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次のプロトコルは、ワシントン大学の施設内倫理委員会のガイドラインに準拠しています。 1。電気生理学的評価<ol><li>セッションの準備<ol><li>部屋の準備：場所マニピュ、それが把握された場合、参加者の範囲を把握内の表に、収集ソフトウェアにタイムスタンプ付きのマーカーを送る取り付けられたセンサ、木製のブロック（ 図1を参照）。脳波取得ソフトウェアをアクティブにして、「新規セッション」（ 図S1）を開始します。 </li><li> NETの準備：104°Fに蒸留水（1リットル）、塩化カリウム（大さじ1）、およびベビーシャンプー（小さじ1杯）の温溶液温めた生理食塩水で128電極高密度のアレイEEGシステムを浸す。 </li><li>参加者の準備：参加者がビデオカムを考慮して快適に約75センチメートル刺激提示モニタから、完全に装着されていることを確認時代。見つけて、肌のマーカーで参加者の頭の上に頂点をマーク。 [後]とイニオンとpreauricularsの中間点間の中間点の交点を見つけることによって、頂点を測定します。 </li><li> Netアプリケーション：頂点電極は頂点マークの上に直接配置されるように、参加者の頭部にEEGキャップを置きます。インピーダンスをチェックして、インピーダンスが閾値以下に、使用中のEEGシステム（ 図S2）のために適切であることを確認。 </li><li>ビデオテーピングのセッションを開始します。 </li></ol></li><li>レコーディングのセットアップ：頂点電極基準信号。 0.1〜100Hzの間のアナログフィルタは、信号を増幅し、そして/秒500サンプルでデジタル化する。 </li><li>刺激提示：3条件を有する本参加者：観察実行し、休息、Muthukumaraswamyと同僚5によって開発パラダイムから適応。 <ol><li>条件を守って静かに座って見て、参加者に指示マニピュランダムを把握する人物の映像。各試行は6秒を持続する必要があります。正確に観測された把握が正確に3秒で起こることを確実にするための時間を観察試験のための事前に記録された映像。タスクの実行中に参加者の視覚的注意を監視し、それらが事後処理中に破棄されるように画面に出席しないその間トライアルをマーク。 </li><li>条件を実行します。ちょうどマニピュ下の右側の休憩で静かに座って、事前に録音された聴覚信号を聞いて、観察条件のビデオクリップからマニピュグラブを模倣するために、参加者に指示します。各試行は6秒を持続する必要があります。聴覚信号を維持し、聴覚トラックを予め記録することにより、正確に3秒で発表されていることを確認し、一貫して、キューと試行間インターバルを実行タイムアウトしました。正確に参加者の把握（ 図S3）が発生した時刻を記録するためにマニピュ上のセンサーを利用する。 </li><li>休止状態：quie座って、参加者に指示TLY目でオープンかつ受動的に刺激モニター上の小さな十字線を観察します。 3分間の休止状態中に記録連続脳波。 </li><li>条件を守って実行の両方について、条件ごとの40の試験の合計10試行の存在無作為化ブロック。マニピュランダムのイメージが試験間を含め、観察し、実行するブロック全体で、画面上に残っていることを確認してください。観察し、実行条件が完了した時点で残りの条件を管理する。 </li></ol></li><li>データ処理<ol><li>データ収集に続いて、インピーダンスを再確認。インピーダンス·レベルの変更に注意してください。エンド収集ソフトウェア記録。 </li><li>後処理：平均にRereference EEG信号。各条件（ 図S4）のための40の6秒の試験にセグメントの連続EEGデータ。 </li><li>自動化されたアーチファクトの検出を行っています。 exceedin速い平均振幅を特定することにより、運動の成果物のためのセグメントを検査する自動化されたアルゴリズムを使用G 200μV、平均100μVを超える振幅、および指定された試験（ 図S5）全体でゼロ分散を差動。 </li><li>データを視覚的に検査し、あらゆる動きアーティファクトと把握ジェスチャーとは関係のないあらゆる動きアーチファクトで汚染実行条件内のすべての試験で汚染観察条件にすべての試験を削除するには、セッションのビデオレビューを確認することで、手動のアーチファクトの検出を行っています。分析からの重要な成果物との比較試験を除外します。出席していないとして、取得の際にフラグが立てられたすべての試験を捨てる。検討し、分析中の各診断グループの試用拒否率に注意してください。 </li></ol></li><li>データ解析<ol><li>あたりのMuthukumaraswamy ら 5両観測（フォトセルによって示された）と（マニピュセンサーによって示された）実行条件の後に把握し、1秒前にデータの1秒からなる2秒のエポックにセグメント洗浄試験。セグメントC静止状態から2秒のエポックを傾け。 </li><li>高速フーリエ変換（FFT）各セグメントを変換する。 Muthukumaraswamy ら 5以下の統計分析のための標準的なC3、C4の位置を（周囲の各半球上の8個の電極のクラスタを選択し、 ベルニエら 3）（ 図2）。各条件について、パワースペクトルを計算するために含ま治験にわたって電力を平均化する。 </li><li> 8月13日Hzの範囲で、安静状態時の平均電力と比較して、モーターのアクションの実行や観察のいずれかの間の平均電力を調べることによって、ムーの減衰を計算します。減衰の度合いを決定するために、この比率のログを使用しています。注：正の値は増加を表している負の値は、実行または観察時の減衰を表す。この方法は、個人のアカウントを横切っ変動を考慮に入れると、比形で発現値の非正規性&lt;BR /&gt;注意：このプロトコルはネットステーションソフトウェアバージョン4.1で128電極高密度のアレイEEGシステムを使用して開発されました。基本的な手順は、EEGシステム間で類似しているが、取得および分析プロトコルが異なる場合があります。 </li></ol></li></ol> 2。サンプルキャラ<ol><li>研究レジストリ、前回の参加者のリスト、または地域の診療所や医師からの紹介を通じてパラダイムへの参加のための潜在的な患者集団を特定します。 </li><li>臨床構築物（ 例えば自閉症スペクトラム障害）の診断基準を満たし、このような頭部外傷、腫瘍、発作歴、または歪むことがあり抗痙攣またはバルビツール薬の使用の有無など任意の除外基準を識別するための可能性のための画面潜在的な参加電気生理学的信号。 </li><li>ゴールドスタンダード診断機器を利用して患者集団の診断状況を確認（ 例えば自閉症診断インタビュー-RE（ADI-R 11）教師ありと自閉症診断観察スケジュール-汎用（ADOS-G、12）の診断と統計マニュアル、次の専門家の臨床医によって投与- 第 5 版 （DSM-5）の基準13を 。 </li><li> など 、年齢、性別、認知能力、のような関心の関連の変数にマッチしたコントロールサンプルを識別</li></ol>

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Psychiatry. 51, 981-988 (2010).</li><li>Bernier, R., Aaronson, B., McPartland, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+imitation+in+the+observed+heterogeneity+in+EEG+mu+rhythm+in+autism+and+typical+development.">The role of imitation in the observed heterogeneity in EEG mu rhythm in autism and typical development.</a> Brain Cogn. 82, 69-75 (2013).</li><li>Pfurtscheller, G., Lopesda Silva, ., H, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Event-related+EEG/MEG+synchronization+and+desynchronization:+basic+principles.">Event-related EEG/MEG synchronization and desynchronization: basic principles.</a> Clin. Neurophysiol. 110, 1842-1857 (1999).</li><li>Marshall, P. J., Young, T., Meltzoff, A. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+correlates+of+action+observation+and+execution+in+14&#8208;month&#8208;old+infants:+An+event&#8208;related+EEG+desynchronization+study.">Neural correlates of action observation and execution in 14&#8208;month&#8208;old infants: An event&#8208;related EEG desynchronization study.</a> Dev. Sci. , 474-480 (2011).</li><li>Marshall, P. J., Meltzoff, A. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+mirroring+systems:+Exploring+the+EEG+mu+rhythm+in+human+infancy.">Neural mirroring systems: Exploring the EEG mu rhythm in human infancy.</a> Dev. Cogn. Neurosci. , 110-123 (2011).</li><li>Oberman, L., McCleery, J., Hubbard, E., Bernier, R., Pineda, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developmental+changes+in+mu+suppression+to+observed+actions+in+individuals+with+autism+spectrum+disorders.">Developmental changes in mu suppression to observed actions in individuals with autism spectrum disorders.</a> Soc. Cogn. Affective Neurosci. 8, 300-304 .</li></ol>

著者らは、競合する経済的利益を宣言していません。

臨床集団にミューリズムとアプリケーションに関連する電気生理学的データを正常に収集、処理、および分析を必要とする1）脳波方法論ツールの適用、2）慎重なアーチファクト検出およびデータ削減、ミューリズムの3）正確な識別、そして4）臨床集団と適切な対照群の同定の正確な特性。 適切な脳波手法は適切に機能して、慎重に統合された機器...

脳波（EEG）は、脳の活動を評価し、記録する、効果的、効率的、かつ非侵襲的な方法です。ニューロンが脳内で発火したように、結果として生じる電圧が、増幅記録し、グラフィカルに表現することができる。 EEGの時間分解能も、簡単な脳の振動パターンの変化、ならびに特定の刺激に対する脳の応答の解析の解析を可能にする。 年代後半19 世紀にまで遡る、最も古い脳のイメージング技術であるにもかかわらず、脳波はまだ幅広い適用性を持っています。機能的磁気共鳴画像法（fMRI）は、優れた空間分解能を有するが、比較的貧弱な時間分解能を有する。これは、脳内で起こるプロセスれる信じられないほどのスピード与えfMRIの評価の主な限界を表している。 EEG電位を与えるiを、ミリ秒レベルで脳の電気的活性を評価する能力を有する脳の処理の段階にnsight。 進化する技術は、脳波の適用を拡大している。記録システムの密度の増加は、空間分解能に関するEEGの制限のいくつかを緩和する、音源定位技術の開発を可能にした。さらに、現代のシステムでは、幼児や臨床サンプル1-3,28-30として以前は利用できなかった集団の評価を可能にし、大幅に個々の参加者のセットアップ時間を削減しました。 優れた時間分解能を考えると、EEGは、特定の行動、状態、または外部刺激に関連した神経応答を調べるために使用することができる。このユーティリティの例では、ヒトにおけるミラーニューロンシステム（MNS）の評価である。ミラーニューロンは、もともとのグループを証明する、単一のニューロン記録4を使ってサルで同定されたモーターアクションの実行と観察の両方に対応したニューロン。脳内に電極を配置するこの直接記録方式はほとんどヒトであり、唯一の悲惨な臨床例で利用されていません。 EEGはEEGミューリズムを監視することにより、MNSを評価するための方法を提供している。 8〜12 Hzの範囲内のこの振動パターンは、実行およびサル5-7で観察された活性化パターンに類似したモータの行動の観察に応答して、EEGパワーを減衰させることが示されている。同様に、経頭蓋磁気刺激（ 例えば下前頭回）を通じて、推定MNS脳領域の刺激が脳波ミューリズム8を廃止し、脳波ミューリズム抑制は、追加のサポートを提供し、被験者9内の推定ミラーニューロンの地域でのfMRIのBOLD信号と相関することが、このリズムインデックスは、少なくとも部分的に、MNS活性。脳波ミューリズムの評価は、ミラーニューロンの行為の非侵襲的評価を可能にしたヒトでivity。 EEGは脳の活動を検査するためのユニークな方法を提供し、行動に基づくアッセイと組み合わせたとき、それは、臨床集団において、そのような模倣として、社会的認知の側面を解明するための強力なツールであることができる。さらに、認知や言語障害を持つ集団で使用するための脳波の適用性は、他の撮像技術や行動パラダイムはあまり成功して利用することがあります誰のための個人の能力への洞察を可能にします。記載されているプロトコルは、理論的にはこのような自閉症スペクトラム障害などの定型および非定型発達と個人におけるミラーニューロンシステムにリンクされた社会的な認知機能を調べるための手段を提供します。

<table border="0" cellpadding="0" cellspacing="0" width="994">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:267px;">Geodesic EEG System</td>
			<td style="width:71px;">EGI</td>
			<td style="width:119px;">N/A</td>
			<td style="width:539px;">Any EEG system, not only EGI based systems, is applicable for the described study</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:267px;">MATLAB software</td>
			<td style="width:71px;">MATLAB</td>
			<td style="width:119px;">N/A</td>
			<td>Any mathematical, statistical software that can work with matrices is applicable</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:267px;">Netstation software</td>
			<td style="width:71px;">EGI</td>
			<td style="width:119px;">N/A</td>
			<td>Any EEG acquisition software is applicable for the described study</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:267px;">Manipulandum</td>
			<td style="width:71px;">custom</td>
			<td style="width:119px;">N/A</td>
			<td>Any object that is co-registered with data acquisition software to signal a successful grasp</td>
		</tr>
	</tbody>
</table>

eeg mu rhythm in typical and atypical development

脳波（EEG）は、脳の活動を評価し、記録する、効果的、効率的、かつ非侵襲的な方法です。優れた時間分解能を考えると、EEGは、特定の行動、状態、または外部刺激に関連した神経応答を調べるために使用することができる。このユーティリティの例は、脳波ミューリズムの検討を通じて、ヒトのミラーニューロンシステム（MNS）の評価である。単に個々の実行、または、目標指向行動を観察するときにEEGミューリズムは、中央に配置された電極から記録された8〜12 Hzの周波数範囲における振動活性が抑制される。このように、MNSの活性を反映することが提案されている。これは、ミラーニューロンシステム（MNS）における機能障害は、自閉症スペクトラム障害（ASD）の社会的障害に寄与する役割を果たしていることが理論化されています。 MNS次いで、非侵襲的にその活性の指標としてEEGミューリズム減衰を用いて、臨床集団において調べることができる。記載されPROTocolは、理論的なASDなどの定型および非定型の開発、を有する個人でMNSにリン​​クされ、社会的な認知機能を調べるための手段を提供します。

典型的な大人、子供や幼児は一貫して、パラダイムと刺激5の様々にわたるアクションの実行と観察の両方の間に14-30をミューリズムを実証している。この周波数帯における減衰量は一貫して、これは他の頭皮の領域に記録されているα電力の低減はないことを示す（ 図3）中央の電極間に局在している。同様に、動きの観察中に、この周波数での減衰量は?...

Watch this Scientific Journal Video about EEG Mu Rhythm in Typical and Atypical Development at JoVE.com

EEG Mu Rhythm in Typical and Atypical Development

脳波ミューリズムの評価は、脳の活動を調べるための独自の方法論を提供し、行動ベースのアッセイと組み合わせたときに、臨床集団では、このような模倣など、社会的認知の側面を解明するための強力なツールとなります。

定型および非定型開発における脳波ムーリズム

Electroencephalography (EEG) is an effective, efficient, and noninvasive method of assessing and recording brain activity. Given the excellent temporal ...

eeg-mu-rhythm-in-typical-and-atypical-development

Research

JoVE Journal

Medicine

25.4K ビュー.  University of Washington. 脳波ミューリズムの評価は、脳の活動を調べるための独自の方法論を提供し、行動ベースのアッセイと組み合わせたときに、臨床集団では、このような模倣など、社会的認知の側面を解明するための強力なツールとなります。 

ビデオ: EEG Mu Rhythm in Typical and Atypical Development

Direct Pressure Monitoring Accurately Predicts Pulmonary Vein Occlusion During Cryoballoon Ablation

Cryoballoon ablation (CBA) is an established therapy for atrial fibrillation (AF). Pulmonary vein (PV) occlusion is essential for achieving antral contact and PV isolation and is typically assessed by contrast injection. We present a novel method of direct pressure monitoring for assessment of PV occlusion.
Transcatheter pressure is monitored during balloon advancement to the PV antrum. Pressure is recorded via a single pressure transducer connected to the inner lumen of the cryoballoon. Pressure curve characteristics are used to assess occlusion in conjunction with fluoroscopic or intracardiac echocardiography (ICE) guidance. PV occlusion is confirmed when loss of typical left atrial (LA) pressure waveform is observed with recordings of PA pressure characteristics (no A wave and rapid V wave upstroke). Complete pulmonary vein occlusion as assessed with this technique has been confirmed with concurrent contrast utilization during the initial testing of the technique and has been shown to be highly accurate and readily reproducible.
We evaluated the efficacy of this novel technique in 35 patients. A total of 128 veins were assessed for occlusion with the cryoballoon utilizing the pressure monitoring technique; occlusive pressure was demonstrated in 113 veins with resultant successful pulmonary vein isolation in 111 veins (98.2%). Occlusion was confirmed with subsequent contrast injection during the initial ten procedures, after which contrast utilization was rapidly reduced or eliminated given the highly accurate identification of occlusive pressure waveform with limited initial training.
Verification of PV occlusive pressure during CBA is a novel approach to assessing effective PV occlusion and it accurately predicts electrical isolation. Utilization of this method results in significant decrease in fluoroscopy time and volume of contrast.

Effective pulmonary vein isolation utilizing a cryoballoon depends on complete pulmonary vein occlusion. The point of occlusion can be effectively predicted by direct analysis of pulmonary vein pressure waveform analysis during balloon inflation using a simple and reproducible technique.

直圧モニタリングは正確Cryoballoonアブレーション中に肺静脈閉塞を予測

Cryoballoon  ablation (CBA) is an established therapy for atrial fibrillation (AF).  Pulmonary vein (PV) occlusion is essential for achieving antral ...

医学

Development of an Audio-based Virtual Gaming Environment to Assist with Navigation Skills in the Blind

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind. Using only audio based cues and set within the context of a video game metaphor, users gather relevant spatial information regarding a building's layout. This allows the user to develop an accurate spatial cognitive map of a large-scale three-dimensional space that can be manipulated for the purposes of a real indoor navigation task. After game play, participants are then assessed on their ability to navigate within the target physical building represented in the game. Preliminary results suggest that early blind users were able to acquire relevant information regarding the spatial layout of a previously unfamiliar building as indexed by their performance on a series of navigation tasks. These tasks included path finding through the virtual and physical building, as well as a series of drop off tasks. We find that the immersive and highly interactive nature of the AbES software appears to greatly engage the blind user to actively explore the virtual environment. Applications of this approach may extend to larger populations of visually impaired individuals.

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind.

ブラインドのナビゲーションスキルを支援するためにオーディオベースのバーチャル·ゲーム環境の開発

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind. Using only audio ...

Clinical Examination Protocol to Detect Atypical and Classical Scrapie in Sheep

The diagnosis of scrapie, a transmissible spongiform encephalopathy (TSEs) of sheep and goats, is currently based on the detection of disease-associated prion protein by post mortem tests. Unless a random sample of the sheep or goat population is actively monitored for scrapie, identification of scrapie cases relies on the reporting of clinical suspects, which is dependent on the individual&#39;s familiarization with the disease and ability to recognize clinical signs associated with scrapie. Scrapie may not be considered in the differential diagnosis of neurological diseases in small ruminants, particularly in countries with low scrapie prevalence, or not recognized&#160;if it presents as nonpruritic form like atypical scrapie. To aid in the identification of clinical suspects, a short examination protocol is presented to assess the display of specific clinical signs associated with pruritic and nonpruritic forms of TSEs in sheep, which could also be applied to goats. This includes assessment of behavior, vision (by testing of the menace response), pruritus (by testing the response to scratching), and movement (with and without blindfolding). This may lead to a more detailed neurologic examination&#160;of reporting animals as scrapie suspects. It&#160;could also be used in experimental TSE studies of sheep or goats to evaluate disease progression or to identify clinical end-point.

A short clinical examination protocol is presented to aid in the detection of classical and atypical scrapie in sheep and goats.

羊の非定型および古典スククレープを検出する臨床検査プロトコル

The diagnosis of scrapie, a transmissible spongiform encephalopathy (TSEs) of sheep and goats, is currently based on the detection of disease-associated ...

Real-time Cytotoxicity Assays in Human Whole Blood

A live cell-based whole blood cytotoxicity assay (WCA) that allows access to temporal information of the overall cell cytotoxicity is developed with high-throughput cell positioning technology. The targeted tumor cell populations are first preprogrammed to immobilization into an array format, and labeled with green fluorescent cytosolic dyes. Following the cell array formation, antibody drugs are added in combination with human whole blood. Propidium iodide (PI) is then added to assess cell death. The cell array is analyzed with an automatic imaging system. While cytosolic dye labels the targeted tumor cell populations, PI labels the dead tumor cell populations. Thus, the percentage of target cancer cell killing can be quantified by calculating the number of surviving targeted cells to the number of dead targeted cells. With this method, researchers are able to access time-dependent and dose-dependent cell cytotoxicity information. Remarkably, no hazardous radiochemicals are used. The WCA presented here has been tested with lymphoma, leukemia, and solid tumor cell lines. Therefore, WCA allows researchers to assess drug efficacy in a highly relevant ex&#160;vivo condition.

The whole blood cytotoxicity assay (WCA) is a cytotoxicity assay developed by incorporating high-throughput cell positioning technology with fluorescence microscopy and automated image processing. Here, we describe how lymphoma cells treated with an anti-CD20 antibody can be analyzed real-time in human whole blood to provide quantitative cellular cytotoxicity analysis.

ヒト全血でのリアルタイム細胞毒性アッセイ

A live cell-based whole blood cytotoxicity assay (WCA) that allows access to temporal information of the overall cell cytotoxicity is developed with ...

Brain Source Imaging in Preclinical Rat Models of Focal Epilepsy using High-Resolution EEG Recordings

Electroencephalogram (EEG) has been traditionally used to determine which brain regions are the most likely candidates for resection in patients with focal epilepsy. This methodology relies on the assumption that seizures originate from the same regions of the brain from which interictal epileptiform discharges (IEDs) emerge. Preclinical models are very useful to find correlates between IED locations and the actual regions underlying seizure initiation in focal epilepsy. Rats have been commonly used in preclinical studies of epilepsy1; hence, there exist a large variety of models for focal epilepsy in this particular species. However, it is challenging to record multichannel EEG and to perform brain source imaging in such a small animal. To overcome this issue, we combine a patented-technology to obtain 32-channel EEG recordings from rodents2 and an MRI probabilistic atlas for brain anatomical structures in Wistar rats to perform brain source imaging. In this video, we introduce the procedures to acquire multichannel EEG from Wistar rats with focal cortical dysplasia, and describe the steps both to define the volume conductor model from the MRI atlas and to uniquely determine the IEDs. Finally, we validate the whole methodology by obtaining brain source images of IEDs and compare them with those obtained at different time frames during the seizure onset.

This video introduces the preparation, recording, and source analysis procedures of high-resolution EEG on sedated rats with a particular preclinical model of focal epilepsy under noninvasive conditions.

高解像度EEGレコーディングを使用して焦点てんかんの前臨床ラットモデルにおける脳ソースイメージング

Electroencephalogram (EEG) has been traditionally used to determine which brain regions are the most likely candidates for resection in patients with ...

Determination of the Relative Potency of an Anti-TNF Monoclonal Antibody (mAb) by Neutralizing TNF Using an In Vitro Bioanalytical Method

This protocol shows the measurement of the apoptotic activity neutralization of TNF&#945; in a mouse fibroblast cell model (WEHI 164) using an anti-TNF&#945; mAb. In addition, this protocol can be used to evaluate other anti-TNF&#945; molecules, such as fusion proteins. The cellular model employed here is sensitive to TNF&#945;-mediated apoptosis when an additional stress factor is induced in cell culture conditions (e.g., serum deprivation). This procedure exemplifies how to execute this analytical assay, highlighting the key operations relating to the sample preparation, cell dilution, apoptosis induction, and spectrophotometric measurements that are critical to ensure successful results. This protocol reveals the best-performance conditions relating to apoptosis induction and efficient signal recording, leading to low uncertainty values.

Determination of the Relative Potency of an Anti-TNF Monoclonal Antibody mAb by Neutralizing TNF Using an In Vitro Bioanalytical Method

A protocol for the determination of the relative anti-apoptotic activity of an anti-TNF&#945; mAb using a neutralization mechanism with WEHI 164 cells is presented here. This protocol is useful for comparing the neutralization strength of different molecules with the same biological functionality.

相対力、抗 TNF モノクローナル抗体 (mAb) 中和体外生物分析法を用いた TNF によって測定

This protocol shows the measurement of the apoptotic activity neutralization of TNF&#945; in a mouse fibroblast cell model (WEHI 164) using an ...

Assessment of Spontaneous Alternation, Novel Object Recognition and Limb Clasping in Transgenic Mouse Models of Amyloid-&#946; and Tau Neuropathology

Here we describe a staged, behavioral testing approach that can be used to screen for compounds that exhibit in vivo efficacy on cognitive and functional motor behaviors in transgenic mouse models of &#946;-amyloidosis and tauopathy. The paradigm includes tests for spontaneous alternation in a Y-maze, novel object recognition, and limb clasping. These tests were selected because they: 1) interrogate function of cognitive or motor domains and the correlate neural circuitry relevant to the human disease state, 2) have clearly defined endpoints, 3) have easily implementable quality control checks, 4) can be run in a moderate throughput format, and 5) require little intervention by the investigator. These methods are designed for investigators looking to screen compounds for activity in short-term and working memory tasks, or functional motor behaviors associated with Alzheimer's disease mouse models. The methods described here use behavioral tests that engage a number of different brain regions including hippocampus and various cortical areas. Investigators that desire cognitive tests that specifically assess cognition mediated by a single brain region could use these techniques to supplement other behavioral tests.

Assessment of Spontaneous Alternation, Novel Object Recognition and Limb Clasping in Transgenic Mouse Models of Amyloid-&amp;#946; and Tau Neuropathology

Described here is a staged, behavioral screening approach that can be used to screen for compounds that exhibit in vivo efficacy on cognitive and functional motor behaviors in transgenic mouse models of &#946;-amyloidosis and tauopathy. These methods are optimized to screen compounds for activity in short-term and working memory tasks.

アミロイド-βおよびタウのトランスジェニックマウスモデルにおける自発的交互作用の評価、新規対象の認識および肢の把握Neuropathology

Here we describe a staged, behavioral testing approach that can be used to screen for compounds that exhibit in vivo efficacy on cognitive and functional ...

The Pilocarpine Model of Temporal Lobe Epilepsy and EEG Monitoring Using Radiotelemetry System in Mice

Temporal lobe epilepsy (TLE) is a common neurological disorder in adulthood. For translational studies of chronic epilepsy, pilocarpine-induced status epilepticus (SE) is frequently selected to recapitulate spontaneous recurrent seizures (SRS). Here we present a protocol of SE induction by intraperitoneal (i.p.) injection of pilocarpine and monitoring of chronic recurring seizures in live animals using a wireless telemetry video and electroencephalogram (EEG) system. We demonstrated notable behavioral changes that need attention after pilocarpine injection and their correlation with hippocampal neuronal loss at 7 days and 6 weeks post-pilocarpine. We also describe the experimental procedures of electrode implantation for video and EEG recording, and analysis of the frequency and duration of chronic recurrent seizures. Finally, we discuss the possible reasons why the expected results are not achieved in each case. This provides a basic overview of modeling chronic epilepsy in mice and guidelines for troubleshooting. We believe this protocol can serve as a baseline for suitable models of chronic epilepsy and epileptogenesis.

This manuscript describes a method of inducing status epilepticus by systemic pilocarpine injection and of monitoring spontaneous recurrent seizures in live animals using a wireless telemetry video and electroencephalogram system. This protocol can be utilized for studying the pathophysiologic mechanisms of chronic epilepsy, epileptogenesis, and acute seizures.

側頭葉てんかんと脳波監視テレメーター システムを使用してマウスのピロカルピン モデル

Temporal lobe epilepsy (TLE) is a common neurological disorder in adulthood. For translational studies of chronic epilepsy, pilocarpine-induced status ...

A Training and Testing System for Performing Vascular Reconstruction In Vitro

Manual vascular reconstruction training is essential for a beginner surgeon. However, an optimal training system for vascular reconstruction in vitro has yet to be developed. In this study, we introduce an in vitro training and testing system using a magnetic anchoring technique with which a trainee can practice manual vascular reconstruction individually. Additionally, this system can also be used to test the quality of the reconstruction. The described system includes a vascular reconstruction training machine, magnetic tractors, and a magnetic suture puller. In this manuscript, we detail an end-to-end vein anastomosis using porcine right and left iliac veins. To identify the potential damage caused by a magnetic suture puller on the suture, we created three groups with six segments of 4-0 polypropylene sutures each: a control group with no intervention on the polypropylene suture, a group in which the polypropylene suture is manually pulled with sterile gloves 20x, and a magnetic puller group in which the magnetic puller pulled the polypropylene suture 20x. These groups were tested by light microscopy and breaking strength tests, and the effect of reconstruction was assessed. In the light microscopy test, the control group was less likely to be damaged (p &#60; 0.05) and the number of damaged points of the manual group and magnetic puller group were similar (p &#62; 0.05). The results of the breaking strength test were compared across groups and no significant difference was observed (p &#62; 0.05). The end-to-end anastomosis of the porcine iliac veins was successfully performed using this training system, and the reconstructed veins could undergo 2.0 kPa perfusion pressure. Using this training and testing system the trainee can practice manual vascular reconstruction in vitro individually with the aid of magnetic tractors and a magnetic suture puller, and the quality of the reconstruction can be tested.

Here we present a training and testing system where a trainee can complete manual vascular reconstruction in vitro individually using a magnetic anchoring technique. The system can also be used to test the quality of reconstruction.

インビトロにおける血管再建を行うためのトレーニングと検査システム

Manual vascular reconstruction training is essential for a beginner surgeon. However, an optimal training system for vascular reconstruction in vitro has ...

Advanced Cardiac Rhythm Management by Applying Optogenetic Multi-Site Photostimulation in Murine Hearts

Ventricular tachyarrhythmias are a major cause of mortality and morbidity worldwide. Electrical defibrillation using high-energy electric shocks is currently the only treatment for life-threatening ventricular fibrillation. However, defibrillation may have side-effects, including intolerable pain, tissue damage, and worsening of prognosis, indicating a significant medical need for the development of more gentle cardiac rhythm management strategies. Besides energy-reducing electrical approaches, cardiac optogenetics was introduced as a powerful tool to influence cardiac activity using light-sensitive membrane ion channels and light pulses. In the present study, a robust and valid method for successful photostimulation of Langendorff perfused intact murine hearts will be described based on multi-site pacing applying a 3 x 3 array of micro light-emitting diodes (micro-LED). Simultaneous optical mapping of epicardial membrane voltage waves allows the investigation of the effects of region-specific stimulation and evaluates the newly induced cardiac activity directly on-site. The obtained results show that the efficacy of defibrillation is strongly dependent on the parameters chosen for photostimulation during a cardiac arrhythmia. It will be demonstrated that the illuminated area of the heart plays a crucial role for termination success as well as how the targeted control of cardiac activity during illumination for modifying arrhythmia patterns can be achieved. In summary, this technique provides a possibility to optimize the on-site mechanism manipulation on the way to real-time feedback control of cardiac rhythm and, regarding the region specificity, new approaches in reducing the potential harm to the cardiac system compared to the usage of non-specific electrical shock applications.

This work reports a method for controlling the cardiac rhythm of intact murine hearts of transgenic channelrhodopsin-2 (ChR2) mice using local photostimulation with a micro-LED array and simultaneous optical mapping of epicardial membrane potential.

マウス心臓における光遺伝学的マルチサイト光刺激の適用による高度な心調律管理

Ventricular tachyarrhythmias are a major cause of mortality and morbidity worldwide. Electrical defibrillation using high-energy electric shocks is ...