Kiyoshi T. Fujii

Termination of early pregnancy by ONO-802 (16,16-Dimethyl-trans-δ2-PGE1 methyl ester)

ONO-802 was infused into the uterine cavity for the termination of early pregnancy in 45 healthy volunteers. Forty two (93%) of the 45 cases had complete abortions and two had incomplete abortions. Nine (20%) of the 45 volunteers complained of nausea, 7 (16%) vomited and 5 (11%) complained of abdominal pain. Neither diarrhea nor weakness was observed. These results suggest that ONO-802 is more acceptable for the termination of early pregnancy than is PGF2alpha.

A rapid molecular diagnosis of posttransfusion graft-versus-host disease by polymerase chain reaction

A woman with recurrent Pagets disease of the vulva developed acute graft‐versus‐host disease (GVHD) 12 days after radical surgery and massive blood transfusion. Molecular diagnosis of lymphocyte chimerism in the peripheral blood was made by polymerase chain reaction (PCR) directed against a Y chromosome‐specific sex‐determining region Y (SRY) gene. PCR with skin biopsy after onset of GVHD also revealed infiltration of SRY‐positive donor lymphocytes. The diagnosis was confirmed by HLA‐DNA typing with PCR‐sequence‐specific oligonucleotide that revealed the presence of complex HLA‐DR chimerism in the peripheral lymphocytes collected after onset of GVHD. The use of SRY‐ directed PCR is a rapid technique for the early diagnosis of acute posttransfusion GVHD in female patients.

The abortifacient effect of 16,16-dimethyl-trans-δ2-PGE1 methyl ester, a new prostaglandin analogue, on mid-trimester pregnancies and long-term follow-up observations

The present clinical trials revealed that 16,16-Dimethyl-trans-delta 2-PGE1 methyl ester in the form of vaginal suppositories is highly effective in inducing mid-trimester termination of pregnancies. It also showed that prior treatment with laminaria and metreurynter may enhance the success rate while reducing the incidence and severity of side effects. It is easy and safe to use clinically, with minimal side effects, and in our series, revealed no deleterious effects on ensuing reproductive physiology. However, the definite mechanism involved in the action of this new analogue to cause myometrial contractions is still not completely understood, and requires further intensive investigation.

A radioimmunoassay of plasma unconjugated and conjugated estetrol

A radioimmunoassay for the measurement of both unconjugated and conjugaged estetrol in plasma has been developed. The antiserum obtained after 6 months of immunization with 6-oxoestetrol-6-(O-carboxy-methyl)oxim-BSA was used at a final dilution of 1:90,000 and showed almost no cross reaction with other steroids except for estriol at 1.24%. Esterol-glucosiduronate was synthesized by incubating with adrenalectomized rat liver homogenate and uridine diphosphoglucuronic acid. Then, plasma estetrol-glucosiduronate was measured in the same manner for unconjugated estetrol after hydrolysis with beta-glucuronidase. Sephadex LH-20 column chromatography (7X110 mm, benzene:methanol, 85:15) was employed for accurate assessment. The sensitivity was 10 pg and the smallest amount measurable was 40 pg/sample. The method bland was consistently negligible. The intra and inter assay precision was 11.8% and 14.2% for unconjugated estetrol and that for estetrol-glucosiduronate was 13.5% and 17.1%.

Expression of the recombinase-activating gene (RAG-1) in murine early embryogenesis

The recombinase activation genes, RAG‐1 and RAG‐2, are expressed together in immature T or B lymphocytes and possess activity to induce V(D)J rearrangement in T cell receptor (TCR) and Ig genes. In vertebrates, only Ig and TCR molecules are reported to have recombination in their development using multiple V, D, J component gene segments. Thus, expression of RAG genes are localized only in lymphoid organs and sites of extrathymic T cell differentiation. In this study, we have used RAG‐1 and RAG‐2 genes as markers of possible genetic recombination in developing murine preimplanation embryos, using the highly sensitive reverse transcriptase polymerase chain reaction (RT‐PCR) technique and in situ hybridization. From 40 preimplantation embryos of various developmental stages we extracted RNA. reverse‐transcribed it into cDNA and used it in RT‐PCR studies. A PCR of 35 cycles disclosed expression of RAG‐1 but not RAG‐2 in morulae and blastocysts. Southern blot hybridization using a specific synthetic oligonucleotide probe for RAG‐1 and RT‐PCR with another primer pair identified RAG‐1 expression in developing embryos. In situ hybridization using a cooled CCD camera also revealed localization of RAG‐1 mRNA in blastocysts. We propose possible genetic recombination during late preimplantation murine embryogenesis which may contribute to the loss of totipotency and differentiation of inner cell mass and trophoectoderm.