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Tongkat Ali (Eurycoma longifolia), traditionally used as an herbal medicine in Southeast Asia, is known to have a variety of effects, including stress reduction. The authors divided 5-month-old pigs (three males and three females) into four groups consisting of a control group, which was not administered Tongkat Ali extract (TAE), and three groups administered TAE at 0.25%, 0.50%, or 0.75% of their feed, respectively, and observed the pigs for a period of eight weeks under conditions of disturbed sleep due to prolonged illumination. In terms of observation items, the pigs' vitality, appetite, fecal characteristics, respiration, emaciation, anxiety, restlessness, body temperature, body weight, and salivary amylase were measured as clinical symptom scores, and the intestinal microbiota in their feces was analyzed using nextgeneration sequencing (NGS). Although members of the control group showed anxiety and restless behaviors; such stress behaviors were clearly inhibited in the groups administered with TAE. Groups of pigs administered with TAE at the ratios of 0.25% and 0.50% showed a decrease in salivary amylase concentration (p<0.05) whereas the results from the control and 0.75% group were not significant. Analyses of intestinal bacteria using NGS showed that Lachnospiraceae, a family of bacteria known for producing butyric acid, increased significantly in the group administered with TAE at 0.50%, whereas it was not changed in the other groups. Based on the above findings, administration of TAE to fattening pigs under high levels of lighting-induced stress is expected to be effective and administration of TAE at 0.25% of the feed intake in particular, has been shown to effectively inhibit behaviors associated with stress. [ABSTRACT FROM AUTHOR]

The sex of the conceived child is a significant concern for parents. To verify whether there women have pregnancy bias toward boys or girls, we investigated whether the history of continuous same-sex pregnancy was associated with the subsequent child's sex. We prospectively analyzed data from the Japan Environment and Children's Study, a birth cohort study. We included all cases of singleton live births (n = 98 412). Women with pregnancy due to infertility treatment were excluded (n = 6255); Similarly, women with a history of miscarriage, artificial abortion, stillbirth, and multiple pregnancies, and those with missing data on the sex of the previous child were excluded. Altogether, 62 718 women were included. For the first live birth, a male-biased sex ratio of 1.055 was observed. Further, no significant difference was found in the sex ratio of the conceived child between women with one boy and those with one girl previously. However, when there were more than two children previously, the subsequently conceived child's male/female sex ratio was significantly higher among boy-only mothers than among girl-only mothers. The results indicated that several pregnant women are biased toward conceiving either boys or girls., Competing Interests: Shuhei So is affiliated with the funded laboratory of Tawara IVF Clinic. Fumiko Tawara, Yu Taniguchi, and Naohiro Kanayama declare that they have no conflict of interest. The funder provided support in the form of salaries for authors [S.S]. There are no patents, products in development or marketed products associated with this research to declare. This does not alter our adherence to PLOS ONE policies on sharing data and materials., (Copyright: © 2023 So et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Purpose: Our aim is to make an ideal embryo culture medium close to human oviduct fluid (HOF) components, and to evaluate the quality of this medium with embryo quality and clinical outcomes in assisted reproductive technology (ART) by a prospective randomized controlled trial (RCT)., Methods: Study I: HOF was collected laparoscopically from patients ( n  = 28) with normal pelvic findings. According to HOF analysis results, the new medium "HiGROW OVIT ® " (OVIT) was designed. Study II: Embryos (2 pronuclei (2PN) = 9633) were assigned from 1435 patients. The blastulation rate (BR), good BR (gBR), utilized (transferred/cryo-preserved) BR (uBR), pregnancy rate (PR), and miscarriage rate (MR) were compared between the OVIT and control groups by RCT., Results: The novel medium 'OVIT' was produced according to 31 HOF components. The concentrations of essential amino acids (e-AAs) were lower in OVIT than in current media, yet the opposite was true for ne-AA concentrations. gBR and uBR were higher in the OVIT group than in the control group. In the older female group, gBT and uBR were significantly higher in the OVIT group., Conclusions: The novel medium 'OVIT' was produced according to HOF data. The OVIT had significantly better embryo quality and clinical outcomes than the current media., Competing Interests: Takafumi Utsunomiya has nothing to disclose. The remaining authors have nothing to disclose., (© 2022 The Authors. Reproductive Medicine and Biology published by John Wiley & Sons Australia, Ltd on behalf of Japan Society for Reproductive Medicine.)

Objective: Hysterosalpingography (HSG) performed with an iodine contrast media can cause thyroid dysfunction, including thyrotoxicosis and hypothyroidism. We investigated the association between the serum levels of thyroid-stimulating hormone receptor antibody (TRAb), an indicator of Graves' disease, and abnormal thyroid function after performing HSG., Methods: The screening of TRAb was conducted in 362 patients who first visited the Tawara IVF Clinic between April and September 2018. The association between TRAb levels and the effects of HSG examinations on thyroid function were evaluated., Results: Of the 362 patients, 2 (0.55%) had high levels (>2.0 IU/L) of TRAb, whereas 18 (5.0%) had intermediate TRAb levels, ranging from 0.3 to 1.9 IU/L. Of the 98 women (including 7 of the 18 women with TRAb level 0.3-1.9 IU/L, and 91 of the 342 women with TRAb level <0.3 IU/L) who had undergone HSG, two women developed overt thyrotoxicosis after HSG, and the frequency was significantly higher ( p  = .0044) in the group with intermediate levels of TRAb (28.6%, 2 of 7) than that in the group with low TRAb levels (<0.3 IU/L; 0.0%, 0 of 91)., Conclusions: These findings indicate that increased serum levels of TRAb are significantly associated with the development of thyrotoxicosis after HSG.

This small-scale randomized controlled study aimed to examine the effect of l-arginine supplementation on the human chorionic gonadotropin (hCG)-positive rate and clinical pregnancy rate (CPR) in women undergoing assisted reproductive technology (ART) treatment for 3 months. From November 2017 to March 2018, 120 patients aged less than 40 years and planning for egg retrieval for embryo transfer were enrolled. The patients were divided into the AS2000 group (n = 36; l-arginine, 2 g; folate, 400 μg; and vitamin E, 10 mg), AS1000 group (n = 37; l-arginine, 1 g; and folate, 200 μg), and control group (n = 36). The main outcome was the hCG-positive rate or CPR in 3 months. The cumulative hCG-positive rates during the administration period were 44.2%, 54.2%, and 52.1%, and the CPRs were 39.5%, 41.7%, and 47.9% in the control, AS1000, and AS2000 groups, respectively. Odds ratios of the hCG-positive rate and CPR in the global l-arginine group (AS1000 and AS2000) versus those in the control group were 1.33 (95% confidence interval [CI], 0.62-2.90) and 1.11 (95% CI, 0.51-2.46), respectively. In the subgroup of women receiving ART because of male infertility, the hCG-positive rate and CPR were significantly increased in the l-arginine groups compared to those in the control group (13.42 [95% CI, 1.42-366.9] and 13.62 (95% CI, 1.42-367.6), respectively). Thus, arginine supplementation may be an option for women who desire pregnancy, especially those undergoing an ART program because of male infertility., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Purpose: We investigated the contribution of subchorionic hematoma (SCH) involvement in early pregnancy to the risk of pregnancy complications in women who underwent frozen-thawed embryo transfer (FET)., Methods: A hypoechogenic area surrounding the gestational sac at early pregnancy on ultrasound was defined as SCH. Simultaneously, the presence of vaginal bleeding was evaluated. We included 1416 women with live births after FET between March 2015 and September 2018 in this study. The frequency of pregnancy complications was compared between the SCH (n = 340) and non-SCH (n = 1076) groups., Results: The adjusted odds ratio of abnormal placental adhesion and placenta previa for the SCH group relative to the non-SCH group was 7.01 [2.96-18.00] and 3.77 [1.24-11.91], respectively. In contrast, hypertensive disorders of pregnancy, non-reassuring fetal status, fetal growth restriction, chorioamnionitis, and premature rupture of the membrane showed no differences between both groups. Furthermore, the frequency of abnormal placental adhesion was higher in the SCH group with vaginal bleeding than in the SCH group without vaginal bleeding., Conclusions: Subchorionic hematoma in early pregnancy may cause abnormal placental adhesion and placenta previa in pregnant women with FET. SCH presence should be carefully noted, particularly in cases with vaginal bleeding during early pregnancy after FET., Competing Interests: S. So and N. Murabayashi are affiliated with the laboratory of Tawara IVF clinic, which funded the study. All other authors declare that they have no conflict of interest., (© 2020 The Authors. Reproductive Medicine and Biology published by John Wiley & Sons Australia, Ltd on behalf of Japan Society for Reproductive Medicine.)

Background: There is no clear cutoff value for thyroid-stimulating hormone (TSH) level that defines subclinical hypothyroidism (SCH). Moreover, TSH levels can be affected by numerous factors. Although mild SCH has been implicated in miscarriage, the relationship between TSH levels and miscarriage remains unelucidated., Methods: We reviewed nine known risk factors affecting TSH levels and 28 studies investigating the potential association between mild SCH and miscarriage, examining whether these factors were considered., Main Findings: Among 28 studies that examined whether mild SCH (TSH > 2.5 mIU/L) contributed to miscarriage, thyroid antibodies were measured in only 15. TSH measurement methods were described in 18 studies. Although the iodinated contrast medium used in hysterosalpingography (HSG) is stored in the body for a long time and is a risk factor for mild SCH, only one study described its potential impact on TSH levels. Nine studies, which concluded that mild SCH contributed to miscarriage, had thyroid status evaluated only after the onset of pregnancy, but not before., Conclusion: TSH levels can be significantly affected by patient demographics and health history, country of origin, and fertility treatment. It is important to consider these factors while evaluating mild SCH. It remains unclear how mild SCH contributes to miscarriage., Competing Interests: Conflict of interest: Shuhei So is affiliated with the funded laboratory of Tawara IVF Clinic. Fumiko Tawara declares that they have no conflict of interest. Human/Animal rights: This article does not contain any studies with human and animal subjects performed by the any of the authors., (© 2020 The Authors. Reproductive Medicine and Biology published by John Wiley & Sons Australia, Ltd on behalf of Japan Society for Reproductive Medicine.)

Purpose: We investigated the clinical results of Japanese men with Y chromosome microdeletions., Methods: This study retrospectively examined 2163 azoospermic or severe oligozoospermic patients. We investigated the frequency of azoospermia factor (AZF) deletions and sperm retrieval rate (SRR) by microTESE in patients with these deletions, then analyzed the ICSI outcomes., Results: Azoospermia factor deletions were found in 201 patients. SRR was significantly higher than that of the control group (74.0% vs 20.4%, P  < .001). Thirty-three couples underwent ICSI using testicular spermatozoa retrieved by microTESE, and eight couples underwent ICSI using ejaculatory spermatozoa. The fertilization rate and clinical pregnancy rate per embryo transfer cycle were significantly higher in the ejaculatory group than that of the testicular group (66.4% vs 43.7%, P  < .001, 53.3% vs 24.7%, P  = .03, respectively). When compared with the control group, the fertilization rate was significantly lower in the testicular group with AZFc microdeletions (43.7% vs 53.6%, P  < .001)., Conclusions: Our study highlights that although microTESE in azoospermic men with AZFc microdeletions led to a higher SRR, ICSI outcomes of these men were worse than that of men without AZF deletions, even if testicular spermatozoa were retrieved., Competing Interests: Conflict of interest: All of the authors declare that they have no conflict of interest. Human rights statements and informed consent: All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1964 and its later amendments. In all participating facilities, strict common informed consent was obtained from all patients for inclusion in the study. All patient data were collected and used following personal information protection such as anonymization. Animal studies: This article does not contain any studies with animal subjects performed by any of the authors. Approval by ethics committee: The protocol for this research project including human patients has been approved by a suitably constituted Ethics Committee., (© 2020 The Authors. Reproductive Medicine and Biology published by John Wiley & Sons Australia, Ltd on behalf of Japan Society for Reproductive Medicine.)

Purpose: To study the effects of mildly elevated thyroid-stimulating hormone (TSH) levels and thyroid antibodies on pregnancy rates among infertile women and their potential contribution to prolonged infertility treatment., Methods: This case-control study included 1479 women who underwent infertility treatment between March 2015 and August 2017. Cumulative pregnancy and miscarriage rates after assisted reproductive technology (ART) or non-ART treatments were compared between women with TSH <2.5 mIU/L and those with TSH 2.5-3.5 mIU/L and between women with and without thyroid antibody positivity., Results: The cumulative pregnancy rate of women with TSH 2.5-3.5 mIU/L was similar to that of women with TSH <2.5 mIU/L in the non-ART (hazard ratio [HR], 0.85; 95% confidence interval [CI], 0.56-1.23) and ART (HR, 1.17; 95% CI, 0.93-1.47) groups. Thyroglobulin antibody (TgAb) and thyroid peroxidase antibody (TPOAb) had no correlation with cumulative pregnancy rates. In the non-ART and ART groups, HRs for TgAb were 0.87 (95% CI, 0.55-1.32) and 1.09 (95% CI, 0.84-1.39) and HRs for TPOAb were 0.88 (95% CI, 0.52-1.39) and 1.29 (95% CI, 0.97-1.68), respectively., Conclusions: Cumulative pregnancy rates and miscarriage rates were similar between women with TSH <2.5 mIU/L and those with TSH 2.5-3.5 mIU/L and were independent of thyroid antibody positivity., Competing Interests: Shuhei So and Nao Murabayashi are affiliated with the laboratory of Tawara IVF Clinic, which funded the study. Wakasa Yamaguchi, Naomi Miyano, and Fumiko Tawara declare that they have no conflict interest., (© 2019 The Authors. Reproductive Medicine and Biology published by John Wiley & Sons Australia, Ltd on behalf of Japan Society for Reproductive Medicine.)

We conducted a single-center cross-sectional study to determine whether scrotal-testicular tissue oxygen saturation (S-T StO 2 ) measured by finger-mounted near-infrared spectroscopy is useful for the evaluation of testicular function. S-T StO 2 was significantly higher in patients with nonobstructive azoospermia (NOA, p< 0.05), and showed a positive correlation with luteinizing hormone levels (LH) even in participants without NOA (r = 0.34, p< 0.05), suggesting that elevated S-T StO 2 is associated with a reduction in testicular function., (Copyright © 2019 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier B.V. All rights reserved.)

Hysterosalpingography (HSG) using iodinated contrast medium is X-ray diagnostic test that examines the inside of the uterus and fallopian tubes in infertile women. In this study, we compared thyroid function (thyroid stimulating hormone: TSH and free-T4: FT4 levels) after HSG with an oil-soluble contrast medium (OSCM) and a water-soluble contrast medium (WSCM). One hundred and sixty-four and 94 patients with normal thyroid function received HSG with OSCM and WSCM, respectively. Approximately 25% of the women in the OSCM group developed subclinical hypothyroidism (SCH), whereas only less than 10% of the patients in the WSCM group developed SCH. Our data clearly indicate that WSCM is safe for thyroid function in women who plan to get pregnant.

We report a case of Klinefelter syndrome with the chief complaint of male infertility. Normal pregnancy and delivery resulting from conventional testicular sperm extraction (TESE) and intracytoplasmic sperm injection (ICSI) was possible by a previous doctor when he was 31 years old. He was referred to our hospital for treatment of infertility when he was 41 years old. Abdominal ultrasonography demonstrated a low echoic region with a relatively uniform content in the left testis and a central low echoic small mass surrounded by a high echoic region in the right testis. In excised tissue of the left testis, only a single seminiferous tubule containing spermatozoa was found and sperm was successfully retrieved by microdissection TESE.

A 46-year-old man who has a child from a previous marriage without artificial reproductive technologies was referred to our hospital with a chief complaint of infertility. He had suffered from bilateral orchitis after parotitis six years ago. On physical examination, both testes were soft and 4 ml in size. Semen analysis showed azoospermia and the serum follicle stimulating hormone value was high (36.9 mIU/ml). Microdissection testicular sperm extraction was performed, and motile sperm were successfully retrieved. The histological examination showed increased thickness of the basement membrane and, peritubular fibrosis in most seminiferous tubules, with few focal areas of normal spermatogenesis.

Purpose: Cervical mucus plays an important role in host defense mechanisms and is involved in the ascendance of sperm. To determine the change of neutrophil mediators contained in cervical mucus, we measured total neutrophil elastase (NE) levels throughout the menstrual cycle., Methods: Cervical NE concentrations in 31 healthy women having regular ovulatory cycles were measured during the follicular, ovulatory and luteal phases., Results: The cervical NE levels during the follicular, ovulatory and luteal phases were 0.73 ± 0.57 (mean ± SD), 0.29 ± 0.33 and 1.32 ± 1.24 μg/ml, respectively. The NE concentrations during the ovulatory phase were significantly lower than the other phases., Conclusions: The decrease in total cervical NE level during the ovulatory phase suggests that neutrophil activity in cervical mucus is depressed around ovulation. The decreased cervical NE levels and the depressed neutrophil activity in cervical mucus at ovulation may be suitable for sperm passage through the cervical canal.