Efficient generation of adenovirus vectors carrying the Clustered regularly interspaced short palindromic repeat (CRISPR)-CRISPR associated proteins (Cas)12a system by suppressing Cas12a expression in packaging cells

Clustered regularly interspaced short palindromic repeat (CRISPR)-CRISPR associated proteins (Cas) 9 system is a powerful tool for genome editing and still being aggressively improved. Cas12a, a recently discovered Cas9 ortholog, is expected to become complementary to Cas9 due to its unique characteristics. Previously we attempted to establish an adenovirus (Ad) vector-mediated delivery of CRISPR-Cas12a system since Ad vector is widely used for gene transfer in basic researches and medical applications. However, we found difficulties preparing of Ad vectors at an adequate titer. In this study, we have developed Ad vectors that conditionally express Cas12a either by a tetracycline-controlled promoter or a hepatocyte specific promoter to avoid putative inhibitory effects of Cas12a. These vectors successfully proliferated in packaging cells, HEK293 cells, and were recovered at high titers. We have also developed packaging cells that express shRNA for Cas12a to suppress expression of Cas12a. Using the cells, the Ad vector directing constitutive expression of Cas12a proliferated efficiently and was successfully recovered at a high titer. Overall, we improved recovery of Ad vectors carrying CRISPR-Cas12a system, thus provided them as a tool in genome editing researches.Tsukamoto T., Sakai E., Nishimae F., et al. Efficient generation of adenovirus vectors carrying the Clustered regularly interspaced short palindromic repeat (CRISPR)-CRISPR associated proteins (Cas)12a system by suppressing Cas12a expression in packaging cells. Journal of Biotechnology 304, 1 (2019); https://doi.org/10.1016/j.jbiotec.2019.08.004

Thick polymer blend organic solar cells fabricated by slow drying

AbstractOrganic solar cells (OSCs) were generally fabricated by two types of deposition method, namely, dry and wet. Many OSCs use very thin films that cannot fully absorb light. Thick film deposition for OSCs has numerous benefits, such as highly efficient harnessing of solar light and low leakage. Thick film fabrication for polymer blend organic solar cells by the wet deposition method was investigated. OSCs were fabricated by spin coating and slow drying using P3HT and bis[60]PCBM in a solvent, respectively. Two-dimensional (2D)-mapping images of the photocurrent of the cells were measured by a laser-beam-induced current method. A low-current density area was not found in all the cells. As a result, we succeeded in fabricating OSCs with a power conversion efficiency of more than 4% by slow drying

大学生の喫煙の実態について : 平成18年度静岡産業大学情報学部新入生を対象として

現在、喫煙行動が健康に害を及ぼすことは広く認識され、社会全体へ浸透している。公共の乗り物内では禁煙は当然、建物内では喫煙場所の限定、もしくは分煙、外に出れば条例で歩きタバコやポイ捨て禁止など、数年前とはうって変わって喫煙者にとって厳しい社会となった。その甲斐あってか、近年の成人喫煙者は全国的に減少傾向をみせている。しかし未成年、特に中・高・大学生の喫煙行動は確実に増加している。本学部においても喫煙学生が年々増加傾向にある一方で、特に女子学生の喫煙者の増加、および喫煙状況も大変気になるところである。女性の喫煙者は男性と比較して、その死亡率は3割～5割高いという報告があるからである。また、学内には所構わずタバコの吸い殻がポイ捨てされ、毎日事務局の職員や大学で契約した清掃会社の職員が拾っているという、高等教育機関とは言い難い光景が見受けられる。昨年度まで学内にあった5ヶ所の喫煙所が、平成18年度からは1ヶ所になったことも原因のひとつであろう。今後は学生への喫煙マナーに関する教育や禁煙指導の必要性が一層高まると考えられる。このプロジェクトを発足するに当たり、まずどのような喫煙経験を持つ学生が入学してきているのか、またその喫煙に関してどのような認識でいるのか、これら現状把握が急務であると考え、入学生全員にアンケート調査を行った。まず、平成18年度情報学部入学生236名全員に喫煙経験の有無、性別、年齢、国籍を調査した。次に喫煙者には、喫煙開始年齢・喫煙のきっかけ・一日の平均喫煙量・喫煙の状態・罪の意識の有無・禁煙への意識・家族の喫煙環境・タバコの害・未成年の喫煙について、また吸わない学生には、家族の喫煙状況・周囲での喫煙に関する意識・今後の喫煙・タバコの害・未成年の喫煙について調査した。その結果、喫煙者は30名(男29名・女1名)、非喫煙者は206名であった。この30名の喫煙者をその環境や意識の違いでいくつかのパターンに分けクロス集計し、さらに深く分析した。また喫煙に関しての認識度や、現在は非喫煙者でも今後喫煙者と成りうることなど、アンケート調査から読み取れる傾向をここに報告する

TICAM-1/TRIF associates with Act1 and suppresses IL-17 receptor–mediated inflammatory responses

TICAM-1 (also called TRIF) is the sole adaptor of TLR3 that recognizes double-stranded RNA. Here, we report that TICAM-1 is involved not only in TLR3 signaling but also in the cytokine receptor IL-17RA signaling. We found that TICAM-1 bound to IL-17R adaptor Act1 to inhibit the interaction between IL-17RA and Act1. Interestingly, TICAM-1 knockout promoted IL-17RA/Act1 interaction and increased IL-17A–mediated activation of NF-κB and MAP kinases, leading to enhanced expression of inflammatory cytokines and chemokines upon IL-17A stimulation. Moreover, Ticam-1 knockout augmented IL-17A–mediated CXCL1 and CXCL2 expression in vivo, resulting in accumulation of myeloid cells. Furthermore, Ticam-1 knockout enhanced delayed type hypersensitivity and exacerbated experimental autoimmune encephalomyelitis. Ticam-1 knockout promoted accumulation of myeloid and lymphoid cells in the spinal cord of EAE-induced mice. Collectively, these data indicate that TICAM-1 inhibits the interaction between IL-17RA and Act1 and functions as a negative regulator in IL-17A–mediated inflammatory responses

A System for Generating Musical Score Indicating the Hitting Hand for Drum Performance

On playing drums, it is important to master the correct stroking order. Drummers are required to learn the drum rudiment to play drums efficiently. Generally, musical scores for drums do not have the annotation that indicates which hand drummers use to stroke each drum, left or right. Although drum teachers handwrite such annotation on the musical score, there is not the system that generates the musical score indicating the hitting hand on playing drums automatically. In this research, we proposed the system that generates the musical score that indicates the hitting hand to stroke each drum. The proposed system recognizes the hitting hand on the basis of the data of a gyro sensor that are embedded in the drum sticks and MIDI message from an electronic drum. We constructed the prototype system and evaluated its effectiveness

Roles of bulk γ(L)-Bi₂MoO₆ and surface β-Bi₂Mo₂O₉ in the selective catalytic oxidation of C₃H₆

γ(L)-Bi₂MoO₆ (L: low temperature phase) catalysts, whose surface compositions have a Mo/Bi ratio above = 0.5, exhibited high selectivity in the partial oxidation of C₃H₆, while catalysts with Mo/Bi surface ratios near or below = 0.5 exhibited low selectivity. γ(L)-phase catalysts which have Mo/Bi surface ratios greater than = 0.5, were demonstrated to form β-Bi₂Mo₂O₉ on their surface. An interaction between the β- and γ(L)-phases was observed in these catalysts’ UV–vis spectra at 430 nm. The new β-phase material seems to grow along b-axis of γ(L)-phase, i.e., perpendicular to MoO₂–Bi₂O₂ layers. Structure visualizations revealed that the α-Bi₂Mo₃O₁₂, β-, and γ(H)-phases, which are selective catalysts, contain twin Mo tetrahedral structures, and that their Mo and Bi ions lie on the same plane. The pure γ(L)-phase does not contain this structure. A model for the very rapid transfer of oxygen between the γ(L)- and β-phases is discussed in relation to the kinetics of C₃H₆ oxidation.ArticleJournal of molecular catalysis. A, Chemical. 318(1-2):94-100 (2010)journal articl