Affinity labeling of binding proteins for the study of endocytic pathways.
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Affinity Labels↗
Biomedical subjects
R Nishimura
Publications and source records attributed to R Nishimura.
Sialic acids, but not their linkage to galactose residues, are required for full expression of the biological activity of human chorionic gonadotropin.
One of the characteristic features of the asparagine-linked sugar chains of human chorionic gonadotropin (hCG) is that their sialic acid residues occur exclusively as the Neu5Ac alpha 2----3Gal group. In order to determine whether this sialic acid linkage is important for the functional role played by the sugar moiety of hCG or not, isomeric hCG containing the Neu5Ac alpha 2----6Gal group was prepared and its hormonal activity in vitro was investigated. On addition of the isomeric hCG to the culture medium of a murine Leydig tumor cell line, it was found that it shows the same hormonal activity as natural hCG.
Adenylyl Cyclases↗
Molecular forms of human chorionic gonadotropin in choriocarcinoma serum and urine.
The molecular forms of human chorionic gonadotropin (hCG) were assessed in parallel serum and urine samples from a choriocarcinoma patient by gel chromatography, isoelectric focusing, and Western blot analysis. There were significant qualitative differences in the molecular forms of hCG between the serum and the urine. The serum hCG had a greater molecular weight and a stronger acidic charge than the urinary hCG. These results may be attributed to differences of sialic acid contents. Affinity chromatography using Ricinus communis agglutinin-conjugated Sepharose showed that the serum hCG was completely sialylated, while the urinary hCG was partially desialylated. In addition to the complete hCG molecule, two kinds of hCGbeta-related fragments were detected in the urine of the patient, but not in the serum. In the Western blot using anti-hCGbeta or anti-hCGbeta-CTP under reducing conditions, the urine presented a new band (CTP') with Mr 23,000 in addition to the beta-subunit, while the serum revealed a single band of the beta-subunit with or without a reducing reagent. The present data revealed striking qualitative differences in the molecular forms of hCG between serum and urine of a patient with choriocarcinoma.
Adult↗
[A case of multiple pulmonary metastases from rectal cancer which responded completely to combination chemotherapy using 5'-DFUR and MMC].
A 68-year-old male, who had advanced rectal cancer with multiple pulmonary metastases, had undergone resection of rectal cancer operatively and was treated with a combination chemotherapy using 5'-DFUR (5'-deoxy-5-fluorouridine) and MMC (mitomycin C) for multiple pulmonary metastases. Nine months from the start of this therapy, pulmonary metastatic lesions disappeared completely on chest X-ray examination and computed tomography. This case corresponded to complete response (CR) according to the response criteria proposed by Koyama-Saitoh. No significant side effects were observed during this chemotherapy. From the results in this case, the combination chemotherapy was considered to be one of the effective antineoplastic therapies available for pulmonary metastasis from large bowel cancer.
Adenocarcinoma↗
[A study of the proliferative activity of DNA polymerase alpha immunoreactivity in breast cancer].
DNA polymerase alpha activity is associated with cell proliferation, independently of the cell cycle phase, and a simple and reproducible method for the immunohistochemical demonstration of DNA polymerase alpha has been developed. In an analysis of a total of 76 human breast cancer tissues using this procedure, the clinico-pathological findings revealed that only the estrogen receptor (ER) significantly correlated with the DNA polymerase alpha activity. Therefore, this suggests that ER-negative tumors had a higher proliferative activity.
Adult↗
Comparative study of the mucin-type sugar chains of human chorionic gonadotropin present in the urine of patients with trophoblastic diseases and healthy pregnant women.
Human chorionic gonadotropins (hCGs) highly purified from the urine of patients with trophoblastic diseases and of healthy pregnant women contain approximately four mucin-type sugar chains in one molecule. The structures of these sugar chains were studied comparatively by using a new sensitive method to obtain mucin-type sugar chains quantitatively as radioactive oligosaccharides from a small amount of glycoproteins. The mucin-type sugar chains of all hCGs include sialylated and nonsialylated Gal beta 1----3GalNAc and Gal beta 1----4GlcNAc beta 1----6(Gal beta 1----3)GalNAc. In the case of normal hCG and hydatidiform mole hCG, oligosaccharides containing the tetrasaccharide core occupy approximately 10% of the total mucin-type sugar chains. The ratio of the tetrasaccharide containing oligosaccharides is increased prominently to approximately 60% in choriocarcinoma hCG. The proportion in invasive mole hCG was also increased, but less than the proportion of choriocarcinoma hCG.
Acetylation↗
Structural differences found in the sugar chains of eutopic and ectopic free alpha-subunits of human glycoprotein hormone.
Free alpha-subunits of human glycoprotein hormone were purified from the urine of a healthy pregnant woman and from that of a patient with adenocarcinoma. Comparative study of their sugar moieties revealed that they have different numbers and different sets of asparagine-linked sugar chains, which are also different from those of alpha-subunit obtained by dissociation of whole hCG molecule. The eutopic free alpha-subunit contained biantennary complex-type sugar chains only. In contrast, the ectopic free alpha-subunit contained tri- and tetraantennary complex-type sugar chains in addition.
Adenocarcinoma↗
Altered glycosylation is induced in both alpha- and beta-subunits of human chorionic gonadotropin produced by choriocarcinoma.
The two subunits of human chorionic gonadotropin (hCG) purified from the urine of a patient with choriocarcinoma were successfully separated by SDS-polyacrylamide gel electrophoresis. A comparative study of the oligosaccharides released from the two subunits by hydrazinolysis revealed that altered glycosylation occurs in both subunits and possibly at all four asparagine sites of the choriocarcinoma hCG molecule.
Asparagine↗
Primary choriocarcinoma of the urinary bladder.
We report a case of choriocarcinoma of the bladder. The initial biopsy of the bladder tumor showed co-existence of choriocarcinoma and transitional cell carcinoma, while the cystectomized bladder tumor and metastatic tumors were composed only of choriocarcinoma. The diagnosis of choriocarcinoma was confirmed morphologically and by the demonstration of hCG in the tumor and in the serum.
Carcinoma, Transitional Cell↗
Calmodulin kinase II in pure cultured astrocytes.
Calcium- and calmodulin-dependent protein kinase activity was studied in pure neuronal and glial cultures. The addition of calcium and calmodulin stimulated 32P incorporation into several neuronal proteins including two in the 50- and 60-kilodalton (kD) region which comigrated with purified forebrain calmodulin kinase II subunits (CaM kinase II). In mature astrocytes, CaM kinase activity was also present, and was inhibited by trifluoroperazine and diazepam. Again in homogenates of these cells, two phosphoproteins of apparent molecular masses of 50 and 60 kD comigrated with purified CaM kinase. CaM kinase activity was absent in immature mixed glia and oligodendrocytes. The presence of CaM kinase in neurons and mature astrocytes was confirmed using monoclonal antibodies specific for the 50-kD subunit of the enzyme. No immunoreactivity was observed in oligodendrocytes. The presence of CaM kinase in astrocytes suggests a more ubiquitous role of this enzyme in regulating cellular processes than was previously recognized.
Animals↗
Fragmentation of the beta-subunit of human chorionic gonadotropin produced by choriocarcinoma.
When human chorionic gonadotropin (hCG) was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions with dithiothreitol (DTT), a smaller weight material (CTP'), in addition to the beta-subunit, could be detected by Western blot analysis using antiserum for hCG beta-carboxy-terminal peptide (CTP). The CTP' band was much more apparent with urinary hCG from a patient with choriocarcinoma than with that from normal pregnant women. Second-dimensional electrophoresis of the choriocarcinoma hCG (c-hCG) after reduction with DTT indicated that the CTP', Mr 25,000, was released from the beta-subunit. The carbohydrate structure of the CTP' was analyzed by affinity with lectin-peroxidase on a nitrocellulose membrane. The CTP' did not interact with Concanavalin A, but exhibited strong interaction with both RCA120 and Arachis hypogaea after removal of sialic acid, indicating that it was released as a fragment containing an O-linked sugar chain as was found in the hCG beta carboxy-terminal portion. Western blot analysis using the antisera for hCG alpha, hCG beta, and hCG beta-CTP showed that the CTP' contains not only the carboxy-terminal portion but also a part of the internal (core) portion of the beta-subunit molecule. This dissociation of the c-hCG beta was further supported by the presence of a faster moving component (FMC) which may correspond to the NH2-terminal side counterpart. The desialylated FMC could be detected by Concanavalin A and RCA120 but not by Arachis hypogaea, indicating that it contains N-linked rather than O-linked sugar chains. The FMC does not contain any of the epitopes for the antisera examined in Western blot. These results indicate that the beta-subunit of the choriocarcinoma urine hCG has an unusual site which is dissociated into two components of Mr 25,000 (CTP') and Mr 18,000 (FMC) by DTT reduction.
Choriocarcinoma↗
[Molecular weight and fragmentation of human chorionic gonadotropin in urine in patients with trophoblastic diseases].
Molecular weights (MW) of urinary human chorionic gonadotropin (hCG) from trophoblastic diseases were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by Western blotting using specific antibodies. Although the MW of hCG beta from hydatidiform mole and invasive mole were identical to that of standard hCG beta, those from choriocarcinoma were individually different. These results indicate that the structural changes of carbohydrates, which may be induced by malignant transformation of trophoblast, can be reflected as differences in MW. In SDS-PAGE under nonreducing conditions. Western blotting using anti-hCG beta-carboxy-terminal peptides (CTP) revealed a single band corresponding to hCG beta. Under reducing conditions with dithiothreitol (DTT), however, a second low MW material (CTP') could be observed with anti-hCG beta-CTP. All hCG samples from invasive mole and choriocarcinoma were much more susceptible than standard hCG to release of CTP' by DTT. Estimation of MW and susceptibility to DTT reducing of urinary hCG by Western blotting are useful in the diagnosis of invasive mole and choriocarcinoma.
Chorionic Gonadotropin↗
[Structural changes in human chorionic gonadotropin in trophoblastic diseases].
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Chorionic Gonadotropin↗
Treatment of pharyngeal gonorrhea with extended oral ampicillin.
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Administration, Oral↗
Structural differences found in the asparagine-linked sugar chains of human chorionic gonadotropins purified from the urine of patients with invasive mole and with choriocarcinoma.
Paper electrophoresis of the oligosaccharide fraction obtained by hydrazinolysis of human chorionic gonadotropin (HCG), which was purified from the urine of two patients with invasive mole, gave fractionation patterns different from those of normal and hydatidiform mole HCGs. Structural study of the oligosaccharides in each fraction revealed that the triantennary complex-type asparagine-linked sugar chains, specifically found in choriocarcinoma HCGs, were included in the two HCG samples. However, the unusual biantennary complex type sugar chains, which are also specific for the tumor HCGs, were not detected at all. This result indicated that abnormal expression of N-acetylglucosaminyl-transferase IV during the development of choriocarcinoma occurs in two steps: (a) ectopic expression of the regular N-acetylglucosaminyltransferase IV; and (b) modification of the substrate specificity of the enzyme.
Adult↗
Combined high-performance liquid chromatography and enzyme immunoassay for active metabolites of a sleep inducer (rilmazafone), a ring-opened derivative of benzodiazepines, in human plasma.
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Chromatography, High Pressure Liquid↗
Effect of intravenous prostaglandin E1 on experimental flaps.
Prostaglandin E1 (PGE1) is a kind of prostaglandin that produces vasodilation of the peripheral vessels and platelet disaggregation, similar to prostacyclin (PGI2). The effects of PGE1 administered intraarterially or intravenously on peripheral vascular disease have been reported by many authors. In this investigation, 2 experiments were designed to assess the effect of intravenous infusion of PGE1 on the experimental skin flaps in rabbits. In the first experiment, the blood flow in the flap was measured before and after the administration of PGE1 by using the technique of clearance of hydrogen gas, which was generated by electrolysis. The result indicated a biphasic effect of PGE1 on the blood flow in the flap. That is, the low dose (3 micrograms per kilogram per hour) increased the blood flow on the critical line of the dye distance, while the high dose (20 micrograms per kilogram per hour) decreased it. The second experiment, in which the survival length was measured, indicated the practical effect of intravenous infusion of PGE1 on flap survival.
Alprostadil↗
Long-term survival of a free split-thickness skin graft on a large seroma.
A free skin graft about 12 cm in diameter transplanted after excision of a Bowen's carcinoma on the back totally survived for a long period on seroma and was confirmed to have revascularization from the host skin margin. Repeated evacuations of the fluid and subsequent pressure dressings failed to cause adherence of the graft to the bed, even on the thirty-ninth postoperative day. Histologic examination of the graft and the bed revealed partial epithelialization on the face-to-face surfaces, to which no adherence was attributed. The incomprehensible phenomenon in this unusual clinical case evokes a new interest in the mechanism of free skin graft survival, particularly in the phase of serum imbibition.
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