Hiroshi Nabeshima

Aberrant behavior of mouse embryo development after blastomere biopsy as observed through time-lapse cinematography

OBJECTIVE To analyze whether blastomere biopsy affects early embryonal growth as observed through time-lapse cinematography. DESIGN Comparative prospective study between embryos in which a blastomere was removed and embryos in which a blastomere was not removed. SETTING An experimental laboratory of the university. MAIN OUTCOME MEASURE(S) We calculated the time between blastocele formation and the end of hatching, the time between the start and end of hatching, the number of contractions and expansions between blastocyst formation and the end of hatching, and the maximum diameter of the expanded blastocyst. RESULT(S) In blastomere removal embryos, compaction began at the six-cell stage instead of at the eight-cell stage. We also found that hatching was delayed in these embryos as compared with matched controls. Moreover, the frequency of contraction and expansion movements after blastocyst formation was significantly higher in the blastomere removal group as compared with the control group. Finally, the maximum diameter of the expanded blastocyst just before hatching was not significantly different between both groups. CONCLUSION(S) These findings suggested that blastomere removal has an adverse effect on embryonic development around the time of hatching. Thus, future developments in preimplantation genetic diagnosis and screening should involve further consideration and caution in light of the influence of blastomere biopsy on embryonal growth.

Genetic significance of skewed X-chromosome inactivation in premature ovarian failure

To determine the relationship between premature ovarian failure (POF) and skewed X‐chromosome inactivation (XCI), karyotype, and XCI status in 43 patients with POF (group I) and 43 age‐matched control women with regular menstrual cycles (group II) were evaluated. Evaluation of XCI status was based on the CAG triplet repeat polymorphism assay in the androgen receptor gene after sodium bisulfite treatment of DNA samples, and XCI patterns were classified as random (XCI < 70% skewing) or skewed (≥70%). Furthermore, skewed XCI was classified under three different thresholds (≥70, ≥80, or ≥90%). Karyotyping by G‐banding and fluorescence in situ hybridization (FISH) on peripheral blood lymphocytes showed that one patient in group I had a deletion of Xq22, and another was 47,XXX. The frequency of low‐level 45,X/46,XX mosaicism was nearly equal in both groups. In women without any X‐chromosomal aberrations, the incidence of skewed XCI in group I was significantly higher than in group II on all threshold levels. Furthermore, extremely skewed XCI (≥90%) was observed only in group I. These results indicate that POF may be caused by some underlying genetic disorders, which may induce skewed XCI.

Analysis of the clonality of ectopic glands in peritoneal endometriosis using laser microdissection

OBJECTIVE To investigate the clonality of ectopic gland cells in peritoneal endometriosis. DESIGN Prospective study. University hospital. PATIENT(S) Seventeen women with surgically diagnosed endometriosis. INTERVENTION(S) Samples of peritoneal endometriotic lesions were obtained from patients during laparoscopic surgery. MAIN OUTCOME MEASURE(S) Clonality analysis used the laser microdissection technique, a phosphoglycerate kinase (PGK) gene polymorphism assay, and an androgen receptor (AR) gene polymorphism assay after digestion of the DNA with methylation-sensitive endonuclease. RESULT(S) Each ectopic gland of the peritoneal endometriotic lesion showed a monoclonal pattern in both the PGK gene and AR gene assays, but the methylation pattern of the PGK gene and/or AR gene was divergent among adjacent glands in the lesion. These data indicate that the peritoneal endometriotic lesions are multicellular in origin, although individual glands of the lesion are derived from single precursor cells. CONCLUSION(S) The colored peritoneal endometriotic lesion in the present study was multicellular in origin. Peritoneal endometriotic lesions may thus be initiated by transplantation of a cluster of eutopic endometrial tissues into the pelvis.

Total Laparoscopic Surgery of Cystic Adenomyoma under Hydroultrasonographic Monitoring

A 19-year-old-woman had a cystic adenomyoma located within the myometrium. She complained of severe dysmenorrhea. Gonadotropin-releasing hormone agonist therapy was administered, but her dysmenorrhea was more pronounced than before treatment. Therefore, total laparoscopic resection of the lesion was performed. The external appearance of the patients uterus was almost normal. A hydroultrasonographic monitoring method was devised (transvaginal ultrasonography with peritoneal hydration of physiologic saline) that proved useful in locating the adenomyoma. The patients dysmenorrhea disappeared postoperatively.

Risk factors for recurrence and re-recurrence of ovarian endometriomas after laparoscopic excision.

Aim:  Since ovarian endometrioma is frequently diagnosed in women of reproductive age, laparoscopic excision of the endometrioma is performed for most cases. However, endometriomas frequently recurs even after repeated surgical procedures. The aim of our study is to identify risk factors for recurrence and re‐recurrence of endometriomas after the first and second laparoscopic excision.

Different embryonic development after blastomere biopsy for preimplantation genetic diagnosis, observed by time-lapse imaging

Using time-lapse imaging, we found different behavior of mouse embryonal development after blastomere biopsy for preimplantation genetic diagnosis. Blastomere removal has effects on the developmental behavior of the mouse embryo, including speed of growth, contraction and expansion movements, and hatching.

Predicting Outcome of One-Step Total Hysteroscopic Resection of Sessile Submucous Myoma

STUDY OBJECTIVE To analyze variables for successful 1-step hysteroscopic myomectomies of sessile submucous myomas. DESIGN Retrospective case-control study. (Canadian Task Force classification II-2). SETTING Single operators practice in a university hospital and its related hospitals. PATIENTS Twenty-eight patients with sessile submucous myomas and menorrhagia, infertility, or both. INTERVENTIONS Our strategy for hysteroscopic myomectomy is as follows. First, we scraped and/or vaporized intrauterine dome of myoma until top of myoma was even with level of wall of cavity. Next, the remnant intramural node was squeezed by uterine contractions induced by prostaglandin F2alpha injection. Finally, the newly raised myoma dome was sectioned or vaporized electrosurgically only within the space of the intrauterine cavity and/or was separated mechanically from healthy myometrium without electrosurgery. MEASUREMENTS AND MAIN RESULTS Submucous myomas in 16 (57.1%) patients were completely removed after 1 surgery. By logistic regression analysis, thickness of outer myometrial layer of myoma node (OR 3.06, p = .02), myoma size (OR 0.86, p = .04), and intramural extension degree (OR 0.91, p = .03) were significantly associated with outcome of complete resection. CONCLUSION Thickness of outer myometrial layer of myoma node, myoma size, and intramural extension degree predicted outcome of 1-step hysteroscopic myomectomy. The chance of performing successful surgery increased with increased thickness of outer myometrial layer of myoma, and decreased with larger myomas and greater degrees of intramural extension.

Total Laparoscopic Conservative Surgery for an Intramural Ectopic Pregnancy

A 38-year-old woman, gravida 3, para 1 with a history of a left salpingectomy for an ectopic pregnancy was admitted for treatment of a presumed ectopic pregnancy. Transvaginal sonography revealed an ill-defined gestational sac and fetal heart beat within the fundal myometrium adjacent to the left cornua. Laparoscopy was performed for a suspected left cornual pregnancy or intramural pregnancy. A cystic mass 3 cm in diameter was visible within the fundal myometrium. Total laparoscopic removal of the gestational sac was performed, and the uterus was preserved. Pathologic evaluation of the excised mass demonstrated chorionic villi involving the myometrium. In the literature, only one other case describing the laparoscopic removal of an intramural pregnancy has been reported. However, in the prior report, the patient still required hysterectomy after conservative surgery. Therefore, this is the first report of the successful treatment of an intramural pregnancy exclusively with laparoscopy.

The shape of the sperm midpiece in intracytoplasmic morphologically selected sperm injection relates sperm centrosomal function

PurposeTo evaluate whether the morphology of the sperm midpiece observed by high magnification microscopy relates to sperm centrosomal function.MethodsSperm selected by conventional microscopy were defined as controls. By high magnification microscopy, sperm with straight midpieces were defined as Group 1, while those with tapering midpieces were defined as Group 2. Heterologous ICSI of human sperm into bovine oocytes was used to assess human sperm centrosomal function and analysis of sperm aster formation.ResultsThe total rate of sperm aster formation was 80.5% in Group 1, which was significantly higher (P < 0.05) than the rate of 33.3% seen for Group 2. Furthermore, sperm aster formation rates tended to be higher for Group 1 than for the controls.ConclusionsThis study demonstrates improvement of sperm aster formation rates by selecting sperm on the basis of midpiece morphology. The injection of selected sperm bearing morphologically straight midpieces may contribute to improved expression of sperm centrosomal function, providing a positive effect on fertilization after ICSI.

Successful Total Laparoscopic Cystic Adenomyomectomy After Unsuccessful Open Surgery Using Transtrocar Ultrasonographic Guiding

A 27-year-old woman with a cystic adenomyoma located within the myometrium underwent laparotomy unsuccessfully, with persistent postoperative heavy dysmenorrhea. Total laparoscopic resection of the cystic adenomyoma was then attempted. Intraoperative transtrocar ultrasonography was used to detect the location and boundaries of the cystic adenomyoma. The cyst was removed laparoscopically, and dysmenorrhea completely disappeared postoperatively. This is the first report of total laparoscopic resection of cystic adenomyoma after unsuccessful laparotomy, a minimally invasive approach that successfully eliminated the patients severe signs and symptoms.