Differentiation of human embryonic stem cells and human induced pluripotent stem cells into steroid-producing cells.

Although there have been reports of the differentiation of mesenchymal stem cells and mouse embryonic stem (ES) cells into steroid-producing cells, the differentiation of human ES/induced pluripotent stem (iPS) cells into steroid-producing cells has not been reported. The purpose of our present study was to establish a method for inducing differentiation of human ES/iPS cells into steroid-producing cells. The first approach we tried was embryoid body formation and further culture on adherent plates. The resultant differentiated cells expressed mRNA encoding the steroidogenic enzymes steroidogenic acute regulatory protein, 3β-hydroxysteroid dehydrogenase, cytochrome P450-containing enzyme (CYP)-11A1, CYP17A1, and CYP19, and secreted progesterone was detected in the cell medium. However, expression of human chorionic gonadotropin was also detected, suggesting the differentiated cells were trophoblast like. We next tried a multistep approach. As a first step, human ES/iPS cells were induced to differentiate into the mesodermal lineage. After 7 d of differentiation induced by 6-bromoindirubin-3'-oxime (a glycogen synthase kinase-3β inhibitor), the human ES/iPS cells had differentiated into fetal liver kinase-1- and platelet derived growth factor receptor-α-expressing mesodermal lineage cells. As a second step, plasmid DNA encoding steroidogenic factor-1, a master regulator of steroidogenesis, was introduced into these mesodermal cells. The forced expression of steroidogenic factor-1 and subsequent addition of 8-bromoadenosine 3',5'-cyclic monophosphate induced the mesodermal cells to differentiate into the steroidogenic cell lineage, and expression of CYP21A2 and CYP11B1, in addition to steroidogenic acute regulatory protein, 3β-hydroxysteroid dehydrogenase, CYP11A1, and CYP17A1, was detected. Moreover, secreted cortisol was detected in the medium, but human chorionic gonadotropin was not. These findings indicate that the steroid-producing cells obtained through the described multistep method are not trophoblast like; instead, they exhibit characteristics of adrenal cortical cells

TNFRSF1B A1466G genotype is predictive of clinical efficacy after treatment with a definitive 5-fluorouracil/cisplatin-based chemoradiotherapy in Japanese patients with esophageal squamous cell carcinoma

<p>Abstract</p> <p>Background</p> <p>Currently definitive 5-fluorouracil (5-FU)/cisplatin (CDDP) -based chemotherapy is recognized as one of the most promising treatments for esophageal cancer. A series of studies performed found genetic polymorphisms and the plasma concentration of 5-FU to be predictive of acute severe toxicities and clinical response. Genetic polymorphisms of <it>tumor necrosis factor (TNF) -α </it>and its surface receptors, <it>TNFRSF1A </it>and <it>TNFRSF1B </it>have been examined in terms of susceptibility to various cancers. In this study, genetic polymorphisms of <it>TNFRSF1B </it>gene were evaluated Japanese esophageal squamous cell carcinoma (ESCC) patients treated with the definitive 5-FU/CDDP-based chemoradiotherapy and their predictive values of prognosis or severe acute toxicities were assessed.</p> <p>Methods</p> <p>Forty-six patients with ESCC were treated with the definitive 5-FU/CDDP-based chemoradiotherapy, one course of which consisted of the continuous infusion of 5-FU for days 1-5 and 8-12, the infusion of CDDP on days 1 and 8, and the radiation at 2 Gy/day on days 1-5, 8-12, and 15-19, with a second course repeated after 2-week interval. Genetic polymorphisms of a TNF-α receptor <it>TNFRSF1B </it>gene were determined by a TaqMan^®MGB probe-based polymerase chain reaction.</p> <p>Results</p> <p>The genotype of <it>TNFSR1B </it>A1466G, but not M196R/T587G or C1493T, was found to be predictive of clinical response, i.e., a complete response or not (p = 0.040). Clinical response was predicted by tumor size (p = 0,002), lymph node metastasis (p = 0.007), distant metastasis (p = 0.001) and disease stage (p < 0.001), but <it>TNFRSF1B </it>A1466G genotype was independent of these factors.</p> <p>Conclusions</p> <p>Genetic polymorphism of <it>TNFRSF1B </it>A1466G was found to be predictive response in Japanese ESCC patients with a definitive 5-FU/CDDP-based chemoradiotherapy. Further clinical investigation with a large number of patients or experiments in vitro should be performed to assess the predictive value of <it>TNFRSF1B </it>A1466G genotype after chemoradiotherapy.</p

A ケン ニ オケル ホウモン カンゴ ステーション ノ 24ジカン オンコール タイセイ ノ ジッタイ : カンリシャ ヘノ アンケート チョウサ カラ

背景 近年, 看護職において, 長く働き続けられる環境づくりを進めるために, ワーク・ライフ・バランスの実現に向けての取り組みが行なわれている. 目的 A県における訪問看護ステーションのオンコール体制の実態を明らかにし, ワーク・ライフ・バランスを検討するための基礎的資料とすることを目的とした. 方法 A県にある70か所の訪問看護ステーションの管理者宛てに, 無記名自記式質問紙調査を送付し, 39か所 (55.7%) から有効回答を得た. 結果 A県にある39か所のステーションにおける1事業所当たりの平均看護師数は6.0±5.6人であり, 全国平均の4.2人と比べると多く, 規模別においては, 中規模事業所が77%を占めていた. 24時間対応体制加算は, 34か所 (87%) が取られているが, オンコール体制に関する規約や手当がない事業所もあった. また, オンコール体制はほとんどが常勤職員で担っており, 管理者の待機日数は, 大規模事業所に比べて小規模および中規模事業所の方が多かった. 考察 オンコール体制が, 在宅看護の重要な機能として捉えている一方で, 訪問看護師の高い離職率や人材確保の難しさの一因となっていることが考えられた. 今後は, 法的根拠に基づいた規約の整備が望まれる. また, 管理者個人が意識化し, 個々のステーションでワーク・ライフ・バランスの取り組みが必要である. 結論 オンコール体制のあり方や職員への指導等を考える資料を得ることができたので, 待機手当のないステーションに手当を提案し, 継続した調査を行う.Background As for the nurse, an action for the realization of the work-life balance is carried out to push forward the making of environment where it is continued acting on for a long time.Purpose To study the actual conditions of home-visit nursing care stations that extend services on an on-call around-the-clock basis, and to make the investigation results available for study of the workers\u27 work-life balance.Methods nvestigations were made by means of sending questionnaires to managers of 70 nursing care stations located in Prefecture A, of which 39 managers, 55.7%, responded on an anonymous basis.Results The average number of nurses per station at 39 nursing care stations in Prefecture A was 6.2, more than the national average of 4.2.Middle-size nursing care stations occupied 77% of the total respondents. Thirty-four stations, or 87%, adopted an additional payment system for around-the-clock services, but there were some that had no internal regulations or additional service charge system for around-the-clock services. Most stations used full-time nurses for around-the-clock services. As for the number of days when managers were on duty for around-the-clock services, small-sized and middle-sized stations had more days than large-sized stations.Discussion The around-the-clock on-call service is an important factor for functioning home-visit nursing care. However, it is also a reason for which home-visit nurses tend to leave the job after a short period and it is getting more difficult to obtain capable nurses. Stipulating legally effective work regulations to improve the working conditions for home-visit nurses is desired in the near future. It is also necessary for each care manager to realize the importance of improving the work-life balance of nurses

Transplantation of vascular cells derived from human embryonic stem cells contributes to vascular regeneration after stroke in mice

<p>Abstract</p> <p>Background</p> <p>We previously demonstrated that vascular endothelial growth factor receptor type 2 (VEGF-R2)-positive cells induced from mouse embryonic stem (ES) cells can differentiate into both endothelial cells (ECs) and mural cells (MCs) and these vascular cells construct blood vessel structures in vitro. Recently, we have also established a method for the large-scale expansion of ECs and MCs derived from human ES cells. We examined the potential of vascular cells derived from human ES cells to contribute to vascular regeneration and to provide therapeutic benefit for the ischemic brain.</p> <p>Methods</p> <p>Phosphate buffered saline, human peripheral blood mononuclear cells (hMNCs), ECs-, MCs-, or the mixture of ECs and MCs derived from human ES cells were intra-arterially transplanted into mice after transient middle cerebral artery occlusion (MCAo).</p> <p>Results</p> <p>Transplanted ECs were successfully incorporated into host capillaries and MCs were distributed in the areas surrounding endothelial tubes. The cerebral blood flow and the vascular density in the ischemic striatum on day 28 after MCAo had significantly improved in ECs-, MCs- and ECs+MCs-transplanted mice compared to that of mice injected with saline or transplanted with hMNCs. Moreover, compared to saline-injected or hMNC-transplanted mice, significant reduction of the infarct volume and of apoptosis as well as acceleration of neurological recovery were observed on day 28 after MCAo in the cell mixture-transplanted mice.</p> <p>Conclusion</p> <p>Transplantation of ECs and MCs derived from undifferentiated human ES cells have a potential to contribute to therapeutic vascular regeneration and consequently reduction of infarct area after stroke.</p

ERRγ enhances cardiac maturation with T-tubule formation in human iPSC-derived cardiomyocytes

ヒトのiPS細胞から新生児レベルまで成熟した心筋細胞を作製する. 京都大学プレスリリース. 2021-06-21.Lowering the cost of heart cell therapies. 京都大学プレスリリース. 2021-06-21.One of the earliest maturation steps in cardiomyocytes (CMs) is the sarcomere protein isoform switch between TNNI1 and TNNI3 (fetal and neonatal/adult troponin I). Here, we generate human induced pluripotent stem cells (hiPSCs) carrying a TNNI1[EmGFP] and TNNI3[mCherry] double reporter to monitor and isolate mature sub-populations during cardiac differentiation. Extensive drug screening identifies two compounds, an estrogen-related receptor gamma (ERRγ) agonist and an S-phase kinase-associated protein 2 inhibitor, that enhances cardiac maturation and a significant change to TNNI3 expression. Expression, morphological, functional, and molecular analyses indicate that hiPSC-CMs treated with the ERRγ agonist show a larger cell size, longer sarcomere length, the presence of transverse tubules, and enhanced metabolic function and contractile and electrical properties. Here, we show that ERRγ-treated hiPSC-CMs have a mature cellular property consistent with neonatal CMs and are useful for disease modeling and regenerative medicine

Three dimensional motion analyses for rehabilitation version of Awa Odori exercise and the expectancy of physical effects

‘Awa Odori Exercise -Rehabilitation version- was developed in 2006 for the new trial of physical exercise for the aging and the impaired person with lower balance performance in Tokushima prefecture, Japan. Public relations of this exercise had been spreading over Tokushima since then. The characteristics of the exercise were highly familiar with most of people in Tokushima because of popularity in original ‘Awa Odori’. This study proposed the efficacies of Awa Odori Exercise as a rehabilitation exercise. This exercise expected the flexible balance reinforcements and the substitution for walking training with prevention of fall, bedridden and participating restriction for the old people, also promoting the health in Tokushima

A Proposal for Practical Diagnosis of Renal Hypouricemia : Evidenced from Genetic Studies of Nonfunctional Variants of URAT1/SLC22A12 among 30,685 Japanese Individuals

Background: Renal hypouricemia (RHUC) is characterized by a low serum uric acid (SUA) level and high fractional excretion of uric acid (FEUA). Further studies on FEUA in hypouricemic individuals are needed for a more accurate diagnosis of RHUC. Methods: In 30,685 Japanese health-examination participants, we genotyped the two most common nonfunctional variants of URAT1 (NFV-URAT1), W258X (rs121907892) and R90H (rs121907896), in 1040 hypouricemic individuals (SUA ≤ 3.0 mg/dL) and 2240 individuals with FEUA data. The effects of NFV-URAT1 on FEUA and SUA were also investigated using linear and multiple regression analyses. Results: Frequency of hypouricemic individuals (SUA ≤ 3.0 mg/dL) was 0.97% (male) and 6.94% (female) among 30,685 participants. High frequencies of those having at least one allele of NFV-URAT1 were observed in 1040 hypouricemic individuals. Furthermore, NFV-URAT1 significantly increased FEUA and decreased SUA, enabling FEUA and SUA levels to be estimated. Conversely, FEUA and SUA data of hypouricemic individuals are revealed to be useful to predict the number of NFV-URAT1. Conclusions: Our findings reveal that specific patterns of FEUA and SUA data assist with predicting the number of nonfunctional variants of causative genes for RHUC, and can also be useful for practical diagnosis of RHUC even before genetic tests