Harumi Kubo

Laparoscopic excision of myometrial adenomyomas in patients with adenomyosis uteri and main symptoms of severe dysmenorrhea and hypermenorrhea

Preoperative magnetic resonance imaging accurately diagnosed adenomyosis uteri in three women. We performed laparoscopic excision of myometrial adenomyomas and localized portions of adenomyosis uteri in all women in whom the disorder was accompanied by severe dysmenorrhea and hypermenorrhea. We used the same procedure as for laparoscopic myomectomy. There were no intraoperative or postoperative complications, and patients were hospitalized only 3 days. The womens dysmenorrhea and hypermenorrhea disappeared by the end of the first postoperative menses.

Weekly docetaxel for patients with platinum/paclitaxel/irinotecan-resistant relapsed ovarian cancer: a phase I study

BackgroundThis study was designed to investigate the dose-limiting toxicity (DLT), maximum tolerated dose (MTD), and recommended dose (RD) of weekly docetaxel treatment in patients with relapsed ovarian cancer after the administration of platinum/paclitaxel/irinotecan.MethodsPatients were enrolled on the basis of inclusion and exclusion criteria. Docetaxel was administered intravenously over a 60-min period on days 1, 8, and 15. Four dosage levels, 30, 35, 40, and 45 mg/m2, were employed, and the dosage was escalated from level 1 to level 4. DLT criteria were established, and the DLT was used as the criterion for deciding the MTD and RD.ResultsTwelve patients were enrolled. No grade 3/4 hematological toxicities were manifested at any dosage level. Grade 3/4 nonhematologic toxicities were manifested at level 4, consisting of fatigue/asthenia in 2 patients and neuropathy/sensory toxicity in 1 patient. Level 4 (45 mg/m2) was thus judged to be the MTD, and the RD was concluded to be one level lower, i.e., level 3 (40 mg/m2).ConclusionsIt was concluded that the RD for weekly docetaxel therapy is 40 mg/m2 per week in patients with relapsed ovarian cancer after the administration of platinum/paclitaxel/irinotecan.

Study of the In Vitro Maturation of Mouse Oocytes Induced by Microinjection of Maturation Promoting Factor (MPF)

AbstractPurpose: Maturation promoting factor (MPF) acts at theresumption of meiosis and nonspecifically throughout theanimal species. There exists a considerable body of literatureon MPF, but little work has been done to study the inductionof maturation of mammalian oocytes by microinjection ofextracted MPF. Methods: Immature (GV-stage) mouse oocytes weremicroinjected MPF extracted from matured Xenopus eggs in thepresense of dbcAMP. Results: The rate of germinal vesicle breakdown (GVBD)induced at 24 hr after MPF injection was significantly higher(90.5%) than that of the control (2.2%), which was injectedwith HTF medium containing dbcAMP (P < 0.0001). Therate of extrusion of the first polar body at 24 hr after MPFinjection was significantly higher (84.1%) than that of thesame control (1.1%) (P < 0.0001). Conclusions: From these results, it is concluded that thematuration of mammalian oocytes can be induced by themicroinjection of MPF extracted from other species.

Preimplantation Diagnosis by Fluorescence In Situ Hybridization Using 13-, 16-, 18-, 21-, 22-, X-, and Y-Chromosome Probes

Purpose:Our purpose was to select the proper chromosomes for preimplantation diagnosis based on aneuploidy distribution in abortuses and to carry out a feasibility study of preimplantation diagnosis for embryos using multiple-probe fluorescence in situ hybridization (FISH) on the selected chromosomes of biopsied blastomeres.Methods:After determining the frequency distribution of aneuploidy found in abortuses, seven chromosomes were selected for FISH probes. Blastomeres were obtained from 33 abnormal or excess embryos. The chromosome complements of both the biopsied blastomeres and the remaining sibling blastomeres in each embryo were determined by FISH and compared to evaluate their preimplantation diagnostic potential.Results:Chromosomes (16, 22, X, Y) and (13, 18, 21) were selected on the basis of the high aneuploid prevalence in abortuses for the former group and the presence of trisomy in the newborn for the latter. Thirty-six (72%) of 50 blastomeres gave signals to permit a diagnosis. Diagnoses made from biopsied blasotmeres were consistent with the diagnoses made from the remaining sibling blastomeres in 18 embryos. In only 2 of 20 cases did the biopsied blastomere diagnosis and the embryo diagnosis not match.Conclusions:If FISH of biopsied blastomere was successful, a preimplantation diagnosis could be made with 10% error. When a combination of chromosome-13, -16, -18, -21, -22, -X, and -Y probes was used, up to 65% of the embryos destined to be aborted could be detected.

Surgery results using different uterine wall incision directions in laparoscopic myomectomy of the intramural myoma

ObjectiveTo study clinical outcomes for different uterine wall incision directions, comparing vertical incision and transverse incision in laparoscopic myomectomy of the intramural myoma.MethodsLaparoscopic myomectomies were performed on 50 women with intramural myomas. Using a table of random numbers, they were randomly divided into a vertical incision group (25 women) and a transverse incision group (25 women) according to the direction of incisions in the uterine wall. The numbers of enucleated myoma, operation duration, amount of bleeding, and numbers of sutures were compared. The Mann-WhitneyU-test was used for analysis.ResultsFor the transverse incision group, the amount of bleeding (137.6 ± 88.1 mL) was a significantly lower value (P = 0.0426) than for the vertical incision group (235.8 ± 169.4 mL). In addition, in cases where the maximum myoma nucleus diameter was 7 cm or larger, operation duration (129.0 ± 32.5 min) and amount of bleeding (158.9 ± 87.1 mL) showed significantly lower values (P = 0.0067 andP = 0.0002, respectively) for the transverse incision group than did operation duration (362.3 ± 147.3 min) and amount of bleeding (362.3 ± 147.3 mL) for the vertical incision group.ConclusionTransverse incision of the uterine wall is useful to reduce the amount of bleeding in the laparoscopic myomectomy of the intramural myoma. Transverse incision also shortens operation duration in cases where the myoma nuclei are large.

Deletion detection for diagnosis of Duchenne muscular dystrophy in the Japanese population—Comparisom between the polymerase chain reaction and the Southern blot analysis

SummaryWe compared the efficacy of the multiplex PCR with that of the cDNA analysis for detection of deletions of the DMD gene in the Japanese patients. Thirty males with DMD from 27 Japanese families were studied by the multiplex PCR, and 24 of them were also investigated by Southern blot analysis. We used five dystrophin cDNA probes for deletion analysis. A total of 19 regions were amplified by the PCR to detect deletions, 9 regions by the method of Chamberlain et al. and another 10 regions by the method of Beggs et al. Deletions were detected in 14 (52%) out of 27 DMD families by the PCR. Southern blot analysis detected deletions in 14 (64%) out of 22 families. Thirteen (93%) of the 14 DMD families with deletions detected by Southern blotting were also confirmed by the multiplex PCR. Provided care is taken in cases where the deletion is limited to a single exon, the multiplex PCR appears to be an efficient and useful alternative to conventional Southern blot analysis for detecting deletions during the prenatal and postnatal diagnosis of DMD.

Quantitative evaluation of the influence of ovarian steroids on plasminogen activators and inhibitors in human endometrial cells and trophoblasts

INTRODUCTION Plasminogen activators and inhibitors were quantitated in cultured human endometrial and trophoblast cells under the influence of ovarian steroids in order to investigate the role of the fibrinolytic system for trophoblast invasion and anchorage. MATERIALS AND METHODS Plasminogen activators (t-PA and u-PA) and their inhibitors (PAI-1 and PAI-2) secretions were assayed in cultures of epithelial, stromal, and trophoblast cells. These cells were also cultured on a fibrin substrate for microscopic examination of the fibrinolytic degradation. RESULTS The u-PA from epithelial cells was predominant among PAs and PAI-1 in endometrial cells. Estradiol (E2) enhanced t-PA production in stromal cells and PAI-1 production in epithelial cells. Progesterone (P4) suppressed u-PA production in epithelial cells and enhanced PAI-1 production in both epithelial and stromal cells. Trophoblasts produced PAI-1, PAI-2, and small quantities of t-PA and u-PA, none of which were notably influenced by E2 or P4. The PAI-1 production in trophoblasts was more than four-fold greater than the u-PA production in epithelial cells. Epithelial and stromal cells initially grew on fibrin substrate but were gradually detached from the substrate with fibrinolytic degradation, with the exception of the stromal cells grown in the presence of P4 (or E2+P4). Trophoblasts grew well on fibrin substrate without fibrinolytic degradation both in the presence and absence of the steroids tested. CONCLUSIONS Fibrinolytic balance seemed to be basically maintained between the endometrial PAs and the relative excess of trophoblasts-derived PAI-1. This balance might be regulated principally by P4 and focally by E2 in the endometrial tissue for placental implantation.

Rescue of mouse embryos from 2-cell blocks by microinjection of maturation promoting factor.

OBJECTIVE To examine the rescue of mouse embryos from 2-cell blocks by the microinjection of maturation promoting factor (MPF) extracted from matured Xenopus eggs into one of the blastomeres of 2-cell stage mouse embryos. DESIGN Controlled laboratory study. SETTING First Department of Obstetrics and Gynecology, Toho University School of Medicine, Tokyo, Japan. ANIMAL(S) Eight- to 10-week-old female Crj:CD-1(ICR) mice. INTERVENTION(S) One of the blastomeres of the mouse 2-cell embryos was injected with MPF (MI group) or mHTF medium (MED group) at 28--32 hours after insemination. MAIN OUTCOME MEASURE(S) The developmental rate to blastocyst. RESULT(S) The developmental rate to blastocyst in the MI group (48.0%) was significantly higher than that in the MED group (0%). CONCLUSION(S) The 2-cell block was specifically rescued by the microinjection of MPF and not by the insertion of pipettes.

Cultrure and Transfer of Embryos as a Testing System for Embryo-toxicity of Chemicals*

ABSTRACT  Culture system of mouse embryos from 2‐cell stage to early post‐implantation stage was established. To assess the growth and development of embryos, morphological, biochemical and cytogenetical endopoints were settled as follows:

Regulating assisted reproductive technologies in Japan.

Japan’s first in vitro fertilization preembryo transfer (IVF-ET) baby was born in 1983 some 5 years behind Louise Brown, the world’s first IVF-ET child (1). Subsequent technological advances have been remarkable, and assisted reproductive technology (ART) has now become firmly established in the treatment of infertility. In 1999, 69,019 cycles of IVF-ET had been performed in Japan, with 11,929 babies born as a result; this equates to 1 in 100 live births. The number of facilities performing IVF-ET has increased annually, reaching 423 in 1999 (2). There are few large-scale IVF facilities in Japan handling in excess of 200 cases a year; instead the country is characterized by its numerous small clinics. To date, however, no legal regulations covering ART have been promulgated in Japan. Since the Japan Society of Obstetrics and Gynecology (JSOG) issued its “Announcements on IVF-ET” in 1983, new statements and revisions have been made in line with the emergence of new technologies. Nevertheless, it is necessary to point out that the development of practical legal regulations has moved slowly in this country. For example, although artificial insemination by donor (AID) was first performed in 1949, for some reason, this date is cited as 1996 in JSOG statements (3). Some couples now travel to countries where IVF is unregulated in order to reap the benefits of ART, including preimplantation genetic diagnosis (PGD),