Hiromi Hashimoto

Fertilization and pregnancy using cryopreserved testicular sperm for intracytoplasmic sperm injection with azoospermia

OBJECTIVES To report fertility for men with obstructive (OA) and nonobstructive (NOA) azoospermia after frozen and thawed spermatozoa recovered from the seminiferous tubules and intracytoplasmic sperm injection (ICSI) and to evaluate the factors other than spermatozoa. DESIGN Retrospective clinical analysis. SETTING Male infertility clinic for testicular sperm extraction and freezing/thawing (cryoTESE) and assisted reproductive technologies clinic for ICSI. PATIENT(S) Seventy-four men with OA and 140 men with NOA undergoing attempted cryoTESE-ICSI. INTERVENTION(S) Seventy-three couples with OA underwent a total of 184 cryoTESE-ICSI cycles, and 46 couples with NOA underwent a total of 75 cryoTESE-ICSI cycles. MAIN OUTCOME MEASURE(S) The numbers of eggs at metaphase II injected, two-pronuclei oocytes, normal cleaved embryos, embryos transferred, transfer cycles, biochemical pregnancies, and clinical pregnancies, as well as the implantation and delivery rates, were examined. RESULT(S) Fertilization rate in NOA was significantly lower than in OA. Neither the pathology, the source, nor the quantity of spermatozoa had any effect on fertilization or pregnancy rates. Maternal age had no effect on fertilization or embryo cleavage, but did dramatically affect the implantation, pregnancy, and delivery rates in NOA and OA. CONCLUSION(S) Good pregnancy rates were achieved without significant differences among the sperm sources. The pregnancy and the delivery rate were dependent strictly on the age of the female partner but not on her ovarian reserve.

Onco-testicular sperm extraction: testicular sperm extraction in azoospermic and very severely oligozoospermic cancer patients

An increased risk of testicular cancer in men with infertility and poor semen quality has been reported. In view of the high cure rates for testicular germ cell tumours, increasing clinical importance is being placed on the protection of fertility. High‐dose cytostatic therapy may be expected to cause long‐term infertility. Thus, the standard procedure for fertility protection is the cryopreservation of ejaculated spermatozoa or testicular tissue before therapy. Four male patients with azoospermia and two patients with very severe oligozoospermia underwent onco‐testicular sperm extraction (TESE). We attempted onco‐TESE in patients with azoospermia and very severe oligozoospermia after orchiectomy. Of the patients with testicular germ cell tumours, four had spermatozoa in their testicular tissues. Sertoli cell‐only syndrome was found in one patient, and one patient showed maturation arrest without the detection of spermatozoa. Three of six showed seminomatous germ cell tumour, two of six had nonseminomatous germ cell tumour and one patient showed no malignancy. Two patients achieved clinical pregnancy. Fertility challenges in men with cancer are the most straightforward because of the relative ease of obtaining and cryopreserving sperm. Testicular sperm extraction is a useful technique for obtaining spermatozoa before cytotoxic therapy in azoospermic and very severely oligozoospermic cancer patients.

Molecular changes induced by bisphenol-A in rat Sertoli cell culture.

Bisphenol-A (BPA) is an industrial chemical and is known to act as an endocrine disrupter. This study was designed to evaluate how BPA regulates Sertoli cell (SC) signal molecules. Purified rat SCs were cultured and treated with BPA (200 µmol/l) at various time points. Western blot analysis was used to determine the activation of extracellular signal-related kinases 1 and 2 (ERK1/2), c-Jun N-terminal kinase (JNK), p38 mitogen activated protein kinase (MAPK), nuclear factor kappa B (NF-κB), cyclooxygenase-1,2 (COX-1, 2), estrogen receptor-α (ER-α), and androgen receptor (AR). The levels of transferrin (TF), prostaglandin E2 (PGE2), and prostaglandin F2α (PGF2α) in culture medium were quantified by ELISA. Interleukin (IL)-1β and IL-6 mRNAs were measured by quantitative real-time PCR (QRT-PCR). Compared with the control, BPA activated the phosphorylation of ERK1/2 (p-ERK1/2) through 30 min to 6 h. TF was down-regulated at 6 and 24 h. Furthermore, IL-1β was up-regulated at 30 min and IL-6 was up-regulated at 1 and 24 h. ERK activity inhibitor (PD98059, 10 µmol/l) inhibited these molecular changes. These results reveal the possibility that BPA may have adverse effects on spermatogenesis via ERK1/2.

Stimulation of endometrium embryo transfer can improve implantation and pregnancy rates for patients undergoing assisted reproductive technology for the first time with a high-grade blastocyst

OBJECTIVE To evaluate whether stimulation of endometrium embryo transfer (SEET) can improve implantation rate and pregnancy rate (PR) for patients undergoing assisted reproductive technology (ART) for the first time by injecting embryo culture supernatant into the uterus before blastocyst transfer (BT). DESIGN Randomized, controlled trial. SETTING Private in vitro fertilization clinic. PATIENT(S) Forty-eight women in the BT group, 48 women in the stimulation group who had culture medium injected into the uterus before BT, and 48 women in the SEET group. INTERVENTION(S) Injection of embryo culture supernatant and injection of culture medium. MAIN OUTCOME MEASURE(S) Implantation rates and PRs. RESULT(S) Odds ratios of successful implantation rate for stimulation and SEET in patients with high-grade blastocysts, having BT as reference, were 2.58 and 6.46 without adjustment, and 5.91 and 9.20 after adjusting for basal FSH levels and period of infertility. Odds ratios of clinical pregnancies were 2.47 and 4.32 without adjustment, and 4.46 and 5.10 with adjustment, respectively. In groups with low-grade blastocysts, such tendencies were not observed. CONCLUSION(S) The SEET may be an effective method for increasing implantation rate and PR for first-time ART patients who have a high-grade blastocyst.

A healthy birth after intracytoplasmic sperm injection using ejaculated spermatozoa from a patient with Kartagener's syndrome

OBJECTIVE To report a healthy birth that was achieved by intracytoplasmic sperm injection (ICSI) with use of ejaculated spermatozoa from a patient with Kartageners syndrome. DESIGN Case report. SETTING Private infertility clinic. PATIENT(S) Couple with male factor infertility due to Kartageners syndrome. INTERVENTION(S) Intracytoplasmic sperm injection with ejaculated sperm. MAIN OUTCOME MEASURE(S) Semen characteristics, sperm motility, fertilization, pregnancy, and birth after ICSI. RESULT(S) With ejaculated sperm, the fertilization rates were 73% in the first stimulation cycle and 100% in the second cycle. Intracytoplasmic sperm injection was successful. The pregnancy resulted in birth of a single healthy child. CONCLUSION(S) With ejaculated sperm, successful pregnancy after ICSI in couples with Kartageners syndrome is possible. Kartageners syndrome is a heterogeneous group of disorders with similar clinical presentations, and treatment should be individualized depending on sperm motility.

Experimental ischaemia-reperfusion injury induces vascular endothelial growth factor expression in the rat testis.

Testicular torsion causes ischaemia‐reperfusion (I‐R) injury of testis and might lead to male infertility. Its injury initiates a pathophysiological cascade, including an activation of inflammatory cytokines and generation of nitric oxide and other reactive oxygen species. Vascular endothelial growth factor (VEGF) mediates angiogenesis and promotes endothelial cell survival. The aim of our study was to investigate the time course expression of VEGF, VEGF‐receptor (R)1, VEGF‐R2, nitric oxide synthases (NOS) in experimental I‐R injury of rat testis. In torsion side testis, the expression of VEGF protein and mRNA significantly increased in a time‐dependent manner (P < 0.001 and P < 0.001, respectively). Although the expression of VEGF‐R1 mRNA was increased in a similar way (P < 0.001), VEGF‐R2 mRNA expression was not detected. In immunohistochemistry, the increase in VEGF protein staining was observed in testicular vascular endothelial cells and germ cells at 24 h after reperfusion. Significant activation of inducible NOS and endothelial NOS was investigated at 12 and 24 h after reperfusion (P < 0.01 and P < 0.001, respectively). This is the first report to show the time course expression of VEGF in experimental I‐R rat testis.

Comparison of the Pregnancy Outcomes in Patients Receiving Hormone Replacement in the Luteal Phase after Transfer of Frozen-Thawed Embryos in the Natural Ovulatory Cycle

ABSTRACT The aim of this study was to determine whether there is a difference in pregnancy outcomes dependent on the kind of hormone replacement treatment in the luteal phase after transfer of frozen-thawed embryos in the natural ovulation cycle. Two hundred and twenty-three cycles were examined in this study. The cycles were divided into three groups. In group A (40 cycles), the luteal phase was supported by human chorionic gonadotropin (hCG). In group B (83 cycles), the luteal phase was supported by hCG and vaginal suppository of progesterone. In group C (100 cycles), the luteal phase was supported by hCG, vaginal progesterone and transdermal estradiol. The pregnancy rate and implantation rate in group C (63.0% and 44.3%) were significantly and 28.0%) and group B (45.8% and 30.3%). In conclusion, administration of vaginal progesterone and transdermal estradiol in addition to hCG in the luteal phase in the natural ovulation cycle is effective for improving pregnancy and implantation rates in patients undergoing transfer of frozen-thawed embryos.

Effectiveness of Buserelin Acetate for Triggering Endogenous LH Surge before Oocyte Retrieval in Clomiphene Citrate Cycle

ABSTRACT The purpose of this study was to evaluate the effectiveness of buserelin acetate (BA) for triggering endogenous LH surge before oocyte retrieval in clomiphene citrate (CC) or CC+hMG cycles. Patients were divided into two groups: a buserelin group, which consisted of patients treated with BA for 73 cycles 34–36 hours before oocyte retrieval, and an hCG group, which consisted of patients treated with hCG for 89 cycles 34–36 hours before oocyte retrieval. Rate of oocyte retrieval, ratio of metaphase II oocytes, fertilization rate in ICSI, fertilization rate in IVF, ratio of good cleaved embryos, rate of blastocyst development and pregnancy rate were 59.1%, 74.3%, 78.8%, 78.4%, 64.6%, 50.0% and 8.3%, respectively in the buserelin group and 63.2%, 71.3%, 80.6%, 67.6%, 58.6%, 66.7% and 12.5%, respectively, in the hCG group. There were no significant differences in those results between the groups. In conclusion, BA is present terse should be used for a conclusion effective for triggering LH surge before oocytes retrieval in CC or CC+hMG cycles.