15,734 research outputs found
Interview with YA mystery author, Valerie Sherrard
No full textAn interview with Valerie Sherrard, a YA author, which focuses on her process of writing the Shelby Belgarden mystery series.
The Role of DAP-kinase in T Cell Activation
No full textDAPK 是一個具有多個功能區域的絲/酥胺酸(serine/threonine)蛋白質激脢，具有抑癌功能，同時也會參與各式的細胞凋亡系統。DAPK 的分子作用機制目前仍不太清楚，其在T 細胞中所扮演的角色也從未被探討過。我們發現DAPK 在T 細胞活化時亦會被活化，然後DAPK 會被分解。我們在T 細胞株和正常的小鼠T 細胞轉殖基因表現活化的DAPK 和會抑制DAPK 活性的變異體，或是利用siRNA 降低DAPK 的表現，發現DAPK 會負向調控T 細胞的活化。DAPK 會影響T 細胞活化後之增殖反應以及IL-2 的產生。同時也發現DAPK 的作用目標之ㄧ是TCR 刺激後引發的NF-κB 活化。在DAPK 被抑制或調降的T 細胞中，PKCθ磷酸化會增加，而進入核中之NF-κBp65 也提升。相對的，IL-1β和TNF-α刺激所引發的NF-κB活化並不受DAPK 影響。我們更進一步發現在T 細胞活化時，DAPK 只會選擇性地影響PKCθ、Bcl-10 和 IKK 等與NF-κB 活化相關的訊息分子進入脂質筏，但對於其他位於脂質筏的分子，例如LAT 和Lck，則沒有影響。們接著探討DAPK 是否藉著抑制T 細胞活化而參與T 細胞耐受性之產生。以A23187 增加鈣離子濃度在體外誘導T 細胞去活化，或以靜脈注射CD3 抗體到小鼠體內直接誘導T 細胞去活化的方式，都可以發現在去活化的T 細胞，DAPKmRNA 的表現增加。DAPK 的表現量與T 細胞去活化的程度成正比，顯示DAPK可能在T 細胞耐受度上扮演角色。此外，我們也發現表現活化的DAPK 會抑制小鼠誘發產生實驗性自身免疫性腦脊髓炎。我們的結果顯示了DAPK 是個新的T 細胞調控分子，會主導T 細胞活化以及TCR 刺激引發的NF-κB 活化。同時，DAPK亦可調節T 細胞去活化而維持活體免疫耐受度以避免自體免疫之產生。Death-associated protein kinase (DAP-kinase, DAPK) is a unique multi-domain kinase acting both as a tumor suppressor and an apoptosis inducer. The molecular mechanism underlying the effector function of DAPK is not fully understood, while the role of DAPK in T lymphocyte activation is mostly unknown. DAPK was activated after TCR stimulation, followed by DAPK degradation. Through the expression of a dominant negative form and a constitutively active form of DAPK in T cell lines and in transgenic mice, we found that DAPK negatively regulated T cell activation. DAPK markedly affected T cell proliferation and IL-2 production. We identified TCR-induced NF-κB activation as a target of DAPK. There was an increased PKCθ phosphorylation and an enhanced NF-κBp65 nuclear translocation in T cells with DAPK inhibited or down-regulated. In contrast, IL-1β- and TNF-α-triggered NF-κB activation was not affected by DAPK. We further found that DAPK selectively modulated the TCR-induced translocation of PKCθ, Bcl-10, and IKK into membrane rafts. Notably, the effect of DAPK on the raft entry was specific for NF-κB pathway, as other raft-associated molecules, such as LAT, were not affected. We further examined the possibility that, by negatively regulating T cell activation, DAPK contributes to the generation of T cell tolerance. DAPK mRNA expression was up-regulated when T cell anergy was induced either in vitro by calcium ionophore, or in vivo by intravenous administration of anti-CD3 antibody. There was a correlation between the amount of DAPK expression and the extent of T cell anergy, suggesting a role of DAPK in T cell tolerance induction. In addition, active DAPK transgene inhibited the induction and progression of autoimmune experimental autoimmune encephalomyelitis (EAE). Our results demonstrate that DAPK is a novel T cell regulator on TCR-activated NF-κB and T cell activation, and DAPK may contribute to maintenance of immune tolerance and avoidance of autoimmunity by promoting T cell anergy.口試委員會審定書……………………………………………………… i謝……………………………………………………………………… ii文摘要………………………………………………………………… iii文摘要………………………………………………………………… iv一章 緒論………………………………………………………..……. 1、 DAPK…………………………………………………………… 1. DAPK 之簡介…………………………………………………… 1. DAPK 之蛋白質結構…………………………………………….. 1. DAPK 的抑癌功能與促細胞凋亡活性…………………………... 3、 T 細胞活化……………………………………………………... 4. T 細胞活化的訊息傳遞…………………………………………... 4. PKCθ…………………………………………………………………6. 脂質筏與T 細胞活化的關係……………………………………. 7、 NF-κB 活化之訊息傳遞路徑………………………………….. 9、 T 細胞去活化…………………………………………………. 10、 研究動機與方向………………………………………………. 11二章 材料與方法…………………………………………………… 12、 細胞株與細胞培養……………………………………………. 12、 藥品與試劑……………………………………………………. 12、 抗體……………………………………………………………. 13、 質體之構築………………………………………………………13. pGC-YFP-DAPK、pGC-YFP-K42A 和pGC-YFP-ΔCAM 之構築…13. pSR-mDAPKsi 和pSR-mDAPKsim 之構築………………………14. pSR-hDAPKsi-1、pSR-hDAPKsi-2 和pSR-hDAPKsim 之構築… 14. CD2-K42A 和CD2-ΔCAM 之構築………………………………… 14、 質體DNA 的轉染……………………………………………… 15. Calcium phosphate 轉染法……………………………………… 15. 反轉錄病毒感染法…………………………………………………15. 電穿孔法……………………………………………………………16、 IL-2 產量分析………………………………………………… 16、 T 細胞增殖分析……………………………………………….. 17、 RNA 的純化…………………………………………………….. 17、 反轉錄與聚合酶連鎖反應…………………………………….. 17、 流式細胞儀分析……………………………………………….. 18. 細胞死亡測定…………………………………………………….. 18. 細胞表面染色分析……………………………………………….. 19一、 細胞萃取液的製備………………………………………….. 19. 全部細胞萃取液的製備………………………………………….. 19. 細胞質及細胞核萃取液的製備………………………………….. 19二、 西方墨點法………………………………………………….. 20三、 免疫沉澱法………………………………………………….. 20四、 電泳移動改變分析法……………………………………….. 20. 探子的製備……………………………………………………….. 21. 電泳移動改變分析……………………………………………….. 21五、 脂質筏的分離與純化……………………………………….. 21六、 X 光底片影像訊號強弱測量……………………………….. 22七、 T 細胞去活化的誘導……………………………………….. 22. 誘導AND 轉殖基因小鼠T 細胞的去活化……………………… 22. 抗小鼠CD3 抗體的靜脈注射誘導T 細胞去活化……………… 22八、 實驗性自身免疫性腦脊髓動物模式…………………………23三章 結果…………………………………………………………… 24、 T 細胞活化時DAPK 亦被活化而後分解……………………... 24、 DAPK 之持續活化變異體抑制T 細胞活化，而顯負性變異體則會增強T 細胞活化……………………………………………... 25、 利用siRNA 降低DAPK 的表現可增強T 細胞活化………..... 26、 [K42A]DAPK 和[ΔCAM]DAPK 兩種轉殖基因鼠的生產與表現………………………………………………………………... 27、 [K42A]DAPK 轉殖基因會增強T 細胞活化，而[ΔCAM]DAPK轉殖基因則會抑制T 細胞活化……………………………….. 28、 DAPK 抑制TCR 刺激所引發的NF-κB 活化……………….... 30、 DAPK 不影響TNF-α和IL-1β刺激所引發的NF-κB 活化…... 32、 抑制DAPK 活性可增加PKCθ、Bcl-10 和IKKα等訊息分子轉移進入脂質筏中的量……………………………………………... 32、 DAPK 在去活化T 細胞的表現增加…………………………... 33. 體外誘發AND 轉殖基因小鼠T 細胞之去活化……………… 33. 抗小鼠CD3 抗體(2C11)的靜脈注射在活體誘發T 細胞之去活化………………………………………………………………... 34、 [ΔCAM]DAPK 轉殖基因則會抑制實驗性自身免疫性腦脊髓炎的發病…………………………………………………………... 35四章 討論…………………………………………………………… 37、 DAPK 參與T 細胞之活化……………………………………… 37、DAPK 與DRAK2 在T 細胞中功能的異同…………………… 38、 DAPK 對NF-κB 活化之調控…………………………………. 39、DAPK 影響NFAT1 及NFAT2 之活化…………………………. 41、 DAPK 與T 細胞去活化的關係………………………………… 42、 DAPK 對自體免疫疾病的調控………………………………… 2表…………………………………………………………………… 45考文獻……………………………………………………………… 85錄……………………………………………………………………10
Interview with YA author Vicki Grant
No full textWe feature an interview with Nova Scotian YA author Vicki Grant. Vicki is a prolific author who has written 14 books and has written over 100 episodes of children‘s TV. Vicki discusses her thoughts on reading positively and the meaning behind positivity, her favorite bad-day books, and how we as a community can promote reading positively. She makes us realize the importance of realism in writing and how important it is to connect with young readers
Author Correction: Cardioprotection induced in a mouse model of neuropathic pain via anterior nucleus of paraventricular thalamus
No full textThe original version of this Article contained an error in the affiliation of the second author, Ya-Ting Chang. The correct affiliations for Ya-Ting Chang are Institute of Biomedical Sciences, Academia Sinica, Taipei, 115, Taiwan and International Graduate Program in Molecular Medicine, National Yang-Ming University and Academia Sinica, Taipei 115, Taiwan.</jats:p
Interview with YA author and Children‘s Editorial Assistant Suzanne Sutherland
No full textAn interview with Young Adult author Suzanne Sutherland. The interview focuses on Toronto, the straight edge scene, music and subculture, Sutherland\u27s first book, When We Were Good and the importance of queer representation in YA books. Sutherland also recommends a number of YA novels
The Tumor Suppressor Death-Associated Protein Kinase Targets to TCR-Stimulated NF-κB Activation
No full textWang Simeng, Illusions et souffrances. Les migrants chinois à Paris, Paris, Éditions Rue d’Ulm, coll. «Sciences sociales» , 2017
No full textChuang Ya-han. Wang Simeng, Illusions et souffrances. Les migrants chinois à Paris, Paris, Éditions Rue d’Ulm, coll. «Sciences sociales» , 2017. In: Études chinoises, vol. 37, n°2,2018. pp. 223-225
- …
