Tom Kiyoshi Fujii

Sex differences in fetal gonadotropins and androgens

Abstract We investigated the hypothalamo-hypophyseo-gonadal system in human fetuses and neonates by serially sampling fetuses obtained either during spontaneous abortions, premature deliveries or under the conditions of the Eugenics Protection Law. Maternal specimens were obtained serially throughout pregnancy, at parturition and in the puerperium. The purpose of our study was to determine the presence or absence of sex differences in fetuses and neonates by determining plasma constituents. The subjects for determination were FSH, LH, hCG and its β-subunit, and androgens. We also attempted to determine the effects of LH-RH priming on the anterior hypophysics. We found fetal sex differences in FSH and testosterone levels, but were unable to detect any differences in the other materials determined. Furthermore we were able to detect an anterior pituitary response to LH-RH priming from the second trimester of pregnancy.

The effects of intramyometrial injection of prostaglandin F2α on severe post-partum hemorrhage

Effectiveness of prostaglandin F2alpha (PGF2alpha) in controlling postpartum uterine hemorrhage was evaluated with the following results. (1) Systemic administration, whether by continuous i.v. infusion or by gluteal i.m. injection, was not a completely adequate method for hemostasis. (2) Local administration by directly injecting into the uterine musculature, whether transabdominally or transvaginally, resulted in a dramatic reduction of the rate of bleeding. Routine clinical application of the direct intramyometrial injection of PGF2alpha for severe post-partum hemorrhage is recommended in view of its easy performance, excellent hemostatic effect, minimal side effects and good prognosis.

Murine fetal resorption and experimental pre-eclampsia are induced by both excessive Th1 and Th2 activation

It has been proposed that immune responses in mammalian normal pregnancy are Th2-like, thereby protecting the fetus and placenta from being rejected. Administration of exogenous Th1 cytokines into pregnant mice is reported to induce feto-placental resorption. However, the effects of exogenous Th2 cytokines and Th2 directed responses in pregnant animals have not been well studied. In this study, we examined IL-4 and IL-12, which play decisive roles in the development of Th2 and Th1 responses, respectively, in the induction of fetal resorption and development of experimental pre-eclampsia. Transfer of either IL-4 and/or IL-12 stimulated splenocytes from BALB/C virgin female mice into BALB/C pregnant mice mated with either C57BL/6 or BALB/C male mice resulted in fetal resorption and glomerular nephritis associated with hypertension and proteinuria. In mice treated with IL-12 stimulated splenocytes, fatty liver degeneration associated with bile retention was observed. These results indicate that both excessive Th1 and Th2 activation contribute to the development of fetal resorption and pre-eclampsia, but that Th1 is critical to the development of liver degeneration.

Effects of Paternal Lymphocyte Immunization on Peripheral Th1/Th2 Balance and TCR Vβ and VΓ Repertoire Usage of Patients with Recurrent Spontaneous Abortions

PROBLEM: The mechanism of immunotherapy for patients with recurrent spontaneous abortions is not well understood. In order to investigate the suppressor mechanism of paternal lymphocyte immunization, we examined peripheral blood lymphocyte subpopulations and the repertoire of T‐cell receptor (TCR) gene segments. METHOD OF STUDY: Twelve patients with recurrent miscarriage were treated with immunization with paternal lymphocyte vaccinations three times during 12–14 weeks. Before and 2 weeks after the final inoculation, lymphocyte subsets and intra‐cellular interferon (IFN)‐Γ and/or interleukin (IL)‐4 production were examined by flow cytometry. TCR Vβ and VΓ repertoires were examined by semi‐quantitative reverse transcription‐polymerase chain reaction (RT‐PCR). RESULTS: We found no significant difference in CD4/CD8 ratios, prevalence of CD56+CD3+ or CD57+CD3+ cells (possible extrathymic T cells), ΓΔT cells, and CD5+CD19+ (B‐1) cells. However, by in vitro activation with phorbol 12‐mytistate 13‐acetate (PMA) and ionomycin, peripheral CD4 cells demonstrated a significant decrease of IFN‐Γ‐producing T helper 1 (Th1) cells and an increase of IL‐4‐producing T helper 2 (Th2) cells after immunotherapy. Seven of nine patients who exhibited remarkable decreases in Th1/Th2 ratios became pregnant within 6 months after three courses of immunotherapy, and four women delivered healthy babies, while none of the three patients who exhibited an increased or unchanged Th1/Th2 ratio had full‐term pregnancies (χ2<0.0001). Further, changes in usage of TCR Vβ and VΓ gene segments were observed after immunotherapy in six patients examined. CONCLUSION: Our findings suggest a shift of Th1‐dominant to Th2‐dominant status by vaccination might play important roles in maintaining successful pregnancies. Induction of some T cells that utilize different TCR repertoires possibly suppresses maternal rejection reactions.

Clinical Evaluation of a Haemostatic and Anti-Adhesion Preparation Used to Prevent Post-Surgical Adhesion

TachoComb® consists of equine collagen in a sponge-like form coated on one side with human fibrinogen and bovine thrombin. This product functions as a haemostatic and physical barrier to inhibit post-surgical adhesion. In this study, we investigated TachoComb® to control oozing in 16 patients who required haemostasis. Evaluation of post-surgical adhesion by second-look laparoscopy was performed at 3 months and 7 months after initial surgery. Observation via laparotomy during Caesarean section was also performed at 13 months, 3 years and 4 years after initial surgery. In all but one patient, no macroscopic evidence of TachoComb® persistence was found. Furthermore, no de novo adhesions were detected at the TachoComb® application site. We have thus demonstrated that TachoComb® can be used to control oozing haemorrhage effectively from surgical sites and can prevent adhesion formation at the application site, and may thus be an effective method of preventing adhesion-induced infertility.

The effect of hyaluronic acid-carboxymethylcellulose in reducing adhesion reformation in rabbits.

The effect of hyaluronic acid-carboxymethylcellulose film (Seprafilm®)* in reducing postoperative adhesion formation was examined in a rabbit induced-adhesion model. During laparotomy, the ileocaecal region was mechanically and chemically abraded to induce lesions. After a 28-day recovery period, adhesions were lysed by microsurgery and Seprafilm® was applied to the lysed lesion in 10 rabbits and six rabbits received physiological saline. A third laparotomy was performed 10 – 14 days later and the area of adhesion reformation was compared to that found prior to application of test materials. The area of adhesion reformation with Seprafilm® decreased to (mean ± standard deviation) 11.71 ± 10.97% of the originally lysed lesion and, the area was significantly reduced compared with controls treated with physiological saline. These results suggest that the use of Seprafilm® may be a valuable new anti-adhesion material for abdominal or pelvic surgery and may be superior to existing anti-adhesion materials and techniques.

Termination of early pregnancy by ONO-802 suppositories (16,16-dimethyl-trans-Δ2-PGE1 methyl ester)

ONO-802 was used in the form of vaginal suppositories for the termination of early pregnancy in 63 healthy volunteers. Fifty-four (86%) of the 63 cases had complete abortions and remaining 9 (14%) had incomplete abortions. One (1.6%) of the 63 cases complained of nausea and vomiting, and 3 (4.8%) complained of headaches. No other side effects were observed. These results suggest that ONO-802 is acceptable in the form of vaginal suppositories for the termination of early pregnancy.

The Effect of Cross-Linked Hyaluronate Hydrogel on the Reduction of Post-Surgical Adhesion Reformation in Rabbits

The effects of cross-linked hyaluronate hydrogel and liquid sodium hyaluronate on post-surgical adhesion reformation were examined using a rabbit model. Primary adhesions in the ileocaecal region of Japanese white rabbits were induced by mechanical and chemical irritants during laparotomy. After 1 month the primary adhesions were lysed by microsurgery and cross-linked hyaluronate hydrogel or liquid sodium hyaluronate was applied to the lysed lesions. After 10 − 14 days the area of adhesion reformation was measured to assess any inhibitory effect of the test materials. Rabbits treated with cross-linked hyaluronate hydrogel showed a significant reduction in adhesion reformation area compared with liquid sodium hyaluronate or physiological saline treatment, and the area reduced to (mean ± standard deviation) 0.6 ± 1.95% of the original lesion. In a separate study, histological evaluation of rabbits treated with cross-linked hyaluronate hydrogel revealed a better healing pattern and a lower inflammatory response compared with controls. All these findings suggest cross-linked hyaluronate hydrogel may be a valuable anti-adhesion material to prevent post-surgical adhesion in abdominal or pelvic surgery.

Outpatient evaluation and treatment of tubal obstruction with selective salpingography and balloon tuboplasty

OBJECTIVE To compare selective salpingography and balloon tuboplasty for the treatment of tubal obstruction. DESIGN A retrospective evaluation of results of women treated for tubal obstruction by outpatient methods at a single center. SETTING Tertiary-care, university-affiliated hospital. PATIENT(S) A total of 3,424 infertile women, of whom 418 had bilateral tubal obstruction by hysterosalpingography, treated at Nihon Medical Center from 1982 to 1997. INTERVENTION(S) Women with tubal obstructions who had visual evidence of an intact uterine tubal ostium at hysteroscopy were treated by selective salpingography. If selective salpingography could not establish patency, then transcervical balloon tuboplasty was performed with one of three catheter systems. Patients were followed expectantly for 1 year after treatment. MAIN OUTCOME MEASURE(S) Postoperative tubal patency and overall pregnancy rates (PRs) at 1-year of follow-up. RESULT(S) The overall patency rate was 67.5%, with 30% of these patients conceiving (20.2% of all subjects). Selective salpingography was associated with a 35. 7% patency rate, and 27.3% of these patients conceived. Of the subjects who failed selective salpingography and underwent balloon tuboplasty, 66.2% achieved patency, of whom 33% spontaneously conceived. Balloon tuboplasty was effective in restoring patency in many cases after selective salpingography had failed. Statistically significant differences were found between selective salpingography and balloon tuboplasty and for the different balloon tuboplasty catheters in terms of patency rates, while a trend was seen for PRs. CONCLUSION(S) Many women diagnosed as having tubal obstruction can be treated by outpatient methods that do not require general anesthesia. Achieving patency by these methods is associated with high PRs and avoids the need for assisted reproductive technologies in some cases. Balloon tuboplasty is a more effective treatment than selective salpingography. The choice of balloon tuboplasty catheter system may affect success rates.

Fimbrial Capture of the Ovum and Tubal Transport of the Ovum in the Rabbit, with Emphasis on the Effects of β2-Adrenoreceptor Stimulant and Prostaglandin F2α on the Intraluminal Pressures of the Tubal Ampullae

Purpose:Our purpose was to elucidate the roles of the ampullar and isthmic portions of the oviduct and the effects of drugs on oviductal contractility.Methods:Prostaglandin F2α (PGF2α; Ono Pharmaceuticals, Osaka) and oxytocin (Atonin-O; Teikoku Hormone Manufacturing Co. Ltd., Tokyo) were used to stimulate oviductal contractility, and ritodrine hydrochloride (Utemerin; Solvay-Duphar Corp., Denmark) to inhibit the contractility.Results:Both PGF2α and Atonin-O were involved in ovum capture by the ampullar oviduct by stimulating contractility, thus altering the intraductal pressures. Utemerin is effective in inhibiting the enhanced contractility induced by PGF2α and Atonin-O.Conclusions:Variations in pressure of the ampullar portion of the oviduct seem necessary for the capture of ova expelled from the ovary. Once in the isthmic portion of the oviduct, transport appears to be under the influence of ciliary activity rather than variations in contractility.