Akira Kuwahara

Purpose: To evaluate the clinical utility of preimplantation genetic testing for aneuploidy and structural rearrangements (PGT‐A/SR) under Advanced Medical Care B in Japan. Methods: This prospective, single‐arm, multicenter benchmarking study included couples with recurrent implantation failure, recurrent pregnancy loss, or structural chromosomal rearrangements who underwent PGT‐A/SR and first transfer of a category A/B embryo. The primary endpoint was ongoing pregnancy rate per transfer (oPR) at 12–13 weeks; clinical pregnancy rate (cPR) was prespecified as supportive because the registry benchmark used cPR. Outcomes were compared with age‐adjusted national registry thresholds using a one‐sample proportion test with a normal approximation. Results: Of 375 couples, 344 underwent PGT‐A/SR and 218 reached first transfer. oPR was 48.6% versus 42.4% ( p = 0.064) and did not reach statistical significance; cPR was 57.8% versus 42.4% ( p < 0.001). Miscarriage rate was 15.9% versus 23.4% ( p = 0.045). In women aged 35–39 and 40–42 years, oPR and cPR exceeded age‐adjusted thresholds, whereas no benefit was demonstrated in women aged < 35 years. Conclusions: On a per‐transfer basis, PGT‐A/SR was associated with higher cPR and lower miscarriage rate, particularly in women aged ≥ 35 years. Interpretation is limited by the single‐arm design and the mismatch between the study endpoint and the registry comparator. Trial Registration: This study was registered in the Japan Registry of Clinical Trials (jRCT) (jRCT1052220197)

Purpose: The Japan Society of Obstetrics and Gynecology conducted a nationwide clinical study to evaluate the pregnancy outcomes of preimplantation genetic testing for aneuploidy or chromosomal structural rearrangement (PGT‐A/SR).Methods: Patients that had experienced recurrent implantation failure, recurrent pregnancy loss, or chromosomal structural rearrangement were recruited from 200 fertility centers in Japan. For patients in whom one or more blastocysts were classified as euploid or euploid with suspected mosaicism, a frozen–thawed single embryo transfer (ET) was performed.Results: A total of 10 602 cycles, maternal age 28–50 years, were enrolled in this study. 42 529 blastocysts were biopsied, and 25.5%, 11.7%, and 61.7% of embryos exhibited euploidy, mosaicism, and aneuploidy, respectively. At least one euploid blastocyst was obtained in 38.3% of egg retrieval cycles with embryo biopsy. A total of 6080 ETs were carried out, and the clinical pregnancy rate per ET, ongoing pregnancy rate per ET, and miscarriage rate per pregnancy were 68.8%, 56.3%, and 10.4%, respectively. The rates of clinical pregnancy and miscarriage remained relatively constant across all maternal ages.Conclusions: Preimplantation genetic testing for aneuploidy or chromosomal structural rearrangement may improve the pregnancy rate per ET and reduce the miscarriage rate per pregnancy, especially in patients of advanced maternal age.

Aim: The actual status of fertility preservation treatments in the startup phase in Japan was investigated as a basis for discussing future directions.Methods: This study was conducted as “Research project to promote support of children and parenting 2016” which was supported by Ministry of Health in Japan with the approval of the institutional review board at St. Marianna University. Subjects of the survey were facilities registered with the Japan Society of Obstetrics and Gynecology as fertility preservation facilities, and facilities belonging to the Japan Association of Private Assisted Reproductive Technology Clinics and Laboratories. We provided questionnaires to survey both the medical care system and cases for which fertility preservation was implemented between 2006 and 2016.Results: Responses were obtained from 68 facilities (of the 64, 59 [92.2%] responded to the questionnaire and 9 clinics cooperated). Many facilities limited the cryopreservation of oocytes and ovaries to patients 40–41 years old and the use of eggs to patients 44–45 years old. In the patient survey, 812 cases of oocyte cryopreservation and 201 cases of ovarian tissue cryopreservation were performed during study period. Breast cancer was the most indicated disease, with oocyte cryopreservation in the late 30s and ovarian tissue cryopreservation in the early 30s. Very few babies were born from fertility preservation, and no live birth cases of ovarian tissue cryopreservation were identified.Conclusions: Even from the early days, fertility preservation was implemented according to certain standards in Japan, but was characterized by a large variety of facilities.

Hypothalamic pro‐inflammatory cytokine expression exhibits a weaker response to lipopolysaccharides (LPS) during the early neonatal period than during the later developmental period. Although toll‐like receptor 4 (TLR4), which recognizes bacterial molecules, activates pro‐inflammatory cytokine responses, the developmental changes in hypothalamic TLR4 expression have not been evaluated. In this study, the hypothalamic TLR4 mRNA levels of saline‐injected and LPS‐injected rats were measured during the neonatal, pre‐pubertal, and post‐pubertal periods. The rats' hypothalamic TLR4 mRNA levels gradually increased from the neonatal to pubertal period and were altered by the injection of LPS at all examined ages (postnatal day (PND) 5, 15, 25, and 42). LPS injection resulted in decreased hypothalamic TLR4 mRNA expression at PND5, whereas it increased hypothalamic TLR4 mRNA expression at PND15, 25, and 42. After the injection of LPS, the hypothalamic mRNA levels of the pro‐inflammatory cytokines interleukin (IL)‐1β, tumor necrosis factor α, and IL‐6 were attenuated during the early developmental period and increased acutely on PND42. The expression profiles of these pro‐inflammatory cytokines exhibited similar, but not entirely consistent, changes to those displayed by TLR4 during the developmental period. Hypothalamic TLR4 mRNA expression gradually increased throughout the developmental period, whereas the mRNA expression levels of the pro‐inflammatory cytokines increased acutely at PND42. Thus, it is assumed that hypothalamic TLR4 hypoactivity contributes to the low sensitivity of pro‐inflammatory cytokines to LPS during the early developmental period.

It has been reported that intrauterine undernutrition is closely associated with the pathogeneses of certain diseases in adulthood; i.e., insulin resistance and diabetes, and that leptin resistance plays a pivotal role in the pathology of such intrauterine growth restriction (IUGR)-related conditions. Therefore, examinations of IUGR-induced leptin resistance in early developmental period are important for protecting against future disease. In this study, the effects of prenatal undernutrition on the serum leptin levels and central leptin responses of rats during the neonatal and/or pre-pubertal period were examined. The 50% food-restricted undernourished dams’ offspring (UNO) exhibited a significantly lower birth weight than the normal nutrition dams’ offspring (NNO). However, the UNO grew rapidly, and their mean body weight had caught up with that of the NNO by postnatal day 8. Thus, there were no significant differences between the body weights of the two groups at postnatal day 12, 16, 20, or 28. The serum leptin levels of the UNO were significantly higher than those of the NNO at postnatal days 20 and 28. At postnatal day 28, no significant difference in the hypothalamic mRNA level of neuropeptide Y, which is the main target of leptin, or that of ObRb, which is the leptin receptor, was detected between the NNO and UNO. The chronic intracerebroventricular injection of leptin attenuated body weight gain in both the NNO and UNO; however, there were no significant differences between the body weights of the two groups at any of the examined postnatal time points, indicating that the UNO and NNO exhibited similar central sensitivity to leptin during the pre-pubertal period. These results suggest that prenatal undernutrition induces leptin resistance until the neonatal to pre-pubertal period and that these alterations might be caused by impaired transportation of leptin to central tissues.

Hyperandrogenemia has become an important factor in the diagnosis of polycystic ovary syndrome (PCOS), in Japan as well as Western countries, since the Japanese PCOS diagnostic criteria were revised in 2007. The revised electrochemiluminescence immunoassay (ECLIA) reagent for total testosterone (T) (ECLusys TESTO II; New T assay), which has lower cross-reactivity with DHEA-S than the previous reagent (ECLusys TESTO I; Old T assay), has recently been developed. We studied the improvement between New and Old T assays in regards to the diagnostic performance of serum T for patients with PCOS. Serum T levels were measured in both normal women (Control, n=92) and PCOS patients (n=86). The rate of elevated T using each kit was significantly higher in the PCOS group than in the Control group, and significantly higher using New T assay (38/86, 44.2%) compared with Old T assay (26/86, 30.2%). A significant correlation was found between T level and body mass index, but not LH level. Furthermore, higher detection rates of hormonal abnormalities were seen in PCOS patients using combined measurement of both T and LH (63/80) compared with single measurement of either T (37/80) or LH (36/80). Therefore, the serum T assay is essential for the diagnosis of PCOS, and New T assay combined with the LH assay may improve the diagnostic process for PCOS. J. Med. Invest. 61: 65-71, February, 2014

Neonatal immune challenges have a long‐lasting influence on immune response. Using male Sprague–Dawley rats, we examined whether neonatal lipopolysaccharide (LPS) challenge alters the sensitivity of male reproductive function to adult LPS challenge and at which level (central or testes) the alteration occurs. We also examined the mRNA expression of proinflammatory cytokines in the hypothalamus and testes because they have a pivotal role in immune stress‐induced suppression of gonadotropin secretion and testosterone synthesis. On day 10 after birth, all the pups were injected with LPS (100 μg/kg, i.p.) or saline. Thereafter, LPS (100 μg/kg, i.p.) or saline was injected in adulthood at 8 weeks of age. The serum LH concentration was decreased by LPS injection during adulthood in the neonatal saline‐injected rats. This suppressive effect was not seen in the neonatal LPS‐injected rats. The serum testosterone concentration was decreased by adult LPS injection in both the neonatal LPS‐injected and neonatal saline‐injected rats. The expression levels of KiSS‐1, which encodes kisspeptin, known to have a crucial role in the regulation of gonadotropin secretion, and GnRH mRNA in the hypothalamus and LHβ mRNA in the pituitary were not influenced by neonatal or adult LPS injection. On the other hand, the expression levels of tumor necrosis factor‐α (TNF‐α) and interleukin‐1β (IL‐1β) mRNA in the hypothalamus and testes were increased by adult LPS injection in both the neonatal LPS‐injected and neonatal saline‐injected rats. Furthermore, the expression levels of these factors in the hypothalamus after adult LPS injection were significantly lower in the neonatal LPS‐injected rats than in the neonatal saline‐injected rats. These findings indicate that neonatal LPS challenge reduces the sensitivity of male reproductive function to the suppressive effects of LPS, mainly at the central level. Attenuation of proinflammatory cytokine synthesis in the hypothalamus might be involved in this alteration.