Self-Organized Formation of Polarized Cortical Tissues from ESCs and Its Active Manipulation by Extrinsic Signals

SummaryHere, we demonstrate self-organized formation of apico-basally polarized cortical tissues from ESCs using an efficient three-dimensional aggregation culture (SFEBq culture). The generated cortical neurons are functional, transplantable, and capable of forming proper long-range connections in vivo and in vitro. The regional identity of the generated pallial tissues can be selectively controlled (into olfactory bulb, rostral and caudal cortices, hem, and choroid plexus) by secreted patterning factors such as Fgf, Wnt, and BMP. In addition, the in vivo-mimicking birth order of distinct cortical neurons permits the selective generation of particular layer-specific neurons by timed induction of cell-cycle exit. Importantly, cortical tissues generated from mouse and human ESCs form a self-organized structure that includes four distinct zones (ventricular, early and late cortical-plate, and Cajal-Retzius cell zones) along the apico-basal direction. Thus, spatial and temporal aspects of early corticogenesis are recapitulated and can be manipulated in this ESC culture

A case of VEXAS syndrome (vacuoles, E1 enzyme, X-linked, autoinflammatory, somatic) with decreased oxidative stress levels after oral prednisone and tocilizumab treatment

VEXAS (vacuoles, E1 enzyme, X-linked, autoinflammatory, somatic) syndrome has recently been described as an autoinflammatory disease associated with severe adult-onset inflammatory manifestations. The various clinical manifestations include recurrent high-grade fever, neutrophilic dermatoses, cutaneous vasculitis, chondritis of the ear and nose, pulmonary infiltrates, cytopenia, uveitis, gastrointestinal pain or inflammation, aortitis, hepatosplenomegaly, and hematological disorders. VEXAS syndrome is caused by somatic mutations of the ubiquitin-like modifier activating enzyme 1 (UBA1) gene in myeloid-lineage cells. It is characterized by vacuolated myeloid and erythroid progenitor cells seen by bone marrow biopsy. We report the case of a 64-year-old Japanese man with VEXAS syndrome. At age 63, he was referred to us with a recurrent erythema on the hands associated with a general fever of 38–40°C that had persisted for 4 or 5 days and had recurred about once a month for a year. The skin rash appeared 2 or 3 days after the onset of each fever episode. Computed tomography (CT) of the chest revealed bilateral hilar lymphadenopathy (BHL), and the mediastinal lymph nodes were swollen. Sarcoidosis was suspected but was ruled out by several tests. Laboratory examinations showed elevated inflammatory markers. Bone marrow examination showed the vacuolization of myeloid precursor cells. A skin biopsy revealed dense dermal, predominantly perivascular, infiltrates. These consisted of mature neutrophils admixed with myeloperoxidase-positive CD163-positive myeloid cells, lymphoid cells and eosinophils. Sequencing analysis identified the somatic UBA1 variant c.122T > C, which results in p.Met41Thr. Treatment with oral prednisone (15 mg/day) and monthly intravenous tocilizumab injections (400 mg) completely resolved the symptoms. Neutrophils are a major source of reactive oxygen species, and the present case demonstrated numerous neutrophilic infiltrates. We hypothesize that the patient might have had elevated derivatives of reactive oxygen metabolites (d-ROMs). d-ROM quantification is a simple method for detecting hydroperoxide levels, and clinical trials have proven it useful for evaluating oxidative stress. In this study, we measured serum d-ROM before and after oral prednisone and tocilizumab treatment. The levels decreased significantly during treatment

コベツ エイヨウ シドウ ニ ジュウジスル カンリ エイヨウシ オ タイショウ トシタ マナビ ナオシ プログラム ノ ケントウ

【目的】病院管理栄養士が「不足していると感じているスキル」や「希望する学び直しプログラム」と、現在各大学で実施されている学び直しプログラムとを比較し、より効果的に個別栄養指導の質を向上させることができる学び直しプログラムを検討すること。【方法】個別栄養指導業務を行っている病院勤務の直営管理栄養士74名(回収率:82.4%、有効回答者数:61名)を対象として、平成23年10月3日～11月15日に実施した選択および自記式のアンケート調査結果を用い、「勤続年数」や「個別栄養指導経験年数」と、「不足していると感じているスキル」の総数や項目とを比較し、解析を行った。また、各大学で行われている学び直しプログラムの状況について、報告書、ホームページ等から調査した。【結果および考察】「不足していると感じているスキル」の総数は、「勤続年数」、「個別栄養指導経験年数」をそれぞれ5年未満群と5年以上群に群分けした場合、5年以上群で有意に低値を示した。不足していると感じているスキルの中で、5年未満群に比べ5年以上群の方が大きく低値を示したスキルは、主にカウンセリングや栄養教育に関するものであった。これらは個別栄養指導を経験することで不足を補いやすいスキルであると考えられる。一方、「運動療法の知識」、「薬の知識」、「論文読解」などの項目においては、両群間に差が認められなかった。これらは、養成施設では学ばないまたは詳しく学ばない、学んだが忘れている、新しい情報を理解できているか不安、科学英語に自信がない等、個別栄養指導を経験するだけでは補いにくいスキルであると考えられる。これらのことから、複数年の個別栄養指導経験を、短期間プログラムで代替でき、かつ、日頃補いきれない知識を充実させることができるような学び直しプログラムの構築が必要であると考えられた。Purpose The present study aimed to investigate registered dietitians\u27 understanding of the problems involved in individual nutritional guidance, and to find ways to improve the effectiveness of their guidance. Methods The subjects were 74 registered dietitians performing individual nutritional guidance work in hospitals(recovery rate: 82.4%, effective number of respondents: 61). We investigated the following using selection and a self administered questionnaire: 1)number of years\u27 work experience; 2)years of experience in individual nutritional guidance; and 3)skills required to give individual nutritional guidance that I lack. We also investigated reports or websites from other universities to determine the current status of the re learning program there. Results The total number of "Skills required to give individual nutritional guidance that I lack" in the "More than 5 years" group was significantly lower than in the "Less than 5 years" group. Particularly, "Skills of counseling" and "Skills of nutritional education" were the most different between the groups. However, no difference between groups was observed in "Knowledge of exercise therapy", "Knowledge of pharmaceutical agents" and "Skills in article reading"

Self-Organization of Polarized Cerebellar Tissue in 3D Culture of Human Pluripotent Stem Cells

During cerebellar development, the main portion of the cerebellar plate neuroepithelium gives birth to Purkinje cells and interneurons, whereas the rhombic lip, the germinal zone at its dorsal edge, generates granule cells and cerebellar nuclei neurons. However, it remains elusive how these components cooperate to form the intricate cerebellar structure. Here, we found that a polarized cerebellar structure self-organizes in 3D human embryonic stem cell (ESC) culture. The self-organized neuroepithelium differentiates into electrophysiologically functional Purkinje cells. The addition of fibroblast growth factor 19 (FGF19) promotes spontaneous generation of dorsoventrally polarized neural-tube-like structures at the level of the cerebellum. Furthermore, addition of SDF1 and FGF19 promotes the generation of a continuous cerebellar plate neuroepithelium with rhombic-lip-like structure at one end and a three-layer cytoarchitecture similar to the embryonic cerebellum. Thus, human-ESC-derived cerebellar progenitors exhibit substantial self-organizing potential for generating a polarized structure reminiscent of the early human cerebellum at the first trimester

Vulnerability of Purkinje Cells Generated from Spinocerebellar Ataxia Type 6 Patient-Derived iPSCs

SummarySpinocerebellar ataxia type 6 (SCA6) is a dominantly inherited neurodegenerative disease characterized by loss of Purkinje cells in the cerebellum. SCA6 is caused by CAG trinucleotide repeat expansion in CACNA1A, which encodes Cav2.1, α1A subunit of P/Q-type calcium channel. However, the pathogenic mechanism and effective therapeutic treatments are still unknown. Here, we have succeeded in generating differentiated Purkinje cells that carry patient genes by combining disease-specific iPSCs and self-organizing culture technologies. Patient-derived Purkinje cells exhibit increased levels of full-length Cav2.1 protein but decreased levels of its C-terminal fragment and downregulation of the transcriptional targets TAF1 and BTG1. We further demonstrate that SCA6 Purkinje cells exhibit thyroid hormone depletion-dependent degeneration, which can be suppressed by two compounds, thyroid releasing hormone and Riluzole. Thus, we have constructed an in vitro disease model recapitulating both ontogenesis and pathogenesis. This model may be useful for pathogenic investigation and drug screening