Comparison of manic switch onset during fluoxetine and trazodone treatment.
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Adult↗
Biomedical subjects
T Terao
Publications and source records attributed to T Terao.
Induction of metallothionein in a human astrocytoma cell line by interleukin-1 and heavy metals.
The effects of cytokines and heavy metals on the expression and localization of metallothioneins (MTs) within U373MG astrocytoma cells were analyzed by using indirect immunofluorescence using a monoclonal anti-MT antibody (MT45). IL-1, CdCl2 (50 microM) or ZnCl2 (500 microM) remarkably augmented intracellular MT levels, whereas IL-6 or 10 microM of ZnCl2 showed no inducing activity. From 24 to 48 h after the addition of CdCl2 or IL-1, immunoreactive MTs were found in the cytoplasm and the nucleus. After 72 h, immunoreactive MTs accumulated in a granular form near the cell surfaces in the presence of CdCl2 (50 microM) or IL-1 plus ZnCl2 (10 microM). However, this accumulation was not observed when only IL-1 was added. Thus, Zn2+ facilitated the appearance of the granular form of immunoreactive MTs at a concentration where they do not induce MTs by themselves.
Astrocytoma↗
Monoclonal antibodies MA54 and MA61 as potential reagents in the prognosis of patients with ovarian cancer.
Monoclonal antibodies (moABs) MA54 and MA61 directed to the O-linked NeuAc alpha 2-6GalNAc epitope were generated by immunization with culture supernatants of lung adenocarcinoma cells. We have examined the correlation between the prognosis of patients (overall survival and progression-free survival) and circulating serum levels of CA54/61 antigen by comparison with CA125 in patients with epithelial ovarian cancer. Circulating serum CA54/61 antigen levels were determined by sandwich enzyme immunoassay kits. Serum antigen levels were elevated in 44.4% of the patients. Survival at 3 years for ovarian cancer patients with CA54/61-negative (serum CA54/61 levels < 15.2 U/ml) versus CA54/61-positive (CA54/61 > or = 15.2 U/ml) tumors was 64% versus 25% (P < 0.05). In contrast, there is no significant difference in the prognosis of patients based upon positive or negative CA125 antigen values (51% vs 67%). The overall survival rate was worse in patients with CA54/61-positive sera, indicating that positive CA54/61 levels in serum is an independent predictor of poor prognosis in ovarian cancer. In addition, CA54/61 status has been shown to be associated with early relapse of this malignancy.
Adult↗
Molecular characteristics of the CA 125 antigen produced by human endometrial epithelial cells: comparison between eutopic and heterotopic epithelial cells.
OBJECTIVE: To characterize the CA 125 antigen purified from the conditioned media using an in vitro system that we have established for the culture of epithelial cells from human uterine endometrium. STUDY DESIGN: CA 125 was purified separately from the conditioned media of eutopic and heterotopic epithelial cells by gel chromatography and OC 125 immunoaffinity column chromatography. RESULTS: Western blot analysis of conditioned media from eutopic epithelial cells revealed a single polydisperse band of 200 kd, whereas two major CA 125 isoforms of 200 and 110 kd, as well as two minor forms of 100 and 70 kd, were observed in heterotopic epithelial cells. The 110 kd CA 125 was more prominent than the 200 kd antigen in heterotopic epithelial cells. CONCLUSION: These results strongly suggest that CA 125 antigens purified by immunoaffinity chromatography have different molecular mass in eutopic and heterotopic epithelial cells even though the OC 125 binding site may remain intact.
Antigens, Tumor-Associated, Carbohydrate↗
Clinical evaluation of new cancer-associated antigen CA125 II in epithelial ovarian cancers: comparison with CA125.
With use of monoclonal antibodies (moABs) M 11 and OC125, a sensitive sandwich immunoradiometric assay (IRMA) for CA125 II was developed. CA125 antigen was adsorbed by moAB OC125 and M 11, but horseradish peroxidase-labelled OC125 did not compete with M 11, indicating that although these two moABs recognized CA125 antigen, they reacted with separate antigenic determinants. We also demonstrate that the dual capture technique with OC125 and M 11 antibodies has a larger dynamic range than OC125 alone. This could potentially increase the sensitivity in terms of low CA125 levels. Forty-six patients with ovarian cancer before therapy and a further 20 cases during follow-up were evaluated for the presence of two serum tumor markers: CA125 II and CA125. The overall positive rate for CA125 II was 97.8%, compared with 91.3% with CA125. Serum CA125 II levels were closely correlated with serum CA125 levels (r = 0.918). The mean CA125 II levels before treatment, at response, and at recurrence were 194.3, 19.6, and 103.5 U/mL, respectively. Normalization of serum CA125 II levels at response does not imply absence of microscopic residual disease. CA125 II changes during follow-up suggest recurrence and can be used to monitor individual patients.
Adult↗
Characterization of CA 125 antigen identified by monoclonal antibodies that recognize different epitopes.
The present study was undertaken to characterize the antigenic determinant of the CA 125 macromolecules recognized by newly developed monoclonal antibodies (M 11, 130-22, 145-9, 602-1, and 602-6), examine their relationship among the epitopes recognized by these antibodies, and develop a series of enzyme-linked immunosorbent assays (ELISAs) in which various combinations of antibodies are used in a double-determinant sandwich mode. The antigenic determinants of CA 125 were characterized by several methods including competitive ELISA and immunoblotting. These antibodies, as well as OC 125, reacted with ovarian cancer (HOC-I) cell extract in a dose-dependent manner. Purified CA 125 antigen with a molecular mass of less than 200 kDa (CA 125 < 200 kDa) had a significant inhibitory effect on the reaction between the cancer cell extract and all these antibodies. The reactivity of M 11, 130-22, 145-9, 602-1, and 602-6 to the cancer cell extract was not significantly inhibited by purified CA 125 > or = 200 kDa, whereas the reactivity of OC 125 was completely inhibited by CA 125 > or = 200 kDa. The antigenic determinants of M 11, 145-9, 602-6, 130-22, and 602-1 were closely related to each other, in this order; whereas OC 125 recognized a different epitope on a structurally identical molecule. The use of these monoclonal antibodies in combination with each other may result in the development of a more specific and sensitive assay for CA 125.
Animals↗
Isolation and characterization of a major allergenic component (gp55) of Aspergillus fumigatus.
IgE class antibodies specific for antigens in a water-soluble extract of Aspergillus fumigatus (strain NHL-5759) were analyzed by immunoblotting with sera from patients with allergic bronchopulmonary aspergillosis. All the sera tested were reactive with a major 50 to 60 kd protein in the extract. This allergen, designated gp55, was purified by gel filtration and preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The antigen was found to be present in the water-soluble extract in the form of a complex composed of approximately eight molecules of gp55. The carbohydrate and phosphate content of the purified antigen were 23.1% and 0.46%, respectively. The molar ratio of mannose to galactose residues was 2.76:1, and the protein was glycosylated predominantly with N-linked oligosaccharides. The serologic activity of the gp55 antigen was abolished by treatment with nonspecific protease (Pronase) but not by treatment with sodium metaperiodate or endoglycosidases. Thus the major antigenic site of the glycoprotein is located within its peptide moiety. The antigen itself displayed no chymotryptic or tryptic activity. The amino acid sequence of the 20 N-terminal residues of the antigen (ATPHEPVFFSWDAGAVTSFP) is different from that of any other protein previously reported.
Amino Acid Sequence↗
Molecular and biochemical analyses of combining sites of monoclonal anti-morphine antibodies.
V region nucleotide sequences were determined by mRNA sequencing for 11 monoclonal anti-morphine antibodies with slightly different specificities for morphine-related opiates. The VH region nucleotide sequences of the antibodies MOR8, MOR33, MOR35, MOR44, MOR83, and MOR76 were classified into the VH-5 (7183) family, while the antibodies MOR39, MOR115, MOR131, MOR158 and MOR180 used VH-1 (J558) family genes. MOR39, MOR115 and MOR131 used the V lambda-1 gene for their L chain V region. MOR158 and MOR180 used the Vk-10 gene. MOR8, MOR33, OR35, MOR44, MOR76 and MOR83 used VK-21D. The antibody sets MOR158 and MOR180; MOR39 and MOR131; and MOR8, MOR33, MOR35, MOR44, MOR76 and MOR83 appeared to be somatic mutants derived from the same clones since they showed the same VH/VL usage and V(D)J recombination pattern. The pH-reactivity profiles for these antibodies revealed that the binding of morphine to the antibodies is highly dependent on the pH value of the assay solution, suggesting the importance of the electrostatic interaction between the positive charge of morphine and the negative charges at or near the combining sites. Direct UV-photoaffinity labeling with 3H-morphine was carried out in order to estimate the orientation of morphine in the combining sites. The H chains were preferentially labeled in MOR8, MOR33, MOR35, MOR76, and MOR83, whereas most of the crosslinked hapten was found in the L chains in MOR39, MOR115, MOR131, MOR158 and MOR180. Thus, these 11 antibodies were classified into two types in terms of reactivity in the photoaffinity labeling.
Affinity Labels↗
A simple, noninvasive, sensitive method for diagnosis of amniotic fluid embolism by monoclonal antibody TKH-2 that recognizes NeuAc alpha 2-6GalNAc.
OBJECTIVE: The sialyl Tn structure (NeuAc alpha 2-6GalNAc alpha 1-O-Ser/Thr) recognized by monoclonal antibody TKH-2 is a characteristic component in meconium and amniotic fluid. The purpose of this study was to determine whether amniotic fluid embolism could be detected by quantification of this antigen in maternal serum by means of an assay using antimucin monoclonal antibody TKH-2. STUDY DESIGN: Sialyl Tn antigen was measured in the serum of women with meconium-stained amniotic fluid and compared with the level in those with clear amniotic fluid, as well as that in women with a clinical picture suggesting amniotic fluid embolism. The concentration of sialyl Tn antigen was determined by an immunoradiometric competitive inhibition assay. RESULTS: Serum sialyl Tn antigen levels in women with meconium-stained amniotic fluid (20.3 +/- 15.4 U/ml) at delivery were slightly higher than those in women with clear amniotic fluid (11.8 +/- 5.6 U/ml). A significantly elevated level of sialyl Tn antigen was observed in serum of patients with amniotic fluid embolism and amniotic fluid embolism-like symptoms (105.6 +/- 59.0 U/ml, p < 0.01). CONCLUSION: The method for detecting sialyl Tn antigen in the serum of patients with amniotic fluid embolism is a direct way to demonstrate the release of meconium- or amniotic fluid-derived mucin into the maternal circulation and is a simple, noninvasive, sensitive method for diagnosis of amniotic fluid embolism.
Adult↗
The effect of salts on the stability of beta-galactosidase in aqueous solution, as related to the water mobility.
The effect of salts (KI, KBr, NaCl, KCl, KF, phosphate, and Na2SO4) on the stability of beta-galactosidase in aqueous solution was studied from the aspect of changes in water mobility. At salt concentrations up to 200 mM, the inactivation rate of beta-galactosidase in all the salt solutions studied increased with increasing salt concentration. At higher concentrations, those salts which had little effect on the spin-lattice relaxation time, T1, of water (KI, KBr, and KCl) continued to increase the inactivation rate of beta-galactosidase with increasing concentration, while those salts which decreased the T1 of water (KF, phosphate, and Na2SO4) decreased the inactivation rate. It appeared that the decrease in water mobility caused by KF, phosphate, and Na2SO4 resulted in stabilization of beta-galactosidase. The results indicate that water mobility is an important factor in the denaturation rate of proteins.
Bromides↗
Stability of beta-galactosidase, a model protein drug, is related to water mobility as measured by 17O nuclear magnetic resonance (NMR).
The inactivation of freeze-dried beta-galactosidase during storage was studied, focusing on the effect of water mobility as measured by the spin-lattice relaxation time, T1, of water using 17O NMR. Inactivation of beta-galactosidase lyophilized from phosphate buffer solution was studied as a function of water content, which in turn affected the T1 of water. An increase in the water content of freeze-dried beta-galactosidase brought about an increase in the T1 of water, as well as a rise in pH. For the freeze-dried enzyme with sufficient water content to be dissolved, the inactivation rate was related to the T1 of water rather than to the pH change. It is suggested that as the water content increases, the mobility of water around the enzyme increases, resulting in enhanced enzyme inactivation. The freeze-dried samples with limited moisture showed inactivation rates faster than those expected from the pH and water mobility, suggesting that the inactivation mechanism is different from that for the freeze-dried enzyme with a larger amount of water. Inactivation of beta-galactosidase in solutions was also studied as a function of phosphate buffer and sodium chloride concentrations, which in turn affected the T1 of water. Because the inactivation rate increased with increasing salt concentrations and the rate extrapolated to zero concentration was negligible, inactivation of the freeze-dried enzyme was apparently induced by the salts used as additives for lyophilization. The enhancing effect of phosphate buffer components, however, was reduced at higher concentrations, an effect related to the decrease in the T1 of water. This result may be ascribed to the decrease in water mobility caused by phosphate buffer components and is consistent with the observation that the inactivation rate of the freeze-dried enzyme with a relatively large amount of water decreased with decreasing T1 of water.
Aspergillus oryzae↗
Aggregates formed during storage of beta-galactosidase in solution and in the freeze-dried state.
Aggregates formed during storage of freeze-dried beta-galactosidase were compared with those formed in solutions. Freeze-dried beta-galactosidase aggregated during storage in the presence of moisture, producing a protein precipitate which was soluble in guanidine hydrochloride solution but not in buffer solution. The protein precipitate dissolved in guanidine solution exhibited a large molecular size by high-performance size exclusion chromatography and converted to proteins of original size in the presence of dithiothreitol. It is suggested that the aggregation involves chemical interaction via covalent disulfide bonding. In contrast, beta-galactosidase in aqueous solution aggregated without formation of protein precipitates. Soluble aggregates were converted to proteins of original size in guanidine solution without dithiothreitol, suggesting noncovalent bonding. The difference in aggregation behavior may be ascribed to the difference in the water:protein ratio. We propose that inactivation of beta-galactosidase is due to formation of thermally denatured (unfolded) protein, which aggregates dependent on the water:protein ratio, either via noncovalent interactions at a high water:protein ratio in solution or via covalent interaction at a low water:protein ratio in the freeze-dried state.
Buffers↗
Decreased protein-stabilizing effects of cryoprotectants due to crystallization.
The stabilizing effects of various additives against inactivation of an enzyme (beta-galactosidase from Aspergillus oryzae) during freeze-drying were studied, with a focus on their crystallinity. The crystalline morphology of mannitol and inositol in freeze-dried cakes depended on the solute concentrations before freezing and the freeze-drying method used. The additives in their amorphous state showed concentration-dependent stabilization of the enzyme, whereas additive crystallization during freeze-drying decreased their effects. Heat treatment before freeze-drying also caused crystallization and diminished the stabilizing effects. Noncovalent soluble aggregates were observed in the inactivated enzyme solution. These results show the importance of maintaining the amorphous state of additives used as stabilizing agents during freeze-drying.
Asparaginase↗
Characterisation and clinical usefulness of CA130 antigen recognised by monoclonal antibodies, 130-22 and 145-9, in ovarian cancers.
A new cancer-associated antigen CA130, recognised by two monoclonal antibodies (moABs) 130-22 and 145-9, was often found to be present at high levels in the sera of patients with ovarian cancer. There was a strong correlation between CA130 and CA125 values. The epitopes recognised by moABs 130-22 and 145-9 were proved to differ from the CA125 epitope, but to exist on the molecule bearing CA125. Unlike OC125, the majority of 130-22/145-9 activity was associated with a much lower molecular mass (less than 200 kDa), indicating that a lower molecular mass immunoreactive determination may be a unique CA130 antigenic determinant within CA125 molecule. Clinical data demonstrate that, (1) elevated levels of CA130 determinant were found in the sera of 91.3% of women with epithelial ovarian cancer, (2) falling or rising levels of CA130 correlated with regression or progression of ovarian cancer in > 95% of cases, (3) normalisation of serum CA130 levels at response does not imply no microscopic residual disease, but CA130 changes during follow-up support the evaluation of recurrence and can be used as a monitoring marker in an individual patient.
Adult↗
Saturation of tumour cell surface receptors for urokinase-type plasminogen activator by amino-terminal fragment and subsequent effect on reconstituted basement membranes invasion.
Single-chain urokinase-type plasminogen activator (pro-uPA) is bound to a specific surface receptor on ovarian cancer HOC-I cells that is incompletely saturated. Saturation of uncovered receptors by uPA polypeptides with intact amino-terminal fragment (ATF) derived from pro-uPA by limited proteolysis (human leucocyte elastase [HLE] or V8 protease) has been studied. HOC-I cells preferentially invaded reconstituted basement membranes in a time- and plasminogen-dependent manner. This process was inhibitable by preincubation with uPA polypeptides in the medium at levels which suggested that complete saturation of cell surface uPA receptors occurred. This result indicates that occupation of uPA receptors by enzymatically inactive uPA fragments or prevention of rebinding of pro-uPA synthesised by tumour cells to the receptors specifically reduces the invasion of the tumour cells through basement membranes in vitro.
Cell Movement↗
Increased cell-surface urokinase in advanced ovarian cancer.
Urokinase-type plasminogen activator (uPA), uPA receptors, and cathepsin B were quantitated by using an immunological method, enzyme-linked immunosorbent assay, and amidolytic activity assays in 15 malignant and 10 benign epithelial ovarian tumors. The levels of uPA and uPA receptors, as well as cathepsin B, were found to be higher in membrane preparations obtained from malignant tumors than in those obtained from benign tumors. Acid-treated membranes acquired the ability to bind uPA, indicating that uPA is bound to a specific surface receptor that is not completely saturated. Levels of single-chain uPA (pro-uPA) and high-molecular-weight uPA in membrane preparations were measured by immunoadsorbent-amidolytic assay. The finding of a significant increase in amidolytic activity following activation of uPAs by plasmin suggested that less than half (30-40%) of all membrane immunoreactive uPAs is present in the enzymatically inactive pro-uPA form. In the membranes of malignant tumors, levels of uPA receptor and cathepsin B did not vary with stage of disease. On the other hand, we found that the level of receptor-bound uPA antigen/activity was significantly increased in advanced malignant tumors. Receptor-bound uPA may play an important role in determining invasive potential of tumor cells. Since ovarian cancer cells produce both pro-uPA and cathepsin B, the possibility of activation of tumor cell-derived pro-uPA by cellular protease cathepsin B must be considered.
Antigens, Neoplasm↗
Magnetic resonance imaging angiography in a case of eclampsia.
We experienced a typical case of eclampsia. After the onset of eclampsia, we performed magnetic resonance imaging angiography (MRI angiography) to estimate the function of the cerebral artery. MRI angiography showed that spasm occurred in many cerebral arteries. The spasm was still observed in some arteries 13 day after the onset of eclampsia. Vasospasm of eclampsia was clearly, easily and noninvasively confirmed in this case by MRI angiography. This observation suggests that MRI angiography is very useful and informative for diagnosis and in treatment of eclampsia.
Adult↗
HELLP syndrome-like biochemical parameters obtained with endothelin-1 injections in rabbits.
We studied the effects of bolus injections and infusion of endothelin-1 (ET-1) in female rabbits by measuring serum glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), lactic dehydrogenase (LDH), antithrombin III (AT-III), thrombin antithrombin (TAT) complexes, platelet counts and indirect bilirubin. Two successive bolus doses of 0.125 and 0.25 nmol/kg of ET-1 with an interval of 30 min were given to conscious non-pregnant female rabbits (n = 8). GOT, GPT and LDH were found to be significantly increased after injections of ET-1 (p < 0.02, p < 0.04 and p < 0.05, respectively). The percent AT-III activity decreased significantly (p < 0.005). Vasospasm of the hepatic artery was demonstrated by angiography with the same bolus doses in rabbits. There was also an increase in GOT (p < 0.003), GTP (p < 0.05), LDH (p < 0.007), indirect bilirubin (p > 0.07) and TAT complexes (p < 0.02) and a decrease in AT-III (p < 0.03) and platelet counts (p < 0.02) in rabbits (n = 10) after 24 h of continuous infusion of ET-1 (0.6 nmol/kg/h). Histological examination of rabbit liver tissues showed varying degrees of ischemic necrosis of hepatocytes. Thus this study suggests that exogenously administered ET-1 causes vasospasm and liver ischemia resulting in HELLP syndrome-like blood parameters in rabbits.
Alanine Transaminase↗