[Production of monoclonal antibodies against factor IX and their immunological characterization].
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Biomedical subjects
Publications and source records attributed to H Fukui.
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Histidine decarboxylases from the stomach and brain of adult rats were purified 380- and 160-fold, respectively, and their properties compared with those of the enzyme from whole bodies of fetal rats (7600-fold purification). The molecular weights (about 90,000) and the apparent Km values for L-histidine (3 X 10(-4) M) of the three enzymes were similar. The pI value of the fetal enzyme was 5.0, and that of the brain enzyme was 5.4. Histidine decarboxylase of the stomach showed two peaks of activity corresponding to those of the fetal and brain enzymes (pI's of 5.0 and 5.4) on isoelectric focusing. Anti-fetal-histidine decarboxylase antiserum inhibited the stomach and fetal enzymes extensively, but the brain enzyme only slightly. These results indicate that there are at least two types of histidine decarboxylase in rat tissue.
Factors II, IX, and X in the plasmas of 10 patients with 'hemorrhagic disease of the newborn' were investigated by means of electroimmunoassay and crossed immunoelectrophoresis (CIE). The biological activity was within 4.2-30 U/dl for factor II, 5.1-20 U/dl for factor IX, and 5.2-24 U/dl for factor X, whereas the immunological antigen was within 33-58, 25-50, and 35-60 U/dl, respectively. Thus, for all factors, 1.7 times more antigen than activity was present. The CIE pattern of factors II and IX in the presence of Ca++ ions clearly showed a biphasic precipitin arc, and in the absence of Ca++ ions, only one precipitin arc was observed. These results implied the presence of precursor proteins PIVKA-II and -IX in the plasma. However, even in the presence of Ca++ ions the CIE pattern of factor X antigen in the patient plasmas only showed a single precipitin arc with a slightly faster than normal electrophoretic mobility. PIVKA-II, -IX, and the abnormal factor X antigen (PIVKA-X) disappeared within 24 h after the patients were treated with vitamin K.
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We investigated the usefulness of enzyme immunoassay (EIA) for pancreatic oncofetal antigen (POA). The crude POA isolated from POA-positive ascitic fluid of patients with pancreatic cancer was injected into rabbits to raise anti-POA serum. The adsorbed antiserum was used for EIA as anti-POA serum. For the establishment of EIA system for POA, anti-POA-Fab' fragment was conjugated to beta-D-galactosidase from Escherichia Coli. Normal subjects (205 controls) and 132 patients (47 with pancreatic cancer, 22 with chronic pancreatitis, and 63 with other malignant disease) were surveyed. The standard serum from patient M with pancreatic cancer was used in quantitatively determining serum POA levels; value was expressed arbitrarily as 1000U/ml. Normal upper limit of POA was defined as less than 400U/ml (mean + 2SD of normal subjects). POA level higher than normal was observed in 72% of patients with pancreatic cancer, 23-44% of patients with other malignant diseases, and 18% of patients with chronic pancreatitis. The susceptibility of the isolated POA to several enzymes and chemical reagents was also studied. These results suggest the usefulness of EIA for POA in diagnosis of pancreatic cancer.
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In order to use FT-207 rectum suppository for a long-term maintenance chemotherapy of squamous cell carcinoma of the uterine cervix, we examined its pharmacokinetics by single administration at a dose of 1 g employing chemical assay. The results were as follows: 1. The blood level of FT-207 showed its peak of 30 mcg/ml at 2 hours after administration, of which half-time was 9 hours. Inter-individual differences were few in changes in the curve. 2. The blood level of 5-FU showed two peaks, 0.25 mcg/ml at 4 hours and 0.017 mcg/ml at 36 hours after administration. There were large inter-individual differences in its alteration. 3. At the time of operation, both FT-207 and 5-FU showed low concentrations in blood. 4. The level of 5-FU in cervical focus was 0.068 mcg/g which was the highest among those in all genital organs, and 2.7 times higher than the level of 5-FU in blood. 5. There was a significantly positive correlation at r = 0.174 (p less than 0.1) between the level of 5-FU in the focus of cervical cancer and the level of FT-207 in blood.
FT-207 rectum suppository at a daily dose of 1 g was repeatedly administered to patients with squamous cell carcinoma of the uterine cervix. Total doses ranged from 7 to 41 g. The following pharmacokinetic results were obtained in comparison with single administration. The blood level of FT-207 showed its peak of 35 to 37 mcg/ml at 4 hours after administration, of which half-time was 11 to 12 hours. The blood level of 5-FU showed its peak of 0.038 to 0.041 mcg/ml at 2 to 4 hours after administration, of which half time was more than 24 hours. Inter-individual differences were less than those of single administration. The level of FT-207 in tumor tissue was 14.2 mcg/g which was 15 times higher than that single administration. The level in healthy uterine cervix was 18.1 mcg/g which was 12 times higher that of single administration. The level of 5-FU in tumor tissue was 0.359 mcg/g which was 5 times than that of single administration. The level in normal tissue was 0.15 mcg/g which was 3 times higher than that of single administration. The level of 5-FU in tumor tissue was 7.8 to 8.4 times higher than that in blood. There was no difference in concentrations of both FT-207 and 5-FU between metastatic and non-metastatic lymph nodes.
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The radicality of Okabayashi's radical hysterectomy for treatment of cervical cancer has been well-known. However, its radicality also brought up the disturbance of fertility, endocrine and sexual function as well as the dysfunction of micturition and defecation. As early cervical cancer cases have increased yearly, and social rehabilitation became more necessary nowadays, functional saving is important and must be considered as the responsibility of the oncologists. This report denotes the preservation of ovarian function, sexual rehabilitation as well as the fertility in the treatment of cervical cancer.
The nature of sugar chain of factor VIII/von Willebrand factor in plasma of normal subjects and patients with von Willebrand's disease (vWd) was examined by crossed affinoimmunoelectrophoresis using anti-human factor VIII rabbit serum, with inserted Ricinus communis agglutinin-120 (RCA-120) agarose layer (RCA - CIE). Molecular weights of factor VIII-related antigen (VIIIR:Ag) were estimated by SDS polyacrylamide gel electrophoresis - crossed affinoimmunoelectrophoresis (SDS PAGE - RCA - CIE). VIIIR:Ag, in normal plasma and in classical form of vWd, showed two precipitin peaks on RCA - CIE. The slower moving component of VIIIR:Ag with molecular weights over 3 x 10(6) daltons from normal subjects and patients with classical form of vWd showed a high affinity for RCA-120. The faster moving component of VIIIR:Ag below 3 x 10(6) daltons from the above-mentioned subjects and patients with a variant form (Type IIA) showed a very weak affinity for RCA-120. These results suggested that all of VIIIR:Ag in these variant cases may have a deficiency of galactose residues reactive with RCA, in addition to an incomplete polymerization of VIIIR:Ag, similar to that of the faster moving component of normal subjects.
A new abnormal hemoglobin, Hb Kariya [alpha 40 (C5) Lys leads to Glu], with an amino acid substitution at the alpha 1 beta 2 contact was discovered in a young Japanese man. This variant migrated to the anode faster than Hb A, being nearly the same as Hb I in electrophoretic mobility. It amounted to about 6% of the total hemoglobin of the hemolysate. This hemoglobin showed an increased oxygen affinity, decreased heme-heme interaction and a lowered 2,3-DPG effect.