A salmon DNA scaffold promotes osteogenesis through activation of sodium-dependent phosphate cotransporters.

We previously reported the promotion of bone regeneration in calvarial defects of both normal and ovariectomy-induced osteoporotic rats, with the use of biodegradable DNA/protamine scaffold. However, the method by which this DNA-containing scaffold promotes bone formation is still not understood. We hypothesize that the salmon DNA, from which this scaffold is derived, has an osteoinductive effect on pre-osteoblasts and osteoblasts. We examined the effects of salmon DNA on osteoblastic differentiation and calcification in MC3T3-E1 cells, mouse osteoblasts, in vitro and bone regeneration in a calvarial defect model of aged mouse in vivo. The salmon DNA fragments (300 bps) upregulated the expression of the osteogenic markers, such as alkaline phosphatase, Runx2, and osterix (Osx) in MC3T3E1 cells compared with incubation with osteogenic induction medium alone. Measurement of phosphate ion concentrations in cultures showed that the DNA scaffold degraded phosphate ions were released to the cell cultures. Interestingly, we found that the inclusion of DNA in osteoblastic cell cultures upregulated the expression of sodium-dependent phosphate (NaPi) cotransporters, SLC20A1 and SLC34A2, in MC3T3-E1 cells in a time dependent manner. Furthermore, the inclusion of DNA in cell cultures increased the transcellular permeability of phosphate. Conversely, the incubation of phosphonoformic acid, an inhibitor of NaPi cotransporters, attenuated the DNA-induced expression and activation of SLC20A1 and SLC34A2 in MC3T3-E1 cells, resulting in suppression of the osteogenic markers. The implantation of a salmon DNA scaffold disk promoted bone regeneration using calvarial defect models in 30-week-old mice. Our results indicate that the phosphate released from salmon DNA upregulated the expression and activation of NaPi cotransporters, resulting in the promotion of bone regeneration.福岡歯科大学2016年

Reactive oxygen species stimulates epithelial mesenchymal transition in normal human epidermal keratinocytes via TGF-beta secretion.

Epithelial to mesenchymal transition (EMT) plays an important role in tumor progression, and is an early step in carcinogenesis. Although reactive oxygen species (ROS) are known to be implicated in EMT in many tumor cell types, its exact role in EMT initiation in normal human cells, especially epidermal keratinocytes (NHEKs), remains unknown. To clarify whether ROS induce EMT in NHEKs, and to establish how ROS regulate EMT, we examined the effect of hydrogen peroxide (H(2)O(2)) on the expression of molecules involved in EMT and cell morphology in NHEKs. H(2)O(2) altered the expression of EMT biomarkers, including downregulation of epithelial cadherin and upregulation of α-smooth muscle actin, through a transcriptional modulator, Snail1. H(2)O(2) also induced epithelial to fibroblast-like morphological changes, together with upregulation of EMT biomarkers, and promoted phosphorylation of ERK1/2 and JNK in a time-dependent manner. Interestingly, H(2)O(2) stimulated the expression and secretion of TGF-β1 in NHEKs. Exogenous TGF-β1 also induced the expression of EMT biomarkers. In contrast, neutralizing antibody anti-TGF-β1 antibody or inhibitor of TGF-β receptor type I suppressed the expression of EMT biomarkers. Our results suggest that ROS stimulated TGF-β1 secretion and MAPK activation, resulting in EMT initiation in NHEKs.福岡歯科大学2013年

Ginsenoside Rb1 Prevents MPP +

Ginsenoside Rb1 shows neuroprotective effects in various neurons, including dopaminergic cells. However, the precise mechanisms of action are uncertain. In this paper, we examine whether Rb1 has a neuroprotective effect on MPP+-induced apoptosis and attempt to clarify the signaling pathway in PC12 cells. Apoptosis of PC12 cells was determined by DNA fragmentation assay, the activation of caspase-3, or by the inactivation of Bcl-xL. Rb1 inhibited MPP+-induced caspase-3 activation and DNA fragmentation and activated Bcl-xL in MPP+-treated PC12 cells. These antiapoptotic effect was abrogated in PC12 cells transfected with estrogen receptor siRNA. Levels of DNA fragmentation were increased by wortmannin or PD 98059, while they were decreased by SB 203580 or SP 600125 in MPP+-treated PC12 cells. Rb1 increased phosphorylation levels of ERK1/2 or Akt in MPP+-treated PC12 cells, while it reduced phosphorylated p38 or SAPK/JNK. The increased phosphorylation of ERK/1/2 or Akt by Rb1 was abrogated by estrogen receptor siRNA. Rb1-induced inhibition of SAPK/JNK or p38 phosphorylation was also abolished by estrogen receptor siRNA. These results suggest that ginsenoside Rb1 protects PC12 cells from caspase-3-dependent apoptosis through stimulation of estrogen receptor with consequent activation of ERK1/2 and Akt and inhibition of SAPK/JNK and p38

Cisplatin-induced programmed cell death ligand-2 expression is associated with metastasis ability in oral squamous cell carcinoma.

Programmed cell death ligands (PD-Ls) are expressed in tumor cells where they bind to programmed cell death-1, an immunocyte co-receptor, resulting in tumor cell evasion from the immune system. Chemotherapeutic drugs have been recently reported to induce the expression of PD-L, such as PD-L1, in some cancer cells. However, little is known regarding PD-L2 expression and its role in oral squamous cell carcinoma (OSCC). In this study, we examined the effect of cisplatin on the expression and regulation of PD-L2 in OSCC cell lines and analyzed malignant behavior in PD-L2-expressing cells using colony, transwell and transformation assays. In addition, we examined PD-L2 expression in the tumor tissues of OSCC patients using cytology and tissue microarray methods. In OSCC cell lines, cisplatin treatment upregulated PD-L2 expression, along with that of the drug efflux transporter ABCG2, via signal transducers and activator of transcription (STAT) 1/3 activation. Moreover, PD-L2-positive or PD-L2-overexpressing cells demonstrated upregulation in both invasion and transformation ability but not in proliferation compared with PD-L2-negative or PD-L2-silencing cells. PD-L2 expression was also observed in OSCC cells of cytology samples and tissue from OSCC patients. The intensity of PD-L2 expression was correlated with more malignant morphological features in the histological appearance and an invasive pattern. Our findings indicate that cisplatin-upregulated PD-L2 expression in OSCC via STAT1/3 activation and the expression of PD-L2 are likely to be associated with malignancy in OSCC. The PD-L2 expression in cisplatin-resistant OSCC cells may be a critical factor in prognosis of advanced OSCC patients.福岡歯科大学2019年

Nutrient-induced FNIP degradation by SCFβ-TRCP regulates FLCN complex localization and promotes renal cancer progression.

Folliculin-interacting protein 1 and 2 (FNIP1 and FNIP2) play critical roles in preventing renal malignancy through their association with the tumor suppressor FLCN. Mutations in FLCN are associated with Birt-Hogg-Dubé (BHD) syndrome, a rare disorder with increased risk of renal cancer. Recent studies indicated that FNIP1/FNIP2 double knockout mice display enlarged polycystic kidneys and renal carcinoma, which phenocopies FLCN knockout mice, suggesting that these two proteins function together to suppress renal cancer. However, the molecular mechanism functionally linking FNIP1/FNIP2 and FLCN remains largely elusive. Here, we demonstrated that FNIP2 protein is unstable and subjected to proteasome-dependent degradation via β-TRCP and Casein Kinase 1 (CK1)-directed ubiquitination in a nutrition-dependent manner. Degradation of FNIP2 leads to lysosomal dissociation of FLCN and subsequent lysosomal association of mTOR, which in turn promotes the proliferation of renal cancer cells. These results indicate that SCFβ-TRCP negatively regulates the FLCN complex by promoting FNIP degradation and provide molecular insight into the pathogenesis of BHD-associated renal cancer.福岡歯科大学2016年

ヨーロッパにおける先進的なデザイン教育方法の調査研究

本研究はデザイン教育研究センターの研究活動、すなわちデザイナーやアーティストを目指す学生のための新たな教育プログラム開発のための調査研究、これを基盤とした公開特別講義を中心とするカリキュラムへの組み込み、研究成果の報告と社会とのコミュニケーションを進めるためのメディアとしての書籍出版、という一連の活動と連動するものである。ここでは、出版された『中山英之/スケッチング』（新宿書房刊）について解説が行われる。はじめに1 本の線があり、それに線が加わり、そのたびごとにそこに現れる空間が変わっていく、このスケッチの方法は中山氏が普段の設計プロセスにおいても行っているスケッチの方法であり、思考の枠組みを広げ、次元を行き来しながら設計を進めていくという独特な方法論を示すものである。このスケッチの描き方を書籍デザインそのものに反映させる試みをおこなった。まるで、絵本のように見える親しみやすい線書きのスケッチには、限りなく奥の深いデザインの思想が隠されている。In early June 2009, Prof. Yuichiro Kodama arranged forNakayama to address our Department of EnvironmentalDesign. At this special guest lecture, he presented anoverview of his projects since going independent fromIto’s office, giving us insight into the rigorous reasoningthat shapes his spaces while yet allowing a remarkablesensitivity toward fragile ephemera: clover growing on aprospective house site, thick weeds in a field designatedfor a kiosk design competition in Hokkaido. Using linedrawings to illustrate how he expands upon a basicframework of spatial concepts, his secret seems to lie inthat wondrous window of consciousness he invokessomewhere between two and three dimensions. InNovember, toward this book, Nakayama gave a privatelecture at his office to only a few people involved with theediting, design and publishing

High Expression of Dihydropyrimidine Dehydrogenase in Lung Adenocarcinoma is Associated With Mutations in Epidermal Growth Factor Receptor: Implications for the Treatment of Non?Small-Cell Lung Cancer Using 5-Fluorouracil

BackgroundIt has been shown that 5-fluorouracil (5-FU) sensitivity in patients with non?small-cell lung cancer (NSCLC) is associated with epidermal growth factor receptor (EGFR) mutation status. However, the relationship between dihydropyrimidine dehydrogenase (DPD), a 5-FU degrading enzyme, and EGFR mutation status is unknown. Here, we focus on clinicopathologic factors and in vitro correlations between DPD expression and EGFR mutation status.Patients and MethodsEGFR mutations and messenger RNA (mRNA) levels of DPD and thymidylate synthase (TS) were analyzed in 47 resected NSCLC tumors by laser-capture microdissection. In addition, relationships between EGFR mutation status and the immunohistochemical expression of DPD and TS in 49 patients with primary NSCLC who were treated with a 5-FU derivative of S-1 postoperatively were examined. Correlations among clinicopathologic factors were evaluated. The effect of epidermal growth factor on DPD expression was also investigated in vitro in various cell lines.ResultsAdenocarcinoma in situ showed significantly higher DPD mRNA levels and more EGFR mutation frequency than other histological types (P < .05). DPD immunopositive cases were more frequently observed in adenocarcinoma, in females, and in nonsmokers. DPD immunopositive cases were correlated with EGFR mutation status (P < .003). The prognoses of wild-type EGFR and mutated EGFR populations were similarly favorable with postoperative S-1 treatment, which overcomes the problem of 5-FU degradation in mutated EGFR. In vitro, EGFR-mutated cell lines showed high DPD mRNA and protein expression.ConclusionHigh DPD expression was shown to be correlated with EGFR mutation in adenocarcinoma cells and tissues. Clinicians should take this finding into consideration when using 5-FU to treat patients with NSCLC

情報化社会における大学のデザイン導入教育に関する研究

神戸芸術工科大学デザイン教育研究センターでは、新しいデザイン教育プログラムに関する研究を継続的におこなっている。新しい社会構造、新しいテクノロジー環境の中では大学教育も変わるべきである。そのような視点から、たとえば情報化社会においてアートとデザインの教育がどのように変わっていくのか、このことは重要な研究テーマのひとつとなる。こうした観点から世界の新たなデザイン教育領域や教育方法を見渡すと、ロンドンの王立芸術学院におけるインタラクション・デザイン教育が際立っている。これまで私たちは数回にわたり同大学院ディグリーショウを訪問し、アンソニー・ダン専攻主任にインタビューをおこなっている。またアンソニー・ダンとフィオナ・レイビイ両氏を招聘し、RCAにおける教育プログラムおよび彼ら自身のデザイン活動について公開特別講義を開催してきた。本年度はフィオナ・レイビイ氏を招いた武蔵野美術大学・IAMAS情報科学芸術大学院大学・神戸芸術工科大学の三大学共同デザインワークショップを開催した。本稿はこの国際デザインワークショップの記録である。The KDU Center for Design Studies continues to further studies into new design education programmes. Higher education can and must change in keeping with new social structures and new technological environments, one crucial aspect being the issue of how art and design education should reflect information society. In this regard, London’s Royal College of Art stands out among design education institutions around the world f or the range and teaching methods of its Design Interactions curriculum.Over the years, we visited RCA degree shows and interviewed AnthonyDunne (currently department Chairman). We also invited Dunne and Raby to give a Special Lecture at KDU on the RCA curriculum and their own design activities.In this year Kobe Design University, together with Musashino Art University and the Institute of Advanced Media Arts and Sciences, hosted a design workshop with Fiona Raby.This is the document of this international design workshop

大学間国際ジョイントワークショップおよび教育ラウンドテーブル開催を基盤とするデザイン教育の新たな可能性に関する実践的研究

研究の目的は、インタラクションデザインに関するこれまでの学術的な交流や研究の基盤に立ち、日本における新しい教育領域・教育方法としてのインタラクションデザインの可能性もしくは問題点などを、実際の教育現場において確認し、導入のための基礎を構築することにある。本研究の中心となったのは、2012年12月17～21日に開催したスプツニ子！客員教授によるインタラクションデザインワークショップである。これは「Body Futures」をテーマとした、未来における身体の在り方を考えるワークショップであり、公開講評会をKIITOで行ない、提案された各グループのプロジェクトは2013年2月22～28日の期間、同じくKIITOにおいて開催された「Body Futures展」において展示された。このワークショップでは、大学院と学部の連携、大学と企業（アシックス）との連携が試みられた。異なる年齢層の参加者間の連携と、異なる組織領域間の連携が新たな発想を生み出し、「未来の身体」を様々な観点からとらえた科学的な提案が行なわれた。The objective of this study is to identify possibilities and problems related to interaction design within actual educational practice, and thus to construct a foundation for its introducation as a new field and methodology of study in Japan. The main part of this study derives from an interaction design workshop conducted 17-21 December 2012 by Visiting Professor Sputniko! (Hiromi Ozaki) entitled "Body Futures" that proposed to consider further dimensions of physicality. An open forum was subsequently held at KIITO and group projects developed from the workshop were exhibited 22-28 February 2013 under the same Body Futures title, again at KIITO. This workshop attempted to create linkages between the graduate school and university departments, as well as between the university and commercial enterprise (ASICS). It spawned connections between participants of different ages, fostered new thinking across diverse organisational sectors, and proposed various scientific ideas about the future of the body

Cardiomyocyte Formation by Skeletal Muscle-Derived Multi-Myogenic Stem Cells after Transplantation into Infarcted Myocardium

BACKGROUND: Cellular cardiomyoplasty for myocardial infarction has been developed using various cell types. However, complete differentiation and/or trans-differentiation into cardiomyocytes have never occurred in these transplant studies, whereas functional contributions were reported. METHODS AND RESULTS: Skeletal muscle interstitium-derived CD34(+)/CD45(-) (Sk-34) cells were purified from green fluorescent protein transgenic mice by flowcytometory. Cardiac differentiation of Sk-34 cells was examined by in vitro clonal culture and co-culture with embryonic cardiomyocytes, and in vivo transplantation into a nude rat myocardial infarction (MI) model (left ventricle). Lower relative expression of cardiomyogenic transcription factors, such as GATA-4, Nkx2-5, Isl-1, Mef2 and Hand2, was seen in clonal cell culture. However, vigorous expression of these factors was seen on co-culture with embryonic cardiomyocytes, together with formation of gap-junctions and synchronous contraction following sphere-like colony formation. At 4 weeks after transplantation of freshly isolated Sk-34 cells, donor cells exhibited typical cardiomyocyte structure with formation of gap-junctions, as well as intercalated discs and desmosomes, between donor and recipient and/or donor and donor cells. Fluorescence in situ hybridization (FISH) analysis detecting the rat and mouse genomic DNA and immunoelectron microscopy using anti-GFP revealed donor-derived cells. Transplanted Sk-34 cells were incorporated into infarcted portions of recipient muscles and contributed to cardiac reconstitution. Significant improvement in left ventricular function, as evaluated by transthoracic echocardiography and micro-tip conductance catheter, was also observed. CONCLUSIONS AND SIGNIFICANCE: Skeletal muscle-derived multipotent Sk-34 cells that can give rise to skeletal and smooth muscle cells as reported previously, also give rise to cardiac muscle cells as multi-myogenic stem cells, and thus are a potential source for practical cellular cardiomyoplasty