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Biomedical subjects

K Taketa

Publications and source records attributed to K Taketa.

At least 73 records · Page 4Linked to original sources
PubMed1990

Lectin-reactive profiles of alpha-fetoprotein characterizing hepatocellular carcinoma and related conditions.

Serum alpha-fetoprotein from 146 patients with hepatocellular carcinoma, other malignancies, and benign liver diseases, was fractionated by lectin-affinity electrophoresis coupled with our sensitive detection method of antibody-affinity blotting. Compared with chronic hepatitis and liver cirrhosis, hepatocellular carcinoma was characterized by the increase in proportions of lentil lectin A-reactive alpha-fetoprotein-L3 and erythroagglutinating phytohemagglutinin-reactive alpha-fetoprotein-P4; the yolk sac tumor was characterized by the increase of concanavalin A-nonreactive alpha-fetoprotein-C1, lentil lectin-A-weakly reactive alpha-fetoprotein-L2, erythroagglutinating phytohemagglutinin-strongly reactive alpha-fetoprotein-P5, and Allomyrina dichotoma lectin-nonreactive, slow-migrating alpha-fetoprotein-Als; and gastrointestinal tumors were characterized by alpha-fetoprotein-C1, alpha-fetoprotein-L2, alpha-fetoprotein-L3, alpha-fetoprotein-P5 and Allomyrina dichotoma-nonreactive alpha-fetoprotein-A1. By combined evaluation of alpha-fetoprotein-L3 and alpha-fetoprotein-P4, hepatocellular carcinoma was discriminated from chronic hepatitis and liver cirrhosis with a sensitivity of 97% at a specificity of 99.7%. Because the alpha-fetoprotein level of the studied cases ranged from 60-1,500,000 ng/mL (60-1,500,000 micrograms/L), mostly greater than 200 ng/mL (200 micrograms/L), additional patients with lower levels of alpha-fetoprotein [16-177 ng/mL (16-177 micrograms/L) for 16 cases of hepatocellular carcinoma with liver cirrhosis and 28-185 ng/mL (28-185 micrograms/L) for 17 cases of liver cirrhosis alone] were analyzed for alpha-fetoprotein-L3 and alpha-fetoprotein-P4. The resulting sensitivity for combined evaluation was still as high as 88% at the same high specificity of 99.7%, indicating that the simultaneous analysis of alpha-fetoprotein-L3 and alpha-fetoprotein-P4 is effective in monitoring the evolution of hepatocellular carcinoma in cirrhotic patients.

Adult↗
PubMed1990

Datura stramonium agglutinin-reactive alpha-fetoprotein isoforms in hepatocellular carcinoma and other tumors.

By means of Datura stramonium agglutinin (DSA) affinity electrophoresis, human alpha-fetoprotein (AFP) was resolved into five bands, AFP-D1, D2, D3, D4 and D5, in order of decreasing mobility. AFP-D1, which had no affinity for DSA, comprised more than 84% of the intensity of total AFP bands. The percentage of AFP-D2 increased marginally in hepatitis and liver cirrhosis with or without hepatocellular carcinoma. AFP-D3 increased characteristically in hepatocellular carcinoma and AFP-D4, which had the highest affinity for DSA, increased up to 12% in other tumors, mostly of gastrointestinal origin. AFP-D5 showed no consistent changes among the benign and malignant diseases. The assay of AFP-D3 and D4 proved useful as a highly specific marker of hepatocellular carcinoma and other tumors, respectively.

Carcinoma, Hepatocellular↗
PubMed1989

Two-dimensional lectin affinity electrophoresis of alpha-fetoprotein: characterization of erythroagglutinating phytohemagglutinin-dependent microheterogeneity forms.

By means of two-dimensional lectin affinity electrophoresis of human alpha-fetoprotein (AFP) from different sources, AFP bands separated with erythroagglutinating phytohemagglutinin (E-PHA) were further characterized with other lectins of known oligosaccharide specificities. The results with a cord serum AFP revealed that not only AFP-P2 (E-PHA-nonreactive) but also AFP-P4 and P5 (E-PHA-reactive) had affinities for Concanavalin A (Con A) and Allomyrina dichotoma lectin (allo A), indicating that the cord serum AFP has nonbisected biantennary complex-type oligosaccharides with the terminal galactose on Man alpha 1----6 residue sialylated at the C-6, but not C-3, position. On the other hand, the results with a hepatoblastoma (HUH-6 C1-5 cell line) AFP showed that not only AFP-P5 but also AFP-P1 (E-PHA-nonreactive) and P3 (E-PHA-less reactive) had Con A-nonreactive AFP and that AFP-P1 had AFP-A1 (allo A-nonreactive) and AFP-A2 (allo A-less reactive), and AFP-P3 and P4 had AFP-A1s (allo A-nonreactive), as main components, in addition to the spots of cord serum AFP. Most of the E-PHA-dependent bands of AFP were further subdivided with Lens culinaris agglutinin (LCA-A) into LCA-A-reactive, weakly reactive and nonreactive spots. Similar results were obtained with AFP preparations from hepatocellular carcinomas and other malignancies, indicating that the bisected bi-(or tri- and tetra-) antennary sugar chains with the exposed terminal galactose of the Man alpha 1----6 arm as well as those with the C-3 sialylated galactose residues could be expressed in AFP upon malignant transformation.(ABSTRACT TRUNCATED AT 250 WORDS)

Concanavalin A↗
PubMed1989

Two distinct cell lines derived from a human osteosarcoma.

Two cell lines were established from a human osteosarcoma transplanted into athymic nude mice after the second (O9N2) and fifth passages (HuO9). Both cell lines expressed 1,25(OH)2D3-responsive alkaline phosphatase activity and produced tumors in the dorsum of nude mice that were histologically similar to the original tumor. However, the morphological and growth characteristics of the two cell lines differed. O9N2 cells were large and polygonal, whereas HuO9 cells showed spindle shapes. HuO9 cells had a higher growth rate and saturation density than O9N2 cells. The c-myc oncogene was amplified 4- to 8-fold in HuO9 cells but not in O9N2 cells. Both cell lines had a homozygous internal deletion, lacking the 7.4-kb HindIII fragment in the Rb gene. The results suggest the importance of the c-myc oncogene in the growth and morphological control of human osteosarcoma cells and of the Rb gene in the pathogenesis of the tumor.

Adolescent↗
PubMed1989

Pancreatic-type hyperamylasemia in end-stage renal disease.

Pancreatic-type isoamylase (P-type) and salivary-type isoamylase (S-type) activities were determined by the wheat protein inhibitor method in 29 patients with end-stage renal disease and in 38 healthy volunteers. Serum levels of total amylase (322 +/- 43 units/liter) and P-type (212 +/- 39 units/liter) in ESRD were significantly higher than those of controls (total: 142 +/- 7 units/liter, P less than 0.01; P-type: 52 +/- 4 units/liter, P less than 0.01). There was no significant difference between S-type activities in ESRD (110 +/- 16 units/liter) and in controls (90 +/- 6 units/liter). The ratios of amylase clearance to creatinine clearance (Cam/Ccr) and S-type clearance to creatinine clearance (Cs-amy/Ccr) rose significantly in ESRD(Cam/Ccr: 5.7 +/- 0.6%; Cs-amy/Ccr: 4.3 +/- 0.55%) compared to controls (Cam/Ccr: 3.2 +/- 0.24%, P less than 0.01; Cs-amy/Ccr: 2.1 +/- 0.17%, P less than 0.01). The ratio of P-type clearance to creatinine clearance (Cp-amy/Ccr) revealed no significant difference between ESRD (5.5 +/- 0.54%) and controls (5.6 +/- 0.42%). The renal excretion of P-type appeared to be more impaired than that of S-type in ESRD.

Adult↗
PubMed1989

Lectin reactivity of alpha-fetoprotein in a case of renal cell carcinoma.

The increased serum level of alpha-fetoprotein (AFP) in a case of renal cell carcinoma, a rare condition of AFP production by mesoderm-derived cells, was evaluated for its lectin reactivity by affinity electrophoresis, followed by the antibody-affinity transfer to nitrocellulose membranes for visualization of separated AFP bands. The AFP of this case was characterized by relative increases of concanavalin A-nonreactive AFP-C1 (60.4%), erythroagglutinating phytohemagglutinin-reactive AFP-P4 (37.8%) and AFP-P5 (46.3%) and Allomyrina dichotoma lectin-nonreactive AFP-A1s (66.7%), and by the total absence of lentil lectin-reactive components, AFP-L2 and AFP-L3. Thus, the lectin-reactive pattern of AFP markedly deviated not only from that of cord serum, but also from those of other malignancies and of fetal kidney cells in culture.

Carcinoma, Renal Cell↗
PubMed1989

An improved sensitive assay method for the heterogeneity of alpha-foetoprotein: possible application for early differential diagnosis.

A combination of lectin-affinity electrophoresis and antibody-affinity blotting was used for the qualitative determination of molecular species of alpha-foetoprotein. Concanavalin A and erythrophytohaemagglutinin were used as lectins. This method was able to quickly discriminate between the molecular species of alpha-foetoprotein of recurrent ovarian yolk sac tumour and those of non-malignant liver diseases and cord sera at term, using only 3 microliters of sera containing 200 micrograms/l of alpha-foetoprotein. The results indicate that this assay method might be useful for the differential diagnosis of recurrent yolk sac tumour from drug- or blood transfusion-induced liver diseases and for the monitoring of the serum alpha-foetoprotein level of patients with yolk sac tumour producing less than 200 micrograms/l of alpha-foetoprotein.

Antibodies↗
PubMed1988

Profiles of carbohydrate-metabolizing enzymes in human hepatocellular carcinomas and preneoplastic livers.

Activities of key carbohydrate-metabolizing enzymes in biopsied human tissues of hepatocellular carcinoma and related conditions were determined by established methods. Among the enzymes analyzed, fetal-type liver enzymes (low-Km hexokinase, glucose 6-phosphate dehydrogenase, and pyruvate kinase-M2) showed increased activities, and adult-type liver enzymes [glucose 6-phosphatase, fructose 1,6-bisphosphatase, high-Km hexokinase (or glucokinase), and pyruvate kinase-L] showed decreased activities, resulting in undifferentiated enzyme patterns not only in fetal livers and hepatocellular carcinomas but also in livers of acute and chronic hepatitis and liver cirrhosis with or without tumors. Hepatocellular carcinomas showed a general tendency of having greater enzyme deviations than hepatitic and cirrhotic livers. The extent of the enzyme deviation in hepatocellular carcinomas varied considerably from one enzyme to another for each tumor tissue as compared with that in the benign liver diseases. Thus, the phenotypic heterogeneity was important for discriminating between the neoplastic and inflammatory changes in differentiation markers. The enzyme patterns of tumors and their corresponding host cirrhotic livers were unrelated, suggesting that the cirrhotic liver has a significance as preneoplastic state only in terms of having a high incidence of evolving hepatocellular carcinoma.

Carbohydrate Metabolism↗
PubMed1987

A primary lung carcinoma producing alpha-fetoprotein, carcinoembryonic antigen, and human chorionic gonadotropin. Immunohistochemical and biochemical studies.

This article documents a patient with lung carcinoma that produced three oncofetal antigens including alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), and human chorionic gonadotropin (hCG). Serum AFP, CEA, and hCG-beta-subunit were extremely high--118,000 ng/ml, 133 ng/ml and 0.9 ng/ml, respectively. Immunohistochemical staining of these tumor markers revealed that these proteins were present in different cells. The pattern of lectin affinity electrophoresis of AFP resembled that of hepatocellular carcinoma. Also investigated was the reactivity of serum CEA to monoclonal antibodies against peptide or sugar moieties. Serum CEA values measured by antipeptide monoclonal antibodies were higher than those measured by antisugar monoclonal antibodies. The demonstration of AFP, CEA, and hCG in different tumor cells suggests that three genomes were not reactivated together in a cell, and the lung carcinoma probably consisted of at least three clones of cancer cells with different phenotypes.

Antibodies, Monoclonal↗
PubMed1987

Production of albumin and alpha-fetoprotein in primary culture of fetal human liver cells on collagenous substrata in the presence of hydrocortisone.

When primary cultures of fetal human liver cells established on type I collagen gels were compared to sister cultures developed on tissue culture plastic, the cells in contact with type I collagen secreted albumin at a higher rate than those without contact. The albumin secretion was dependent on the presence of hydrocortisone (HC) in the medium. Also, alpha-fetoprotein (AFP), of which the level decreased gradually and became undetectable after 6 d regardless of the presence or absence of HC in the cells cultured on plastic, was maintained for longer periods of time by plating the cells on type I collagen gels in the presence of HC. Different secretion rates of albumin and AFP were observed after Day 13 and Day 16, respectively, between cells maintained on type I collagen gels and those on film plastic. The cells secreted larger amounts of both albumin and AFP in plates coated with type IV or I collagens than with fibronectin after Day 10. The cells cultured on type I collagen gels were cuboidal in shape, whereas those on plastic were flattened in cultures with HC. These data indicate that the secretion of human albumin and AFP is facilitated by synergies between HC and collagenous substrata.

Albumins↗
PubMed1987

Immunohistochemical and affinity electrophoretic studies of alpha-fetoprotein of a patient with hepatocellular carcinoma in the intrathoracic space.

Immunohistochemical and affinity electrophoretic studies were carried out on alpha-fetoprotein components produced by a tumor of hepatocellular origin located on the chest wall and originating from the intrathoracic and extrapleural space tissue. The major components of alpha-fetoprotein were consistent with those produced by the hepatocellular carcinoma. Accordingly, this seems to be the first reported case of hepatocellular carcinoma that may have developed from accessory liver tissue seemingly isolated from the liver and located in the intrathoracic space.

Angiography↗
PubMed1986

A tetrazolium method for staining peroxidase labels in blotting assays.

A sensitive staining method of horseradish peroxidase-labeled immunoglobulins on nitrocellulose membrane was established by employing a reaction chain leading to formazan formation with phenol as a substrate of peroxidase and NADH as a hydrogen donor to reduce nitro blue tetrazolium. Higher concentrations of NADH relative to phenol were necessary to increase the intensity of staining and to ensure a wide dose-response range of color production with respect to the applied enzyme activities. By an optimized tetrazolium method in combination with antibody-affinity blotting, as low as 4 ng/ml alpha-fetoprotein was detected and 3-4-fold greater color intensities in a working assay range as compared with those of existing methods were obtained. The present technique of peroxidase staining may prove to have a wide application for the enzyme immunoassay using blotting modalities.

Immunoenzyme Techniques↗
PubMed1986

Microheterogeneity of Kasahara isozyme.

The microheterogeneity of Kasahara isozyme was investigated by affinity electrophoresis with Con A as the affinity ligand in combination with polyacrylamide gradient gel electrophoresis. On two-dimensional Con A-containing agarose gel electrophoresis, the Kasahara isozyme was separated into three molecular species. Kasahara isozyme electrophoresed as two distinct bands with enzyme activity on polyacrylamide gradient gel, but liver, intestinal or placental alkaline phosphatase showed only one distinct spot or band on both electrophoreses. One of the three molecular species of Kasahara isozyme separated by Con A-containing agarose gel electrophoresis was extracted from the gel and applied to the polyacrylamide gradient gel electrophoresis again, resulting in the same electrophoretic pattern as that of the original Kasahara isozyme. These findings indicated that the Kasahara isozyme consists of at least four molecular species. The same analysis was conducted with alkaline phosphatase of the HuH-6 cl-5 cell line, which has been reported to release an alkaline phosphatase closely resembling the Kasahara isozyme, and the results were compared with those obtained with the Kasahara isozyme.

Alkaline Phosphatase↗
PubMed1986

The effect of conditioned medium on fetal human liver cells in primary culture.

A medium conditioned by rat embryo cultures (RCM) promoted the adhesion of liver cells from human fetuses to plastic dish. Colonies were also formed in primary cultures of the same cells in the presence of RCM. The majority of the colonies formed were composed of large polygonal cells with a few colonies composed of both clear epithelial-like cells and fibroblast-like cells. RCM was superior to the rat embryo feeder layer for promotion of colony formation of cells. A number of colonies were formed from fetal human livers when a conditioned medium from human hepatoma cells (HCM) was used, but most of the colonies formed were composed of fibroblast-like cells. The cells derived from the polygonal cell colonies, which were formed in the presence of RCM, have been passaged four times and they are still growing with albumin-producing capacity. The effect of RCM was reduced by various physico-chemical treatments.

Animals↗
PubMed1986

Allomyrina dichotoma lectin-nonreactive alpha-fetoprotein in hepatocellular carcinoma and other tumors: comparison with Ricinus communis agglutinin-I.

Allomyrina dichotoma lectin (allo A) with a specificity to beta-D-galactose was used to fractionate human alpha-fetoprotein (AFP) by affinity electrophoresis. AFP from cord sera and serum of a patient with fulminant hepatitis showed single bands with a high affinity for allo A. Some patients with hepatocellular carcinoma and patients with gastric cancer and yolk sac tumor had two additional AFP bands, one weakly reactive and the other nonreactive with allo A. Patterns of AFP bands obtained with Ricinus communis agglutinin-I (RCA-I) and erythroagglutinating phytohemagglutinin from Phaseolus vulgaris were entirely different from those obtained with allo A. Of the two common bands reactive with RCA-I, the weakly reactive one was relatively intense in some malignant patients and the strongly reactive one was detected in patients with extrahepatic tumors. Thus, affinity electrophoresis with those lectins provides a potentially useful adjunct for the discrimination between benign and malignant conditions with increased serum levels of AFP.

Carcinoma, Hepatocellular↗