Studies have shown that some electrolytes, including Na+ and K+, play important roles in embryonic development. However, these studies evaluated these electrolytes by using inhibitors or knockout mice, with no mention on the changes in the intracellular electrolyte concentrations during embryogenesis. In this study, we used the electrolyte indicators CoroNa Green AM and ION Potassium Green-2 AM to directly visualise intracellular concentrations of Na+ and K+, respectively, at each embryonic developmental stage in mouse embryos. We directly observed intracellular electrolyte concentrations at the morula, blastocyst, and hatching stages. Our results revealed dynamic changes in intracellular electrolyte concentrations; we found that the intracellular Na+ concentration decreased, while K+ concentration increased during blastocoel formation. The degree of change in intensity in response to ouabain, an inhibitor of Na+/K+ ATPase, was considered to correspond to the degree of Na+/K+ ATPase activity at each developmental stage. Additionally, after the blastocyst stage, trophectoderm cells in direct contact with the blastocoel showed higher K+ concentrations than in direct contact with inner cell mass, indicating that Na+/K+ ATPase activity differs depending on the location in the trophectoderm. This is the first study to use CoroNa Green AM and ION Potassium Green-2 AM in mouse embryos and visualise electrolytes during embryonic development. The changes in electrolyte concentration observed in this study were consistent with the activity of Na+/K+ ATPase reported previously, and it was possible to image more detailed electrolyte behaviour in embryo cells. This method can be used to improve the understanding of cell physiology and is useful for future embryonic development studies.

Anti-glutamic acid decarboxylase antibody-positive gestational diabetes mellitus with autoimmune type 1 diabetes mellitus in the early postpartum period: a case report and literature review

Background: Approximately 10% of all Graves' disease cases are triiodothyronine (T3)-predominant. T3predominance is characterized by higher T3 levels than thyroxine (T4) levels. Thyroid stimulating hormone receptor autoantibody (TRAb) levels are higher in T3-predominant Graves' disease cases than in non-T3-predominant Graves' disease cases. Treatment with oral drugs is difficult. Here, we report a case of fetal goiter in a pregnant woman with T3-predominant Graves' disease. Case presentation: A 31-year-old woman had unstable thyroid function during the third trimester of pregnancy, making it impossible to reduce her dosage of antithyroid medication. She was admitted to our hospital at 34 weeks of gestation owing to hydramnios and signs of threatened premature labor, and fetal goiter (thyromegaly) was detected. The dose of her antithyroid medication was reduced, based on the assumption that it had migrated to the fetus. Subsequently, the fetal goiter decreased in size, and the hydramnios improved. The patient underwent elective cesarean delivery at 36 weeks and 5 days of gestation. The infant presented with temporary symptoms of hyperthyroidism that improved over time. Conclusions: The recommended perinatal management of Graves' disease is to adjust free T4 within a range from the upper limit of normal to a slightly elevated level in order to maintain the thyroid function of the fetus. However, in T3-predominant cases, free T4 levels may drop during the long-term course of the pregnancy owing to attempts to control the mother's symptoms of thyrotoxicosis. Little is known about the perinatal management and appropriate therapeutic strategy for T3-predominant cases and fetal goiter. Therefore, further investigation is necessary.

Study question In Japan, how much of freezing cost and the yearly storage cost of medical and social egg freezing for fertility preservation is clearly disclosed on the institution's website? Summary answer Of the 621 infertility treatment institutions, 146 listed egg freezing on their websites, 46 specified the freezing cost, and 71 specified the yearly storage cost. What is known already In Japan, from April 2022, standard assisted reproductive technology will be covered by insurance, but medical and social egg freezing will continue to be privately funded. Therefore, it is unclear how many facilities in Japan perform egg freezing. Furthermore, while the cost of freezing and yearly storage varies significantly from facility to facility, there has been no data comparing egg freezing among facilities in Japan or other countries. Therefore, we extracted data about freezing and storage cost from each institution's website and compared them for the first time. Study design, size, duration This study is a web-based survey in which a single board-certified specialist by Japan Society for Reproductive Medicine exhaustively browsed the websites of all 621 fertility treatment institutions in Japan in October 2021 and analyzed the implementation of egg freezing, freezing costs, and yearly storage costs of egg are clearly stated on the websites. The costs of drugs used for egg retrieval and ovarian stimulation were excluded. Participants/materials, setting, methods Of the 621 institutions, 88 institutions that clearly stated on their websites that they offer egg freezing for medical reasons were divided into a medical group, and 58 institutions that clearly stated that they offer egg freezing for only social reasons were divided into social group for statistical analysis. We compared the freezing and the yearly storage cost, respectively, whether it varies depending on the number of eggs or whether it is a fixed cost. Main results and the role of chance Of the 621 institutions, 146 (23.5%), 88 in the medical group and 58 in the social group, specified that they offer egg freezing. The percentages of clarification for freezing cost and yearly storage cost on the website were 27.3% and 30.7% in the medical group, 72.4%, and 75.9% in the social group, so the social group was higher in both cases (p < 0.01). The percentage of fixed costs for egg freezing was significantly lower in the social group (6/42, 14.3%) than in the medical group (9/24, 37.5%) (p < 0.05). The percentage of fixed costs for yearly storage cost was samely significantly lower in the social group (30/44, 68.2%) than in the medical group (25/27, 92.6%) (p < 0.05). The mean freezing cost/yearly storage cost was as follows for 1, 5, and 10 eggs. The medical group was (JPY45,527/JPY22,327, JPY65,179/JPY22,530, JPY90,311/JPY22,770), and the social group was (JPY56,822/JPY40,178, JPY88,429/JPY56,502, JPY136,550/JPY79,090), so the social group had the significantly highest amount of costs in each case. Note that JPY 10,000 is about EUR77.7. The most expensive clinic to store ten eggs was JPY500,000 (EUR3,883) per year, and the least university hospital was JPY 4,600 (EUR35.7) per year. Limitations, reasons for caution This survey was conducted by carefully browsing the facility's website, but since we did not confirm the information through questionnaires or e-mail, there is the possibility that the information may have been updated. Another limitation is that less than half of the medical groups list cost details on their websites. Wider implications of the findings This is the first report to analyze the cost of egg freezing in Japan using the institution's website. The rate of cost disclosure on the website is still low. Moreover, the costs of social freezing are significantly higher than medical reasons, but this may change with the insurance of ART. Trial registration number not applicable

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