プロトコルの提案や代表的なデータへの貢献のためにジェシカオットに感謝。このプロトコルを立証するためのサブリナBlackledge、ローレンコホーテク彗星とKerisaシェルトンに感謝。

1。設定<ol><li>すべての研究は、適切な治験審査委員会またはデータ収集の前に、ヒトを対象に審査委員会によって承認されるべきである。 </li><li>理想的には、2つの部屋がTSSTを実施できるようにする必要があります。快適な待合室にはTSSTを開始し、回復期間中に前に参加者が利用できるはずです。別のインタビュールームは、新規性とuncontrolabilityを紹介するスピーチの準備、スピーチ性能とTSSTの数学の部分で使用する必要があります。 </li><li>参加者は南軍の面接と参加者の視野内の位置のビデオまたはオーディオ記録装置が直面されるようにインタビュールームを手配する。面接はTSST時の応力を増加させるTSST中に着用するために白衣が利用可能になります。参加者とアイコンタクトを維持し、TSSTの間に感情的な表情を慎む面接官に指示します。 </li><li>慎重に目を考慮するeの研究の質問（複数可）に依存する測定値を選択する。 ：ストレス反応の種々の測定の分析と公開のためのガイドラインは、いくつかのソース（精神生理学的指標の公表および分析のためのガイドラインは、精神生理学研究協会によって提供されることで提供されています<a href="http://www.sprweb.org/journal/index.cfm#guidelines" target="_blank">http://www.sprweb.org/journal/index。 CFM＃のガイドライン</a> ）。実験の設計時にこれらのリソースの5の相談が推奨されます。 </li><li> TSSTへの生理的、心理的または内分泌応答を記録するために必要なすべての機器をセットアップします。この装置は、心拍数、心血管反応性、皮膚コンダクタンスまたは応答、瞬目、筋収縮/トーン、呼吸数を記録する（ただし、これらに限定されるものではない）電極と電子機器および/またはソフトウェアが含まれる場合があります、の紙と鉛筆または電子版をこのようなInv状態特性不安など心理的な楽器entory、6管や尿、血液や唾液サンプルの他のコレクションの材料;またはビデオの録画機器では、 このデモでは、我々は（ECG）TSST時の心拍数を測定するための電極、および反復投与3リード心電図を使用することになりますストレッサーの前と後の心理的反応を測定するために、状態特性不安インベントリー。私たちは、心拍数を記録するために生理学的反応を測定するために設計された基本的な記録システム（BIOPAC MP36システム）を活用していきます。 </li><li>血液サンプル（ではなく、唾液）が実験中に収集される場合は、収集時に痛みや苦痛を最小限にする努力がなされるべきである。採血の手順は、追加の物理的または心理的ストレス、TSSTに交絡応答を導入することができます。 </li><li>参加者が到着すると、研究室にそれらを歓迎する。参加者は快適にインフォームドコンセントと席を入手。 </li><li>人口統計情報、スクリーニング情報やadditioを収集するあなたの実験で必要とされるNAL到着対策。コルチゾールの測定値を収集した場合、参加者が過去1時間で、食品または飲料の（水以外の）消費していないことを確認します。 空腹時の短い期間は、分析のための唾液のサンプルを収集する場合は推奨されます。酸性の食べ物や飲み物や目に見える汚染に伴うpHの変化は、唾液マーカーの正確な測定の妨げになることがあります。興味のある措置が薬や他の代謝物によって汚染される恐れがある場合に尿の薬物中毒のスクリーニング検査が必要になることがあります。時スクリーニングの参加者はストレス応答（例えば、薬、病気、睡眠、最近のストレスの多い出来事、社会恐怖）に影響を与える可能性のある要因についての情報も考慮する必要があります。 </li></ol> 2。プレストレスの測定<ol><li>待合室で45分間快適に待つよう参加者に指示します。あなたはエモを含む雑誌など、資料を読んで参加者を提供することもできますtionally中立的なコンテンツ、またはこの期間中に別の感情を伴わない活動。 </li><li> 45分の待機期間の後、プレストレス不安を評価するために状態特性不安インベントリーを管理する6 また、ストレス、不安やリラクゼーションの状態の変化を評価するために設計されている他の機器（例えば視覚的アナログ尺度）に投与することができる。 </li><li> &quot;プレストレス&quot;唾液のサンプルを収集する。水で口をすすぐために参加者に指示します。参加者にストローの5-10 cmのセクションでクリーンな、使い捨て2.0ミリリットルのポリプロピレンバイアルを与える。その後バイアル瓶にストローで唾液を通過口の床にプールに唾液を可能にするために前方に頭を傾ける参加者に指示します。参加者からの唾液のおよそ1.5-2.0 mlを収集する。また、唾液を吸収歯の芯を（または類似の材料）を使用し、唾液と材料をかむまたは飽和する参加者を指示することで収集できます。 POSの場合すい回路プライバシーと参加者を提供するために部屋を残す。ラベルと、できるだけ早く-20℃ですべての唾液のサンプルを格納する。 </li><li>プレストレスの生理学的測定値の5分を記録します。 </li></ol> 3。 TSST <ol><li>別のインタビュールーム（TSSTのストレスの効果を高めるために ）使用する場合は、面接の部屋に待っている部屋からの参加者をもたらす。 </li><li> TSSTスピーチの準備期間を開始するには、参加者に次のスクリプトを読んで：&quot;これはタスクのスピーチの準備の部分である、あなたは精神的に、あなたの理想的な仕事のための良い候補となる理由を説明する5分間のスピーチを準備することです。あなたのスピーチは人前で話すの訓練を受けた裁判官のパネルによって録画して検討されます。あ​​なたは準備に10分を持っているとあなたの時間が今始まる。&quot; </li><li> 10分のスピーチの準備期間中に生理的測定値​​を記録します。 10分間のデジタルタイマーを設定し、部屋を残す。 </LI&gt; <li> 10分後、白​​衣を着て部屋に戻る。参加者に次のスクリプトを読む：&quot;これはタスクの音声部分であるあなたは、あなたの理想的な仕事のための良い候補となる理由を説明する音声を配信するために使われるかは、全体の5分間の期間のために話してください。。。あなたの時間が始まります。&quot;プロップのビデオカメラは、性能/評価的なストレスを高めるために、現在オンにする必要があります。 </li><li> 5分間のスピーチのパフォーマンス期間中に生理的測定値​​を記録します。参加者が演説中に話して停止した場合、彼または彼女が20秒間沈黙することができます。彼または彼女が話して再開されない場合は、それらを指示することで言えば継続して参加者を促す：&quot;あなたは、まだ残り時間を持っている。&quot; </li><li> 5分間のスピーチのパフォーマンス期間の終わりに、参加者に次のスクリプトを読んで：&quot;あなたは順番に1022から番号13を減算するように求められます、このタスクの最後の5分間の数学の部分であなたがなる言葉。LYは声を出してあなたの答えを報告し、ミスをした場合は1022からやり直すように頼まれる。あなたの時間が今始まる&quot;参加者がミスをした場合、それらをプロンプト：&quot;。。それは間違っている、1022からやり直すください&quot;5分間のデジタルタイマーを設定します。 </li><li> 5分の数学の性能の期間中に生理的測定値​​を記録します。 </li><li> TSSTの数学の業績期間の終了時に、&quot;ポストストレス&quot;唾液のサンプルを収集する。 </li><li>後ストレス状態特性不安インベントリーを管理する。 </li></ol> 4。後ストレス回復測定<ol><li>オリジナルの待合室に参加者を返します。最初の回復期間中に、参加者は20分間快適に待つように指示。 20分間のデジタルタイマーを設定し、部屋を残す。 </li><li> 20分後に待合室に戻り、&quot;回復1&quot;唾液のサンプルを収集する。 </li><li> 2番目の回収期間中、参加者が再び20分間快適に待つように指示。 20分間のデジタルタイマーを設定し、部屋を残す。 </li><li> 20分後に待合室に戻ると、&quot;回復2&quot;唾液のサンプルを収集する。 </li><li> &quot;回復&quot;状態特性不安インベントリーを完了するために参加者に指示します。 </li><li>実験の真の性質として報告を受ける参加者。彼らのパフォーマンスが記録されていないことと、その音声または演算性能のない分析が実施されないことを参加者に通知します。彼らが提示されたときのタスクが不当に困難であったことを参加者に説明し、参加者の適性や能力に反映するものではありません。 </li></ol> 5。データ解析<ol><li>心理的な対策を分析するには、プレストレス、後のストレスと回復時間の点から各状態特性不安インベントリーを獲得。6 </li><li>生理的な対策を分析するために、&quot;、&quot;&quot;、プレストレス&quot;&quot;スピーチのパフォーマンス&quot;と&quot;mをスピーチの準備を5分ごとの平均心拍数のスコアを計算する&quot;TSSTの期間をATHを。また、TSSTの間に、離散時間の点でTSSTまたは測定心拍数の毎分平均スコアを計算する。 </li><li>神経内分泌対策、市販のEIAキットを使用して、ホルモン濃度のアッセイ唾液のサンプルを分析することができます。 </li></ol> 6 オプション：追加の唾液中コルチゾールアッセイプロトコル情報（Salimetrics唾液中コルチゾールの測定キットのインサートから再生） <ol><li>室温にすべての唾液アッセイキットの試薬をもたらす。 </li><li>キットの指示にしたがって洗浄バッファーを準備します。 </li><li>重複の適切なウェルにプレートレイアウトと標準とコントロールのピペット25μLを決定します。 </li><li> 15分、1,500 XG（@ 3000 RPM）で融解唾液のサンプルを、ボルテックス、遠心。 </li><li>プレートの適当なウェル（重複の）に遠心分離試料から、透明な液体のピペット25μL。 </li><li>すべてのウェルに酵素複合体とピペット200μLを準備します。 </li&gt; <li> 500rpmで5分間プレートをミックスし、さらに55分間インキュベートする。 </li><li>洗浄バッファーで4回洗浄し、プレートを覆い隠す。 </li><li>マルチチャンネルピペットを用いて各ウェルに200μLのテトラメチルベンジジン（TMB）溶液を加える。 </li><li> 500rpmで5分間プレートをミックスし、さらに25分間暗所でインキュベートする。 </li><li>反応を停止するには、500rpmで3分間各ウェルとのミックスに50μLの反応停止液を追加します。 </li><li>プレートの底が停止溶液を添加して10分以内に450nmでプレートリーダーでプレートを洗浄し、読んで拭いてください。 </li><li>コルチゾールの濃度を計算するには、すべてのウェルの平均光学濃度を計算する。ゼロ、標準、コントロールとサンプルウェルの平均光学濃度から非特異的結合のウェルの平均光学濃度を引きます。 </li><li> AVERでこれらのウェルの平均光学濃度（B）を分周して各標準、コントロールとサンプルのためにバインドされたパーセント（B / Boを）計算するゼロ井戸（ボー）の年齢光学密度。物流が可能なソフトウェアを使用して補間することによって、コントロールとサンプルの濃度を決定する。 </li></ol> 7。代表的な結果： ここで紹介する代表的なデータは、文献の見直し、健康な成人男性の参加者との午後の間に私たちのラボで実施された二つの研究からコンパイルされます。これらのデータは、約この記事で紹介したプロトコルを使用して検出される可能性があります結果を示すことを意図したものである。 、心拍数（図3）、および唾液中コルチゾール濃度（図4）。このTSSTプロトコール（図1）自己申告不安（図2のような状態特性不安インベントリーによる測定）の増加を誘発する。図1に示す実験プロトコルは、TSSTでデータ収集セッション中に行われる可能性がありますどのように種々の測定の一例を提供することを意味する。 measuremenのタイミングTSと異なる措置を含めることは、特定の研究課題によって決定されます。不安および/またはストレス応答を軽減するために意図されている介入を使用している場合、これらの対策の鈍化や減衰の増加が予測されています。 <img alt="図1" src="/files/ftp_upload/3238/3238fig1.jpg" /> 図1。実験的なプロトコル （注）。トリーアの社会ストレステスト（TSST）、状態特性不安インベントリー（STAI）、5分前ストレスの期間（PS）、10分の予期ストレス期間（スピーチの準備）（AS）、5分間のスピーチの期間（S）、5分の数学の期間（M）。 <img alt="図2" src="/files/ftp_upload/3238/3238fig2.jpg" /> 図2。代表的な状態不安の得点注：状態不安スコアは状態特性不安インベントリー（STAI）の20 itenの状態不安の指標から算出した代表的なスコアが私たちの研究室で収集し、文献で ​​公開されたデータを反映。。 図3。代表的な心拍数のデータ 注 ：。。分間の拍数（BPM）の心拍代表値は、私たちの研究室で収集し、文献で ​​公開されたデータを反映する。 <img alt="図4" src="/files/ftp_upload/3238/3238fig4.jpg" /> 図4。代表的な唾液中コルチゾールのデータ 注 ：。。UL / DLのコルチゾール濃度代表値は、私たちの研究室で収集し、文献で ​​公開されたデータを反映している。

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J., Joober, R., Lupien, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Education+modulates+cortisol+reactivity+to+the+Trier+Social+Stress+Test+in+middle-aged+adults.">Education modulates cortisol reactivity to the Trier Social Stress Test in middle-aged adults.</a> Psychoneuroendocrinology. 32, 1158-1163 (2007).</li><li>Oswald, L. M., Zandi, P., Nestadt, G., Potash, J. B., Kalaydjian, A. E., Wand, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relationship+between+cortisol+responses+to+stress+and+personality.">Relationship between cortisol responses to stress and personality.</a> Neuropsychopharmacology. 31, 1583-1591 (2006).</li><li>Fries, E., Hellhammer, D. H., Hellhammer, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Attenuation+of+the+hypothalamic-pituitary-adrenal+axis+responsivity+to+the+Trier+Social+Stress+Test+by+the+benzodiazepine+alprazolam.">Attenuation of the hypothalamic-pituitary-adrenal axis responsivity to the Trier Social Stress Test by the benzodiazepine alprazolam.</a> Psychoneuroendocrinology. 31, 1278-1288 (2006).</li><li>Kudielka, B. M., Fischer, J. E., Metzenthin, P., Helfricht, S., Preckel, D., von K&#228;nel, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=No+effect+of+5-day+treatment+with+acetylsalicylic+acid+(aspirin)+or+the+beta-blocker+propranolol+(Inderal)+on+free+cortisol+responses+to+acute+psychosocial+stress:+A+randomized+double-blind,+placebo-controlled+study.">No effect of 5-day treatment with acetylsalicylic acid (aspirin) or the beta-blocker propranolol (Inderal) on free cortisol responses to acute psychosocial stress: A randomized double-blind, placebo-controlled study.</a> Neuropsychobiology. 56, 159-166 (2007).</li><li>Gaab, J., Bl&#228;ttler, N., Menzl, T., Pabst, B., Stoyer, S., Ehlert, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Randomized+controlled+evaluation+of+the+effects+of+cognitive-behavioral+stress+management+on+cortisol+responses+to+acute+stress+in+healthy+subjects.">Randomized controlled evaluation of the effects of cognitive-behavioral stress management on cortisol responses to acute stress in healthy subjects.</a> Psychoneuroendocrinology. 28, 767-779 (2003).</li><li>Khalfa, S., Bella, S. D., Roy, M., Peretz, I., Lupien, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+relaxing+music+on+salivary+cortisol+level+after+psychological+stress.">Effects of relaxing music on salivary cortisol level after psychological stress.</a> Ann. NY Acad. Sci. 999, 374-376 (2003).</li><li>Balodis, I. M., Wynne-Edwards, K. E., Olmstead, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+other+side+of+the+curve:+Examining+the+relationship+between+pre-stressor+physiological+responses+and+stress+reactivity.">The other side of the curve: Examining the relationship between pre-stressor physiological responses and stress reactivity.</a> Psychoneuroendocrinology. 35, 1363-1373 (2010).</li><li>Shultheiss, O. C., Stanton, S. J., Harmon-Jones, E., Beer, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+salivary+hormones.">Assessment of salivary hormones.</a> Methods in Social Neuroscience. , 17-44 (2009).</li><li>Kudielka, B. M., Hellhammer, D. H., W&#252;st, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Why+do+we+respond+so+differently?+Reviewing+determinants+of+human+salivary+cortisol+responses+to+challenge.">Why do we respond so differently? Reviewing determinants of human salivary cortisol responses to challenge.</a> Psychoneuroendocrinology. 34, 2-18 (2009).</li><li>Buske-Kirschbaum, A., Jobst, S., Wustmans, A., Kirschbaum, C., Rauh, W., Hellhammer, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Attenuated+free+cortisol+response+to+psychosocial+stress+in+children+with+atopic+dermatitis.">Attenuated free cortisol response to psychosocial stress in children with atopic dermatitis.</a> Psychosom. Med. 59, 419-426 (1997).</li></ol>

利害の衝突は宣言されません。

この記事では、我々は健康なボランティアのトリーアの社会ストレステストを実施する方法を示しました。 TSSTは、心理学的、生理学的および神経内分泌対策に堅牢で信頼性の増加を誘導する標準化された実験室社会的ストレス因子です。 TSSTは、コールド押えテストまたはトレッドミル歩行における物理的なストレス因子への便利な代替である、と評価的聴衆の存在下で性能のより自然な心...

<table border="1"><tbody><tr><th>機器/サプライの名前</th><th>会社</th><th>カタログ番号</th></tr><tr><td> BSL精神生理学システム、MP36データアクイジションユニット三誘導心電図の電極など、Mac OS </td><td>モンテシステム株式会社</td><td> BSLPSY - M </td></tr><tr><td>状態特性不安インベントリー</td><td>マインドガーデン、（株） </td><td>落ち着いた- B </td></tr><tr><td>唾液中コルチゾールの酵素免疫アッセイキット</td><td> Salimetrics </td><td> 1から3002 </td></tr><tr><td> 2.0ミリリットルポリプロピレンバイアル</td><td>フィッシャーサイエンティフィック</td><td> 05 40B 146 </td></tr><tr><td> BioRad社マイクロプレートリーダー、マイクロManagerソフトウェアとモデル680、 </td><td> BioRad社</td><td>プレートリーダー：168 1000ソフトウェア：1706800 </td></tr></tbody></table>

the trier social stress test protocol for inducing psychological stress

この記事では、実験室の設定で使用するための心理的ストレスのプロトコルを示しています。研究者が健康と病気におけるストレス応答の生物学的経路を研究するためのプロトコルは、ストレスや不安の研究の進展の基本です1。多数のプロトコルが研究室でのストレス応答を誘導するために存在しますが、多くの放置は自然な文脈を提供したり、社会的、心理的ストレスの側面を組み込むことができます。心理的ストレスのプロトコルのうち、メタアナリシスでは、トリーアの社会ストレステスト（TSST）がストレスホルモンの反応性の研究のための最も有用かつ適切な標準化されたプロトコルである。TSSTのオリジナルの説明2、研究者が手順を設計し、評価するために求められることを示唆している健康なボランティアの大部分で信頼性の高いストレス反応を誘発することができる。3これらの研究は、心拍数、血圧およびいくつかの内分泌ストレスmの標高を発見TSSTへの応答でarkers（心理的ストレッサー）は生理食塩水注入（物理的ストレッサー）に比べて3を TSSTは、様々な研究グループのニーズに合わせて変更されているが、それは一般的には到着時に待機期間、先行発話で構成されています一つ以上の回復期間が続くの準備、スピーチのパフォーマンス、および口頭で演算パフォーマンス期間、。 TSSTは、参加者がスピーチを準備し、提供する必要があり、口頭で社会的に評価的な聴衆の存在下で挑戦的な算術の問題に対応する。3社会の評価と制御不能の状態がTSSTによるストレス誘導の主要コンポーネントとして同定されている。4をするために使用で十年間、TSSTの目的は、体系的にタスクの実行中に視床下部 - 下垂体 - 副腎（HPA）または交感神経 - 副腎髄質-（SAM）軸の反応性、不安と活性化の違いを測定するために、ストレス応答を誘導することです。 1 Researc彼女は一般的にTSSTに反応して自己申告不安、生理的な措置（例えば、心拍数）、および/または神経内分泌指標（ストレスホルモンのコルチゾールなど）の変化を評価する。多くの研究者らは、血液収集技術の交絡ストレスを軽減するために血液採取に代わるものとして、コルチゾールおよびα-アミラーゼ（自律神経系の活性化のマーカー）などのストレスマーカーのために唾液のサンプリングを採用している。 TSSTを完了する個々が経験する変化に加えて、研究者は、異なる治療群（例えば、臨床対健康な対照試料）または介入のストレス軽減の効果との間の変化を比較することができます。1

Watch this Scientific Journal Video about The Trier Social Stress Test Protocol for Inducing Psychological Stress at JoVE.com

The Trier Social Stress Test Protocol for Inducing Psychological Stress

この記事では、単一の参加者内やグループ間のストレスに対する心理学的、生理学的および神経内分泌反応を測定する研究者を可能にする参加者、で精神的ストレスを誘導するためのプロトコルについて説明します。

心理的ストレスを誘導するためのトリーアの社会ストレステストプロトコル

This article demonstrates a psychological stress protocol for use in a laboratory setting.  Protocols that allow researchers to study the biological ...

the-trier-social-stress-test-protocol-for-inducing-psychological

Research

JoVE Journal

Medicine

55.6K ビュー.  Northern Arizona University. この記事では、単一の参加者内やグループ間のストレスに対する心理学的、生理学的および神経内分泌反応を測定する研究者を可能にする参加者、で精神的ストレスを誘導するためのプロトコルについて説明します。

ビデオ: The Trier Social Stress Test Protocol for Inducing Psychological Stress

Using Visual and Narrative Methods to Achieve Fair Process in Clinical Care

The Institute of Medicine has targeted patient-centeredness as an important area of quality improvement. A major dimension of patient-centeredness is respect for patient's values, preferences, and expressed needs. Yet specific approaches to gaining this understanding and translating it to quality care in the clinical setting are lacking. From a patient perspective quality is not a simple concept but is best understood in terms of five dimensions: technical outcomes; decision-making efficiency; amenities and convenience; information and emotional support; and overall patient satisfaction. Failure to consider quality from this five-pronged perspective results in a focus on medical outcomes, without considering the processes central to quality from the patient's perspective and vital to achieving good outcomes. In this paper, we argue for applying the concept of fair process in clinical settings. Fair process involves using a collaborative approach to exploring diagnostic issues and treatments with patients, explaining the rationale for decisions, setting expectations about roles and responsibilities, and implementing a core plan and ongoing evaluation. Fair process opens the door to bringing patient expertise into the clinical setting and the work of developing health care goals and strategies. This paper provides a step by step illustration of an innovative visual approach, called photovoice or photo-elicitation, to achieve fair process in clinical work with acquired brain injury survivors and others living with chronic health conditions. Applying this visual tool and methodology in the clinical setting will enhance patient-provider communication; engage patients as partners in identifying challenges, strengths, goals, and strategies; and support evaluation of progress over time. Asking patients to bring visuals of their lives into the clinical interaction can help to illuminate gaps in clinical knowledge, forge better therapeutic relationships with patients living with chronic conditions such as brain injury, and identify patient-centered goals and possibilities for healing. The process illustrated here can be used by clinicians, (primary care physicians, rehabilitation therapists, neurologists, neuropsychologists, psychologists, and others) working with people living with chronic conditions such as acquired brain injury, mental illness, physical disabilities, HIV/AIDS, substance abuse, or post-traumatic stress, and by leaders of support groups for the types of patients described above and their family members or caregivers.

This paper illustrates an innovative visual approach (photovoice or photo-elicitation) to achieve fair process in clinical care for patients living with chronic health conditions, illuminate gaps in clinical knowledge, forge better therapeutic relationships, and identify patient-centered goals and possibilities for healing.

臨床ケアのフ​​ェアプロセスを達成するために、Visualと物語メソッドの使用

The Institute of Medicine has targeted patient-centeredness as an important area of quality improvement. A major dimension of patient-centeredness is ...

医学

Development of a Unilaterally-lesioned 6-OHDA Mouse Model of Parkinson's Disease

The unilaterally lesioned 6-hyroxydopamine (6-OHDA)-lesioned rat model of Parkinson's disease (PD) has proved to be invaluable in advancing our understanding of the mechanisms underlying parkinsonian symptoms, since it recapitulates the changes in basal ganglia circuitry and pharmacology observed in parkinsonian patients1-4. However, the precise cellular and molecular changes occurring at cortico-striatal synapses of the output pathways within the striatum, which is the major input region of the basal ganglia remain elusive, and this is believed to be site where pathological abnormalities underlying parkinsonian symptoms arise3,5.
In PD, understanding the mechanisms underlying changes in basal ganglia circuitry following degeneration of the nigro-striatal pathway has been greatly advanced by the development of bacterial artificial chromosome (BAC) mice over-expressing green fluorescent proteins driven by promoters specific for the two striatal output pathways (direct pathway: eGFP-D1; indirect pathway: eGFP-D2 and eGFP-A2a)8, allowing them to be studied in isolation. For example, recent studies have suggested that there are pathological changes in synaptic plasticity in parkinsonian mice9,10. However, these studies utilised juvenile mice and acute models of parkinsonism. It is unclear whether the changes described in adult rats with stable 6-OHDA lesions also occur in these models. Other groups have attempted to generate a stable unilaterally-lesioned 6-OHDA adult mouse model of PD by lesioning the medial forebrain bundle (MFB), unfortunately, the mortality rate in this study was extremely high, with only 14% surviving the surgery for 21 days or longer11. More recent studies have generated intra-nigral lesions with both a low mortality rate &gt;80% loss of dopaminergic neurons, however expression of L-DOPA induced dyskinesia11,12,13,14 was variable in these studies. Another well established mouse model of PD is the MPTP-lesioned mouse15. Whilst this model has proven useful in the assessment of potential neuroprotective agents16, it is less suitable for understanding mechanisms underlying symptoms of PD, as this model often fails to induce motor deficits, and shows a wide variability in the extent of lesion17, 18.
Here we have developed a stable unilateral 6-OHDA-lesioned mouse model of PD by direct administration of 6-OHDA into the MFB, which consistently causes &gt;95% loss of striatal dopamine (as measured by HPLC), as well as producing the behavioural imbalances observed in the well characterised unilateral 6-OHDA-lesioned rat model of PD. This newly developed mouse model of PD will prove a valuable tool in understanding the mechanisms underlying generation of parkinsonian symptoms.

Development of a Unilaterally-lesioned 6-OHDA Mouse Model of Parkinson&#039;s Disease

A protocol for performing unilateral 6-OHDA lesions of the medial forebrain bundle in mice is described. This method has a low mortality rate (13.3 %) with 89% of the surviving animals showing &#62;95% loss of striatal dopamine and 90.63&#177;-4.02 % ipsiversive rotational bias towards the side of the lesion.

パーキンソン病の単方向、病変6-OHDAモデルマウスの開発

The unilaterally lesioned 6-hyroxydopamine (6-OHDA)-lesioned rat model of Parkinson's disease (PD) has proved to be invaluable in advancing our ...

Implantation of a Carotid Cuff for Triggering Shear-stress Induced Atherosclerosis in Mice

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce atherosclerosis (reviewed in 1 and 2). The role of shear stress has been extensively studied in vitro investigating the influence of flow dynamics on cultured endothelial cells 1,3,4 and in vivo in larger animals and humans 1,5,6,7,8. However, highly reproducible small animal models allowing systematic investigation of the influence of shear stress on plaque development are rare. Recently, Nam et al. 9 introduced a mouse model in which the ligation of branches of the carotid artery creates a region of low and oscillatory flow. Although this model causes endothelial dysfunction and rapid formation of atherosclerotic lesions in hyperlipidemic mice, it cannot be excluded that the observed inflammatory response is, at least in part, a consequence of endothelial and/or vessel damage due to ligation.
In order to avoid such limitations, a shear stress modifying cuff has been developed based upon calculated fluid dynamics, whose cone shaped inner lumen was selected to create defined regions of low, high and oscillatory shear stress within the common carotid artery 10. By applying this model in Apolipoprotein E (ApoE) knockout mice fed a high cholesterol western type diet, vascular lesions develop upstream and downstream from the cuff. Their phenotype is correlated with the regional flow dynamics 11 as confirmed by in vivo Magnetic Resonance Imaging (MRI) 12: Low and laminar shear stress upstream of the cuff causes the formation of extensive plaques of a more vulnerable phenotype, whereas oscillatory shear stress downstream of the cuff induces stable atherosclerotic lesions 11. In those regions of high shear stress and high laminar flow within the cuff, typically no atherosclerotic plaques are observed.
 
In conclusion, the shear stress-modifying cuff procedure is a reliable surgical approach to produce phenotypically different atherosclerotic lesions in ApoE-deficient mice.

The constricting cuff presented in this article is designed to induce atherosclerosis in the murine common carotid artery. Due to the conical shape of its inner lumen the implanted cuff generates well-defined regions of low, high and oscillatory shear stress triggering the development of atherosclerotic lesions of different inflammatory phenotypes.

マウスでのせん断応力誘発アテローム性動脈硬化症をトリガするための頸動脈カフの注入

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce ...

Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for objective diagnosis but also for the evaluation of therapeutic efficacy and resilience to trauma. Ongoing research is directed at identifying molecular biomarkers for PTSD, including traumatic stress induced proteins, transcriptomes, genomic variances and genetic modulators, using biologic samples from subjects' blood, saliva, urine, and postmortem brain tissues. However, the correlation of these biomarker molecules in peripheral or postmortem samples to altered brain functions associated with psychiatric symptoms in PTSD remains unresolved. Here, we present an animal model of PTSD in which both peripheral blood and central brain biomarkers, as well as behavioral phenotype, can be collected and measured, thus providing the needed correlation of the central biomarkers of PTSD, which are mechanistic and pathognomonic but cannot be collected from people, with the peripheral biomarkers and behavioral phenotypes, which can.
Our animal model of PTSD employs restraint and tail shocks repeated for three continuous days - the inescapable tail-shock model (ITS) in rats. This ITS model mimics the pathophysiology of PTSD 17, 7, 4, 10. We and others have verified that the ITS model induces behavioral and neurobiological alterations similar to those found in PTSD subjects 17, 7, 10, 9. Specifically, these stressed rats exhibit (1) a delayed and exaggerated startle response appearing several days after stressor cessation, which given the compressed time scale of the rat's life compared to a humans, corresponds to the one to three months delay of symptoms in PTSD patients (DSM-IV-TR PTSD Criterian D/E 13), (2) enhanced plasma corticosterone (CORT) for several days, indicating compromise of the hypothalamopituitary axis (HPA), and (3) retarded body weight gain after stressor cessation, indicating dysfunction of metabolic regulation.
The experimental paradigms employed for this model are: (1) a learned helplessness paradigm in the rat assayed by measurement of acoustic startle response (ASR) and a charting of body mass; (2) microdissection of the rat brain into regions and nuclei; (3) enzyme-linked immunosorbent assay (ELISA) for blood levels of CORT; (4) a gene expression microarray plus related bioinformatics tools 18. This microarray, dubbed rMNChip, focuses on mitochondrial and mitochondria-related nuclear genes in the rat so as to specifically address the neuronal bioenergetics hypothesized to be involved in PTSD.

We describe a rat model of post traumatic stress disorder (PTSD) that reveals the persistent alterations in neuroendocrine function and the delayed long-term, exaggerated fear response, characteristic of PTSD patients. The animal model and methods described here are useful for correlating biomarkers in brain nuclei, which are mechanistic but cannot be measured in patients, with biomarkers in peripheral white blood cells, which can.

PTSDの、ニューラル血液学的および行動的相関関係を明らかにするための動物モデルにおけるバイオマーカー

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for ...

Cardiac Stress Test Induced by Dobutamine and Monitored by Cardiac Catheterization in Mice

Dobutamine is a &beta;-adrenergic agonist with an affinity higher for receptor expressed in the heart (&beta;1) than for receptors expressed in the arteries (&beta;2). When systemically administered, it increases cardiac demand. Thus, dobutamine unmasks abnormal rhythm or ischemic areas potentially at risk of infarction. 
Monitoring of heart function during a cardiac stress test can be performed by either ecocardiography or cardiac catheterization. The latter is an invasive but more accurate and informative technique that the former.
Cardiac stress test induced by dobutamine and monitored by cardiac catheterization accomplished as described here allows, in a single experiment, the measurement of the following hemodynamic parameters: heart rate (HR), systolic pressure, diastolic pressure, end-diastolic pressure, maximal positive pressure development (dP/dtmax) and maximal negative pressure development (dP/dtmin), at baseline conditions and under increasing doses of dobutamine.
As expected, in normal mice we observed a dobutamine dose-related increase in HR, dP/dtmax and dP/dtmin. Moreover, at the highest dose tested (12 ng/g/min) the cardiac decompensation of high fat diet-induced obese mice was unmasked.

We describe the protocol to perform a cardiac stress test induced by dobutamine and monitored by cardiac catheterization in normal mice. Also we show its application to unmask subclinical cardiac disease in high fat diet-induced obese mice.

心臓ストレステストドブタミンによって誘導され、マウスにおける心臓カテーテル検査によって監視される

Dobutamine is a &beta;-adrenergic agonist with an affinity higher for receptor expressed in the heart (&beta;1) than for receptors expressed in the ...

Reduction in Left Ventricular Wall Stress and Improvement in Function in Failing Hearts using Algisyl-LVR

Injection of Algisyl-LVR, a treatment under clinical development, is intended to treat patients with dilated cardiomyopathy. This treatment was recently used for the first time in patients who had symptomatic heart failure. In all patients, cardiac function of the left ventricle (LV) improved significantly, as manifested by consistent reduction of the LV volume and wall stress. Here we describe this novel treatment procedure and the methods used to quantify its effects on LV wall stress and function.
Algisyl-LVR is a biopolymer gel consisting of Na+-Alginate and Ca2+-Alginate. The treatment procedure was carried out by mixing these two components and then combining them into one syringe for intramyocardial injections. This mixture was injected at 10 to 19 locations mid-way between the base and apex of the LV free wall in patients.
Magnetic resonance imaging (MRI), together with mathematical modeling, was used to quantify the effects of this treatment in patients before treatment and at various time points during recovery. The epicardial and endocardial surfaces were first digitized from the MR images to reconstruct the LV geometry at end-systole and at end-diastole. Left ventricular cavity volumes were then measured from these reconstructed surfaces.
Mathematical models of the LV were created from these MRI-reconstructed surfaces to calculate regional myofiber stress. Each LV model was constructed so that 1) it deforms according to a previously validated stress-strain relationship of the myocardium, and 2) the predicted LV cavity volume from these models matches the corresponding MRI-measured volume at end-diastole and end-systole. Diastolic filling was simulated by loading the LV endocardial surface with a prescribed end-diastolic pressure. Systolic contraction was simulated by concurrently loading the endocardial surface with a prescribed end-systolic pressure and adding active contraction in the myofiber direction. Regional myofiber stress at end-diastole and end-systole was computed from the deformed LV based on the stress-strain relationship.

This article describes procedures for implanting a novel hydrogel in failing hearts and quantifying its effect on left ventricular wall stress and function. These procedures have been successfully applied in dogs and humans.

Algisyl-LVRを使っハーツ失敗関数内で左心室壁応力と改善の削減

Injection of  Algisyl-LVR, a treatment under clinical development, is intended to treat  patients with dilated cardiomyopathy. This treatment was recently ...

A Mouse Tumor Model of Surgical Stress to Explore the Mechanisms of Postoperative Immunosuppression and Evaluate Novel Perioperative Immunotherapies

Surgical resection is an essential treatment for most cancer patients, but surgery induces dysfunction in the immune system and this has been linked to the development of metastatic disease in animal models and in cancer patients. Preclinical work from our group and others has demonstrated a profound suppression of innate immune function, specifically NK cells in the postoperative period and this plays a major role in the enhanced development of metastases following surgery. Relatively few animal studies and clinical trials have focused on characterizing and reversing the detrimental effects of cancer surgery. Using a rigorous animal model of spontaneously metastasizing tumors and surgical stress, the enhancement of cancer surgery on the development of lung metastases was demonstrated. In this model, 4T1 breast cancer cells are implanted in the mouse mammary fat pad. At day 14 post tumor implantation, a complete resection of the primary mammary tumor is performed in all animals. A subset of animals receives additional surgical stress in the form of an abdominal nephrectomy. At day 28, lung tumor nodules are quantified. When immunotherapy was given immediately preoperatively, a profound activation of immune cells which prevented the development of metastases following surgery was detected. While the 4T1 breast tumor surgery model allows for the simulation of the effects of abdominal surgical stress on tumor metastases, its applicability to other tumor types needs to be tested. The current challenge is to identify safe and promising immunotherapies in preclinical mouse models and to translate them into viable perioperative therapies to be given to cancer surgery patients to prevent the recurrence of metastatic disease.

A mouse tumor model of surgical stress is used to explore how postoperative immune suppression promotes metastatic disease and to evaluate immunostimulatory perioperative therapies.

術後免疫抑制のメカニズムを探索し、新しい周術期免疫療法を評価するための手術侵襲のマウス腫瘍モデル

Surgical resection is an essential treatment for most cancer patients, but surgery induces dysfunction in the immune system and this has been linked to ...

A Modified In vitro Invasion Assay to Determine the Potential Role of Hormones, Cytokines and/or Growth Factors in Mediating Cancer Cell Invasion

Blood serum serves as a chemoattractant towards which cancer cells migrate and invade, facilitating their intravasation into microvessels. However, the actual molecules towards which the cells migrate remain elusive. This modified invasion assay has been developed to identify targets which drive cell migration and invasion. This technique compares the invasion index under three conditions to determine whether a specific hormone, growth factor, or cytokine plays a role in mediating the invasive potential of a cancer cell. These conditions include i) normal fetal bovine serum (FBS), ii) charcoal-stripped FBS (CS-FBS), which removes hormones, growth factors, and cytokines and iii) CS-FBS + molecule (denoted &#8220;X&#8221;). A significant change in cell invasion with CS-FBS as compared to FBS, indicates the involvement of hormones, cytokines or growth factors in mediating the change. Individual molecules can then be added back to CS-FBS to assay their ability to reverse or rescue the invasion phenotype. Furthermore, two or more factors can be combined to evaluate the additive or synergistic effects of multiple molecules in driving or inhibiting invasion. Overall, this method enables the investigator to determine whether hormones, cytokines, and/or growth factors play a role in cell invasion by serving as chemoattractants or inhibitors of invasion for a particular type of cancer cell or a specific mutant. By identifying specific chemoattractants and inhibitors, this modified invasion assay may help to elucidate signaling pathways that direct cancer cell invasion.

A Modified In vitro Invasion Assay to Determine the Potential Role of Hormones, Cytokines and/or Growth Factors in Mediating Cancer Cell Invasion

This video article describes a modified in vitro method to examine the role of hormones, cytokines and/or growth factors in driving cancer cell invasion.

変更された<em&gt;インビトロ</em媒介する癌細胞浸潤におけるホルモン、サイトカインおよび/または成長因子の潜在的な役割を決定するために&gt;浸潤アッセイ

Blood serum serves as a chemoattractant towards which cancer cells migrate and invade, facilitating their intravasation into microvessels. However, the ...

Glutamate and Hypoxia as a Stress Model for the Isolated Perfused Vertebrate Retina

Neuroprotection has been a strong field of investigation in ophthalmological research in the past decades and affects diseases such as glaucoma, retinal vascular occlusion, retinal detachment, and diabetic retinopathy. It was the object of this study to introduce a standardized stress model for future preclinical therapeutic testing.
Bovine retinas were prepared and perfused with an oxygen saturated standard solution, and the ERG was recorded. After recording stable b-waves, hypoxia (pure N2) or glutamate stress (250 &#181;m glutamate) was exerted for 45 min. To investigate the effects on photoreceptor function alone, 1 mM aspartate was added to obtain a-waves. ERG-recovery was monitored for 75 min.
For hypoxia, a decrease in a-wave amplitude of 87.0% was noted (p &#60;0.01) after an exposition time of 45 min (decrease of 36.5% after the end of the washout p = 0.03). Additionally, an initial decrease in b-wave amplitudes of 87.23% was recorded, that reached statistical significance (p &#60;0.01, decrease of 25.5% at the end of the washout, p = 0.03).
For 250 &#181;m glutamate, an initial 7.8% reduction of a-wave amplitudes (p &#62;0.05) followed by a reduction of 1.9% (p &#62;0.05). A reduction of 83.7% of b-wave amplitudes (p &#60;0.01) was noted; after a washout of 75 min the reduction was 2.3% (p = 0.62). In this study, a standardized stress model is presented that may be useful to identify possible neuroprotective effects in the future.

With this study, we introduce a standardized stress model for the isolated superfused bovine retina for future preclinical therapeutic testing. The effect of either hypoxia (pure N2) or glutamate stress (250 &#181;M glutamate) on retinal function represented by a- and b-wave amplitudes was evaluated.

単離された灌流脊椎動物の網膜のためのストレスモデルとしてグルタミン酸および低酸素症

Neuroprotection has been a strong field of investigation in ophthalmological research in the past decades and affects diseases such as glaucoma, retinal ...

Induction and Analysis of Oxidative Stress in Sleeping Beauty Transposon-Transfected Human Retinal Pigment Epithelial Cells

Oxidative stress plays a critical role in several degenerative diseases, including age-related macular degeneration (AMD), a pathology that affects ~30 million patients worldwide. It leads to a decrease in retinal pigment epithelium (RPE)-synthesized neuroprotective factors, e.g., pigment epithelium-derived factor (PEDF) and granulocyte-macrophage colony-stimulating factor (GM-CSF), followed by the loss of RPE cells, and eventually photoreceptor and retinal ganglion cell (RGC) death. We hypothesize that the reconstitution of the neuroprotective and neurogenic retinal environment by the subretinal transplantation of transfected RPE cells overexpressing PEDF and GM-CSF has the potential to prevent retinal degeneration by mitigating the effects of oxidative stress, inhibiting inflammation, and supporting cell survival. Using the Sleeping Beauty transposon system (SB100X) human RPE cells have been transfected with the PEDF and GM-CSF genes and shown stable gene integration, long-term gene expression, and protein secretion using qPCR, western blot, ELISA, and immunofluorescence. To confirm the functionality and the potency of the PEDF and GM-CSF secreted by the transfected RPE cells, we have developed an in vitro assay to quantify the reduction of H2O2-induced oxidative stress on RPE cells in culture. Cell protection was evaluated by analyzing cell morphology, density, intracellular level of glutathione, UCP2 gene expression, and cell viability. Both, transfected RPE cells overexpressing PEDF and/or GM-CSF and cells non-transfected but pretreated with PEDF and/or GM-CSF (commercially available or purified from transfected cells) showed significant antioxidant cell protection compared to non-treated controls. The present H2O2-model is a simple and effective approach to evaluate the antioxidant effect of factors that may be effective to treat AMD or similar neurodegenerative diseases.

Induction and Analysis of Oxidative Stress in Sleeping Beauty Transposon-Transfected Human Retinal Pigment Epithelial Cells

We present a protocol for the development and use ofan oxidative stress-model by treating retinal pigment epithelial cells with H2O2, analyzing cell morphology, viability, density, glutathione, and UCP-2 level. It is a useful model to investigate the antioxidant effect of proteins secreted by transposon-transfected cells to treat neuroretinal degeneration.

睡眠美容トランスポゾントランスポゾントランスフェクトヒト網膜色素上皮細胞における酸化ストレスの誘導と分析

Oxidative stress plays a critical role in several degenerative diseases, including age-related macular degeneration (AMD), a pathology that affects ~30 ...