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Biomedical subjects

K Taketa

Publications and source records attributed to K Taketa.

At least 91 records · Page 5Linked to original sources
PubMed1986

Tissue of origin of elevated alpha-fetoprotein in ataxia-telangiectasia.

Although only a single gene exists for alpha-fetoprotein (AFP), differential glycosylation generates several different forms and these are associated with different tissues of origin, namely, liver, gastrointestinal tract, and yolk sac. This microheterogeneity of serum AFP was studied in seven patients with ataxia-telangiectasia (AT) in order to determine the tissue of origin of their elevated AFP levels. Concanavalin A (Con A), Lens culinaris agglutinin A (LCA-A), and erythroagglutinating phytohemagglutinin (E-PHA) affinity electrophoresis and antibody-affinity blotting were used to fractionate AFP. It was found that serum AFP in AT patients was composed mainly of Con-A band 2 (AFP-C2), LCA-A band 1 (AFP-L1), and E-PHA band 2 (AFP-P2). This profile of AFP species in AT patients is similar to those seen in neonates and patients with chronic hepatitis, but clearly different from AFP originating in hepatocellular carcinoma, gastric carcinoma, and yolk sac tumour cells. These data are compatible with a hepatic origin for the elevated AFP in AT patients. Since no evidence exists for ongoing liver damage in these patients, we suggest that the AFP gene in the AT liver may be under aberrant transcriptional control, perhaps secondary to a defect of DNA regulatory proteins which are necessary for hepatic maturation.

Ataxia Telangiectasia↗
PubMed1985

Responses of plasma cyclic AMP, serum immunoreactive insulin, C-peptide immunoreactivity and blood sugar levels to glucagon in patients with liver diseases.

Levels of plasma cyclic AMP, serum immunoreactive insulin (IRI), serum c-peptide immunoreactivity (CPR) and blood sugar (BS) were determined 0, 15, 30, 45 and 60 min after a glucagon injection (0.01 mg per kg body weight) in normal controls, patients with acute hepatitis and liver cirrhosis. Plasma cyclic AMP responses to glucagon in liver disease patients varied widely in peak value, and only in patients with fulminant hepatitis and decompensated liver cirrhosis with poor prognosis was the response suppressed. The peak response of BS was found significantly later in liver cirrhosis patients than in normal controls. IRI and CPR responses to glucagon were lower in acute hepatitis patients than in normal controls and liver cirrhosis patients. IRI levels and their sum were also lower in acute hepatitis patients, although CPR levels were not significantly different. Thus, the ratio of the sum of CPR from 0 to 60 min to that of IRI was significantly higher in acute hepatitis, indicating impaired pancreatic secretion of insulin to glucagon stimulation as well as increased uptake of insulin by the liver in acute hepatitis.

Adult↗
PubMed1985

Hormonal control of alpha-fetoprotein secretion in human hepatoma cell lines proliferating in chemically defined medium.

The regulation of alpha-fetoprotein (AFP) secretion and growth rate by various hormones in established human hepatoma (HuH-7, PLC/PRF/5, huH-1, huH-4, and KIM-1/c-4) and hepatoblastoma (HUH-6 Clone 5) cell lines was studied. These 6 cell lines replicated continuously in a chemically defined medium and secreted 84 ng (HuH-7) to 23 pg (huH-4) AFP per 24 h per 1 X 10(4) cells into the culture medium. The addition of insulin increased the growth rate of all examined cell lines and partially inhibited the AFP secretion in those cell lines except KIM-1/c-4, while the addition of dexamethasone inhibited the growth and stimulated the AFP secretion in all of the cell lines. The addition of 3,3',5-triiodothyronine inhibited the growth of all cell lines; however, different effects on the AFP secretion were observed depending on the cell lines used. Obviously, the AFP secretion was unrelated to the change in growth rate. When dexamethasone and N6-O2-dibutyryl cyclic AMP were added together, the AFP secretion was further stimulated. On the other hand, when dexamethasone and insulin were added simultaneously, the dexamethasone-mediated stimulation of AFP secretions was diminished. The data indicated that the regulatory mechanisms of AFP secretion by the hormones in the established human hepatoma and hepatoblastoma cell lines cannot be deduced according to the results of one cell line.

Animals↗
PubMed1985

Further resolution of human alpha-fetoprotein by affinity electrophoresis with erythroagglutinating phytohemagglutinin of Phaseolus vulgaris lectin.

Major molecular species of human alpha-fetoprotein(AFP), which were separated as single components by serial affinity chromatography with concanavalin A(Con-A) and Lens culinaris agglutinin, were further resolved into several bands by affinity electrophoresis with erythroagglutinating phytohemagglutinin of Phaseolus vulgaris lectin(E-PHA). Among the newly separated main molecular species, both Con-A- and E-PHA-reactive AFP(AFP-1X1) was demonstrated, contrary to the known sugar specificity of Con-A and E-PHA, in addition to molecular species of AFP reacting with Con-A but not with E-PHA(AFP-1X0) and of AFP reacting with E-PHA but not with Con-A(AFP-0X1). AFP-0X1 was formed from AFP-0X0, and AFP-1X1 from AFP-1X0 by neuraminidase treatment; thus, AFP-0X1 and AFP-1X1 represent asialylated and AFP-0X0 and AFP-1X0 sialylated molecular species. AFP-1X1' and AFP-0X0' were present as minor components. AFP-0X0' had no affinity for E-PHA, and the affinity increased in the order of AFP's-0X0(or 0X1), -1X1', -1X1 and -0X1. Proportions of those components varied depending on the pathophysiological conditions of AFP production.

Asialoglycoproteins↗
PubMed1985

Increased asialo-alpha-fetoprotein in patients with alpha-fetoprotein-producing tumors: demonstration by affinity electrophoresis with erythroagglutinating phytohemagglutinin of Phaseolus vulgaris lectin.

Affinity electrophoresis of human alpha-fetoprotein (AFP) with the erythroagglutinating phytohemagglutinin of Phaseolus vulgaris (E-PHA) gave up to five resolved bands (y, h, i, l and a; given in the order of decreasing affinity for E-PHA); band a having no affinity and bands y and h representing asialo-AFP's. The proportion of band y increased in extrahepatic tumors producing AFP, including yolk sac tumor, and of band h, in addition, in hepatocellular carcinoma. The proportion of either band y or h (or y + h) increased, over the means plus 2 standard deviations of the respective bands of cord serum AFP, in 20 out of 25 cases (80%) of hepatocellular carcinoma, including cell lines, and in all the patients with extrahepatic malignancy. Band i was detected in more than half the cases with malignancy, although the extent of its increases was much less. Band a appeared only in limited cases. None of the hepatitis and cirrhotic patients showed increased proportions of band y or h (or y + h), indicating the usefulness of the determination of asialo-AFP for the discrimination between benign and malignant liver diseases.

Adenocarcinoma↗
PubMed1984

Electrophoresis and electro-affinity transfer with specific antibodies to alpha-fetoprotein for detection of circulating immune complexes of alpha-fetoprotein.

A combination of agarose gel electrophoresis and a newly developed technique of electro-affinity transfer was applied to the detection of circulating immune complexes of human alpha-fetoprotein (AFP) and anti-AFP. After electrophoretic transfer to nitrocellulose membrane, to which affinity-purified polyclonal horse antibodies to human AFP were bound, the membranes were treated with or without rabbit immunoglobulins to human AFP, followed by overlaying with horseradish peroxidase-labeled goat anti-rabbit IgG for color development. Artificial complexes formed in vitro from human AFP and rabbit anti-AFP were clearly separated from free AFP by the agarose electrophoresis. The complexes were stained 20-40% as dark as the equivalent amount of free AFP by treatment with rabbit anti-AFP, and 10-20% as dark without the antibody treatment over a wide range of antigen-antibody ratios.

Animals↗
PubMed1984

Association of gamma-glutamyltransferase with plasma lipoprotein and lipid-protein complex in cholestasis.

Distribution of plasma gamma-glutamyltransferase (GGT) activity among lipoproteins was studied by ultracentrifugation and polyacrylamide gradient gel electrophoresis of plasma from 2 normal controls, 7 patients with intrahepatic cholestasis, and 5 patients with biliary obstruction. Half the total plasma activity was associated with lipoproteins, and more than 90 per cent of the residue activity with macromolecular lipid-protein complex. Healthy subjects had the highest GGT activity in HDL, intermediate activity in LDL, and the lowest in VLDL. In cholestasis, GGT activity increased mainly by the following two mechanisms: 1) the appearance of cholestatic lipoproteins carrying GGT, such as lipoprotein X or Slow-Migrating HDL, or increase in concentration of lipid-protein complex with GGT activity and 2) increase in GGT activity per unit weight of HDL.

Cholestasis↗
PubMed1984

Phenotypical stability of a human hepatoma cell line, HuH-7, in long-term culture with chemically defined medium.

A chemically defined medium containing selenium was further improved, and the improved medium showed excellent ability to support growth and colony formation of a human hepatoma cell line, HuH-7. The improved defined medium (designated as IS-RPMI) contained small amounts of unsaturated fatty acids, inorganic trace elements including selenium, and HEPES buffer. Many of these additions exert only a small individual effect on the growth of cells, but their cumulative effects were substantial. HuH-7 cells replicated continuously with a doubling time of 47-51 hr in IS-RPMI. A conditioned medium from a semiconfluent population increased the colony-forming ability of HuH-7 cells in IS-RPMI. The addition of insulin to the medium gave good colony growth, thus duplicating the effect of the conditioned medium. Although some alterations of morphological and growth characteristics of HuH-7 cells were observed, the differentiated liver functions, as revealed by production of plasma proteins including high levels of alpha 1-fetoprotein, were maintained for a period of more than 3 years and through 70 passages under the above culture conditions.

Animals↗
PubMed1983

Distinct molecular species of human alpha-fetoprotein due to differential affinities to lectins.

Resolution of human alpha-fetoprotein (AFP) into four distinct molecular species was demonstrated by a combination of two affinity chromatographies with crossed-immuno-affino-electrophoresis (CIAE) using concanavalin A (Con A) and Lens culinaris hemagglutinin (LcH)-A and LcH-B as affinity media. Of the four AFPs, AFP1 had no affinity for Con A, LcH-A, or LcH-B; AFP2 showed a high affinity for Con A, a low affinity for LcH-A, and an intermediate affinity for LcH-B (or a low affinity, depending on the lot of LcH-B preparations used); AFP3 revealed strong affinities for all of the three lectins; and AFP4, a trace component of hepatoma AFP in the present study, showed no interaction with Con A, but a definite interaction with LcH-A or LcH-B. These results were based on the determination of dissociation constants (Kd) of AFP-lectin complex by CIAE on isolated preparations of the three major hepatoma AFPs. These AFPs had identical electrophoretic mobilities of 0.86-0.87 (relative to human albumin) in the absence of lectins. The calculated mobilities of AFP2 and AFP3 were both reduced to 0.50-0.58 by saturation with lectins, but these two AFPs were clearly separated by 1 mg/ml LcH-A or LcH-B because of their large differences in Kd.

Carcinoma, Hepatocellular↗
PubMed1983

Separation and quantitation of hepatoma-associated gamma-glutamyltransferase by affinity chromatography with Affi-Gel blue and Con A-Sepharose.

Isozymes of serum gamma-glutamyltransferase (GGT) in patients with hepatocellular carcinoma (HCC) and other liver diseases were separated into two groups by double-affinity column chromatography with Affi-Gel blue and Con A-Sepharose, one recovered in the unbound fraction and the other in the bound fraction. Upon electrophoresis with polyacrylamide gradient gel slabs, the unbound fraction gave a GGTI1 band and a faint II1 band and the bound fraction gave a GGT I band and faint bands of GGT I", II' and X, when the original serum contained hepatoma-associated GGT (I1, I" and II') and high-molecular-weight lipid-protein complex, GGT(X). GGT I was present in all cases as a common isozyme. Other lipoprotein-associated GGT isozymes, III-IX, were removed by passing through Affi-Gel blue. GGT activities of unbound fraction in patients with HCC were generally higher than those in patients with non-HCC liver diseases, although the difference was not significant. When the percent of GGT activity of unbound (unbound + bound) was taken, 54% of patients with HCC had a ratio greater than 22%, whereas none of the healthy subjects or patients with other liver diseases gave values greater than this. The present technique may prove to be a useful clinical test for the diagnosis of HCC.

Carcinoma, Hepatocellular↗
PubMed1983

Polyacrylamide gradient gel electrophoretic studies of residual catalase in acatalasemia.

Erythrocyte catalse in a Japanese-type acatalasemia and a normal control subject was separated by chromatofocusing with or without prior partial purification with DEAE-cellulose. Fractions were analyzed by polyacrylamide gradient gel electrophoresis for catalse activity and protein stain. Chromatofocusing revealed no marked difference in pI values between normal and acatalasemic catalases with or without partial purification. In the gel electrophoresis, molecular weights were also similar; two bands of catalase activity with molecular weights of 290,000 and 350,000 for the acatalasemia and of 280,000 and 360,000 for the normal control were found in the partially purified preparations. The molecular weight of normal catalase in untreated hemolysate was 250,000. Normal catalse was identified as protein bands on polyacrylamide gradient gel after fractionation of hemolysate by chromatofocusing. A more sensitive method for protein stain is still required for demonstration of residual catalse protein on the gel.

Acatalasia↗
PubMed1982

Increased excretion of urinary cyclic GMP in primary hepatoma and preneoplastic liver.

Urinary excretion of cyclic GMP (cGMP) and the plasma level of cyclic AMP (cAMP) were determined in patients with liver diseases. The urinary excretion of cGMP, expressed on the basis of creatinine excreted per day, was at significantly higher levels not only in primary hepatoma but also in liver cirrhosis, while the plasma level of cAMP was higher only in liver cirrhosis. Thus, the ratio of urinary cGMP excretion to plasma cAMP level in primary hepatoma was significantly higher than that in liver cirrhosis. In cirrhotic patients studied by catheterization, the level of cGMP in the hepatic vein was significantly lower than that in the superior mesenteric or portal vein, indicating the uptake of cGMP by the liver. Since cGMP excretion correlated with KICG both in liver cirrhosis and primary hepatoma, the increased cGMP excretion appeared to be explained by a reduced uptake of cGMP by the liver.

Adolescent↗
PubMed1982

Growth of human hepatoma cells lines with differentiated functions in chemically defined medium.

A human hepatoma cell line, HuH-7, which was established from a hepatocellular carcinoma, was found to replicate continuously in a chemically defined medium when the medium was supplemented with Na2SeO3. The cells grew better in this medium than in serum-containing medium without any adaptation period. Other established human hepatoma and hepatoblastoma cell lines, HuH-6 cl-5, PLC/PRF/5, huH-1, and huH-4, also grew in the defined medium. Although HLEC-1 cells failed to proliferate continuously with Na2SeO3 alone, they grew if a cell-free conditioned medium from HuH-7 cells was added to the medium. These cell lines, except the HLEC-1 cell line, produced the following human plasma proteins among those examined: albumin, prealbumin, alpha 1-antitrypsin, ceruloplasmin, fibrinogen, fibronectin, haptoglobin, hemopexin, beta-lipoprotein, alpha 2-macroglobulin, beta 2-microglobulin, transferrin, lipoprotein, alpha 2-macroglobulin, beta 2-microglobulin, transferrin, Complement Components 3 and 4, and alpha 1-fetoprotein. Beside plasma proteins, the media from HuH-7, HuH-6 cl-5, PLC/PRF/5, and huH-1 contained anti-carcinoembryonic antigen-reactive proteins, and those from PLC/PRF/5, huH-1, and huH-4 medium contained hepatitis B surface antigen. These proteins were detected during periods of serial cultivation over 9 months under the above culture conditions. The hepatoma cell lines grown in the fully defined synthetic medium may provide a new approach for investigating the growth and metabolism of human hepatoma cells in vitro.

Blood↗
PubMed1981

Effect of nicomol on high density lipoprotein (HDL) subfractions, HDL2e and HDL3e, separated by electrophoresis.

High density lipoprotein (HDL) fractions were separated by dextran sulfate-Mg++ from sera of 5 healthy males before and after oral administration of a daily 1200 mg dose of Nicomol (2,2,6,6-tetrakis (nicotinoyloxymethyl) cyclohexanol; Cholexamin, Kyorin Pharmaceutical Co. Ltd.) for 2 weeks. The HDL fractions were further subdivided into HDL2e and HDL3e by Utermann's electrophoretic method on polyacrylamide gel containing lauric acid. These subfractions roughly corresponded to HDL2 and HDL3, respectively, which were separated by ultracentrifugation. Nicomol treatment increased the HDL2e/HDL3e ratio as is observed for the HDL2/HDL3 ratio, in addition to the increase in total HDL cholesterol concentration. The electrophoretic determination of HDL2e/HDL3e ratio in combination with HDL cholesterol level seemed to be clinically important in assessment of the alteration of HDL2 and HDL3 under physiological and pathological conditions.

Adult↗
PubMed1980

Cathodic isozyme of serum creatine kinase in a case of stomach cancer complicated by disseminated intravascular coagulation.

A rare isozyme of serum creatine kinase (CK) migrating cathodic to CK-MM on electrophoresis was found in a 30-year-old male with stomach cancer complicated by disseminated intravascular coagulation leading to massive upper gastrointestinal bleeding and marked anemia. Serum CK activity rose to a maximum of 374 U/l without detectable CK-MB isoenzyme. The patient was also characterized by a marked increase in serum lactate dehydrogenase (all isozymes elevated) and by preferential leakage of mitochondrial aspartate aminotransferase and glutamate dehydrogenase, indicating the presence of extensive tissue damage involving mitochondria. Skeletal muscle mitochondria were considered the most likely source of the additional CK isozyme.

Adenocarcinoma↗
PubMed1980

Hypoglycemic effect of disopyramide in a case of diabetes mellitus under insulin treatment.

In a 41-year-old male with diabetes mellitus well controlled with insulin (50 units/day), hypoglycemia developed after starting disopyramide treatment (200 mg/day) for correction of tachycardia. Lower levels of blood glucose and smaller amounts of urinary sugar persisted until disopyramide was withdrawn even after reducing insulin to a minimum dose of 20 units/day. The insulin requirement increased again thereafter to the original dose. These results indicate that disopyramide had a hypoglycemic effect in this patient.

Adult↗
PubMed1980

Effect of streptococcal preparation (picibanil) on the postoperative rise in serum alanine aminotransferase activity in patients with urogenital cancer.

The effect of Picibanil, a streptococcal agent, on the development of liver injury after operations for urogenital cancer was studied retrospectively in the light of serum alanine aminotransferase (ALT) activity. The series comprised 32 cases receiving Picibanil and 33 controls with otherwise comparable clinical backgrounds. Picibanil reduced the incidence of postoperative ALT rise over 50 U/l within 6 weeks but increased it thereafter. The increase in ALT activity after 6 weeks was relatively small and was seen more often in patients given blood transfusions. It was interpreted as retardation and suppression of ALT rise and as being related to the induction of interferon or to immunopotentiation. Other antihepatotoxic effects of Picibanil, due to its antioxidant activity, for example, may also account for the prevention of the early postoperative rise in ALT activity.

Adult↗