Effect of transdermal estrogen dose regimen for endometrial preparation of frozen-thawed embryo transfer on reproductive and obstetric outcomes.

PURPOSE: Previous studies have reported different methods of estrogen administration during endometrial preparation for frozen-thawed embryo transfer (FET). This study aimed to investigate a beneficial regimen of transdermal estrogen administration for FET. METHODS: We investigated the reproductive and obstetric outcomes of FET by comparing the increasing dose (ID) group that mimics changes in serum estradiol during the menstrual cycle and the constant dose (CD) group. Transdermal patches were used for estrogen administration in both groups. In our hospital, we targeted 315 cycles of the ID group in which FET was performed in 2017 and 324 cycles of the CD group in which FET was performed in 2018. In all cases, single embryo transfer was performed. RESULTS: All were singleton pregnancies. There was no difference in clinical pregnancy rate (28.9% vs 28.2%, P =.837) and live birth rate (17.3% vs 21.4%, P =.201) between the ID and CD groups. Spontaneous abortion rate was significantly lower in the CD group than in the ID group (37.2% vs 23.0%, P =.041). There was no difference in obstetrical outcomes. CONCLUSIONS: It was considered that the simple CD regimen may be more beneficial than the complicated ID regimen.

Follicle-stimulating Hormone-secreting Pituitary Adenoma Accompanied by Painful Thyroiditis.

A 30-year-old woman with multiple ovarian cysts presented with high serum estradiol levels. She had a pituitary adenoma, but the follicle-stimulating hormone (FSH) concentration was within the normal range. The patient complained of neck pain and palpitations during the disease course, and laboratory results revealed thyrotoxicosis and a systemic inflammatory response with negative findings for anti-thyroid stimulating hormone (TSH) receptor antibody and positive findings for anti-thyroglobulin and anti-thyroid peroxidase antibodies. Prednisolone improved the symptoms and the thyroid function and was discontinued after two months. A histological examination of the pituitary tumor confirmed it to be FSH-producing pituitary adenoma, with subsequent normalization of the serum estradiol concentration.

Correlation of abnormal mitochondrial distribution in mouse oocytes with reduced developmental competence.

Selection of good quality oocytes is important for improvement of assisted reproductive technology. Here, we studied the relationship of the mitochondrial distribution in metaphase II stage (MII) oocytes with fertility, since mitochondria in ooplasm are essential for energy production required for fertilization and embryo development. To observe mitochondria non-invasively, we used oocytes from a transgenic mouse, in which enhanced green fluorescent protein is targeted to the mitochondrial matrix and thus fluorescence is observed exclusively in the mitochondria. Control oocytes with mitochondria distributed around the nucleus showed normal embryo developmental competence, whereas oocytes with abnormal diffuse and fragmented mitochondria showed a significantly lower rate of embryo development after activation by intracytoplasmic sperm injection or strontium, which is a very effective agent for activation of mouse oocytes. Also, we showed that the reduced developmental competence of oocytes with diffuse and fragmented mitochondria caused by vitrification and thawing is similar to that of oocytes with abnormal mitochondrial foci obtained naturally. These findings suggest that abnormal mitochondrial distribution in oocytes at MII is a cause of developmental retardation and therefore normal mitochondrial distribution could be used as a criterion for selection of good oocytes.

Cytoplasmic transfer in the mouse in conjunction with intracytoplasmic sperm injection.

Cytoplasmic transfer (CT) from a donor to a recipient oocyte has been used clinically to facilitate human pregnancies. Data reported here describe the first characterization of CT coincident with intracytoplasmic sperm injection in the mouse system. Sibling oocytes were used to transfer 2, 4, or 6 pl of ooplasm to a recipient egg along with a sperm head using piezo-actuated injection. Survival and fertilization after CT were comparable to controls at 2 pl and 4 pl, but survival was significantly reduced with 6 pl volumes. Development to the blastocyst stage was also inversely related to CT volume, with some decline beginning with the 4 pl CT group. However, some blastocysts did develop in all of the groups. The results are in contrast with human eggs, which tolerate larger CT volumes. Results indicate that the mouse system can be used to characterize the transfer of exogenous materials concomitant with sperm injection, provided that the CT volume is not excessive.

Life-table analysis of artificial insemination pregnancy rates for couples with male factor and idiopathic infertility.

Background: In the summer of 2002, standard guidelines for the application of assisted reproductive technology were reported by a research group of the Ministry of Health, Labor and Welfare. The present study aimed to examine the relationship between the number of cycles of artificial insemination and the cumulative pregnancy rates according to the cause of infertility. Methods: Patients who experienced their first cycle of artificial insemination during the period of January 1999-December 2002 were included in the study and were divided into a male factor infertility group and an idiopathic infertility group. Cumulative pregnancy rates resulting from artificial insemination with the husband's semen were calculated by the life-table approach. Results: During the study period, 139 couples entered the assisted reproduction program and underwent 581 cycles. Significant differences were observed in cumulative pregnancy rates between the two groups. Conclusion: It is recommended that couples with male factor infertility and who fail to conceive within six or seven cycles of intrauterine insemination, consider a modification of treatment strategy such as in vitro fertilization, because cumulative pregnancy rates of this group were reached at a plateau within six or seven cycles. In contrast, patients with idiopathic infertility, the cumulative pregnancy rates appeared to increase constantly with each subsequent cycle. It is important to consider modifications of treatment strategy in the light of the cause of infertility. (Reprod Med Biol 2004; 3: 27-31).

Effect of repeated administration of clomiphene citrate at two different times on the endometrium in patients undergoing intrauterine insemination.

Background, Aims and Methods: Clomiphene citrate (CC) has been widely used for induction of ovulation; however, despite the high rate of ovulation, the pregnancy rate is only 30%. The anti-estrogenic effect of CC on the endometrium is one explanation for this finding. It is well known that repeated administration of CC enhances its anti-estrogenic effect. To overcome this adverse affect, a number of techniques have been used. One technique is the early administration of CC, in an attempt to decrease the anti-estrogenic effects of CC on the endometrium. The aim of the present study was to retrospectively evaluate if repeated administration of CC at varying times may affect the endometrium during preovulatory period; pregnancy rates were compiled for patients with unexplained infertility undergoing intrauterine insemination (IUI). The patients were divided into four groups based on the number of repeat administrations and the time of CC administration. Results: The endometrial thickness at the day of human chorionic gonadotropin administration was greater in early administration within three consecutive CC cycles than the others. Conclusion: When the endometrium is thin during classical administration of CC, it is worth attempting the early administration of CC in the CC/IUI treatment for patients with unexplained infertility. However, this effect disappeared over the duration of the three consecutive CC cycles. (Reprod Med Biol 2004; 3: 153-157).

Oocyte mitochondria: strategies to improve embryogenesis.

Mitochondria play a central role to provide ATP for fertilization and preimplantation embryo development in the ooplasm. The mitochondrial dysfunction of oocyte has been proposed as one of the causes of high levels of developmental retardation and arrest that occur in preimplantation embryos generated using Assisted Reproductive Technology. Cytoplasmic transfer (CT) from a donor to a recipient oocyte has been applied to infertility due to dysfunctional ooplasm, with resulting pregnancies and births. However, neither the efficacy nor safety of this procedure has been appropriately investigated. In order to improve embryogenesis, we observed the mitochondrial distribution in ooplasma under the several conditions using mitochondrial GFP-transgenic mice (mtGFP-tg mice) in which the mitochondria are visualized by GFP. In this report, we will present our research about the mitochondrial distribution in ooplasm during early embryogenesis and the fate of injected donor mitochondria after CT using mtGFP-tg mice. The mitochondria in ooplasm from the germinal vesicle stage to the morula stage were accumulated in the perinuclear region. The mitochondria of the mtGFP-tg mouse oocyte transferred into the wild type mouse embryo could be observed until the blastocysts stage, suggesting that the mtGFP-tg mice oocyte is very useful for visual observation of the mitochondrial distribution in the oocyte, and that the aberrant early developmental competences due to the oocyte mitochondrial dysfunction may be overcome by transferring the "normal" mitochondria.

Relationship between sperm mitochondrial membrane potential, sperm motility, and fertility potential.

AIM: To analyze the relationship between sperm mitochondrial membrane potential and sperm motility parameters by means of a computer-assisted sperm analyzer (CASA) and in-vitro fertilization rate(%FR). METHODS: Semen samples were obtained from 26 men undergoing in vitro fertilization-embryo transfer (IVF-ET). Informed consent was obtained from all men prior to the study. Samples were prepared using wash and swim-up method in HEPES-HTF medium. The sperm motility (%MOT), progressive motility (%PMOT), average path velocity (VAP) microm/s), straight line velocity (VSL) (micro m/s), curvilinear velocity (VCL) (microm/s) and %hyperactivated sperm (%HA), and the %FR were assessed. The samples were incubated in the presence of 2.0 mciromol/L of 5,5',6,6'-tetra-chloro-1,1',3,3'-tetraethylbenzimidazolyl-carbocyanine iodide (JC-1) for 30 min at 37 degrees C in air and washed in PBS before flow cytometry (FACSCalibur: Becton Dickinson) analysis. The mitochondrial probe JC-1 was used to identify the mitochondrial membrane potential. The sperm was divided into three populations according to the fluorescence pattern as follows: the high mitochondrial membrane potential group (n=8), the moderate group (n=5), and the low group (n=13). Statistical analysis was performed using unpaired t-test. RESULTS: Significant differences were found between the high and the low groups in %MOT (91.1+/-8.5 vs 63.0+/-32.7, mean+/-SD), VAP (73.0+/-14.2 vs 52.1+/-12.5), VCL (127.0+/-28.1 vs 87.0+/-22.6), %HA (27.3+/-23.6 vs 7.2+/-9.0) and %FR [73.2 (48/56) vs 59.0 (69/117)]. No significant differences were found in other CASA parameters. CONCLUSION: When the sperm mitochondrial membrane potential increases, sperm motility parameters and fertility potential will also increase. The JC-1 dye method is useful to predict sperm fertility potential.

Oxidative state and zona-binding ability in mouse spermatozoa treated with reduced glutathione.

Background and Aims: Nuclear proteins in mature mammalian spermatozoa nuclei are oxidized to form numerous disulfide bonds. Reduced glutathione (GSH) in the oocyte has been linked to spermatozoan nuclear decondensation after fertilization. In this study, we analyzed whether GSH reduced protamines in sperm nuclei in vitro, and examined the zona-binding ability of treated nuclei. Methods: Three groups of mouse cauda epididymal spermatozoa were prepared. The first group was cultured in Chatot-Tasca-Ziomek (CZB; control group), the second in 10 mmol/L GSH (GSH group), and the third group was the GSH group re-cultured in CZB (re-cultured group). Each sperm was stained with acridine orange, and the oxidative and reductive state of nuclei was analyzed by using fluorescence microscopy. Furthermore, we examined the zona-binding ability for each group by insemination to mouse oocytes after exposure to hyaluronidase. Results: All sperm nuclei from the control group displayed an oxidized pattern (green), and those from the GSH group displayed a reduced pattern (red), attributable to reduced protamines. Sperm nuclei from the re-cultured group displayed the oxidized pattern. Although the zona-binding ability of the GSH group was deteriorated compared with the control group sperm, no significant differences were observed between the control and re-cultured groups. Conclusion: From these results, in vitro reduced sperm were shown to be oxidized in CZB. A reduction of mouse spermatozoa in vitro by GSH is, therefore, reversible, and the oxidative state of sperm nuclei exerts an effect on zona-binding ability. (Reprod Med Biol 2002; 1: 55-58).