マクロファージはクロストーク機構により子宮内膜細胞を酸化ストレスから保護する

This aim of this study was to investigate whether macrophages protect endometriotic cells from oxidative injury and to elucidate the underlying mechanisms of any protection. Endometriotic cells cultured with or without differentiated macrophages (dTHP-1 cells) were treated with hydrogen peroxide (H2O2) or methemoglobin, a major component of hemoglobin species in endometriotic cyst fluid. Co-culture experiments, microarray analysis, screening and validation of differentially expressed genes (DEGs), cell proliferation and viability assays, and experiments using a specific inhibitor were conducted to investigate the functional cross-talk between endometriotic cells and macrophages. Microarray analysis revealed that endometriotic cells co-cultured with dTHP-1 differentially express several genes compared with monoculture. Quantitative enzyme-linked immunosorbent assay (ELISA) and Western blotting analysis identified TGF-β1 as a promising candidate gene expressed in endometriotic cells co-cultured with dTHP-1 cells. TGF-β1 stimulated the expression of heme oxygenase-1 (HO-1) in dTHP-1 cells. HO-1 expression was increased in dTHP-1 cells co-cultured with endometriotic cells compared with the dTHP-1 monoculture. Both H2O2 and methemoglobin upregulated the expression of the HO-1 protein in the dTHP-1 monoculture; moreover, co-culture with endometriotic cells further enhanced HO-1 production. The co-culture with dTHP-1 protected endometriotic cells against oxidative injury. Blockade of HO-1 abolished the protective effects of macrophages. In an oxidative stress environment, TGF-β1 produced by endometriotic cells may protect against oxidative injury through the upregulation of macrophage-derived HO-1. The cross-talk between endometriotic cells and macrophages may contribute to the progression and pathogenesis of endometriosis.博士（医学）・甲第849号・令和4年9月28日Copyright © 2022, Society for Reproductive Investigation.© 2022 Springer Nature Switzerland AG.The version of record of this article, first published in Reproductive Sciences, is available online at Publisher's website: https://doi.org/10.1007/s43032-022-00890-6.発行元が定める登録猶予期間終了の後、本文を登録予定（2023.08

HNF-1β過剰発現を呈する卵巣明細胞癌においてGSK-3βは新たなシグナル伝達経路を介在する

Deubiquitinase USP28 is a target gene of the transcription factor HNF1 homeobox β (HNF-1β), which promotes the survival of ovarian clear cell carcinoma (OCCC) cell lines. However, the pharmacological inhibition of HNF-1β can cause several adverse effects as it is abundantly expressed in numerous organ systems, including the kidney, liver, pancreas and digestive tract. Therefore, small interfering RNA (siRNA) screening was performed in the current study to identify other potential downstream targets of the HNF-1β-mediated pathway. The results revealed that glycogen synthase kinase-3β (GSK-3β) may be a potential downstream target affecting cell viability. To further clarify the effects of GSK-3β, two human OCCC cell lines, TOV-21G (HNF-1β overexpressing line) and ES2 (HNF-1β negative) were transfected with siRNA targeting GSK-3β or control vectors. Loss-of-function studies using RNAi-mediated gene silencing indicated that HNF-1β facilitated GSK-3β expression, resulting in the loss of phosphorylated nuclear factor-κB (p-NFκB) and the reduction of TOV-21G cell proliferation. The cell proliferation assay also revealed that GSK-3β inhibitors rescued the effects of HNF-1β silencing on cell viability in a dose-dependent manner. Furthermore, the GSK-3β inhibitor, AR-A014418, effectively inhibited tumor cell proliferation in a xenograft mouse model. In conclusion and to the best of our knowledge, the current study was the first to determine that GSK-3β is a target gene of HNF-1β. In addition, the results of the present study revealed the novel HNF-1β-GSK-3β-p-NFκB pathway, occurring in response to DNA damage. Targeting this pathway may therefore represent a putative, novel, anticancer strategy in patients with OCCC.博士（医学）・甲第785号・令和3年3月15日Copyright: © Kawaharaet al. This is an open access article distributed under theterms of CreativeCommons Attribution License(https://creativecommons.org/licenses/by-nc-nd/4.0/)

Determination of the genetic structure of remnant Morus boninensis Koidz. trees to establish a conservation program on the Bonin Islands, Japan

BACKGROUND: Morus boninensis, is an endemic plant of the Bonin (Ogasawara) Islands of Japan and is categorized as "critically endangered" in the Japanese red data book. However, little information is available about its ecological, evolutionary and genetic status, despite the urgent need for guidelines for the conservation of the species. Therefore, we adopted Moritz's MU concept, based on the species' current genetic structure, to define management units and to select mother tree candidates for seed orchards. RESULTS: Nearly all individuals of the species were genotyped on the basis of seven microsatellite markers. Genetic diversity levels in putative natural populations were higher than in putative man-made populations with the exception of those on Otouto-jima Island. This is because a limited number of maternal trees are likely to have been used for seed collection to establish the man-made populations. A model-based clustering analysis clearly distinguished individuals into nine clusters, with a large difference in genetic composition between the population on Otouto-jima Island, the putative natural populations and the putative man-made populations. The Otouto-jima population appeared to be genetically differentiated from the others; a finding that was also supported by pairwise F(ST )and R(ST )analysis. Although multiple clusters were detected in the putative man-made populations, the pattern of genetic diversity was monotonous in comparison to the natural populations. CONCLUSION: The genotyping by microsatellite markers revealed strong genetic structures. Typically, artificial propagation of this species has ignored the genetic structure, relying only on seeds from Otouto-jima for replanting on other islands, because of a problem with inter-specific hybridization on Chichi-jima and Haha-jima Islands. However, this study demonstrates that we should be taking into consideration the genetic structure of the species when designing a propagation program for the conservation of this species

Quantitative Evaluation of Osteocyte Morphology and Bone Anisotropic Extracellular Matrix in Rat Femur

Osteocytes are believed to play a crucial role in mechanosensation and mechanotransduction which are important for maintenance of mechanical integrity of bone. Recent investigations have revealed that the preferential orientation of bone extracellular matrix (ECM) mainly composed of collagen fibers and apatite crystallites is one of the important determinants of bone mechanical integrity. However, the relationship between osteocytes and ECM orientation remains unclear. In this study, the association between ECM orientation and anisotropy in the osteocyte lacuno-canalicular system, which is thought to be optimized along with the mechanical stimuli, was investigated using male rat femur. The degree of ECM orientation along the femur longitudinal axis was significantly and positively correlated with the anisotropic features of the osteocyte lacunae and canaliculi. At the femur middiaphysis, there are the osteocytes with lacunae that highly aligned along the bone long axis (principal stress direction) and canaliculi that preferentially extended perpendicular to the bone long axis, and the highest degree of apatite c-axis orientation along the bone long axis was shown. Based on these data, we propose a model in which osteocytes can change their lacuno-canalicular architecture depending on the mechanical environment so that they can become more susceptible to mechanical stimuli via fluid flow in the canalicular channel.Ishimoto T., Kawahara K., Matsugaki A., et al. Quantitative Evaluation of Osteocyte Morphology and Bone Anisotropic Extracellular Matrix in Rat Femur. Calcified Tissue International, 109, 4, 434. https://doi.org/10.1007/s00223-021-00852-1

卵巣子宮内膜症性嚢胞の嚢胞液鉄濃度が不妊に及ぼす影響について

The causes of infertility in women with endometriosis may range from anatomical distortions to various pathophysiological disturbances. The aim of the present study was to examine the effects of the cyst fluid (CF) concentration of iron on infertility in patients with ovarian endometrioma (OMA). Patients with histologically confirmed OMA were enrolled at the Department of Obstetrics and Gynecology, Nara Medical University Hospital between 2013 and 2019. The patients were divided into 2 groups, namely women experiencing current infertility (infertile group) and those without complaints of infertility (non‑infertile group). There were 2 types of patients in the infertile group: Patients who failed to achieve a clinical pregnancy following ≥12 months of regular unprotected sexual intercourse and those who had already been treated at fertility clinics. The CF concentration of iron was measured by the inductively coupled plasma‑optical emission spectrometry (ICP‑OES) method. The clinical data were analyzed retrospectively. A total of 77 patients were enrolled in the present study. Among these, 32 (41.6%) patients had infertility. When compared with the non‑infertile women, the infertile women were significantly younger (median age, 35 years; range, 24‑47 years; vs. median age, 40 years; range, 21‑53 years; respectively; P=0.003). The CF concentrations of iron (median, 324.8 mg/l; range, 71.3‑1046.3 mg/l; vs. median, 226.5 mg/l; range, 65.3‑737.5 mg/l; respectively; P=0.019) were significantly higher in the infertile group compared with the non‑infertile group. Multivariate logistic regression analysis indicated that age at diagnosis (≤38 years), the CF concentrations of iron (>326.6 mg/l) and the infertility index (iron/age ratio, >8.37) were independent risk factors for endometriosis‑related infertility. Multivariate analysis revealed that age (HR, 6.44; 95% CI, 2.06‑20.12) and iron (HR, 4.90; 95% CI, 1.48‑16.22) were independent factors for the identification of patients with OMA who presented with a complaint of infertility. In addition, the infertility index (iron/age ratio, >8.37; HR, 4.85; 95% CI, 1.01‑23.27) was an important predictor of infertility. ROC curve analysis also revealed that the areas under the ROC (AUC) for age, iron and infertility index were 0.699, 0.666 and 0.731, respectively. On the whole, the findings of the present study demonstrate that age at diagnosis and the CF concentration of iron may be potentially effective markers for predicting infertility in women with OMA.博士（医学）・乙第1500号・令和3年3月15日Copyright © Nagayasuet al. This is an open access article distributed under theterms of CreativeCommons Attribution License(https://creativecommons.org/licenses/by-nc-nd/4.0/)

Adaptive action supervision in reinforcement learning from real-world multi-agent demonstrations

Modeling of real-world biological multi-agents is a fundamental problem in various scientific and engineering fields. Reinforcement learning (RL) is a powerful framework to generate flexible and diverse behaviors in cyberspace; however, when modeling real-world biological multi-agents, there is a domain gap between behaviors in the source (i.e., real-world data) and the target (i.e., cyberspace for RL), and the source environment parameters are usually unknown. In this paper, we propose a method for adaptive action supervision in RL from real-world demonstrations in multi-agent scenarios. We adopt an approach that combines RL and supervised learning by selecting actions of demonstrations in RL based on the minimum distance of dynamic time warping for utilizing the information of the unknown source dynamics. This approach can be easily applied to many existing neural network architectures and provide us with an RL model balanced between reproducibility as imitation and generalization ability to obtain rewards in cyberspace. In the experiments, using chase-and-escape and football tasks with the different dynamics between the unknown source and target environments, we show that our approach achieved a balance between the reproducibility and the generalization ability compared with the baselines. In particular, we used the tracking data of professional football players as expert demonstrations in football and show successful performances despite the larger gap between behaviors in the source and target environments than the chase-and-escape task.Comment: 14 pages, 5 figure

母体血中における分娩形式による分娩前後の、羊水塞栓症のバイオマーカーとなりうるSCC 抗原の大きな変化

Amniotic fluid embolism (AFE) is a serious disease in which amniotic fluid components enter the maternal systemic circulation, causing cardiopulmonary collapse in pregnant women. It has been previously reported that amniotic fluid contains extremely high levels of squamous cell carcinoma antigen (SCCA), and that pregnant women who do not survive due to AFE have high blood SCCA levels. The aim of the present study was to determine the possible mechanisms through which SCCA in amniotic fluid enters the maternal blood, as well as the potential origin of SCCA. The prospective study included a cohort of 464 women (339 normal vaginal deliveries, 97 cesarean deliveries without labor, and 28 cesarean deliveries with labor). The dynamic changes in maternal serum SCCA levels were determined before and after delivery in relation to the mode of delivery, and SCCA levels were measured in the placenta, fetal skin, amniotic fluid cell components, amniotic fluid and neonatal urine. Serial serum samples collected at the time of admission, at 2 h postpartum, and on postpartum day 3 were quantitatively measured for SCCA by enzyme‑linked immunosorbent assay. Amniotic fluid and neonatal urine SCCA levels were also measured. The protein expression of SCCA in the placenta and fetal skin was assessed by immunohistochemistry. In vaginal deliveries, there was a significant increase in serum SCCA levels from admission to 2 h postpartum, and SCCA levels decreased on postpartum day 3. In cesarean deliveries, the SCCA levels at the time of admission and 2 h postpartum did not differ. The SCCA levels were significantly higher in women who underwent vaginal deliveries compared to those that underwent cesarean deliveries without labor (P=0.033). Immunohistochemical staining revealed no SCCA expression in the placenta and fetal skin. The SCCA levels in neonatal urine immediately after birth were as high as those in the amniotic fluid, suggesting that SCCA may originate from fetal urine. The present study thus suggests that amniotic fluid SCCA levels, which may be derived from fetal urine, can enter the maternal circulation during vaginal delivery. The onset of labor and full cervical dilatation are the main causes of entry of amniotic fluid components into the maternal circulation.博士（医学）・甲第796号・令和3年6月25日Copyright: © Nakano et al. This is an open access article distributed under the terms of Creative Commons Attribution License (https://creativecommons.org/licenses/by-nc-nd/4.0/)

A Case of Bilateral Adrenal and Pleural Metastases from Prostate Cancer

Our case was 65 years old. At check-up, a high PSA level of 515 ng/ml was observed, the patient was diagnosed with having clinical stage D prostate cancer and a Maximum Androgen Blockade (MAB therapy) was started. In response to the exacerbated prostate cancer, we started a therapy involving the administration of 8 mg/kg body weight of dexamethasone and 55 mg/m2 of docetaxel every 3 weeks. After completing 8 courses, an enlargement of the bilateral adrenal tumor was observed, and after completing 12 courses, a pleural tumor was discovered and the PSA level was also increased. The patient was therefore diagnosed with having bilateral adrenal metastasis and pleural metastasis of prostate cancer through diagnostic imaging. So far, there have been no reports of multiple occurrences of prostate cancer in the adrenal glands and the pleura, thus making this case the first such case