis of six naturally contaminated barley samples for NIV revealed that radioimmunoassay results agreed well with gas chromatographic analyses.

Chromatography, Gas↗
PubMed1990

Congenital factor XIII deficiency with treatment of factor XIII concentrate and normal vaginal delivery.

The outcome of all pregnancies complicated by congenital factor XIII (F XIII) deficiency has resulted in abortion without replacement therapy. We experienced a case with this disease and succeeded in normal vaginal delivery after treatment with weekly F XIII concentrate (Fibrogammin P, Behringwerke AG) during pregnancy. This 20-year-old woman, gravida 1, para 0, was found to have an F XIII level of 0% at age 6 and was treated with F XIII concentrate occasionally when she suffered from massive bleeding. In 1986 she became pregnant and was hospitalized at 6 weeks' gestation because of genital bleeding. Subsequent to this episode, F XIII concentrate was administered every week. At 37 weeks' gestation a 2,646-gram girl with a 1-min Apgar score of 9 was delivered. Postpartum blood loss was 260 ml. One year after delivery neither the mother nor the infant were found to have hepatitis B, nonA, nonB hepatitis, ATL or HIV. F XIII concentrate proved effective in such cases without any risk of viral infection.

Adult↗
PubMed1990

Production and characterization of monoclonal antibodies to nivalenol tetraacetate and their application to enzyme-linked immunoassay of nivalenol.

Three monoclonal antibodies were obtained by the fusion of mouse myeloma cells with splenocytes isolated from BALB/c mice that had been immunized with 8-hydroxy-3,4,7,15-tetraacetyl-nivalenol hemiglutarate covalently bound to bovine serum albumin. These anti-nivalenol tetraacetate monoclonal antibodies were of the IgG type and highly specific to nivalenol tetraacetate, with an apparent association constant of about 10(8)M-1. The relative cross-reactivities of one monoclonal antibody with nivalenol tetraacetate, acetyl T-2 toxin, and scirpenol triacetate were found to be 1.0, 0.02 and 0.03, respectively. Other derivatives showed no cross-reactivity at all. An indirect enzyme-linked immunosorbent assay (ELISA) based on the competitive binding principle was developed using the antibody from clone D18.102.59. The sensitivity of the system was about 0.1 ng of nivalenol tetraacetate per assay. Comparison of nivalenol levels detected in naturally contaminated barley samples by competitive indirect ELISA and gas chromatography (GC) showed good agreement, indicating that the antibody is useful for the measurement of nivalenol in naturally contaminated cereals and grains.

Animals↗
PubMed1990

Production and characterization of monoclonal and polyclonal antibodies against digoxin.

Three hybridoma cell lines (DIG 64. 2B. 5, DIG 104. H10.1 and DIG 222. 4D. 5) producing monoclonal antibodies against digoxin were established, and the properties of these monoclonal antibodies were characterized and compared with three polyclonal rabbit anti-digoxin antisera. Both polyclonal and monoclonal antibodies gave association constants ranging from 10(9) to 10(10) (M-1). The monoclonal antibodies were of the IgG1(kappa) or IgG2b(kappa) subclass. Cross-reactivities of these monoclonal antibodies and polyclonal antisera with various related cardenolides and their aglycones were determined by competitive radioimmunoassay. The monoclonal antibodies were specific for digoxigenin-containing glycosides such as lanatoside C and deslanoside, but not for digitoxigenin-containing glycosides. On the other hand, the specificities of the polyclonal antisera were less strict than the monoclonal antibodies and relatively cross-reactive with digitoxin. It was shown that the RIA for digoxin using DIG 64. 2B. 5 is sensitive enough to detect 0.2-0.3 nM (30-60 pg/tube) of digoxigenin-containing cardenolides.

Animals↗