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Biomedical subjects

H Fukui

Publications and source records attributed to H Fukui.

At least 559 records · Page 31Linked to original sources
PubMed1976

Enzyme-linked sandwich immunoassay of macromolecular antigens using the rabbit antibody-coupled glass rod as a solid phase.

A highly sensitive sandwich immunoassay of macromolecular antigens using the rabbit antibody Fab' - beta-D-galactosidase complex and the rabbit antibody immunoglobulin-G-coupled glass rod as a solid phase is described. The Fab' fragments of rabbit antibody IgG are conjugated with beta-D-galactosidase from Escherichia coli using N,N'-o-phenylenedimaleimide. Rabbit antibody IgG is coupled to the aminoalkylsilyl glass rods (3 mm in diameter and 5 mm in length) using glutaraldehyde. A wide range of the concentrations of rabbit IgG fraction (20-2000 mug/ml) is effective for coupling, and the amount of rabbit immunoglobulin G coupled can be controlled. The smallest amounts of ornithine delta-aminotransferase from rat liver, human immunoglobulin G and 2,4-dinitrophenyl human immunoglobulin G that can be determined are 0.03, 0.3 and 0.04 fmol, respectively. The sensitivity of the assay for these antigens is affected mainly by the non-specific binding of the complexes to the solid phase and the ability of antigen molecules, adsorbed on the solid phase, to bind specifically the complexes. The assay with the rabbit antibody immunoglobulin-G-coupled glass rods is simpler and more sensitive than that with the rabbit antibody immunoglobulin-G-coupled Sepharose 4B.

Adsorption↗
PubMed1976

Enzyme-linked immunoassay. Conjugation of rabbit anti-(human immunoglobulin G) antibody with beta-D-galactosidase from Escherichia coli and its use for human immunoglobulin G assay.

Rabbit immunoglobulin G (IgG) was reduced by incubating with 10 mM 2-mercaptoethylamine and then treated with excess amounts of N,N'-o-phenylenedimaleimide. As a result, maleimide residues were introduced into rabbit IgG molecules. Rabbit IgG containing maleimide residues could be coupled to beta-D-galactosidase from Escherichia coli which has sulfhydryl groups in the molecule. The resulting rabbit IgG (antibody)-enzyme complex may be useful for immunoassay of antigens. As an example, human IgG was assayed by the sandwich method using the rabbit anti-(human IgG) IgG-beta-D-galactosidase complex and amounts of human IgG as small as 3 fmoles were measurable.

Animals↗
PubMed1976

A technique for the induction of labor by the extraovular administration of prostaglandin F2alpha.

Prostaglandin F2alpha (PG F2alpha) was administered to pregnant women at a dosage of 50 microng or 100 microng by extraovular route around the expected time of childbirth in order to induce labor. (1) PG F2alpha 50 microng or 100 microng dissolved in 10 ml of physiological saline solution was infused by extraovular administration through the cervical canal of the uterus with the use of Nelaton catheter No. 9. The catheter was then immediately withdrawn and the patient allowed to return home. (2) Instilation of 10ml of physiological saline solution was carried out for control purposes. (3) Those who reported labor contractions within 24 hours after the infusion were regarded as cases of excellent response, and those who reported labor contractions more than 24 hours after the infusion, or those who showed dilatation and improvement in effacement of the uterine cervical canal though without the occurrence of labor contractions were deemed to be cases of good response. The overall positive response rate (excellent plus good) was 78.9% for primiparae and 85.7% for multiparae in the group receiving 50 microng and 89.5% for primiparae and 100% for multiparae in the group receiving 100 microng. (4) Labor contraction were induced at a rate of almost 100% in those who showed dilatation of the cervical canal of the uterus to the extent of 3 cm or more and effacement to the extent of 50% or more. (5) It is advisable to perform reinfusion of PG F2alpha or instilation of PG F2alpha as a post-treatment in patients with good responses. (6) The infusion of PG F2alpha is effective for the induction of labor in older primiparae, cases of breech presentation and suspected cases of CPD. (7) There were some cases of enhanced response when PG F2alpha was given as posttreatment.

Cervix Uteri↗
PubMed1976

Intraocular irrigating solutions and lens clarity.

By in vitro incubation of rhesus monkey lenses at 37degreesC, we determined that solutions superior to intraocular irrigating solutions in common clinical use could be prepared by aspetically adding commerically available solutions of dextrose 5% in water and sodium bicarbonate to either Ringer's injection or Ringer's lactate solutions. While not as effective as glutathione-bicarbonate-Ringer's solution (G-B-R) in maintaining lens clarity, these mixtures can be prepared easily in the operating room (in contrast to G-B-r) and may improve surgical results in procedures requiring prolonged intraocular irrigation.

Animals↗
PubMed1976

Enzyme-linked sandwich immunoassay of ornithine delta-aminotransferase from rat liver using antibody-coupled glass rods as solid phase.

A macromolecular antigen, ornithine delta-aminotransferase [EC 2.6.1.13] from rat liver (OAT) was assayed by the sandwich procedure using rabbit (anti-OAT) Fab'-beta-D-galactosidase complex and rabbit (anti-OAT) IgG-coupled glass rods as a solid phase. The Fab' fragments of the rabbit (anti-OAT) IgG were conjugated with beta-D-galactosidase [EC 3.2.1.23] from Escherichia coli using N, N'-o-phenylenedimaleimide. The rabbit (anti-OAT) IgG was coupled to the aminoalkylsilyl glass rods using glutaraldehyde. The rabbit (anti-OAT) IgG-coupled glass rods were incubated with OAT and then with the rabbit (anti-OAT) Fab'-beta-D-galactosidase complex. The amount of OAT was determined from the activity of beta-D-galactosidase bound to the glass rods. A minimum of 0.03 fmoles of OAT could be determined by this method and use of the glass rods gave greater reproducibility, and was more sensitive and simpler than use of Sepharose 4B.

Animals↗
PubMed1976

Enzyme-linked immunoassay: conjugation of the Fab' fragment of rabbit IgG with beta-D-galactosidase from E. coli and its use for immunoassay.

1. A method for the conjugation of the Fab' fragment of rabbit IgG with beta-D-galactosidase from Escherichia coli is described. The method consists of two main steps: treatment of the Fab' fragments containing sulfhydryl groups with excess N,N'-o-phenylenedimaleimide, to introduce maleimide residues into the fragments, and then incubation of the dimaleimide-treated Fab' fragments with beta-D-galactosidase, which also contains sulfhydryl groups, to form the rabbit Fab'-beta-D-galactosidase complex. More than 90% of the enzyme used can be converted to the Fab'-enzyme complex, and the complex is readily separated from free Fab' fragments by chromatography on a Sepharose 6B column. 2. The application of the rabbit Fab'-beta-D-galactosidase complex for immunoassay of macromolecular antigens is shown by measuring human IgG by the sandwich method. The rabbit (anti-human IgG) IgG-coupled Sepharose 4B is incubated with human IgG and then with the rabbit (anti-human IgG) Fab'-enzyme complex, and the enzyme activity bound to the Sepharose is measured. In this way it is possible to determine as little as 0.3 fmoles of human IgG.

Animals↗
PubMed1975

Enzyme-linked immunoassay. II. A simple method for synthesis of the rabbit antibody-beta-D-galactosidase complex and its general applicability.

Rabbit anti-human IgG antibody was mildly reduced in the presence of 10 mM 2-mercaptoethylamine and then coupled to beta-D-galactosidase [EC 3.2.1.23] from Escherichia coli using N,N'-o-phenylenedimaleimide. Human IgG and the anti-human IgG antibody-beta-D-galactosidase complex were successively adsorbed on Sepharose 4B binding rabbit anti-human IgG antibody. In this way amounts of human IgG as small as 5X10(-15) moles could be measured by determining the activity of beta-D-galactosidase bound to the Sepharose.

Animals↗
PubMed1975

Enzyme-linked immunoassay. I. Novel method for synthesis of the insulin-beta-D-galactosidase conjugate and its applicability for insulin assay.

Pork insulin was subjected to mercaptosuccinylation and then coupled to beta-D-galactosidase [EC 3.2.1.23] from Escherichia coli using N,N'-o-phenylenedimaleimide. The competitive binding of the conjugate and insulin to anti-insulin antibody was tested. Results showed that formation of an insulin-beta-D-galactosidase conjugate could be used for immunoassay of insulin.

Binding, Competitive↗