Dysregulation of intracellular pH is a cause of impaired capacitation in Slc22a14-deficient mice.

Solute carrier 22a member 14 (SLC22A14) plays a critical role in male infertility in mice. We previously revealed that one of the causes of infertility is impaired capacitation. However, the molecular mechanism remained unclear. Here, we show that the influx of HCO3-, a trigger of capacitation, is impaired and intracellular pH (pHi) is decreased in the sperm of Slc22a14 knockout (KO) mice. While intracellular cAMP concentration did not increase during capacitation in Slc22a14 KO spermatozoa, HCO3--dependent soluble adenylate cyclase activity was normal, and the addition of 8-bromo cAMP rescued the decreased protein tyrosine phosphorylation. In addition, the pHi of Slc22a14 KO sperm was lower than that of WT sperm and did not increase after the addition of HCO3-. Although its relationship to the regulation of pHi is unknown, transmembrane protein 225, a possible protein phosphatase inhibitor, was found to be decreased in Slc22a14 KO sperm. The decreased in vitro fertilization rate of Slc22a14 KO sperm was partially rescued by an increase in the pHi and the addition of 8-bromo cAMP. These results suggest that SLC22A14 is involved in capacitation through the regulation of HCO3- transport and pHi.

Serum thyroid-stimulating hormone receptor antibody levels and thyroid dysfunction after hysterosalpingography: a case-control study.

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.

Risk factors of subclinical hypothyroidism and the potential contribution to miscarriage: A review.

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.

Impact of subchorionic hematoma in early pregnancy on obstetric complications: A retrospective cohort study in women who had live births after frozen-thawed embryo transfer.

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.

Effect of moderately increased thyroid-stimulating hormone levels and presence of thyroid antibodies on pregnancy among infertile women.

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.

The effect of oil and water-soluble contrast medium in hysterosalpingography on thyroid function.

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.

Pregnancy bias toward boys or girls: The Japan Environment and Children's Study.

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.

Control of body size in C. elegans dependent on food and insulin/IGF-1 signal.

The body size of an organism is governed by genetic and environmental factors. As an environmental factor, food appears to be the most important for body size control in animals. C. elegans worms are usually grown on an E. coli strain OP50. We show that the wild-type worms fed on another E. coli strain HB101 grow 1.6 times as large as those fed on OP50. The regression line representing the relationship between the sizes of worms grown on each food for over 30 mutants was drawn, indicating that small mutants tend to be more affected by the change in food. Mutants for the DAF-2 insulin/IGF-1 receptor and downstream SGK-1, a homolog of the serum- and glucocorticoid-inducible kinase, grow less or little larger on HB101, indicating control of body size by these factors. Results on the suppression of mutations in these factors by a mutation in the DAF-16/FOXO transcription factor indicate both DAF-16-dependent and DAF-16-independent control. Furthermore, we show that the food-dependent body size change is because of a change in cell size that is closely related to the protein content per cell.

Microscopy and biomimetics: the NanoSuit® method and image retrieval platform.

This review aims to clarify a suitable method towards achieving next-generation sustainability. As represented by the term 'Anthropocene', the Earth, including humans, is entering a critical era; therefore, science has a great responsibility to solve it. Biomimetics, the emulation of the models, systems and elements of nature, especially biological science, is a powerful tool to approach sustainability problems. Microscopy has made great progress with the technology of observing biological and artificial materials and its techniques have been continuously improved, most recently through the NanoSuit® method. As one of the most important tools across many facets of research and development, microscopy has produced a large amount of accumulated digital data. However, it is difficult to extract useful data for making things as biomimetic ideas despite a large amount of biological data. Here, we would like to find a way to organically connect the indispensable microscopic data with the new biomimetics to solve complex human problems.

Identification and evolution of venom phospholipase A2 inhibitors from Protobothrops elegans serum.

The cDNAs encoding venom phospholipase A(2) (PLA(2)) inhibitors (PLIs), named Protobothrops elegans (Pe)γPLI-A, PeγPLI-B, PeαPLI-A, and PeαPLI-B, were cloned from the P. elegans liver cDNA library. They were further divided into several constituents due to nucleotide substitutions in their open reading frames. For PeαPLI-A, two constituents, PeαPLI-A(a) and PeαPLI-A(b), were identified due to three nonsynonymous substitutions in exon 3. Far-western blot and mass-spectrometry analysis of the P. elegans serum proteins showed the presence of γPLIs, and αPLIs, which can bind venom PLA(2)s. In αPLIs from Protobothrops sera, A or B subtype-specific amino acid substitutions are concentrated only in exon 3. A comparison of γPLIs showed that γPLI-As are conserved and γPLI-Bs diversified. Mathematical analysis of the nucleotide sequences of Protobothrops γPLI-B cDNAs revealed that the particular loops in the three-finger motifs diversified by accelerated evolution. Such evolutionary features should have made serum PLIs acquire their respective inhibitory activities to adapt to venom PLA(2) isozymes.

Beneficial effect of l-arginine in women using assisted reproductive technologies: a small-scale randomized controlled trial.

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.

Noninvasive evaluation of testicular function by measuring oxygen saturation of scrotal-testicular tissue.

We conducted a single-center cross-sectional study to determine whether scrotal-testicular tissue oxygen saturation (S-T StO2) measured by finger-mounted near-infrared spectroscopy is useful for the evaluation of testicular function. S-T StO2 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 StO2 is associated with a reduction in testicular function.

Suppression of severe lesions, myonecrosis and hemorrhage, caused by Protobothrops flavoviridis venom with its serum proteins.

Protobothrops flavoviridis serum proteins precipitated with ammonium sulfate were chromatographed on a DEAE-Toyopearl 650M column at pH 7.5 with stepwise increase or with linear gradient of NaCl concentration. Peaks 3 and 4 serum proteins, obtained by linear gradient elution and named Fr(de3) and Fr(de4), contained Habu serum factors (HSF) and phospholipase A2 (PLA2) inhibitors (PfPLI), respectively. The serum proteins eluted at 0.2 M NaCl by stepwise elution, named Fr(0.2NaCl), effectively suppressed myonecrosis and hemorrhage caused by P. flavoviridis venom in rat or mouse thigh muscles. The Fr(0.2NaCl) were fractionated by HPLC and the fractions, after SDS-PAGE, underwent far-western blot analysis with PLA2 ([Asp(49)]PLA2) and BPI ([Lys(49)]PLA2) as the probes. Four PfPLIs, namely, PfαPLI-A, PfαPLI-B, PfγPLI-A and PfγPLI-B, were identified together with their selective binding specificities to PLA2 species. In addition, a new 9 kDa protein, which is specifically bound to BPI, was found. Suppression of P. flavoviridis venom-induced severe lesions, such as myonecrosis, hemorrhage and edema, with its serum proteins was histopathologically observed in the present work for the first time. The cooperative use of P. flavoviridis antivenom and its serum proteins as medication for P. flavoviridis snake bites is discussed.

Body size change in various nematodes depending on bacterial food, sex and growth temperature.

We previously reported significant body size change in the nematode Caenorhabditis elegans, depending on the food strain of E. coli. Here, we examined this body size change in 11 other nematode species as well, and found that it is common to most of these nematodes. Furthermore, this food-dependent body size change is influenced by sex and growth temperature.

Control of lifespan by food bacteria, nutrient limitation and pathogenicity of food in C. elegans.

The increased lifespan caused by food limitation has been observed in a wide range of animals including the nematode Caenorhabditis elegans. We show here that the lifespans of eat-2 and eat-5 feeding-defective mutants and a mutant of dbl-1 encoding a TGFß ligand significantly change between the cultures fed on Escherichia coli strain OP50 or a more nutrient-rich strain HB101. On HB101 food, the eat-2, eat-5 and dbl-1 mutants show increased lifespan compared to that of the wild type. This result is probably due to nutrient limitation because the eat mutations reduce food uptake and the mutation of dbl-1 that regulates expression of several digestive enzymes leads to nutrient limitation. In contrast, the lifespans of the eat-2 and dbl-1 mutants decreased from that of the wild type on OP50 food. We found that live OP50 cells within a worm were markedly more in these mutants than in the wild type, which suggests that impaired digestion of pathogenic OP50 decreased lifespan in the eat-2 and dbl-1 mutants.

Identification of the B subtype of gamma-phospholipase A2 inhibitor from Protobothrops flavoviridis serum and molecular evolution of snake serum phospholipase A2 inhibitors.

A cDNA encoding a novel phospholipase A(2) (PLA(2)) inhibitor (PLI) was isolated from a Protobothrops flavoviridis snake (Tokunoshima island, Japan) liver cDNA library. This cDNA encoded a signal peptide of 19 amino acids followed by a mature protein of 181 amino acids. Its N-terminal amino acid sequence was completely in accord with that of a PLI, named PLI-II, previously found in P. flavoviridis serum. PLI-II showed a high similarity in sequence to the B subtype of gammaPLI, denoted gammaPLI-B, isolated from Agkistrodon blomhoffii siniticus serum. Thus, PLI-II is P. flavoviridis serum gammaPLI-B. Since PLI-I, previously isolated from P. flavoviridis serum, can be assigned as gammaPLI-A, P. flavoviridis serum contains both A and B subtypes of gammaPLI. Phylogenetic analysis of gammaPLIs from the sera of various kinds of snakes, Elapinae, Colubrinae, Laticaudinae, Acanthophiinae, Crotalinae, and Pythonidae, based on the amino acid sequences revealed that A and B subtypes of gammaPLIs are clearly separated from each other. It was also found that phylogenetic topologies of gammaPLIs are in good agreement with speciation processes of snakes. The BLAST search followed by analyses with particular Internet search engines of proteins with Cys/loop frameworks similar to those of PLI-II and PLI-I revealed that gammaPLI-Bs, including PLI-II and PLI-II-like proteins from mammalian sources, form a novel PLI-II family which possesses the common Cys/loop frameworks in the anterior and posterior three-finger motifs in the molecules. Several lines of evidence suggest that PLI-II is evolutionarily ancestral to PLI-I.