The 1986 McCollum award lecture. Fuel-mediated teratogenesis during early organogenesis: the effects of increased concentrations of glucose, ketones, or somatomedin inhibitor during rat embryo culture.

Whole rat embryos were explanted at head-fold, late pre-somite stage (day 9.5 of gestation) and cultured in rat sera varyingly supplemented with glucose (3, 6, 9, or 12 mg/mL), D,L sodium beta-hydroxybutyrate (2, 4, 8, or 16 mM), or both (6 mg/mL D-glucose plus 8 mM beta-hydroxybutyrate). During 48 h culture, increasing glucose alone or beta-hydroxybutyrate alone effected growth retardation and faulty neural and extraneural organogenesis in dose-dependent fashion.

An enzyme-linked immunosorbent assay for antibodies against saline-soluble muscle components in myasthenia gravis.

An enzyme-linked immunosorbent assay (ELISA) system has been developed for measuring antibodies against rat skeletal muscle components solubilized with phosphate-buffered saline. With this assay, 53.8% (50/93) of sera from patients with myasthenia gravis (MG) was positive (the values over the mean plus 3 SD of 256 healthy individuals were considered significant).

Mitogenic heparin-binding lectin-like protein from cloned thymic myoid cells.

A mitogenic heparin-binding (reactive) lectin-like protein (HBP) was purified from the extract of a cloned rat thymic myoid cell R615B2 by a one-step procedure of affinity chromatography on a heparin--Sepharose CL-6B column. Four distinct peptide bands with molecular weights of 10,000, 13,000, 13,700, and 14,600 were detected on SDS-polyacrylamide gel electrophoresis. This protein is mitogenic at concentrations of as low as 1.

The cytotoxicity of 5-fluorouracil is due to its incorporation into RNA not its inhibition of thymidylate synthase as evidenced by the use of a mouse cell mutant deficient in thymidylate synthase.

The biochemical basis for the cytotoxicity of 5-fluorouracil is still controversial; it is not clear whether the mechanism involves interference with DNA metabolism or incorporation of the drug into RNA. To distinguish between these two possibilities, we took advantage of a mutant strain of the mouse mammary tumor cell line FM3A that is deficient in thymidylate synthase, which is widely believed to be the target of 5-fluorouracil. We demonstrated that the target of the cytotoxicity of 5-fluorouracil was not thymidylate synthase; instead, the cytotoxicity was positively correlated with drug incorporation into RNA under conditions where thymidine increased the incorporation of 5-fluorouracil into RNA concentration-dependently.

A new method of detection of islet cell antibodies (ICA) using peroxidase-labeled protein A, and incidence of ICA in type 1 (insulin-dependent) diabetes.

We have developed a new method of detecting islet cell antibodies using peroxidase-labeled protein A, and have determined the incidence of ICA in Type 1 (insulin-dependent) diabetes in Japan. In our method, fresh frozen sections of human pancreas and serum samples were incubated and then treated with peroxidase-labeled protein A at room temperature. Conjugates of peroxidase and protein A were subjected to Sephadex G-200 column chromatography, and only the 80,000 dalton peak was employed.

Relationships between glucose and mannose during late gestation in normal pregnancy and pregnancy complicated by diabetes mellitus: concurrent concentrations in maternal plasma and amniotic fluid.

Enzymatic assays were modified to permit sensitive and highly reproducible simultaneous measurements of D-mannose and D-glucose in biological fluids during weeks 34-40 of human pregnancy. Plasma mannose and glucose averaged 9.8 +/- 0.

[Biochemical modulation of 5-FU by thymidine and N-phosphoacetyl-L-aspartate: the relation between 5-FU incorporation into RNA and its cytocidal effect].

We attempted to study the relation between 5-FU incorporation into RNA or DNA and the cytocidal effect of 5-FU using thymidine (TdR) and the effect of N-phosphoacetyl-L-aspartate (PALA) on the cytotoxicity of 5-FU. A mouse mammary carcinoma cell line FM3A was used in all of the experiments. The results showed that: addition of TdR to cell cultures resulted in an increase of 5-FU incorporation into RNA and although 5-FU incorporation into nucleic acid decreased, addition of 10(-3) M TdR induced of complete incorporation of 5-FU into only RNA as revealed by alkaline hydrolysis and cesium gradient analysis; no addition of TdR caused 5-FU to be completely incorporated into DNA only; cytocidal effect of 5-FU, in a clonogenic assay, occurred only during the time when TdR was added to the cell culture; PALA possessed the capacity to enhance the cytocidal effect of 5-FU in a clonogenic assay.

Fuel-mediated teratogenesis. Use of D-mannose to modify organogenesis in the rat embryo in vivo.

The unique embryotoxic properties of D-mannose have been used as the basis for a new technique to secure precise temporal correlations between metabolic perturbations during organogenesis and subsequent dysmorphogenesis. Conscious, pregnant rats were infused with D-mannose or equimolar amounts of D-glucose by "square wave" delivery during the interval in which the neural plate is established and early fusion of neural folds takes place, that is, days 9.5-10.

[Diagnostic values of serum type III procollagen N-terminal peptide in type IV gastric cancer].

Since increased synthesis of collagen has been demonstrated in tissue of type IV gastric cancer, we attempted to distinguish type IV gastric cancer from other cancers by measuring serum levels of type III procollagen N-terminal peptide (type III-N-peptide). Mean serum levels in type IV gastric cancer patients without metastasis were found to be elevated above normal values and developed a tendency to be higher than those in types I, II and III gastric cancer patients without metastasis. Highly positive ratios were found in patients with liver diseases including hepatoma and colon cancer, biliary tract cancer, and esophageal cancer patients with liver, lung or bone metastasis, but only 2 out of 14 of these cancer patients without such metastasis showed positive serum levels of type III-N-peptide.

[Therapeutic effect of sequential doses of methotrexate (MTX) and 5-fluorouracil (5-FU) in advanced gastric cancer: comparison of intermediate-dose MTX with high-dose MTX].

Twenty-one patients were treated with sequential doses of MTX and 5-FU so as to be classified by MTX dosage into an intermediate MTX-dose group and a high MTX-dose group. In the intermediate-dose MTX group, the drug was given at a dosage of 100 mg/m2 intravenously (i.v.

Radioimmunoassay of human plasma apolipoprotein A-1: pretreatment of plasma with guanidine hydrochloride.

A sensitive and specific double antibody radioimmunoassay for apolipoprotein A-1 was first reported by Schonfeld and Pfleger (1). Their procedure required delipidation of the serum since direct radioimmunoassay of apolipoprotein A-1 in untreated sera resulted in much lower values than those obtained from the corresponding delipidated samples. In an attempt to find a rapid and simple procedure for radioimmunoassay of apolipoprotein A-1 without using organic solvents, the serum was subjected to various physical and chemical treatments which disrupt or alter high density lipoproteins (HDL).

A practical enzyme immunoassay for anti-acetylcholine receptor antibodies in myasthenia gravis.

A practical enzyme immunoassay (EIA) has been developed for the measurement of anti-acetylcholine receptor (AChR) antibodies in sera from patients with myasthenia gravis (MG). This system is based on the double antibody technique, using denervated rat muscle AChR labeled with horseradish peroxidase-linked alpha-bungarotoxin (HRP-alpha BGT). This method has the following advantages compared to conventional radioimmunoassay (RIA): (1) HRP-alpha BGT is more stable than [125I]alpha BGT and can be used for at least one year without any loss of the binding activity to AChR and enzymatic activity, (2) the procedure avoids the use of radioactive isotopes, and (3) the equipment for our EIA is more economical than that for RIA.

[Diagnostic values of type III Procollagen N-terminal peptide and combination assay of type III procollagen N-terminal peptide with CEA and CA 19-9 in gastric cancer].

It is known that interstitial collagens are initially synthesized as precursors (procollagen), which possess extra peptide segments at both ends of the molecules. The authors attempted to detect the aminoterminal peptide of type III procollagen (type III-N-peptide) and also to measure the carcinoembryonic antigen (CEA) and carbohydrate antigen (CA 19-9) together in sera of patients with gastric cancer. The results showed that: (1) mean serum levels and positive ratios of the type III-N-peptide increased as the clinical stage of the patients with gastric cancer advanced; (2) serum levels of the type III-N-peptide were not correlated either with those of CEA or CA 19-9; (3) positive ratios of type III-N-peptide, CEA and CA 19-9 were 51.

[Combination chemotherapy of CIS-diamminedichloroplatinum (II) and fluoropyrimidine derivatives for advanced gastric cancer: a preliminary report].

A total of thirteen patients with histologically-proved adenocarcinoma of stomach took part in this study at Saitama Cancer Center between July 1982 and September 1983; nine patients were considered evaluable. There were 7 male and 2 female patients with a median age of 52 yrs (range 42-75) and a median performance status of 3 (range 2-4). Patients were treated with a two drug combination of cis-diammine dichloroplatinum (II) and fluoropyrimidine derivatives (tegafur, cormobur ).

[Effect of cis-diammine dichloroplatinum, vindesine sulfate combination chemotherapy for advanced esophageal carcinoma].

Eleven patients with advanced squamous cell carcinoma of the esophagus were treated with a two-drug combination of cis-diammine dichloroplatinum and vindesine sulfate at Saitama Cancer Center between July 1982 and September 1983. Median age was 71 years old (range: 47-48) and 8 patients were greater than 70 years old. Median performance status was 3 (range: 1-4) by Koyama -Saito Criteria.

[Clinical trial on the effect of tegafur (SF-SP)].

SF-SP capsules containing sustained release granules of tegafur were orally administered 800 mg, b.i. d.

[Combination chemotherapy of nicardipine and vindesine sulfate, cis-diammine dichloroplatinum (II) for patients with advanced esophageal carcinoma].

Since calcium (Ca) antagonist enhances the antitumor effect of vinca alkaloid in vitro, The authors attempted to combine nicardipine (Ca antagonist) with vindesine sulfate (VDS) and cis-diammine dichloroplatinum (II)(CDDP) for treatment of two patients with advanced esophageal carcinoma who were considered to be resistant to two courses of combination chemotherapy of VDS and CDDP. In the first case nicardipine was given orally and for only one day at a dose of 60 mg followed by 40 mg two and a half hours later. Thirty minutes after 60 mg of nicardipine, 3 mg of VDS on day 1, and 50 mg of CDDP (repeated for 3 days) were given.