約 5,271,641 件
https://w.atwiki.jp/kumicit/pages/1056.html
Kumicitのコンテンツ Who s Who GRISDAの創造論者Ariel A. Roth 創造論を教義とするキリスト教宗派のひとつ Seventh Day Adventists (SDA)の傘下の Seventh-day Adventist Loma Linda Universityの中に、研究人員10名程度の小さな創造論研究所 Geoscience Research Institute（GRI） というのがある。ここの 元所長であり、機関紙Originの編集人を23年間つとめたのがジュネーブ生まれの創造論者であり動物学者であるDr. Ariel A. Roth である。"若い地球の創造論"ミニストリAnswers in Genesisによれば、生物学でPh.DをUniversity Michiganで取得しており、反進化論者としては珍しく生物系である。ネタとして楽しめるような珍説も出しておらず、創造論者の中では無名に近い。 そのAriel A. Rothが1998年に出版した"ORIGINS"という創造論本の日本語版、2006年に日本のSeventh Day Adventists傘下の出版社である福音社が出した。価格はなんと6000円である。そんな本を、誰かが間違って買って古書店に売り払ったのか、保存状態の良いものが、神田の古書店のひとつ明倫館に3000円であったので、買ってみた。 1996年という出版時点を考えてもネタ的に目新しいものはなく、オリジナルの珍説もなく、基本的には、「Mark Isaakの創造論者の主張」が対応済みな主張の寄せ集め。 Geoscience Research Institute（GRI） がほとんど存在も知られていない理由のわかる一冊だった。 一応、お約束ネタについての立場だけは見ておくと.... 「太陽系は6000歳だが、宇宙は135億歳」あるいは「宇宙は恒星からの光も含めて6000年前に創造された」の両論併記： 創造週に関してしばしば提起される問題の一つは、遠い星からの光が届くのにかかる時間の長さに関するものです。晴れた夜、望遠鏡なしでも、かすかに見ることができるアンドロメダ星雲は、その光が我々のもとに届くのにはおおよそ200万年かかります。もし星が数千年前の第4日目に造られたのなら。どのようにして我々はその光をすでに見ることができるのでしょうか。光が我々のもとに届くのに10億年もかかる程遠くの星もあるというのに？ 星は創造週のはるか以前に造られたと提案することがその問題を解く一つの方法です。別の提案に、神は数千年前の最近に星を造られたのであろうが、すでに燦然と輝きを放つ光が地球に達している状態で完成しており、人は始めからその光を見て楽しうことができたのだ、という考え方もあります。[p.338] 「光も含めて創造」は昔は流行っていたが、今は破棄された説。哲学的には合理的だが、気持ちが悪いらしい。しかし、Dr. Ariel A. Rothは気にしないようだ。 創造論者は一般に最初の概念（小進化や小突然変異）は受け入れますが、第二の概念（大進化や大突然変異）は否定します。[p.98] これは標準的な創造論者。Answers in Genesisが「見た目ではなく、遺伝子レベルの変化の大きさ」という表現をとるのに対して、シンプル。 聖書は現在とは幾分異なっていた洪水前の地球を記しています。おそらく雨がなかったようです。しかし河川を含めて十分な湿気がありました。[p.223] どうでもいいネタで、Answers in Genesisは放棄したネタだが、Institute for Creation Researchなど老いたる創造論ミニストリには残っている。 創造論者は洪水時には種の数は少なかったであろうと仮定します。洪水以降の制約限定された変異が理由で、種レベルで最もありそうですが、箱舟に保存された種の数よりももっと多くの種の変種が現在の世界に存在するでしょう。[p.224] これは標準的な"若い地球の創造論者"のポジション。これがあるので、小進化がありになり、自然選択による種形成も受け入れることになる。 大洪水を起こした水は、洪水前の地球上にすでに存在していたようです。その大部分が洪水前の海に。「淵の源」にいくらか、そして大気中に少量あったのでしょう。[p.229] 創造論者は大洪水説のための多くのモデルを提案しています。そして、注意すべきことは、それぞれのモデルが仮のものと考えられていると述べることです。一般に、モデルは3つの広範囲のカテゴリの中に入ります。1) 洪水時における大陸と大洋の交換、2) 地球の収縮と膨張、3) 洪水中の大陸の沈下と続いて起こる大陸の隆起。これらを組み合わせたモデルと、また別の他のモデルも可能です。[p.226] Austin SA et al "Catastrophic plate tectonics a global flood model of earth history", Proceedings of the third International Conference on Creationism, pp.609-621, 1994. Baumgardner JR "Computer modeling of the large-scale tectonics assicuated with the Genesis flood", Proceedings of the third International Conference on Creationism, pp.49-62, 1994. Baumgardner JR "Runaway subduction as the driving mechanism for the Genesis flood", Proceedings of the third International Conference on Creationism, pp.62-75, 1994. 地球外に水源を求めない説のみを列挙。 これらのうち、最後の"ノアの洪水"の水源については、少なくとも2007年10月まではGeoscience Research Institute（GRI）は彗星を衝突させて水源にしようとしていた： It is possible that more water was added during the flood by collisions of one or more comets, which appear to be made largely of water. 洪水の期間にひとつかそれ以上の彗星が衝突して、水を追加したかもしれない。彗星はほとんどが水である。 [ FREQUENTLY ASKED QUESTIONS ABOUT THE GENESIS FLOOD (Archive.org 2007/10/22) ] もちろん、笑えることに、地球全体を厚さ1cmで覆い尽くすとすると、5兆トン、直径20kmくらいの彗星核が必要なこと。同一質量の岩石だと直径10km以上になる。衝突速度に依存するが、このサイズだと6500万年前と推定されるChicxulub Craterを形成した隕石衝突クラスの被害が出そう。すなわち、全地球的地震・急激な気温上昇・数年にわたり闇に閉ざされる地球など。 その他、全体としてDr. Ariel R. Rothの本は1998年という出版時期を考えても、かなり古びている。それを今更、日本語にしても、ネタにもならない。役に立つとしたら、Geoscience Research Institute（GRI）がカビが生えたネタを抱えていることの確認のみ。
https://w.atwiki.jp/aias-jsdoctoolkit/
このサイトについて 「JsDoc Toolkitを使う！」は、JavaScriptドキュメンテーションツールであるJsDoc Toolkitの使い方を紹介するサイトです。JsDoc Toolkitは非常に優れたツールですが、まとまった日本語の情報が少ないようなので何となく作ってみました。 このサイトの内容はあくまで管理人個人の理解に基づくものです。ご利用は自己責任でお願いします。 記載されている情報はバージョン 2.4.0 時点のものです。 JsDoc Toolkitとは JsDoc Toolkit は、JavaScriptソースコードに記述されたコメントを元に、HTML形式のドキュメントを生成するオープンソースのフリーソフトウェアです。作者はMichael Mathews、ライセンスはMITライセンスです。 JavaScriptのドキュメント作成ツールとしては JSDoc が良く知られていますが、JsDoc ToolkitとJSDocは全く別のものです。（ちなみにMichael MathewsはJSDocの主要な開発者の一人だったようです）なおJSDocは既に開発を停止しており、JsDoc Toolkitはその後継プロジェクトとされています。 JsDoc Tooikitはそれ自身がJavaScriptで記述されており、JavaによるJavaScriptの実装である Rhino 上で動作します。従ってJsDoc Tooikitを使用するにはJavaの実行環境(JRE)が必要です。 JsDoc Toolkitのインストールについては、こちらを参照してください。 ドキュメント生成のためのコメント（ドックコメントと呼びます）記法はJavaDocをベースにしており、比較的容易に習得することができます。ドックコメント付きのソースコードと、そこからJsDoc Toolkitによって生成されたドキュメントのスクリーンショットを下に示します。 + クリックするとソースコードを展開します... /** * 新しいStringBufferオブジェクトを作成する。 * @class 文字列をバッファリングし、文字列の効率的な結合を行うクラス * @param {String} [str] バッファリングする文字列 * @example alert(new StringBuffer("このように").append("文字列を").append("結合します。").toString()); */ function StringBuffer(str) { /** * バッファされている文字列の配列 * @type String[] * @private */ this._str = str?[str] []; } /** * バッファに文字列を追加する。 * @param {String} str バッファリングする文字列 * @return {StringBuffer} このオブジェクトへの参照 */ StringBuffer.prototype.append = function(str) { this._str.push(str); return this; }; /** * バッファリングされた文字列を結合して返す。 * @return {String} 結合された文字列 */ StringBuffer.prototype.toString = function() { return this._str.join(""); }; - クリックするとスクリーンショットを展開します... ドックコメントの書き方については、こちらを参照してください。 ドキュメントの作成手順については、こちらを参照してください。 JsDoc Tooikitの特徴のひとつは、独自のテンプレートシステムによるドキュメント表示形式の柔軟性です。テンプレートはスキンのようなもので、JsDoc Tooikitのコア機能である構文解析機能から完全に独立しています。このためユーザはテンプレートを切り替えることで、JsDoc Tooikitのソースコードに手を入れることなく独自のドキュメントスタイルを利用できるようになっています。 例えば上に挙げたソースコードを管理人が作成したテンプレートaias-frameを使って出力すると、次のようになります。 - クリックするとスクリーンショットを展開します... テンプレートなどによるJsDoc Toolkitのカスタマイズについては、こちらを参照してください。 管理人が作成したテンプレートなどをこちらからダウンロードできます。 コンテンツ トップページ インストール ドックコメントの書き方 タグリファレンス ネームパス インライン・ドックコメント メタタグ ドキュメントの作成 コマンドラインオプション JsDoc Toolkit Ant Task カスタマイズ テンプレート publish.js JsPlateファイル プラグインの追加 組み込みオブジェクト リファレンス 組み込みオブジェクト リファレンス JsDoc Toolkit版 ダウンロード リンク JsDoc Toolkit official sitehttp //code.google.com/p/jsdoc-toolkit/ JsDoc Toolkit Users Grouphttp //groups.google.com/group/jsdoc-2
https://w.atwiki.jp/xbox360score/pages/893.html
STORMRISE 項目数：43 総ポイント：1000 難易度： ※PC版は別実績 ストーリーをHARDでクリアしても下位難易度の実績は解除されない。 オンラインの勝利実績は、負ける側が途中抜けしてもカウントされる。 Epic Saga Complete Story Mode on Hard 50 Scrap Metal Destroy 27 Stalkers 20 Decked Out Win 10 ranked matches fully equipped 35 Totaled Eclipse Destroy 9 Eclipses 25 Veteran s Affair Win 99 ranked matches 40 Eradicator Beat the AI on all Skirmish maps 30 Epic Campaign Complete Story Mode on Easy or Normal 30 The Other Side Complete Act 1 of Story Mode 25 The Other Path Complete Act 2 of Story Mode 25 Qualify for Duty Complete the Tutorial 15 Clean Sweep Destroy all hiding Sai insurgents in the Tutorial 10 Ulterior Motives Use the playbook to switch weapons 10 First Assault Complete Mission 1 Domestic Disturbance 20 In Transit Secure the Control Node powering the turrets in Mission 2 Assault Charge 20 Telepathic Establish a portal in Mission 3 Double Jeopardy 20 H.Y.D.R.A. Defeat the Hydra in Mission 4 Multiple Counts 20 Hammer Time Take out the Arc-Hammers in Mission 5 Self Defense 20 Evacuation Rescue Vantage in Mission 6 Innocent Victim 20 A Bridge Not Too Far Secure all 3 bridges in Mission 7 Unfair Surprise 20 Power Up Secure all 3 power junctions in Mission 8 Reasonable Doubt 20 Fur Coat Defeat Sable in Mission 9 Shock Verdict 20 Rite of Passage Rescue Eona in Mission 10 Prison Break 25 In Pursuit Secure the AA Turrets in Mission 11 Hot Pursuit 25 Reverse Polarity Recalibrate the airfield station in Mission 12 Natural Justice 25 Spray and Pray Use the Matriarch s Acid Rain ability 15 Demolisher Destroy 55 Control Nodes with the Demo Bomb ability 25 Puppet Master Destroy an enemy unit with one you have Mind-Controlled 20 Unleashed Use the Rage Smash ability 15 Playing Both Sides Win 8 ranked matches with each faction 15 Demoraliser Destroy 11 enemy units in the first 5 minutes of a ranked match 20 First Down and Ten Win a ranked match in under 10 minutes 20 Winning Streak Win 6 consecutive ranked matches 25 Feeding the Habit Play 16 ranked matches 15 Calling all Units Recruit one of every unit type in any game mode 15 Gone Shopping Lose a multiplayer match while the deployment queue is open 20 Maximum Firepower Upgrade a Control Node to have level 3 turrets (multiplayer or skirmish) 20 Slice and Dice Use the Spectre s Rapid Slice ability to kill an enemy commander (multiplayer or skirmish) 15 False Sense of Security Attack 24 cloaked Spectres while using the Infiltrator s Thermal Vision 25 Appetite for Destruction Destroy 77 units using the Stalker s SAMs 25 Pest Control Kill 80 Broodlings 20 Full Repertoire You have used all unit abilities at least once 50 Formidable You destroyed 500 units 50 Whip it Good You whipped 250 times during a map or match. 20
https://w.atwiki.jp/tribeswar/pages/22.html
Description Min. Max. Exp. Energy Avr./Eng Exp./Eng Loot Req.1 Req.2 Req.3 Req.4 Patrol the border 125 158 5 5 28.3 1.00 Fon Shield African Shield ×5 Axe ×5 Horse ×1 Locate water source 158 225 5 5 38.3 1.00 Scout ×1 African Shield ×4 Axe ×4 Spy foe actions 158 173 6 6 27.6 1.00 Scout ×1 African Shield ×5 Axe ×5 Spy the foe 181 218 12 9 22.2 1.33 Scout ×1 Executioner Sword ×1 Fipa Helmet ×1 Sukumu Shield ×1 Raid the watch tower 169 196 9 7 26.1 1.29 Executioner Sword ×1 Fipa Helmet ×1 Sukumu Shield ×1 Elephant ×1 Ambush enemies 186 197 10 7 27.4 1.43 Kuba Sword Executioner Sword ×2 Fipa Helmet ×2 Sukumu Shield ×1 Horse ×2 Kill warriors 193 298 25 17 14.4 1.47 Executioner Sword ×2 Fipa Helmet ×2 Sukumu Shield ×2 Flowers（Collection Bonus + 2 Defense） Rose Marigold Tulip Sunflower African Violet African Daisy Lilies 戦士階級 このクラスのJobsではいよいよ資金力が重要になり、初めてAnimalsが必要になります。Scoutが要求される三つのJobsは後回しにして、やはり経験値指数の高いJobsを優先的に選択しましょう。 Patrol the border（国境警備）☆★★ Horseを必要とする初めてのJobsです。が、悲しいかな経験値指数は最低の1。全く優先すべきJobsではありません。獲得できるFon Shieldも性能も高くない上に直ぐに代替品を入手できます。Level3 masteredになるまでに3枚は引けるでしょう。 Locate water source（水源確保）★★★ Spy foe actions（敵情偵察）★★★ Spy the foe（反抗勢力の調査）☆★★ まとめてしまいますが、消耗するScoutを大量に必要とするこれら3つのJobsはWorkerを幾度もしなければならない苦痛が伴ないます。しかもSpy the foe(1.33)を除いて経験値効率も最低。レベル100を超えて、やる事がなくなったら改めてScout獲得からチャレンジすれば良いのではないでしょうか。 ちなみにこれまでに獲得したScoutはSend undercovers to enemyで消費するのが効率が良いので、今は我慢してキープしておくのが賢明です。 Raid the watch tower（監視塔奇襲作戦）☆☆★ Ambush enemies（待ち伏せ）☆☆☆ Kill warriors（戦士を倒せ）☆☆☆ こちらもまとめてしまいますが、このクラス中では優先すべきJobsです。経験値効率も高い上にリクエストは全てコインが解決してくれるので思う存分集中出来ます。 Ambush enemiesはElite Warrior、Commanderの2つのクラスに跨って合計6つのJobsで必要になるKuba Swordを獲得する重要なJobsです。Level3 masteredになるまでに最低でも2本は引き当ててください。 レベル13になると次のElite Warriorクラスがアンロックされますが、経験値効率が高いJobsがアンロックされるまでは、ここに留まるのも良いでしょう。
https://w.atwiki.jp/net-tools/pages/53.html
前ページ次ページWireshark 目次 目次 Tshark を使ってみるTshark使用例１（普通にキャプチャ、キャプチャしたデータをコマンドプロンプトに表示）キャプチャインタフェース番号の調べ方 Tshark使用例２（キャプチャしたデータをpcap形式で保存） Tshark使用例３（保存したデータをTsharkで参照） Tshark使用例４（詳細を表示） Tshark＋grepの使い方 Tshark を使ってみる Tshark使用例１（普通にキャプチャ、キャプチャしたデータをコマンドプロンプトに表示） Tsharkを使って４番のインタフェースのパケットをキャプチャするには、-i 4を指定します。このようにすることで、コマンドプロンプト上にキャプチャデータが表示されます。 以下の例では、192.168.11.1に関するarpリクエストとリプライがキャプチャできました。 C \Program Files\Wireshark tshark -i 4 Capturing on ORINOCO PC Card (Microsoft s Packet Scheduler) 1199370775.557633 00 02 2d 65 4e 5d - ff ff ff ff ff ff ARP Who has 192.168.11.1? Tell 192.168.11.2 1199370775.559342 00 07 40 aa 5b 54 - 00 02 2d 65 4e 5d ARP 192.168.11.1 is at 00 07 40 aa 5b 54 （Ctrl+Cでキャプチャを停止できます。） キャプチャインタフェース番号の調べ方 ところで、-i 4で指定した4というインタフェースの番号は、どのように調べればいいのでしょうか。。。 以下のように -D で調べることができます。 C \Program Files\Wireshark tshark -D 1. \Device\NPF_GenericDialupAdapter (Adapter for generic dialup and VPN capture) 2. \Device\NPF_{6BA966AC-2889-44F1-9162-B4CE57FFDE6F} (VMware Virtual Ethernet Adapter) 3. \Device\NPF_{8C55A32F-392C-40D4-A3ED-3BF84158C0CB} (VMware Virtual Ethernet Adapter) 4. \Device\NPF_{CE53EEB8-2CA7-41F7-8AE6-F9A2F12DFC26} (ORINOCO PC Card (Microsoft s Packet Scheduler) ) 5. \Device\NPF_{DD071C3E-0216-42C0-833F-F57EE11F57BE} (Intel 8255x-based Integrated Fast Ethernet (Microsoft s Packet Scheduler) ) Tshark使用例２（キャプチャしたデータをpcap形式で保存） キャプチャしたデータをコマンドプロンプトではなく、ファイルにpcap形式で保存することも可能です。 以下の例では、-w tshark1.pcapでキャプチャしたデータはtshark1.pcapに保存されます。実際にキャプチャされたデータ数が、一番下に表示されます。以下の例では、このとき14パケットがキャプチャされていることがわかります。 C \Program Files\Wireshark tshark -i 4 -w tshark1.pcap Capturing on ORINOCO PC Card (Microsoft s Packet Scheduler) 14 （Ctrl+Cでキャプチャを停止できます。） Tshark使用例３（保存したデータをTsharkで参照） 保存したファイルをTsharkで開くことも可能です。 開きたいファイルを-r tshark1.pcapのように指定します。 C \Program Files\Wireshark tshark -r tshark1.pcap 1 0.000000 192.168.11.2 - 192.168.11.1 ICMP Echo (ping) request 2 0.002041 192.168.11.1 - 192.168.11.2 ICMP Echo (ping) reply 3 1.000839 192.168.11.2 - 192.168.11.1 ICMP Echo (ping) request 4 1.002712 192.168.11.1 - 192.168.11.2 ICMP Echo (ping) reply 5 2.002034 192.168.11.2 - 192.168.11.1 ICMP Echo (ping) request 6 2.003898 192.168.11.1 - 192.168.11.2 ICMP Echo (ping) reply 7 3.003227 192.168.11.2 - 192.168.11.1 ICMP Echo (ping) request 8 3.005079 192.168.11.1 - 192.168.11.2 ICMP Echo (ping) reply 9 4.004419 192.168.11.2 - 192.168.11.1 ICMP Echo (ping) request 10 4.006289 192.168.11.1 - 192.168.11.2 ICMP Echo (ping) reply 11 5.000804 00 07 40 aa 5b 54 - 00 02 2d 65 4e 5d ARP Who has 192.168.11.2? Tell 192.168.11.1 12 5.000840 00 02 2d 65 4e 5d - 00 07 40 aa 5b 54 ARP 192.168.11.2 is at 00 02 2d 65 4e 5d 13 5.005591 192.168.11.2 - 192.168.11.1 ICMP Echo (ping) request 14 5.007366 192.168.11.1 - 192.168.11.2 ICMP Echo (ping) reply Tshark使用例４（詳細を表示） -Vプション でパケットの詳細が表示されます。 以下の例では、保存したファイルを読み込む際に-Vを付けていますが、実際に流れているデータを-i 4のようにライブキャプチャする際にも-Vは使用可能です。 CygwinかLinuxでgrepでフィルタしながら見ると便利なことがあるかもしれませんね。 C \Program Files\Wireshark tshark -V -r tshark1.pcap | more Frame 1 (74 bytes on wire, 74 bytes captured) Arrival Time Jan 3, 2008 23 39 38.232860000 [Time delta from previous captured frame 0.000000000 seconds] [Time delta from previous displayed frame 0.000000000 seconds] [Time since reference or first frame 0.000000000 seconds] Frame Number 1 Frame Length 74 bytes Capture Length 74 bytes [Frame is marked False] [Protocols in frame eth ip icmp data] Ethernet II, Src 00 02 2d 65 4e 5d (00 02 2d 65 4e 5d), Dst 00 07 40 aa 5b 54 (00 07 40 aa 5b 54) Destination 00 07 40 aa 5b 54 (00 07 40 aa 5b 54) Address 00 07 40 aa 5b 54 (00 07 40 aa 5b 54) .... ...0 .... .... .... .... = IG bit Individual address (unicast) .... ..0. .... .... .... .... = LG bit Globally unique address (factory default) Source 00 02 2d 65 4e 5d (00 02 2d 65 4e 5d) Address 00 02 2d 65 4e 5d (00 02 2d 65 4e 5d) .... ...0 .... .... .... .... = IG bit Individual address (unicast) .... ..0. .... .... .... .... = LG bit Globally unique address (factory default) Type IP (0x0800) （以下省略） Tshark＋grepの使い方 Tshark＋grepの使い方 前ページ次ページWireshark
https://w.atwiki.jp/atachi/pages/58.html
EntityFrameworks4(以降、EF4）はVS2010と共にリリースされ、POCOエンティティやモデルファーストなどの開発手法が取り入れられた先進的で斬新なアップデートとなりましたが、CTP4にはさらにコードファーストによるEntityFrameworkの新しい開発アプローチが提供されました。 (EF4はCTP4からCTP5にバージョンアップしました(ReleaseNote) Microsoft ADO.NET Entity Framework Feature Community Technology Preview 4 コードファースト [#j958e6a0] EF4.1 EF4 CTP5 EF4 CTP4 [#o94d1795] コードファーストによるアプローチの結論 [#lcdc1bbe] コードファースト コードファーストからのアプローチで永続化エンティティを扱う開発手法は、既存の開発手法を駆逐するようなものではありません。 実際の開発現場に於いて、コードファーストを使用して開発を行うケースは限られており、次のコードファーストが持つ特徴が開発現場の生産性とのトレードオフとマッチした場合に非常に斬新なアプローチとして取り入れることができるかもしれません。 PlainOldなクラスをデータモデルクラスに使用できる(POCOと同じ) CoC(convention over configuration) (Wikipedia 設定より規約) IDEが持つデザイナやXMLによる、マッピング記述が不必要 特にCoCでエンティティの永続化を実現できる点が、開発生産性の向上に大きく貢献しています。(*1) 多くの規約によりエンティティとオブジェクトとのマッピングはソースコード上に特別な記述なしに設定されます(*2) EF4.1 EntityFrameworkのバージョン4.1です。 主キーの作成をデータベースが自動的につくることができるなどのオプションが追加されました。 EF4.1ダウンロード EF4.1オフィシャルブログ EF4 CTP5 Microsoft ADO.NET Entity Framework Feature Community Technology Preview 5 CTP4からバージョンアップされCTP5となりました。 変更点がいくつかあるため、CTP4のコードをそのままCTP5でコンパイルできなくなっています。 アセンブリ名に変更があります。 コードファーストでのデータベース構造の初期化コードがより良いものになっています コードファーストにおいて、既存のデータベースからEdmMetadataテーブルを除外してマッピングを行います。 クラス名に変更があります DbSetがObservableCollection型のコレクションを返すことができるようになりました。WPFのListItem系のコントロール(ListControlやListView、DataGridなど)のItemsSourceにバインディングすることができます。 CTP5のプロジェクト EF4 CTP4 CTP4はMicrosoft ADO.NET Entity Framework Feature Community Technology Preview 4 からダウンロードできます。 セットアップファイルでの配布ですが、セットアップには「Microsoft.Data.Entity.CTP.DLL」しか含まれていません。(Project Filesフォルダに放り込まれるので、このDLLを直接参照するか、参照したいプロジェクトにコピーして参照して使用する) CTP4に含まれている名前空間は次の通り。 System.ComponentModel.DataAnnotations System.Data.Entity System.Data.Entity.Infrastructure System.Data.Entity.ModelConfiguration 余談ですが、System.Data.Entity.DLLにはSystem.Data.Entityという名前空間は含まれていません。 CTP4のプロジェクト コードファーストによるアプローチの結論 すべてコードによってモデルの設計からデータベースへのマッピングまで行えるため、コードファーストによるアプローチでのエンティティ永続化は非常に使いやすいことがわかります。バックグラウンドにSQLCEを使用することで、外部にサーバーを用意したり、エンドユーザーにデータベースを稼働させる必要もなくなります。 さらに、SQLCEを使うことでオブジェクトのシリアライズデータをバイナリファイルで保存するより確実にデータの永続化が実現できます。 パフォーマンスについてはそれほどよくはありません。といっても、すでに埋め込み型データベースの地位を確立したSQLiteと同じかそれ以上のパフォーマンスは発揮できます。 もちろん、サーバーデーモン型データベースサーバーであるのSQLServerやMySQLやPostgreSQLを使用するよりもパフォーマンスは圧倒的に落ちますし、機能もかなり限定的です。(SQLiteやSQLCEにはストアドプロシージャをはじめとするおおくの機能が実装されていない) 「コードファースト手法＋EF4＋SQL Server Compact Edition」の組み合わせは、小規模な商用ソフトやツールなどで使用する分には最高の生産性を提供できると思われます。
https://w.atwiki.jp/pikopedia/pages/37.html
提供：狐 softbank218179204025.bbtec.net(????年？月？日-)は、結菜スレに突然現れて 「馬鹿の集まりかｗ」と、失礼な書き込みをした非常識人。 愛知県に住んでおり、常時みそカツを所持している。 その他 IP：218.179.204.25、接続回線はxDSLである。
https://w.atwiki.jp/tiger/pages/4.html
TEC-/- BTK-/- double mutant T cells exhibit severely impaired T cell activitity. RLK-/-ITK-/- double mutant celles exhibit severely imparired Th2 responses. Grb2(+/-) mice disrupt T cell signaling networks and development. Dendric cells and macrophages of MEK3 deficient mice have impaired IL12 production. Bam32(-/-) B cell develop normally but have impaired T-independent antibody responses in vivo. T-cell and B-cell of RAP1A deficient mice impair integrin-mediated cell adhesion. T-cell of WASP deficient mice impair the proliferaction and antigen receptor cap formation in response to anti-CD3zeta stimulation. T-cell of SHB defective mice impair the phosphorylation of LAT and consequently the activation of MAP kinase pathways. B-cell of 3BP2 (-/-) deficient mice have defective in proliferation, cell cycle progression, PLC-gamma2 phosphorylation, calcium mobilization, NF-ATp dephosphorylation, and Erk and Jnk activation in response to BCR ligation. B-cell of Vav2(-/-) deficient mice are defective in the ability to switch immunoglobulin class. T-cell of Vav1(-/-) deficient mice exhibit impaired antigen receptor signaling. Vav1(-/-)Vav2(-/-) mice exhibit greatly reduced the mature B-cells. Vav-1-/-Vav-2-/- B cells were unresponsive to BCR-driven proliferation in vitro and to thymus-indepen-dent antigen in vivo. Fyn-deficient mice exhibit a remarkably specific lymphoid defect thymocytes are refractile to stimulation through the TCR with mitogen or antigen. Lck-deficient mice show a pronounced thymic atrophy, with a dramatic reduction in the double-positive (CD4+CD8+) thymocyte population. T cell from mice deficient in LCK is required for normal signal transduction through the TCR. T cells from mice deficient in SLAP-130/Fyb show markedly impaired proliferation. B cell of chicken deficient ITK reduce IP3 generation and phospholipase C gamma 2 tyrosine phosphorylation. T cell of mice deficient ITK reduce IP3 generation and phospholipase C gamma 1 tyrosine phosphorylation. T cell of mice deficient ITK have failure of Th2 development. Mice deficient in ITK have reduced proliferative responses to MHC stimulation and to anti-TCR cross-linking Mutations in Btk cause X-linked immunodeficiency. Gads(GRAP2) has a role in thymocyte proliferaction for maturation of T-cells. Gads(GRAP2) has a role for homeostatic proliferaction in B cells. Grap negatively regulates T-cell proliferation. Gab2 is a substrate of ZAP-70 and functions as a switch molecule toward inhibition of TCR signal transuduction. B cell signaling causes tyrosine phosphorylation of Gab1, and in turn SHP2 bind to Gab1 Gab1 phosophorylation potentiate the phosphorylation of Akt, PI3K-dependent response. RasGRP1 mediates Ras activation following TCR stimulatioin. RasGRP1 and RasGRP3 induces RAS activation in B-cell to response to T-cell stimulation. Grb2-hSos1-PLCgamma1-p36/p38-ZAP70 complexes localize in the vicinity of TCR-zeta Gads(Grap2) plays an important role in T-cell signaling via its association with SLP-76 and LAT. Lck is required for normal signal transduction through the TCR. ZAP-70 plays crucial roles in T-cell activation and development. Syk triggers cellular activation in T-cell. TEC-/- BTK-/- double mutant T cells exhibit severely impaired T cell activitity. 1 J Exp Med. 2000 Dec 4;192(11) 1611-24. Severe B cell deficiency in mice lacking the tec kinase family members Tec and Btk. Ellmeier W, Jung S, Sunshine MJ, Hatam F, Xu Y, Baltimore D, Mano H, Littman DR. Molecular Pathogenesis Program, Skirball Institute of Biomolecular Medicine. wilfried.ellmeier@univie.ac.at The cytoplasmic protein tyrosine kinase Tec has been proposed to have important functions in hematopoiesis and lymphocyte signal transduction. Here we show that Tec-deficient mice developed normally and had no major phenotypic alterations of the immune system. To reveal potential compensatory roles of other Tec kinases such as Bruton s tyrosine kinase (Btk), Tec/Btk double-deficient mice were generated. These mice exhibited a block at the B220(+)CD43(+) stage of B cell development and displayed a severe reduction of peripheral B cell numbers, particularly immunoglobulin (Ig)M(lo)IgD(hi) B cells. Although Tec/Btk(null) mice were able to form germinal centers, the response to T cell-dependent antigens was impaired. Thus, Tec and Btk together have an important role both during B cell development and in the generation and/or function of the peripheral B cell pool. The ability of Tec to compensate for Btk may also explain phenotypic differences in X-linked immunodeficiency (xid) mice compared with human X-linked agammaglobulinemia (XLA) patients. Publication Types Research Support, Non-U.S. Gov t PMID 11104803 [PubMed - indexed for MEDLINE] RLK-/-ITK-/- double mutant celles exhibit severely imparired Th2 responses. 1 Nat Immunol. 2001 Dec;2(12) 1183-8. Mutation of Tec family kinases alters T helper cell differentiation. Schaeffer EM, Yap GS, Lewis CM, Czar MJ, McVicar DW, Cheever AW, Sher A, Schwartzberg PL. National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA. The Tec kinases Rlk and Itk are critical for full T cell receptor (TCR)-induced activation of phospholipase C-gamma and mitogen-activated protein kinase. We show here that the mutation of Rlk and Itk impaired activation of the transcription factors NFAT and AP-1 and production of both T helper type 1 (TH1) and TH2 cytokines. Consistent with these biochemical defects, Itk-/- mice did not generate effective TH2 responses when challenged with Schistosoma mansoni eggs. Paradoxically, the more severely impaired Rlk-/-Itk-/- mice were able to mount a TH2 response and produced TH2 cytokines in response to this challenge. In addition, Rlk-/-Itk-/- cells showed impaired TCR-induced repression of the TH2-inducing transcription factor GATA-3, suggesting a potential mechanism for TH2 development in these hyporesponsive cells. Thus, mutations that affect Tec kinases lead to complex alterations in CD4+ TH cell differentiation. Publication Types Research Support, Non-U.S. Gov t Research Support, U.S. Gov t, P.H.S. PMID 11702066 [PubMed - indexed for MEDLINE] Grb2(+/-) mice disrupt T cell signaling networks and development. 1 Nat Immunol. 2001 Jan;2(1) 29-36. Disruption of T cell signaling networks and development by Grb2 haploid insufficiency. Gong Q, Cheng AM, Akk AM, Alberola-Ila J, Gong G, Pawson T, Chan AC. Howard Hughes Medical Institute, Washington University School of Medicine, St. Louis, MO 63110, USA. The developmental processes of positive and negative selection in the thymus shape the T cell antigen receptor (TCR) repertoire and require the integration of multiple signaling networks. These networks involve the efficient assembly of macromolecular complexes and are mediated by multimodular adaptor proteins that permit the functional integration of distinct signaling molecules. We show here that decreased expression of the adaptor protein Grb2 in Grb2+/- mice weakens TCR-induced c-Jun N-terminal kinase (JNK) and p38, but not extracellular signal-regulated kinase (ERK), activation. In turn, this selective effect decreases the ability of thymocytes to undergo negative, but not positive, selection. We also show that there are differences in the signaling thresholds of the three mitogen-activated protein kinase (MAPK) families. These differences may provide a mechanism by which quantitative differences in signal strength can alter the balance of downstream signaling pathways to induce the qualitatively distinct biological outcomes of proliferation, differentiation or apoptosis. PMID 11135575 [PubMed - indexed for MEDLINE] Dendric cells and macrophages of MEK3 deficient mice have impaired IL12 production. 1 EMBO J. 1999 Apr 1;18(7) 1845-57. Defective IL-12 production in mitogen-activated protein (MAP) kinase kinase 3 (Mkk3)-deficient mice. Lu HT, Yang DD, Wysk M, Gatti E, Mellman I, Davis RJ, Flavell RA. Howard Hughes Medical Institute and Section of Immunobiology, Yale University School of Medicine, New Haven, CT 06520, USA. The p38 mitogen-activated protein kinase (MAPK) pathway, like the c-Jun N-terminal kinase (JNK) MAPK pathway, is activated in response to cellular stress and inflammation and is involved in many fundamental biological processes. To study the role of the p38 MAPK pathway in vivo, we have used homologous recombination in mice to inactivate the Mkk3 gene, one of the two specific MAPK kinases (MAPKKs) that activate p38 MAPK. Mkk3(-/-) mice were viable and fertile; however, they were defective in interleukin-12 (IL-12) production by macrophages and dendritic cells. Interferon-gamma production following immunization with protein antigens and in vitro differentiation of naive T cells is greatly reduced, suggesting an impaired type I cytokine immune response. The effect of the p38 MAPK pathway on IL-12 expression is at least partly transcriptional, since inhibition of this pathway blocks IL-12 p40 promoter activity in macrophage cell lines and IL-12 p40 mRNA is reduced in MKK3-deficient mice. We conclude that the p38 MAP kinase, activated through MKK3, is required for the production of inflammatory cytokines by both antigen-presenting cells and CD4(+) T cells. PMID 10202148 [PubMed - indexed for MEDLINE] Bam32(-/-) B cell develop normally but have impaired T-independent antibody responses in vivo. 1 Immunity. 2003 Oct;19(4) 621-32. Bam32 links the B cell receptor to ERK and JNK and mediates B cell proliferation but not survival. Han A, Saijo K, Mecklenbrauker I, Tarakhovsky A, Nussenzweig MC. Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 10021, USA. Bam32 is an adaptor protein recruited to the plasma membrane upon B cell receptor (BCR) crosslinking in a phosphoinositol 3-kinase (PI3K)-dependent manner; however, its physiologic function is unclear. To determine its physiologic function, we produced Bam32-deficient mice. Bam32(-/-) B cells develop normally but have impaired T-independent antibody responses in vivo and diminished responses to BCR crosslinking in vitro. Biochemical analysis revealed that Bam32 acts in a novel pathway leading from the BCR to MAPK/ERK Kinases (MEK1/2), MAPK/ERK Kinase Kinase-1 (MEKK1), extracellular signal-regulated kinase (ERK), and c-jun NH2-terminal kinase (JNK), but not p38 mitogen-activated protein kinase (p38). This pathway appears to be initiated by hematopoietic progenitor kinase-1 (HPK1), which interacts directly with Bam32, and differs from all previously characterized BCR signaling pathways in that it is required for normal BCR-mediated proliferation but not for B cell survival. PMID 14563325 [PubMed - indexed for MEDLINE] T-cell and B-cell of RAP1A deficient mice impair integrin-mediated cell adhesion. 1 Mol Cell Biol. 2006 Jan;26(2) 643-53. Rap1A-deficient T and B cells show impaired integrin-mediated cell adhesion. Duchniewicz M, Zemojtel T, Kolanczyk M, Grossmann S, Scheele JS, Zwartkruis FJ. Department of Computational Molecular Biology, Max Planck Institute for Molecular Genetics, Ihnestrasse 73, D-14195 Berlin, Germany. Studies in tissue culture cells have demonstrated a role for the Ras-like GTPase Rap1 in the regulation of integrin-mediated cell-matrix and cadherin-mediated cell-cell contacts. To analyze the function of Rap1 in vivo, we have disrupted the Rap1A gene by homologous recombination. Mice homozygous for the deletion allele are viable and fertile. However, primary hematopoietic cells isolated from spleen or thymus have a diminished adhesive capacity on ICAM and fibronectin substrates. In addition, polarization of T cells from Rap1-/- cells after CD3 stimulation was impaired compared to that of wild-type cells. Despite this, these defects did not result in hematopoietic or cell homing abnormalities. Although it is possible that the relatively mild phenotype is a consequence of functional complementation by the Rap1B gene, our genetic studies confirm a role for Rap1A in the regulation of integrins. PMID 16382154 [PubMed - indexed for MEDLINE] T-cell of WASP deficient mice impair the proliferaction and antigen receptor cap formation in response to anti-CD3zeta stimulation. 1 Immunity. 1998 Jul;9(1) 81-91. Wiskott-Aldrich syndrome protein-deficient mice reveal a role for WASP in T but not B cell activation. Snapper SB, Rosen FS, Mizoguchi E, Cohen P, Khan W, Liu CH, Hagemann TL, Kwan SP, Ferrini R, Davidson L, Bhan AK, Alt FW. Howard Hughes Medical Institute, Children s Hospital, Boston, Massachusetts 02115, USA. The Wiskott-Aldrich syndrome (WAS) is a human X-linked immunodeficiency resulting from mutations in a gene (WASP) encoding a cytoplasmic protein implicated in regulating the actin cytoskeleton. To elucidate WASP function, we disrupted the WASP gene in mice by gene-targeted mutation. WASP-deficient mice showed apparently normal lymphocyte development, normal serum immunoglobulin levels, and the capacity to respond to both T-dependent and T-independent type II antigens. However, these mice did have decreased peripheral blood lymphocyte and platelet numbers and developed chronic colitis. Moreover, purified WASP-deficient T cells showed markedly impaired proliferation and antigen receptor cap formation in response to anti-CD3epsilon stimulation. Yet, purified WASP-deficient B cells showed normal responses to anti-Ig stimulation. We discuss the implications of our findings regarding WASP function in receptor signaling and cytoskeletal reorganization in T and B cells and compare the effects of WASP deficiency in mice and humans. PMID 9697838 [PubMed - indexed for MEDLINE] T-cell of SHB defective mice impair the phosphorylation of LAT and consequently the activation of MAP kinase pathways. 9 Sep 24;274(39) 28050-7. Requirement of the Src homology 2 domain protein Shb for T cell receptor-dependent activation of the interleukin-2 gene nuclear factor for activation of T cells element in Jurkat T cells. Lindholm CK, Gylfe E, Zhang W, Samelson LE, Welsh M. Department of Medical Cell Biology, Box 571, Biomedicum, Uppsala University, S-75123 Uppsala, Sweden. Stimulation of the T cell antigen receptor (TCR) induces tyrosine phosphorylation of numerous intracellular proteins. We have recently investigated the role of the adaptor protein Shb in the early events of T cell signaling and observed that Shb associates with Grb2, linker for activation of T cells (LAT) and the TCR zeta-chain in Jurkat cells. We now report that Shb also associates with phospholipase C-gamma1 (PLC-gamma1) in these cells. Overexpression of Src homology 2 domain defective Shb caused diminished phosphorylation of LAT and consequently the activation of mitogen-activated protein kinases was decreased upon TCR stimulation. In addition, the Shb mutant also blocked phosphorylation of PLC-gamma1 and the increase in cytoplasmic Ca(2+) following TCR stimulation. Nuclear factor for activation of T cells is a major target for Ras and calcium signaling pathways in T cells following TCR stimulation, and the overexpression of the mutant Shb prevented TCR-dependent activation of the nuclear factor for activation of T cells. Consequently, endogenous interleukin-2 production was decreased under these conditions. The results indicate a role for Shb as a link between the TCR and downstream signaling events involving LAT and PLC-gamma1 and resulting in the activation of transcription of the interleukin-2 gene. PMID 10488157 [PubMed - indexed for MEDLINE] B-cell of 3BP2 (-/-) deficient mice have defective in proliferation, cell cycle progression, PLC-gamma2 phosphorylation, calcium mobilization, NF-ATp dephosphorylation, and Erk and Jnk activation in response to BCR ligation. 1 Mol Cell Biol. 2006 Jul;26(14) 5214-25. 3BP2 deficiency impairs the response of B cells, but not T cells, to antigen receptor ligation. de la Fuente MA, Kumar L, Lu B, Geha RS. Division of Immunology, Children s Hospital, 300 Longwood Ave., Boston, MA 02115, USA. The adapter protein 3BP2 is expressed in lymphocytes; binds to Syk/ZAP-70, Vav, and phospholipase C-gamma (PLC-gamma); and is thought to be important for interleukin-2 gene transcription in T cells. To define the role of 3BP2 in lymphocyte development and function, we generated 3BP2-deficient mice. T-cell development, proliferation, cytokine secretion, and signaling in response to T-cell receptor (TCR) ligation were all normal in 3BP2(-/-) mice. 3BP2(-/-) mice had increased accumulation of pre-B cells in the bone marrow and a block in the progression of transitional B cells in the spleen from the T1 to the T2 stage, but normal numbers of mature B cells. B-cell proliferation, cell cycle progression, PLC-gamma2 phosphorylation, calcium mobilization, NF-ATp dephosphorylation, and Erk and Jnk activation in response to B-cell receptor (BCR) ligation were all impaired. These results suggest that 3BP2 is important for BCR, but not for TCR signaling. PMID 16809760 [PubMed - indexed for MEDLINE] B-cell of Vav2(-/-) deficient mice are defective in the ability to switch immunoglobulin class. 1 Nat Immunol. 2001 Jun;2(6) 542-7. Comment in Nat Immunol. 2001 Jun;2(6) 482-4. Signal transduction through Vav-2 participates in humoral immune responses and B cell maturation. Doody GM, Bell SE, Vigorito E, Clayton E, McAdam S, Tooze R, Fernandez C, Lee IJ, Turner M. Laboratory of Lymphocyte Signaling and Development, Molecular Immunology Programme, The Babraham Institute, Babraham, Cambridge CB2 4AT, UK. B and T lymphocytes develop normally in mice lacking the guanine nucleotide exchange factor Vav-2. However, the immune responses to type II thymus-independent antigen as well as the primary response to thymus-dependent (TD) antigen are defective. Vav-2-deficient mice are also defective in their ability to switch immunoglobulin class, form germinal centers and generate secondary immune responses to TD antigens. Mice lacking both Vav-1 and Vav-2 contain reduced numbers of B lymphocytes and display a maturational block in the development of mature B cells. B cells from Vav-1(-/-)Vav-2(-/-) mice respond poorly to antigen receptor triggering, both in terms of proliferation and calcium release. These studies show the importance of Vav-2 in humoral immune responses and B cell maturation. PMID 11376342 [PubMed - indexed for MEDLINE] T-cell of Vav1(-/-) deficient mice exhibit impaired antigen receptor signaling. 1 Nature. 1995 Mar 30;374(6521) 474-7. Defective T-cell receptor signalling and positive selection of Vav-deficient CD4+ CD8+ thymocytes. Fischer KD, Zmuldzinas A, Gardner S, Barbacid M, Bernstein A, Guidos C. Program in Molecular Biology and Cancer, Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada. During lymphocyte development, cellular proliferation and positive and negative selection events ensure the production of T and B lymphocytes bearing highly diverse, but self-tolerant, repertoires of antigen receptors. These processes are initiated when engagement of growth-factor receptors, or the T and B lymphocyte antigen receptors, induces tyrosine phosphorylation of specific SH2- and SH3-domain-containing cytoplasmic proteins, including Vav. Here we show that vav-/- embryonic stem cells generate only limited numbers of immature and mature T and B lymphocytes in the RAG-2 blastocyst complementation assay. Furthermore, Vav-deficient T lymphocytes showed severely impaired antigen receptor signalling. Finally, we demonstrate that Vav-dependent signalling pathways regulate maturation, but not CD4/CD8 lineage commitment, during T-cell-receptor-mediated positive selection of immature CD4+ CD8+ precursors into mature CD4+ CD8- or CD4- CD8+ T cells. PMID 7700360 [PubMed - indexed for MEDLINE] Vav1(-/-)Vav2(-/-) mice exhibit greatly reduced the mature B-cells. Vav-1-/-Vav-2-/- B cells were unresponsive to BCR-driven proliferation in vitro and to thymus-indepen-dent antigen in vivo. 1 Nat Immunol. 2001 Jun;2(6) 548-55. Comment in Nat Immunol. 2001 Jun;2(6) 482-4. Compensation between Vav-1 and Vav-2 in B cell development and antigen receptor signaling. Tedford K, Nitschke L, Girkontaite I, Charlesworth A, Chan G, Sakk V, Barbacid M, Fischer KD. Abteilung Physiologische Chemie, Universitat Ulm, Albert-Einstein-Allee 11, D-89069 Ulm, Germany. Vav-1 and Vav-2 are closely related Dbl-homology GTP exchange factors (GEFs) for Rho GTPases. Mutation of Vav-1 disrupts T cell development and T cell antigen receptor-induced activation, but has comparatively little effect on B cells. We found that combined deletion of both Vav-1 and Vav-2 in mice resulted in a marked reduction in mature B lymphocyte numbers. Vav-1(-/-)Vav-2(-/-) B cells were unresponsive to B cell antigen receptor (BCR)-driven proliferation in vitro and to thymus-independent antigen in vivo. BCR-stimulated intracellular calcium mobilization was greatly impaired in Vav-1(-/-)Vav-2(-/-) B cells. These findings establish a role for Vav-2 in BCR calcium signaling and reveal that the Vav family of GEFs is critical to B cell development and function. PMID 11376343 [PubMed - indexed for MEDLINE] Fyn-deficient mice exhibit a remarkably specific lymphoid defect thymocytes are refractile to stimulation through the TCR with mitogen or antigen. 1 Cell. 1992 Sep 4;70(5) 751-63. Defective T cell receptor signaling in mice lacking the thymic isoform of p59fyn. Appleby MW, Gross JA, Cooke MP, Levin SD, Qian X, Perlmutter RM. Howard Hughes Medical Institute, Department of Immunology, University of Washington, Seattle 98195. Considerable evidence supports the hypothesis that the nonreceptor protein tyrosine kinase p59fyn participates in signal transduction from the T cell receptor (TCR). To examine this hypothesis in detail, we have produced mice that lack the thymic isoform of p59fyn but retain expression of the brain isoform of the protein. fynTnull mice exhibit a remarkably specific lymphoid defect thymocytes are refractile to stimulation through the TCR with mitogen or antigen, while peripheral T cells, following what appears to be a normal maturation sequence, reacquire significant signaling capabilities. These data confirm that p59fynT plays a pivotal role in TCR signal transduction and demonstrate that additional developmentally regulated signaling components also contribute to TCR-induced lymphocyte activation. PMID 1516132 [PubMed - indexed for MEDLINE] Lck-deficient mice show a pronounced thymic atrophy, with a dramatic reduction in the double-positive (CD4+CD8+) thymocyte population. 1 Nature. 1992 May 14;357(6374) 161-4. Comment in Nature. 1993 Jan 21;361(6409) 213. Profound block in thymocyte development in mice lacking p56lck. Molina TJ, Kishihara K, Siderovski DP, van Ewijk W, Narendran A, Timms E, Wakeham A, Paige CJ, Hartmann KU, Veillette A, et al. Ontario Cancer Institute, University of Toronto, Canada. The protein Lck (p56lck) has a relative molecular mass of 56,000 and belongs to the Src family of tyrosine kinases. It is expressed exclusively in lymphoid cells, predominantly in thymocytes and peripheral T cells. Lck associates specifically with the cytoplasmic domains of both CD4 and CD8 T-cell surface glycoproteins and interacts with the beta-chain of the interleukin-2 receptor, which implicates Lck activity in signal transduction during thymocyte ontogeny and activation of mature T cells. Here we generate an lck null mutation by homologous recombination in embryonic stem cells to evaluate the role of p56lck in T-cell development and activation. Lck-deficient mice show a pronounced thymic atrophy, with a dramatic reduction in the double-positive (CD4+CD8+) thymocyte population. Mature, single-positive thymocytes are not detectable in these mice and there are only very few peripheral T cells. These results illustrate the crucial role of this T-cell-specific tyrosine kinase in the thymocyte development. PMID 1579166 [PubMed - indexed for MEDLINE] T cell from mice deficient in LCK is required for normal signal transduction through the TCR. 1 Cell. 1992 Aug 21;70(4) 585-93. Genetic evidence for the involvement of the lck tyrosine kinase in signal transduction through the T cell antigen receptor. Straus DB, Weiss A. Howard Hughes Medical Institute, Department of Medicine, University of California, San Francisco 94143. Signaling through the T cell antigen receptor (TCR) results both in rapid increases in tyrosine phosphorylation on a number of proteins and in the activation of the phosphatidylinositol pathway. It is not clear how stimulation of the TCR leads to these signaling events. Mutants of the Jurkat T cell line have been previously isolated that fail to show increases in calcium following receptor stimulation. Analysis of one of these mutants, JCaM1, which is defective in the induction of tyrosine phosphorylation, revealed a defect in the expression of functional lck tyrosine kinase. The lack of lck activity was caused in part by a splicing defect. Expression of the lck cDNA in JCaM1 restores the ability of the cell to respond to TCR stimulation. These results indicate that lck is required for normal signal transduction through the TCR. PMID 1505025 [PubMed - indexed for MEDLINE] T cells from mice deficient in SLAP-130/Fyb show markedly impaired proliferation. 1 Science. 2001 Sep 21;293(5538) 2263-5. Coupling of the TCR to integrin activation by Slap-130/Fyb. Peterson EJ, Woods ML, Dmowski SA, Derimanov G, Jordan MS, Wu JN, Myung PS, Liu QH, Pribila JT, Freedman BD, Shimizu Y, Koretzky GA. The Abramson Family Cancer Research Institute, Department of Medicine, School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA. SLAP-130/Fyb (SLP-76-associated phosphoprotein or Fyn-binding protein; also known as Fyb/Slap) is a hematopoietic-specific adapter, which associates with and modulates function of SH2-containing leukocyte phosphoprotein of 76 kilodaltons (SLP-76). T cells from mice lacking SLAP-130/Fyb show markedly impaired proliferation following CD3 engagement. In addition, the T cell receptor (TCR) in SLAP-130/Fyb mutant cells fails to enhance integrin-dependent adhesion. Although TCR-induced actin polymerization is normal, TCR-stimulated clustering of the integrin LFA-1 is defective in SLAP-130/Fyb-deficient cells. These data indicate that SLAP-130/Fyb is important for coupling TCR-mediated actin cytoskeletal rearrangement with activation of integrin function, and for T cells to respond fully to activating signals. PMID 11567141 [PubMed - indexed for MEDLINE] B cell of chicken deficient ITK reduce IP3 generation and phospholipase C gamma 2 tyrosine phosphorylation. 1 J Exp Med. 1996 Jul 1;184(1) 31-40. A role for Bruton s tyrosine kinase in B cell antigen receptor-mediated activation of phospholipase C-gamma 2. Takata M, Kurosaki T. Department of Oncology and Immunology, Wyeth-Ayerst Research, Pearl River, New York 10965, USA. Defects in the gene encoding Bruton s tyrosine kinase (Btk) result in a disease called X-linked agammaglobulinemia, in which there is a profound decrease of mature B cells due to a block in B cell development. Recent studies have shown that Btk is tyrosine phosphorylated and activated upon B cell antigen receptor (BCR) stimulation. To elucidate the functions of this kinase, we examined BCR signaling of DT40 B cells deficient in Btk. Tyrosine phosphorylation of phospholipase C (PLC)-gamma 2 upon receptor stimulation was significantly reduced in the mutant cells, leading to the loss of both BCR-coupled phosphatidylinositol hydrolysis and calcium mobilization. Pleckstrin homology and Src-homology 2 domains of Btk were required for PLC-gamma 2 activation. Since Syk is also required for the BCR-induced PLC-gamma 2 activation, our findings indicate that PLC-gamma 2 activation is regulated by Btk and Syk through their concerted actions. PMID 8691147 [PubMed - indexed for MEDLINE] T cell of mice deficient ITK reduce IP3 generation and phospholipase C gamma 1 tyrosine phosphorylation. 1 J Exp Med. 1998 May 18;187(10) 1721-7. T cell receptor-initiated calcium release is uncoupled from capacitative calcium entry in Itk-deficient T cells. Liu KQ, Bunnell SC, Gurniak CB, Berg LJ. Program of Immunology, Division of Medical Sciences, Harvard University, Boston, Massachusetts 02115, USA. Itk, a Tec family tyrosine kinase, plays an important but as yet undefined role in T cell receptor (TCR) signaling. Here we show that T cells from Itk-deficient mice have a TCR-proximal signaling defect, resulting in defective interleukin 2 secretion. Upon TCR stimulation, Itk-/- T cells release normal amounts of calcium from intracellular stores, but fail to open plasma membrane calcium channels. Since thapsigargin-induced store depletion triggers normal calcium entry in Itk-/- T cells, an impaired biochemical link between store depletion and channel opening is unlikely to be responsible for this defect. Biochemical studies indicate that TCR-induced inositol 1,4,5 tris-phosphate (IP3) generation and phospholipase C gamma1 tyrosine phosphorylation are substantially reduced in Itk-/- T cells. In contrast, TCR-zeta and ZAP-70 are phosphorylated normally, suggesting that Itk functions downstream of, or in parallel to, ZAP-70 to facilitate TCR-induced IP3 production. These findings support a model in which quantitative differences in cytosolic IP3 trigger distinct responses, and in which only high concentrations of IP3 trigger the influx of extracellular calcium. PMID 9584150 [PubMed - indexed for MEDLINE] T cell of mice deficient ITK have failure of Th2 development. 1 Immunity. 1999 Oct;11(4) 399-409. Impaired NFATc translocation and failure of Th2 development in Itk-deficient CD4+ T cells. Fowell DJ, Shinkai K, Liao XC, Beebe AM, Coffman RL, Littman DR, Locksley RM. Department of Medicine, University of California San Francisco 94143, USA. Naive Itk-deficient CD4+ T cells were unable to establish stable IL-4 production, even when primed in Th2-inducing conditions. In contrast, IFNgamma production was little affected. Failure to express IL-4 occurred even among cells that had gone through multiple cell divisions and was associated with a delay in the kinetics and magnitude of NFATc nuclear localization. IL-4 production was restored genetically by retroviral reconstitution of Itk or biochemically by augmenting the calcium flux with ionomycin. In vivo, Itk-deficient mice were unable to establish functional Th2 cells. Development of protective Th1 cells was unimpeded. These data define a nonredundant role for Itk in modulating signals from the TCR/CD28 pathways that are specific for the establishment of stable IL-4 but not IFNgamma expression. PMID 10549622 [PubMed - indexed for MEDLINE] Mice deficient in ITK have reduced proliferative responses to MHC stimulation and to anti-TCR cross-linking 1 Immunity. 1995 Dec;3(6) 757-69. Altered T cell receptor signaling and disrupted T cell development in mice lacking Itk. Liao XC, Littman DR. Department of Microbiology and Immunology, University of California, San Francisco 94143-0414, USA. Itk is a T cell protein tyrosine kinase (PTK) that, along with Btk and Tec, belongs to a family of cytoplasmic PTKs with N-terminal pleckstrin homology domains. Btk plays a critical role in B lymphocyte development. To determine whether Itk has an analogous role in T lymphocytes, we used gene targeting to prepare mice lacking expression of Itk. Such animals had decreased numbers of mature thymocytes, an effect most clearly observed in mice expressing T cell receptor (TCR) transgenes. Mature T cells from Itk-deficient mice had reduced proliferative responses to allogeneic MHC stimulation and to anti-TCR cross-linking, but responded normally to stimulation with phorbol ester plus ionomycin or with IL-2. These results provide genetic evidence that Itk is involved in T cell development and also suggest that Itk has an important role in proximal events in TCR-mediated signaling pathways. PMID 8777721 [PubMed - indexed for MEDLINE] Mutations in Btk cause X-linked immunodeficiency. 1 Semin Immunol. 1998 Aug;10(4) 309-16. Btk function in B cell development and response. Satterthwaite AB, Li Z, Witte ON. Department of Microbiology and Molecular Genetics, University of California, Los Angeles 90095-1662, USA. Mutations in Bruton s tyrosine kinase (Btk) result in the B cell immunodeficiencies XLA in humans and Xid in mice. Both the maintenance of peripheral B cell numbers and their response to B cell antigen receptor (BCR) crosslinking depend on Btk. Btk integrates signals from multiple cell surface receptors, including BCR and G-protein coupled receptors. These Btk dependent signals control B cell proliferation and survival by mediating Ca2+ flux, activating JNK and p38 and inducing cell cycle regulatory genes. Publication Types Review PMID 9695187 [PubMed - indexed for MEDLINE] Gads(GRAP2) has a role in thymocyte proliferaction for maturation of T-cells. 1 Science. 2001 Mar 9;291(5510) 1987-91. Requirement for the SLP-76 adaptor GADS in T cell development. Yoder J, Pham C, Iizuka YM, Kanagawa O, Liu SK, McGlade J, Cheng AM. Medical Scientist Training Program, Washington University School of Medicine, St. Louis, MO 63110, USA. GADS is an adaptor protein implicated in CD3 signaling because of its ability to link SLP-76 to LAT. A GADS-deficient mouse was generated by gene targeting, and the function of GADS in T cell development and activation was examined. GADS- CD4-CD8- thymocytes exhibited a severe block in proliferation but still differentiated into mature T cells. GADS- thymocytes failed to respond to CD3 cross-linking in vivo and were impaired in positive and negative selection. Immunoprecipitation experiments revealed that the association between SLP-76 and LAT was uncoupled in GADS- thymocytes. These observations indicate that GADS is a critical adaptor for CD3 signaling. PMID 11239162 [PubMed - indexed for MEDLINE] Gads(GRAP2) has a role for homeostatic proliferaction in B cells. 1 Eur J Immunol. 2005 Apr;35(4) 1184-92. Expression and function of the adaptor protein Gads in murine B cells. Yankee TM, Draves KE, Clark EA. Department of Immunology, University of Washington, Seattle, USA. tyankee@kumc.edu Nearly all hematopoietic receptors are dependent on adaptor proteins for the activation of downstream signaling pathways. The Gads adaptor protein is expressed in many hematopoietic tissues, including bone marrow, lymph node, and spleen. Using intracellular staining, we detected Gads protein in a number cells, including B cells, T cells, NK cells, monocytes, and plasmacytoid DC, but not in macrophages, neutrophils, or monocyte-derived DC. In the B cell compartment, Gads was first expressed after immature B cells leave the bone marrow and was down-regulated after B cell antigen receptor (BCR) ligation. Female Gads(-/-) mice had increased numbers of splenic B cells, as compared to female Gads(+/+) mice, suggesting a role for Gads in B cell homeostasis. Although B cell production and turnover of splenic B cell subsets appeared normal in Gads(-/-) mice, homeostatic proliferation was significantly impaired in Gads(-/-) B cells. Whereas BCR ligation can induce apoptosis in wild-type transitional stage 1 (T1) B cells, Gads(-/-) T1 B cells were resistant to BCR-induced apoptosis. Gads(-/-) B cells also showed increased BCR-mediated calcium mobilization. We conclude that Gads may have a negative regulatory role in signaling through survival pathways, and is necessary for normal homeostatic proliferation in B cells. PMID 15761845 [PubMed - indexed for MEDLINE] Grap negatively regulates T-cell proliferation. 1 Mol Cell Biol. 2002 May;22(10) 3230-6. Grap negatively regulates T-cell receptor-elicited lymphocyte proliferation and interleukin-2 induction. Shen R, Ouyang YB, Qu CK, Alonso A, Sperzel L, Mustelin T, Kaplan MH, Feng GS. Program in Signal Transduction Research, The Burnham Institute, La Jolla, California 92037, USA. Grb-2-related adaptor protein (Grap) is a Grb2-like SH3-SH2-SH3 adaptor protein with expression restricted to lymphoid tissues. Grap(-/-) lymphocytes isolated from targeted Grap-deficient mice exhibited enhanced proliferation, interleukin-2 production, and c-fos induction in response to mitogenic T-cell receptor (TCR) stimulation, compared to wild-type cells. Ectopic expression of Grap led to a suppression of Elk-1-directed transcription induced by the Ras/Erk pathway, without having effects on gene expression mediated by Jnk and p38 mitogen-activated protein kinases. Together, these data suggest that Grap, unlike Grb2, acts as a negative regulator of TCR-stimulated intracellular signaling by downregulating signal relay through the Ras/Erk pathway. PMID 11971956 [PubMed - indexed for MEDLINE] Gab2 is a substrate of ZAP-70 and functions as a switch molecule toward inhibition of TCR signal transuduction. 1 J Biol Chem. 2001 Nov 30;276(48) 45175-83. Epub 2001 Sep 25. Docking protein Gab2 is phosphorylated by ZAP-70 and negatively regulates T cell receptor signaling by recruitment of inhibitory molecules. Yamasaki S, Nishida K, Hibi M, Sakuma M, Shiina R, Takeuchi A, Ohnishi H, Hirano T, Saito T. Molecular Genetics, Chiba University Graduate School of Medicine, Chiba 260-8670, Japan. To maintain various T cell responses and immune equilibrium, activation signals triggered by T cell antigen receptor (TCR) must be regulated by inhibitory signals. Gab2, an adaptor protein of the insulin receptor substrate-1 family, has been shown to be involved in the downstream signaling from cytokine receptors. We investigated the functional role of Gab2 in TCR-mediated signal transduction. Gab2 was phosphorylated by ZAP-70 and co-precipitated with phosphoproteins, such as ZAP-70, LAT, and CD3zeta, upon TCR stimulation. Overexpression of Gab2 in Jurkat cells or antigen-specific T cell hybridomas resulted in the inhibition of NF-AT activation, interleukin-2 production, and tyrosine phosphorylation. The structure-function relationship of Gab2 was analyzed by mutants of Gab2. The Gab2 mutants lacking SHP-2-binding sites mostly abrogated the inhibitory activity of Gab2, but its inhibitory function was restored by fusing to active SHP-2 as a chimeric protein. A mutant with defective phosphatidylinositol 3-kinase binding capacity also impaired the inhibitory activity, and the pleckstrin homology domain-deletion mutant revealed a crucial function of the pleckstrin homology domain for localization to the plasma membrane. These results suggest that Gab2 is a substrate of ZAP-70 and functions as a switch molecule toward inhibition of TCR signal transduction by mediating the recruitment of inhibitory molecules to the TCR signaling complex. PMID 11572860 [PubMed - indexed for MEDLINE] B cell signaling causes tyrosine phosphorylation of Gab1, and in turn SHP2 bind to Gab1 Gab1 phosophorylation potentiate the phosphorylation of Akt, PI3K-dependent response. 1 J Biol Chem. 2001 Apr 13;276(15) 12257-65. Epub 2001 Jan 22. The Gab1 docking protein links the b cell antigen receptor to the phosphatidylinositol 3-kinase/Akt signaling pathway and to the SHP2 tyrosine phosphatase. Ingham RJ, Santos L, Dang-Lawson M, Holgado-Madruga M, Dudek P, Maroun CR, Wong AJ, Matsuuchi L, Gold MR. Departments of Microbiology and Immunology and Zoology, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada. B cell antigen receptor (BCR) signaling causes tyrosine phosphorylation of the Gab1 docking protein. This allows phosphatidylinositol 3-kinase (PI3K) and the SHP2 tyrosine phosphatase to bind to Gab1. In this report, we tested the hypothesis that Gab1 acts as an amplifier of PI3K- and SHP2-dependent signaling in B lymphocytes. By overexpressing Gab1 in the WEHI-231 B cell line, we found that Gab1 can potentiate BCR-induced phosphorylation of Akt, a PI3K-dependent response. Gab1 expression also increased BCR-induced tyrosine phosphorylation of SHP2 as well as the binding of Grb2 to SHP2. We show that the pleckstrin homology (PH) domain of Gab1 is required for BCR-induced phosphorylation of Gab1 and for Gab1 participation in BCR signaling. Moreover, using confocal microscopy, we show that BCR ligation can induce the translocation of Gab1 from the cytosol to the plasma membrane and that this requires the Gab1 PH domain as well as PI3K activity. These findings are consistent with a model in which the binding of the Gab1 PH domain to PI3K-derived lipids brings Gab1 to the plasma membrane, where it can be tyrosine-phosphorylated and then act as an amplifier of BCR signaling. PMID 11278704 [PubMed - indexed for MEDLINE] RasGRP1 mediates Ras activation following TCR stimulatioin. RasGRP1 and RasGRP3 induces RAS activation in B-cell to response to T-cell stimulation. 1 Immunol Lett. 2006 May 15;105(1) 77-82. Epub 2006 Feb 20. The role of RasGRPs in regulation of lymphocyte proliferation. Coughlin JJ, Stang SL, Dower NA, Stone JC. Department of Biochemistry, University of Alberta, Edmonton, Alta., Canada T6G 2H7. RasGRP1 links TCR signaling to Ras in T cells, while both RasGRP1 and RasGRP3 link BCR signaling to Ras in B cells. T cells deficient in RasGRP1 have defective proliferative responses as do B cells deficient in both RasGRP1 and RasGRP3, confirming the importance of Ras activation in lymphocyte proliferation. While aged Rasgrp1-/- mice develop late-onset autoimmunity characterized by splenomegaly and the presence of anti-nuclear antibodies (ANA), the additional loss of RasGRP3 expression inhibits this phenotype. We show here that the autoimmunity in Rasgrp1-/- mice is T cell dependent. Compared to wildtype, Rasgrp1-/- T cells induce greater in vitro B cell proliferation that is due, at least in part, to increased production of interleukin-4 (IL-4). Rasgrp1 Rasgrp3 double mutant B cells are less responsive to this T cell stimulation. The reduced double mutant B cell proliferative response was paralleled by decreased induction of cyclin D2 upon stimulation with IL-4 and anti-IgM. Taken together these results suggest that double mutant mice fail to generate autoimmunity due to their decreased B cell cyclin D2 accumulation, and thus proliferation, in response to the elevated levels of IL-4 produced by mutant T cells. PMID 16530850 [PubMed - indexed for MEDLINE] Grb2-hSos1-PLCgamma1-p36/p38-ZAP70 complexes localize in the vicinity of TCR-zeta 1 J Biol Chem. 1995 Aug 4;270(31) 18428-36. Ligation of the T-cell antigen receptor (TCR) induces association of hSos1, ZAP-70, phospholipase C-gamma 1, and other phosphoproteins with Grb2 and the zeta-chain of the TCR. Nel AE, Gupta S, Lee L, Ledbetter JA, Kanner SB. Department of Medicine, UCLA School of Medicine 90024, USA. Signaling by the T-cell antigen receptor (TCR) involves both phospholipase C (PLC)-gamma 1 and p21ras activation. While failing to induce Shc/Grb2 association, ligation of the TCR/CD3 receptor in Jurkat T-cells induced hSos1-Grb2 complexes. In addition to hSos1, Grb2 participates in the formation of a tyrosine phosphoprotein complex that includes 145-, 95-, 70-, 54-, and 36-38-kDa proteins. p145 was identified as PLC-gamma 1 and p70 as the protein tyrosine kinase, ZAP-70. Although of the same molecular weight, p95 was not recognized by an anti-serum to p95 Vav. The SH2 domains of Grb2 and PLC-gamma 1 were required for the formation of this protein complex. In anti-CD3-treated cells, Grb2 redistributed from the cytosol to a particulate cell compartment along with p36/p38, ZAP-70, and PLC-gamma 1. Part of the Grb2 complex associated with the particulate compartment could be extracted with Nonidet P-40, while the rest was Nonidet P-40 insoluble. In both the detergent-soluble and -insoluble fractions, Grb2 coimmunoprecipitated with the zeta-chain of the TCR. Taken together, these results indicate that anti-CD3 induces Grb2-hSos1-PLC-gamma 1-p36/p38-ZAP70 complexes, which localize in the vicinity of TCR-zeta. PMID 7629168 [PubMed - indexed for MEDLINE] Gads(Grap2) plays an important role in T-cell signaling via its association with SLP-76 and LAT. 1 Curr Biol. 1999 Jan 28;9(2) 67-75. The hematopoietic-specific adaptor protein gads functions in T-cell signaling via interactions with the SLP-76 and LAT adaptors. Liu SK, Fang N, Koretzky GA, McGlade CJ. Department of Medical Biophysics, University of Toronto, The Arthur and Sonia Labatt Brain Tumour Research Centre, Hospital for Sick Children, Research Institute, 555 University Ave, Toronto, Ontario M5G 1X8, Canada. BACKGROUND The adaptor protein Gads is a Grb2-related protein originally identified on the basis of its interaction with the tyrosine-phosphorylated form of the docking protein Shc. Gads protein expression is restricted to hematopoietic tissues and cell lines. Gads contains a Src homology 2 (SH2) domain, which has previously been shown to have a similar binding specificity to that of Grb2. Gads also possesses two SH3 domains, but these have a distinct binding specificity to those of Grb2, as Gads does not bind to known Grb2 SH3 domain targets. Here, we investigated whether Gads is involved in T-cell signaling. RESULTS We found that Gads is highly expressed in T cells and that the SLP-76 adaptor protein is a major Gads-associated protein in vivo. The constitutive interaction between Gads and SLP-76 was mediated by the carboxy-terminal SH3 domain of Gads and a 20 amino-acid proline-rich region in SLP-76. Gads also coimmunoprecipitated the tyrosine-phosphorylated form of the linker for activated T cells (LAT) adaptor protein following cross-linking of the T-cell receptor; this interaction was mediated by the Gads SH2 domain. Overexpression of Gads and SLP-76 resulted in a synergistic augmentation of T-cell signaling, as measured by activation of nuclear factor of activated T cells (NFAT), and this cooperation required a functional Gads SH2 domain. CONCLUSIONS These results demonstrate that Gads plays an important role in T-cell signaling via its association with SLP-76 and LAT. Gads may promote cross-talk between the LAT and SLP-76 signaling complexes, thereby coupling membrane-proximal events to downstream signaling pathways. PMID 10021361 [PubMed - indexed for MEDLINE] Lck is required for normal signal transduction through the TCR. 1 Cell. 1992 Aug 21;70(4) 585-93. Genetic evidence for the involvement of the lck tyrosine kinase in signal transduction through the T cell antigen receptor. Straus DB, Weiss A. Howard Hughes Medical Institute, Department of Medicine, University of California, San Francisco 94143. Signaling through the T cell antigen receptor (TCR) results both in rapid increases in tyrosine phosphorylation on a number of proteins and in the activation of the phosphatidylinositol pathway. It is not clear how stimulation of the TCR leads to these signaling events. Mutants of the Jurkat T cell line have been previously isolated that fail to show increases in calcium following receptor stimulation. Analysis of one of these mutants, JCaM1, which is defective in the induction of tyrosine phosphorylation, revealed a defect in the expression of functional lck tyrosine kinase. The lack of lck activity was caused in part by a splicing defect. Expression of the lck cDNA in JCaM1 restores the ability of the cell to respond to TCR stimulation. These results indicate that lck is required for normal signal transduction through the TCR. PMID 1505025 [PubMed - indexed for MEDLINE] ZAP-70 plays crucial roles in T-cell activation and development. Syk triggers cellular activation in T-cell. 1 Mol Cell Biol. 1998 Mar;18(3) 1388-99. Genetic evidence for differential coupling of Syk family kinases to the T-cell receptor reconstitution studies in a ZAP-70-deficient Jurkat T-cell line. Williams BL, Schreiber KL, Zhang W, Wange RL, Samelson LE, Leibson PJ, Abraham RT. Department of Immunology, Mayo Clinic, Rochester, Minnesota 55905, USA. T-cell antigen receptor (TCR) engagement activates multiple protein tyrosine kinases (PTKs), including the Src family member, Lck, and the Syk-related PTK, ZAP-70. Studies in ZAP-70-deficient humans have demonstrated that ZAP-70 plays crucial roles in T-cell activation and development. However, progress toward a detailed understanding of the regulation and function of ZAP-70 during TCR signaling has been hampered by the lack of a suitable T-cell model for biochemical and genetic analyses. In this report, we describe the isolation and phenotypic characterization of a Syk- and ZAP-70-negative somatic mutant derived from the Jurkat T-cell line. The P116 cell line displays severe defects in TCR-induced signaling functions, including protein tyrosine phosphorylation, intracellular Ca2+ mobilization, and interleukin-2 promoter-driven transcription. These signaling defects were fully reversed by reintroduction of catalytically active versions of either Syk or ZAP-70 into the P116 cells. However, in contrast to ZAP-70 expression, Syk expression triggered a significant degree of cellular activation in the absence of TCR ligation. Transfection experiments with ZAP-70-Syk chimeric proteins indicated that both the amino-terminal regulatory regions and the carboxy-terminal catalytic domains of Syk and ZAP-70 contribute to the distinctive functional properties of these PTKs. These studies underscore the crucial role of ZAP-70 in TCR signaling and offer a powerful genetic model for further analyses of ZAP-70 regulation and function in T cells. PMID 9488454 [PubMed - indexed for MEDLINE]
https://w.atwiki.jp/borris/
霊が住んでいる？ よく私の家に霊が住んでいます。という話しを耳にする。 これっていつも思う事がある。 本当に霊なのか？いや、勝ってにテレビがつくんです。 寝ていて気配を感じる。視線を感じる。 カーテンが動く。おもちゃが買ってに鳴り出す。などなど。 内容をみてみるとこれって、、、小動物の仕業ではなかろうかといつも思う。 人間は五感があるから小動物が動くと気配を感じるはずだ。 もうはっきりいう。これは霊ではありません。 ねずみちゃんと、ごきぶりちゃんなのです。 どうしたのかな？ 毎朝、通勤の時に見掛けていた彼。 数日前からその姿を見掛ける事が無くなってしまいました。 運転する車の窓越しに写っていた自転車で爆走している姿。 少し長い髪と何時も黒いTシャツ姿で、何時も後ろ姿ばかりで顔は見た事がありませんが、 その姿がとても印象的で、何時の頃からか、彼が私の車を通り過ぎる事が楽しみになっていました。 どうしたのかな？風邪引いたのかな？？ 転職をしてしまったのかな？？？ 彼の姿が見られなくなってから、毎朝、彼の姿を探す様になってしまっています。 また逢えるのかな... http //www.setuyaku.jp.net/useful/deco-cup.html 久しぶりに出した靴 だいぶ前に「クラークス」の靴が流行りました。 男も女もクラークスの靴を履いていて、私も欲しくなって買いました。 当時の私にとってブーツの次に高い靴でした。 私は貧乏性のところもあるので、もったいなくて痛むのが嫌で、あんまり履かなかったんです。 その靴を最近、久しぶりに、げた箱から出しました。 靴や服もある程度は着たり履いたりして、休ませてあげながらも使わないと意味がないですよね。 ペタンコなので歩く時に履きたいと思います。
https://w.atwiki.jp/iyasinoseika/pages/13.html
まずは来てくださった事を感謝します。 Wikiなのでご利用簡単