Shozo Kurotsuchi

PTEN and Akt expression during growth of human ovarian follicles

PurposeTo assess the expression of PTEN and total and phosphorylated Akt in human ovarian follicles during follicular growth.MethodsImmunohistochemistry of ovarian tissues and Western blotting and immunofluorescence of primary cultured luteinized granulosa cells for PTEN and Akt.ResultsImmunoreactivity of Akt was found in the oocytes, granulosa cells and theca cells in primordial follicles, follicles at each growing stage and luteal cells. As the follicles grew, staining for PTEN became intense in the granulosa cells, whereas the intensity of phospho-Akt became weak. Western blotting and immunofluorescence analysis using primary cultured granulosa-lutein cells showed Akt and PTEN expression, and phosphorylation of Akt in vitro.ConclusionsPTEN and Akt are present in the granulosa cells during folliculogenesis. An increase in PTEN may lead to changes in proliferation and/or differentiation of granulosa cells during follicular growth via regulation of Akt phosphorylation.

Successful delivery derived from vitrified-warmed spermatozoa from a patient with nonobstructive azoospermia

OBJECTIVE To report the clinical outcomes following intracytoplasmic sperm injection (ICSI) with vitrified sperm from patients with severe male factor infertility. DESIGN Retrospective case series. SETTING IVF unit of a medical center. PATIENT(S) Three patients with severe oligozoospermia or nonobstructive azoospermia (NOA). INTERVENTION(S) Cryopreservation of limited numbers of spermatozoa with the use of Cryotop and Cell Sleeper as nonbiologic containers. MAIN OUTCOME MEASURE(S) Four cycles underwent intracytoplasmic sperm injection (ICSI) with vitrified sperm. RESULT(S) A total of 148 spermatozoa in 18 containers (8.2 sperm per container) were vitrified and 36 of them (5 containers) were warmed. Thirty-three sperm (92%) were retrieved successfully and injected individually into 17 mature oocytes. Fertilization was observed in 12 oocytes (71%), and all zygotes (100%) cleaved. A couple with NOA achieved a singleton pregnancy and concluded with full-term delivery of a healthy boy (2,632 g). CONCLUSION(S) A successful delivery was achieved after transfer of a blastocyst derived from vitrified-warmed spermatozoa. A small number of vitrified sperm cells were used for ICSI to fertilize oocytes with predictable timing.

Long-term administration of dienogest reduces recurrence after excision of endometrioma

Background Pain-relieving effects of dienogest against endometriosis are comparable to leuprolide acetate for 24 weeks. We assessed whether long-term dienogest administration reduces recurrence after endometrioma excision. Methods In this retrospective cohort study, 568 women with MRI-based diagnosis of ovarian endometrioma, who underwent laparoscopic stripping between 2008 and 2013, were studied. Recurrence rates and side effects over 5 years were investigated in 417 without postoperative medication and 151 who received dienogest postoperatively at 2 mg. Transvaginal sonography was performed every 3 months, and when cystic lesions ≥2 cm were observed, diagnostic MRI was conducted. Recurrence was defined as a lesion previously diagnosed as endometrioma by MRI, equal in size or larger 3 months later on ultrasonography. Cumulative recurrence rates were calculated with the Kaplan-Meier method, and group comparison involved log-rank tests. Blood examinations were completed every 3 months, and bone mineral density was measured with DEXA every 6 months. Results Cumulative recurrence rates at the 5th postoperative year in the no-postoperative-medication and 2-mg dienogest groups were 69 and 4%, respectively, showing significant decreases (odd ratio [OR] = 0.09, 95% confidence interval, 0.03-0.26). Anemia occurred in 4% due to metrorrhagia directly after administration, metrorrhagia including spotting was observed in 20% at 1 year and decreases in bone mineral density and depression were observed in 4 and 2.6%, respectively, in the dienogest group. Conclusions Dienogest significantly prevented postoperative endometrioma recurrence. However, side effects such as metrorrhagia and a decreased bone mineral density were observed. Therefore, careful long-term follow-up is necessary.

Psychological stress during in vitro fertilization and embryo transfer is influenced by the patients’ background and gender

PurposeThis study evaluated the changes in psychological stress during in vitro fertilization and embryo transfer (IVF–ET) and the relationship of such stress to the patients’ background and gender.MethodsSixty couples undergoing IVF–ET were administered the State–Trait Anxiety Inventory-JYZ (STAI) test at six different points during IVF–ET procedures. Anxiety scores at each time point were recorded and analyzed according to gender, fertility status, and duration of treatment.ResultsThe median state anxiety score for women increased following induction until oocyte collection, after which it temporarily declined and then increased again until the pregnancy test. No such changes were noted in men. Scores for women who had undergone a shorter period of IVF treatments were higher while state and trait anxiety in men increased with a prolonged treatment period. Unsuccessful treatment increased the state and trait anxiety of women.ConclusionsPsychological stress changed periodically depending on the duration of the patients’ treatment and fertility status also influenced anxiety levels. These findings will prove helpful in guiding psychological therapy and counseling for couples attempting to conceive by in vitro fertilization.