すべてのプロトコルは、機関動物ケアおよび使用委員会 (IACUC)、ニューヨーク州立大学バッファローによって承認されています。侵襲を描いた方法論と非侵襲的モニタリングの図は図 1に示します。
1. 動物
<ol><li>使用時間付、Q 熱陰性、妊娠羊 (雑種、サフォーク ドーセット カターディン品種) 127-143 日間の妊娠で胎仔羊と。 注: 羊的妊娠は 145 日間と 127 妊娠羊が極端な未熟児のように動作します。</li>
</ol>2. 術前準備
<ol><li>雌 12 時間、手術前から、食べ物を与えない。</li>
	<li>物静かで静脈内 (IV) ジアゼパムとエウェ (0.25 – 1.5 mg/kg) とケタミン (4 mg/kg)。</li>
	<li>リフト カートに横方向の臥床にエウェをガイドします。</li>
	<li>V トップ外科テーブルの上に背の臥床にエウェを移動します。</li>
</ol>3. 麻酔
注: 前に帝王切開でエウェはジアゼパムとケタミンと鎮静、挿管イソフルラン (1-4%) の連続的な吸入が続きます。麻酔の妥当性は、パルス酸素濃度計と心拍数血中酸素濃度の連続監視と共に筋肉のトーンと目の反射の評価によって監視されます。機器の中に羊母体麻酔の影響の下で、計装のサイトへ局所麻酔薬を受け取ります。
<ol><li>
10.0 mm かふ付け気管内チューブ (ETT) と雌を挿管します。事前の声帯を通して気管チューブ カフを超えて 1-2 cm のコードします。気管内チューブのカフをふくらませ、安全な場所に羊の顎周り ETT。
	<ol>
<li>眼瞼の応答がない場合は、イソフルランを調整 (2-3% で通常十分です)。</li>
		<li>0.21 の促された酸素 (FIO2) の割合を従来の機械的人工換気 (CMV) による人工呼吸を提供します。10-15 mL/kg の換気量を提供するために人工呼吸器を設定します。</li>
		<li>羊の舌は、パルスオキシメータによるオキシ ヘモグロビン飽和度を継続的に監視します。</li>
		<li>カプノグラフ、終末呼気炭酸ガス (EtCO2) を測定します。</li>
		<li>FIO2と CMV それぞれ 90 97% と呼気 CO2間 35-45 mm Hg、オキシ ヘモグロビン飽和度を維持するために調整します。</li>
	</ol>
</li>
	<li>
羊の手術タイダウンで手術台に手足を固定します。
	<ol>
<li>逆に胎児への圧力を和らげるためにトレンデレンブルグ体位に手術台を調整します。</li>
	</ol>
</li>
	<li>解凍し、胃内容を排出する orogastric チューブを配置します。</li>
	<li>補足輸液 (生理食塩水入りまたは一日リンガーズ ソリューション) 10 mL/kg/h。</li>
	<li>連続コア温度を取得する直腸温度プローブを配置します。</li>
</ol>4. 手術
注: 帝王と胎児の手順は、動物がナトリウム ペントバルビ タール 100 mg/kg の IV の投与による安楽死は急性の手術と見なされます。羊が羊たちの配信、次を安楽死させ、羊たちは実験終了後安楽死させた。安楽死は、心停止によって確認されます。両側開胸下や急激な多量の二次的な方法も採用されています。この場合、無菌技術は、手術中に実践されていません。担当者はまだ動物の接触の間に保護の服装を着用する必要があります。
<ol><li>腹側腹部から毛を剃る。</li>
	<li>
白線上 15-18 cm の腹部皮一重切開をするため、焼灼を使用して筋を公開するアルバ。
	<ol>
<li>圧力を用いた鈍ティップド止血腹部に小さな開口部を作成します。</li>
		<li>腹壁に鈍い先端のハサミをスライドさせながらこの開口部を維持します。</li>
		<li>リネアの切開を完了するはさみを使用してアルバ。</li>
	</ol>
</li>
	<li>
外在の腹部から子宮内胎児の頭を見つけて。
	<ol>
<li>片方の手で胎児の頭を押し、焼灼を使用 (子羊の額) 以上子宮壁を 10 cm の切開をしています。任意の子葉を回避するを確認します。</li>
		<li>バブコック鉗子を使用して、4 つの反対のコーナーで子宮と胎盤の層を一緒にクランプします。</li>
		<li>雌の腹部外子宮で子羊の頭を公開します。</li>
		<li>胎児手術中に腹部の外敷設バブコック鉗子を残します。</li>
	</ol>
</li>
</ol>5. 胎児気道
<ol><li>
部分的に露出した挿管 3.5-4.5 mm 胎仔羊かふ付け気管内チューブ。声帯カフを超えて 1 cm を通して気管チューブを進めます。
	<ol>
<li>カフを膨らませるし、チューブと、頭の周囲の臍帯テープを結ぶことによって、気管チューブを固定します。</li>
		<li>側に頭を傾けることによって受動的に排出する ETT の過剰な胎児肺液を許可します。これは分娩中に肺液の出口をシミュレートします。</li>
		<li>仮死の期間にあえいでいる時のガス交換を防ぐために気管内チューブを閉塞します。</li>
	</ol>
</li>
</ol>6. 首容器計装
<ol><li>
毛を剃るし、切開のすべてのサイトに 1-2 mL の皮下ブピバカイン塩酸塩 (0.25%) に潜入します。
	<ol>
<li>首 (約 6 cm 頭の遠位) の各側に 2 つの 3 cm の皮膚切開を行う気管に隣接します。切開は横首を必要があります。</li>
		<li>深部組織のタンコブを作ったを避けるために、皮膚をテンティング中、焼灼を使用して切開を実行します。</li>
	</ol>
</li>
	<li>湾曲したケリーやモスキート鉗子を用いた筋膜を分離します。</li>
	<li>
右の内頚静脈と右総頚動脈に分離します。
	<ol>
<li>場所 2 つ 20 cm、両船の下 0 絹糸。</li>
		<li>シルク縫合糸の間に 1 cm の (近位遠位に) スペースを許可します。</li>
		<li>アイリス外科はさみを使用して容器に 1-2 mm 横カットをして血流を制限する、表面的に、自船の周囲は各縫合をそっと持ち上げます。</li>
	</ol>
</li>
	<li>
頚動脈の血圧測定や採血の大動脈弓尾の方に向かって右頸動脈の 1 つ前フラッシュ カテーテル (15-17 G) を挿入します。
	<ol>
<li>完全に動脈を結紮するのに近位のネクタイを使用します。</li>
		<li>6.3.2 6.3.3 の手順を繰り返します。</li>
		<li>あらかじめフラッシュ頚動脈カテーテルを挿入し、動脈、3 ノットでカテーテルにネクタイに遠位絹縫合糸を使用します。</li>
	</ol>
</li>
	<li>頸静脈、手順 6.3.2–6.3.3 に 1 つ前フラッシュ カテーテル (14-16 G) を挿入するのには、同様の手法を使用して事前に尾側 (向かって右のアトリウム) 胸郭入口内で残りの部分に 7-10 cm。流体および薬物の管理だけでなく、静脈圧測定に使用されます。<ol>
<li>開存とヘパリン生食液のためのカテーテルをチェックし、縫合ネクタイを完了容器周辺カテーテル (近位の縫合のみ) で 2-3 ノットを使用しています。</li>
		<li>(14-16 G) の 2 番目のカテーテルを挿入脳循環から血を収集するために吻方約 5 cm。両方のカテーテルの配置で同じカットを使用します。</li>
		<li>遠位部の縫合糸やカテーテルに関連した手順 6.5.1 とねじれを防ぐためにそれらを確保するため反対のカテーテル ライン周り絹縫合糸を結びます。</li>
	</ol>
</li>
	<li>
以前作った左側に T 形 (1-2 cm 切開を拡張します。
	<ol>
<li>開いた曲線の止血剤を使用して左の頚動脈を昇格させます。</li>
		<li>血流を測定する容器の周り 2 mm の血管の流れ-プローブを配置します。</li>
		<li>位置にプローブを安定させるために柔軟なポリマーで 1 cm 袖付きフロー プローブをカバーします。</li>
		<li>T 字切開、船と並列実行するケーブルにフロー プローブ ケーブル線を配置します。</li>
		<li>皮膚切開部を閉じる。そしてプルした場合の流れを制限することを避けるために、ケーブルのループ、ケーブルまわりを結ぶ。</li>
	</ol>
</li>
</ol>7. 大血管計測
<ol><li>胸を公開する羊胎仔を抽出し、熱損失を防ぐためにポリエチレン ラップで露出した皮膚をカバーします。</li>
	<li>右横ずれの臥床でラムを配置します。</li>
	<li>手術部位を公開する首左前肢の位置。</li>
	<li>
4th肋に沿ってブピバカイン塩酸塩 (0.25%) の 3 mL に潜入し、焼灼を使用して 6 cm の皮膚切開を行います。
	<ol>
<li>慎重に鈍カーブタイプ鉗子、3rdの肋骨の下の円と肋間筋を貫通、次の肋の間に出てくる。肺や心臓を傷つけないようにことを確認します。</li>
		<li>0 シルクのカット済み部分のクランプを開き、肋骨を取り囲むようにそれもたらします。</li>
		<li>4thの下で縫合糸リブをセキュリティで保護する手順 7.4.1–7.4.2 を繰り返します。</li>
		<li>各リブ周り関係をクランプします。そっと引き上げ、胸から下の 4番目の肋間スペースに綿の先端アプリケータを挿入します。</li>
		<li>リブ関係に巻き込まれる、焼灼を使用して最終的なオープニング、肺癌ではないことを確認 6-8 cm。 の 1 cm の肋間スペースを開き、胸の内容を保護するためにアプリケーター ・ スティックを使用します。</li>
		<li>肋骨の拡散機を胸に開口部に配置します。胸を開くと、動脈管開存、肺動脈 (PA) 肺左上葉が明らかになります。</li>
	</ol>
</li>
	<li>
2 x 2 インチの金網を使用して、肺を包む、外科の領域から任意の露出した組織がゆっくりとします。
	<ol>
<li>小さな手動のリトラクターを使用すると、(必要に応じて) ビューのフィールドを向上させます。</li>
		<li>(1/4 インチ幅、6 インチの長さ) 臍帯テープの場所は、大血管の周りの羊膜にあらかじめ浸してください。臍帯テープを浸漬容器壁に摩擦・損傷が減少します。</li>
	</ol>
</li>
	<li>鉗子を使用して、心膜を持ち上げて、迷走神経を切らないように世話主肺動脈に沿ってカットします。迷走神経に沿って心膜の外側のカットと干渉から右心房を維持するのにアプリケーター スティックを使用します。</li>
	<li>
ジェミニ鉗子を使用すると、鉗子ゆっくりと移動/PA 膵切欠きからの周りと「囲み」、左 PA の反対側に出現する左 PA の背後にあります。多くの繊細な組織は、この領域の下を実行し、抵抗がない場合にのみ進みます。
	<ol>
<li>鉗子の先端を観察するとき、事前に湿らせた臍帯テープの片方の端をつかむところだけ開きます。場所でクリックして 1 つだけの位置にクランプします。それは周囲の組織がキャッチされること鉗子でも、可能な限りしっかりとクランプとその組織可能性があります血管損傷の結果臍帯テープでキャッチされると。</li>
		<li>所定の位置にテープをプルする鉗子を優しくバックアップします。これはフロー プローブの空間を開く機能、フロー プローブ (図 2) の挿入を指示することができます。切り込みを保護する気に任意の容器の部分は、これは組織の最も脆弱な部分が不可欠です。</li>
	</ol>
</li>
	<li>
優しく臍帯テープを持ち上げてプローブ設立の開口部に沿って船での (L-ブラケット) の金属の部分を導く 4-6 mm の血管の流れプローブを配置します。
	<ol>
<li>L-ブラケットのスライドの閉鎖をように保護する終わりを視覚化して座席の場所で容器、プローブの背面に向かってテープを直接します。</li>
		<li>船が > 75% 確実のタイトなフィット感と正確な流量計測プローブ サイズを確認します。</li>
		<li>プローブに近い臍帯テープの端を慎重に切り取って容器からテープを削除するもう一方の端を軽く引いてください。テープは、削除されていない場合、測定の妨げに。</li>
		<li>超音波ゲルを適用し、信号強度と品質を改善するためにプローブとケーブルの位置を調整します。</li>
	</ol>
</li>
	<li>動脈管開存の周り 6-8 mm の血管の流れ-プローブの配置に関連した 7.6-7.7.4 の手順を繰り返します。</li>
	<li>任意の保護の 2 x 2 のガーゼを胸から外し、再配置する肺を許可します。</li>
	<li>肺を保護するためにフロー プローブの各金属ブラケットの周りの新しい 2 x 2 ガーゼを配置します。</li>
	<li>テーパー針と 2.0 合成モノフィラメント縫合糸を使用してレイヤーの胸部を閉じます。切削針と 2.0 の絹糸を使用してループの外側の皮膚にフロー プローブ ケーブルを固定します。</li>
</ol>8. 非侵襲計測
<ol><li>右腋窩、左腋窩、右鼠径部 (3 鉛 EKG) で心電図リードを取り付けます。前と後膵動脈オキシ ヘモグロビン飽和度 (スポ2) は、いずれかの後肢と右前肢にそれぞれ配置パルスオキシメータで監視されます。縫合糸と不透明ドレッシング額以上近赤外分光装置 (NIRS) モニターを固定します。</li>
</ol>9. データの収集
<ol><li>収集し、データ集録ソフトウェアを使用してデータを記録します。図 1などの侵襲的、非侵襲的なパラメーターを示しています: 動脈、中心静脈圧、左一般的な頸動脈、左肺、膵動脈の血の流れ、心電図、SpO2、放医研、ETCO2。 注: データ収集ソフトウェア ・機器セットアップ変わることができる、この資料の範囲を超えています。</li>
</ol>10 実験的プロトコル
<ol><li>閉塞し、へその緒を切るし、母体の腹部から放射ウォーマーに子羊を移動します。</li>
	<li>仮死の期間中にエピネフリン投与の臍静脈カテーテル挿入 (0.01-0.03 mg/kg IV、NRP の推奨事項/)。</li>
	<li>後膵動脈血ガスを収集する臍動脈カテーテルを挿入します。</li>
	<li>間近に迫った心静止、検出された頸動脈の流れと圧力の欠如として定義することができますのための羊を監視します。</li>
	<li>人員が利用可能なことを確認し、次の作業を一人一人を捧げる: (1) 気道と換気の管理、(2) 胸部圧迫を行う、(3) 薬の管理、(4) 血液サンプルを描画、(5) コードのリードします。</li>
	<li>35/5 cm H2o. 開始蘇生実験的プロトコルの 20 分タイマーの圧力下で T ピース蘇生で肯定的な圧力換気 (PPV) を提供することにより蘇生を始めます。</li>
	<li>
30 後の換気、s は親指と 1 つの手の指を胸の反対側に配置することによって胸部圧迫を開始します。
	<ol>
<li>指と一緒に約 1/3 の深さに親指を絞ることによって胸部圧迫を提供します。</li>
	</ol>
</li>
	<li>3 分間隔でエピネフリンを管理します。</li>
	<li>自発の循環のリターンまで蘇生努力を継続または 20 分まで。</li>
	<li>自然の循環、連続静脈内輸液 (5 cc/kg/hr) とフェンタニルのリターンで (0.5 2.0 マイクログラム/kg/hr; 効果に滴定された)、以前に確立された頸静脈カテーテルを介して与えられています。20 分蘇生期間、動物に安楽死です。子羊は、20 分で ROSC を達成しない場合、は、安楽死を続行します。</li>
</ol>

<ol>
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B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiovascular+transition+at+birth:+a+physiological+sequence.">Cardiovascular transition at birth: a physiological sequence.</a> Pediatr Res. 77 (5), 608-614 (2015).</li><li>Vali, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+Timing+and+Route+of+Epinephrine+in+a+Neonatal+Model+of+Asphyxial+Arrest.">Evaluation of Timing and Route of Epinephrine in a Neonatal Model of Asphyxial Arrest.</a> J Am Heart Assoc. 6 (2), (2017).</li><li>Vali, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Continuous+Chest+Compressions+During+Sustained+Inflations+in+a+Perinatal+Asphyxial+Cardiac+Arrest+Lamb+Model.">Continuous Chest Compressions During Sustained Inflations in a Perinatal Asphyxial Cardiac Arrest Lamb Model.</a> Pediatr Crit Care Med. , (2017).</li><li>Vali, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemodynamics+and+gas+exchange+during+chest+compressions+in+neonatal+resuscitation.">Hemodynamics and gas exchange during chest compressions in neonatal resuscitation.</a> PLoS One. 12 (4), e0176478 (2017).</li><li>Tana, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+Lung+Volume+and+Hemodynamic+Changes+During+High-Frequency+Ventilation+Recruitment+in+Preterm+Neonates+With+Respiratory+Distress+Syndrome.">Determination of Lung Volume and Hemodynamic Changes During High-Frequency Ventilation Recruitment in Preterm Neonates With Respiratory Distress Syndrome.</a> Crit Care Med. 43 (8), 1685-1691 (2015).</li><li>Morin, F. 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Simulated correction permits fetal lung growth with survival at birth.</a> Surgery. 88 (2), 260-268 (1980).</li><li>Turley, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intrauterine+cardiothoracic+surgery:+the+fetal+lamb+model.">Intrauterine cardiothoracic surgery: the fetal lamb model.</a> Ann Thorac Surg. 34 (4), 422-426 (1982).</li><li>Reddy, V. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+utero+placement+of+aortopulmonary+shunts.+A+model+of+postnatal+pulmonary+hypertension+with+increased+pulmonary+blood+flow+in+lambs.">In utero placement of aortopulmonary shunts. A model of postnatal pulmonary hypertension with increased pulmonary blood flow in lambs.</a> Circulation. 92 (3), 606-613 (1995).</li><li>Hemway, R. 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著者がある利害の衝突を宣言するには

ラム モデルはひと新生児に匹敵するサイズで、侵襲血行力学的変数を測定する簡単な計測を可能します。子羊胎児/新生児モデルは、豊かな移行の循環として新生児の呼吸・循環生理学の理解に貢献している貴重な研究ツールをされています。いくつかの実験的ラム モデルは、早産における最適な換気戦略羊13,14,21, 永続?...

周産期仮死はアメリカ合衆国用語出産 1,000 回あたり約 4 で発生し、400 万新生児死亡世界中1,2の約 25% があります。胎児の自然な発展の中では、内から子宮外の環境へのシームレスな移行を可能にする肺は、ガス交換器官として胎盤の役割を担う場合に出産、分娩中にいくつかの適応を行う必要があります。適切に移行さらに出生時に新生児の蘇生の努力が損なわれます。胎児肺クリアランスが不完全または遅延3,4と永続的な高肺血管抵抗 (PVR)5影響最も残っている換気の効果は、状況のインスタンス6asphyxiated の新生児の蘇生法における重要な介入。さらに、即時クランプ臍帯と低抵抗胎盤の除去、心筋機能不全7、8を引き起こす可能性のある心臓出力の急激な変化につながります。
積極的な蘇生法 (胸骨圧迫やエピネフリン投与の必要性)1,9の間欠的な必要性により現在をサポートする大規模な無作為化臨床試験から強力な証拠の欠乏があります。新生児蘇生プログラム (NRP) のガイドライン。新生児蘇生における多くのトランスレーショナルリサーチ研究移行を適切に表現する失敗した産後の動物モデル (特に子豚) を用いて、胎児循環や液体で満たされた肺の配信で新生児に固有お部屋です。新生児循環に胎児循環から遷移に関連するユニークな課題を与え、周産期の窒息心不全羊胎仔モデルは新生児蘇生の生理学を勉強する最適です。
1930 年代には早くも胎児にジョゼフ ・ バークロフトによる研究羊胎児・新生児の生理10の基礎を築いた。20 世紀の後半では、子羊の胎児のジェフリー ・ ドーズの革新的なきめ細やかな実験モデルと後でアブラハム ルドルフによってそれらは途方もなく胎児の心血管および肺生理学の知識に貢献しています。11,12. 近年、胎児・新生児の子羊モデルに関する研究を提供しているの理解を深める血行動態13,14換気の影響で PVR15,16酸素の効果コードの中に発生する循環の変更だけでなく、7,17をクランプします。最後に、過去 1 年間の新生児の子羊は蘇生18,19,20時血行動態の影響を研究する新たなモデルとして役立った。何の順を追って物語は手術機器の詳細な説明と同様、ラム実験の実施に関与し、実験の方法が表示されます。

<table border="0" cellpadding="0" cellspacing="0" width="819">
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;">Babcock forceps</td>
			<td>Miltex</td>
			<td>16-44</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Blood pressure transducer</td>
			<td style="width:192px;">Becton Dickinson</td>
			<td style="width:211px;">P23XL-1</td>
			<td style="width:200px;">Used with saline filled diaphragm domes</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Blunt tipped scissors</td>
			<td style="width:192px;">Miltex</td>
			<td style="width:211px;">98SCS50-56</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Capnograph</td>
			<td style="width:192px;">Philips</td>
			<td style="width:211px;">7900</td>
			<td style="width:200px;">Used with Neonatal Flow Sensors</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Cautery pencil</td>
			<td style="width:192px;">Valley Lab</td>
			<td style="width:211px;">287879</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Cautery unit</td>
			<td style="width:192px;">Valley Lab</td>
			<td style="width:211px;">SSE2K</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Curved Forceps</td>
			<td style="width:192px;">Everost</td>
			<td style="width:211px;">711714</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Data acquisition software</td>
			<td style="width:192px;">Biopac Systems Inc.</td>
			<td style="width:211px;">ACK100W</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">EKG</td>
			<td style="width:192px;">Biopac Systems Inc.</td>
			<td style="width:211px;">ECG100C</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Endotracheal tube -cuffed</td>
			<td style="width:192px;">Rusch</td>
			<td style="width:211px;">111780035</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Flow modulator</td>
			<td style="width:192px;">Transonic Systems Inc.&#160;</td>
			<td style="width:211px;">T403</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Flow-probe</td>
			<td style="width:192px;">Transonic Systems Inc.&#160;</td>
			<td style="width:211px;">MC4PSS-LS-WC100-CM4B-GA</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Gastric tube</td>
			<td style="width:192px;">Jorgensen Labs Inc.</td>
			<td style="width:211px;">J0106LE</td>
			<td style="width:200px;">To decompress and drain ewe stomach</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Hair clipper</td>
			<td style="width:192px;">Andis Company</td>
			<td style="width:211px;">65340</td>
			<td style="width:200px;">&#160;# 40 Clipper Blade</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Infant radiant warmer</td>
			<td>GE healthcare</td>
			<td>7810</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Intravenous catheters</td>
			<td>Becton Dickinson</td>
			<td>381234</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Iris surgical scissors</td>
			<td style="width:192px;">Patterson</td>
			<td style="width:211px;">510585</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Kelly Foreceps</td>
			<td style="width:192px;">Patterson</td>
			<td style="width:211px;">510535</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Mosquito Forceps</td>
			<td style="width:192px;">RICA Surgical Products INC</td>
			<td style="width:211px;">1-74</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Near-infrared spectroscopy</td>
			<td style="width:192px;">Nonin Medical Inc.&#160;</td>
			<td style="width:211px;">X-100M</td>
			<td style="width:200px;">Sensmart Equanox &#38; PureSAT</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">RSO2 Sensor</td>
			<td style="width:192px;">Nonin Medical Inc.&#160;</td>
			<td style="width:211px;">8004CB-NA</td>
			<td style="width:200px;">Neonatal&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Scalpel handle and blade</td>
			<td style="width:192px;">Everost</td>
			<td>707203</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Sutures -silk 2-0</td>
			<td style="width:192px;">Covidien</td>
			<td style="width:211px;">SS-695</td>
			<td style="width:200px;">Used for tying catheters to vessels</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Sutures -vicryl&#160; 2-0</td>
			<td style="width:192px;">Ethicon</td>
			<td style="width:211px;">J269H</td>
			<td style="width:200px;">Used for closing thoracotomy&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">T-piece resuscitator</td>
			<td>Neo-Tee</td>
			<td>MCM1050812</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Umbilical ties</td>
			<td style="width:192px;">Jorgensen Labs Inc.</td>
			<td>J0025UA</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Weitlander Retractor</td>
			<td style="width:192px;">Miltex</td>
			<td style="width:211px;">11-625</td>
			<td style="width:200px;"></td>
		</tr>
	</tbody>
</table>

the perinatal asphyxiated lamb model: a model for newborn resuscitation

出生時仮死は世界中ほぼ 100 万の死のために毎年をアカウント、早期新生児死亡率と死亡率の主要な原因の一つです。現在の新生児蘇生法ガイドラインの多くの側面のまま無作為化の臨床試験、不定期のため、しばしば豊富な蘇生に必要な予測を行う上での難しさを与え物議を醸す。新生児蘇生法に関するほとんどの研究は、本当に生理学的な変更を反映するように失敗するマネキン モデルやピグレット モデル彼らの肺の液体をクリアするし、胎児から新生児循環への移行を完了しているから幹します。この議定書は、周産期窒息羊胎仔モデルを作成する方法の詳細なステップバイ ステップの説明を提供します。提案モデルは移行の循環と流体で満たされた肺は、以下の配信、人間の新生児を模倣し、したがって、新生児の生理学を研究する優秀な動物モデルであります。ラム実験の重要な制限事項が関連付けられているコストが高いです。

胎仔羊のインストルメンテーションは、次動態変数は記録された (図 3および図 4)、することができますし、分析し、解釈 (図 5)。頻繁に血液サンプルを集めることができると図 6 pH と実験のいずれかからパコ2データを示します。時折、カテーテルまたは流量が誤動作や?...

Watch this Scientific Journal Video about The Perinatal Asphyxiated Lamb Model: A Model for Newborn Resuscitation at JoVE.com

The Perinatal Asphyxiated Lamb Model: A Model for Newborn Resuscitation

羊胎仔の侵襲的な計測では、生まれたばかりの乳児によく似たモデルで処理中の循環の生理学的計測を提供します。

周産期に窒息ラム モデル: 新生児蘇生のためのモデル

Birth asphyxia accounts for nearly one million deaths worldwide each year, and is one of the primary causes of early neonatal morbidity and mortality. ...

the-perinatal-asphyxiated-lamb-model-a-model-for-newborn-resuscitation

Research

JoVE Journal

Medicine

10.7K ビュー.  University of California Davis. 羊胎仔の侵襲的な計測では、生まれたばかりの乳児によく似たモデルで処理中の循環の生理学的計測を提供します。

ビデオ: The Perinatal Asphyxiated Lamb Model: A Model for Newborn Resuscitation

Normothermic Cardiac Arrest and Cardiopulmonary Resuscitation: A Mouse Model of Ischemia-Reperfusion Injury

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused by whole-body hypoperfusion.5,6 Successful reproduction of whole-body hypoperfusion in rodent models has been fraught with 
difficulty.7-9,9,10 Models which employ focal ischemia have repeatedly demonstrated results which do not translate to the clinical setting, and larger 
animal models which allow for whole body hypoperfusion lack access to the full toolset of genetic manipulation possible in the mouse.11,12 However, in recent 
years a mouse model of cardiac arrest and cardiopulmonary resuscitation has emerged which can be adapted to model AKI.13 This model reliably reproduces 
physiologic, functional, anatomic, and histologic outcomes seen in clinical AKI, is rapidly repeatable, and offers all of the significant advantages of a murine surgical 
model, including access to genetic manipulative techniques, low cost relative to large animals, and ease of use. Our group has developed extensive experience 
with use of this model to assess a number of organ-specific outcomes in AKI.14,15

A powerful model for perioperative and critical care related acute kidney injury is presented. Using whole body hypoperfusion induced by cardiac arrest it is possible to nearly replicate the histologic and functional changes of clinical AKI.

正常体温心停止と心肺蘇生法：虚血再灌流障害のマウスモデル

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused ...

医学

A Rat Model of Ventricular Fibrillation and Resuscitation by Conventional Closed-chest Technique

A rat model of electrically-induced ventricular fibrillation followed by cardiac resuscitation using a closed chest technique that incorporates the basic components of cardiopulmonary resuscitation in humans is herein described. The model was developed in 1988 and has been used in approximately 70 peer-reviewed publications examining a myriad of resuscitation aspects including its physiology and pathophysiology, determinants of resuscitability, pharmacologic interventions, and even the effects of cell therapies. The model featured in this presentation includes: (1) vascular catheterization to measure aortic and right atrial pressures, to measure cardiac output by thermodilution, and to electrically induce ventricular fibrillation; and (2) tracheal intubation for positive pressure ventilation with oxygen enriched gas and assessment of the end-tidal CO2. A typical sequence of intervention entails: (1) electrical induction of ventricular fibrillation, (2) chest compression using a mechanical piston device concomitantly with positive pressure ventilation delivering oxygen-enriched gas, (3) electrical shocks to terminate ventricular fibrillation and reestablish cardiac activity, (4) assessment of post-resuscitation hemodynamic and metabolic function, and (5) assessment of survival and recovery of organ function. A robust inventory of measurements is available that includes &#8211; but is not limited to &#8211; hemodynamic, metabolic, and tissue measurements. The model has been highly effective in developing new resuscitation concepts and examining novel therapeutic interventions before their testing in larger and translationally more relevant animal models of cardiac arrest and resuscitation.

This article describes a rat model of electrically-induced ventricular fibrillation and resuscitation by chest compression, ventilation, and delivery of electrical shocks that simulates an episode of sudden cardiac arrest and conventional cardiopulmonary resuscitation. The model enables gathering insights on the pathophysiology of cardiac arrest and exploration of new resuscitation strategies.

従来のクローズド·チェスト法による心室細動と蘇生のラットモデル

A rat model of electrically-induced ventricular fibrillation followed by cardiac resuscitation using a closed chest technique that incorporates the basic ...

A Piglet Model of Neonatal Hypoxic-Ischemic Encephalopathy

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the world&#8217;s 2.9 million neonatal deaths. Animal models of HIE have contributed to the understanding of the pathophysiology in HIE, and have highlighted the dynamic process that occur in brain injury due to perinatal asphyxia. Thus, animal studies have suggested a time-window for post-insult treatment strategies. Hypothermia has been tested as a treatment for HIE in pdiglet models and subsequently proven effective in clinical trials. Variations of the model have been applied in the study of adjunctive neuroprotective methods and piglet studies of xenon and melatonin have led to clinical phase I and II trials1,2. The piglet HIE model is further used for neonatal resuscitation- and hemodynamic studies as well as in investigations of cerebral hypoxia on a cellular level. However, it is a technically challenging model and variations in the protocol may result in either too mild or too severe brain injury. In this article, we demonstrate the technical procedures necessary for establishing a stable piglet model of neonatal HIE. First, the newborn piglet (&#60; 24 hr old, median weight 1500 g) is anesthetized, intubated, and monitored in a setup comparable to that found in a neonatal intensive care unit. Global hypoxia-ischemia is induced by lowering the inspiratory oxygen fraction to achieve global hypoxia, ischemia through hypotension and a flat trace amplitude integrated EEG (aEEG) indicative of cerebral hypoxia. Survival is promoted by adjusting oxygenation according to the aEEG response and blood pressure. Brain injury is quantified by histopathology and magnetic resonance imaging after 72 hr.

Hypoxic-ischemic encephalopathy following perinatal asphyxia can be studied using animal models. We demonstrate the procedures necessary for establishing a piglet model of neonatal hypoxic-ischemic encephalopathy.

新生児低酸素性虚血性脳症の子豚モデル

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the ...

Breast Milk Enhances Growth of Enteroids: An Ex Vivo Model of Cell Proliferation

Human small intestinal enteroids are derived from the crypts and when grown in a stem cell niche contain all of the epithelial cell types. The ability to establish human enteroid ex vivo culture systems are important to model intestinal pathophysiology and to study the particular cellular responses involved. In recent years, enteroids from mice and humans are being cultured, passaged, and banked away for future use in several laboratories across the world. This enteroid platform can be used to test the effects of various treatments and drugs and what effects are exerted on different cell types in the intestine. Here, a protocol for establishing primary stem cell-derived small intestinal enteroids derived from neonatal mice and premature human intestine is provided. Moreover, this enteroid culture system was utilized to test the effects of species-specific breast milk. Mouse breast milk can be obtained efficiently using a modified human breast pump and expressed mouse milk can then be used for further research experiments. We now demonstrate the effects of expressed mouse, human, and donor breast milk on the growth and proliferation of enteroids derived from neonatal mice or premature human small intestine.

Breast Milk Enhances Growth of Enteroids: An Ex Vivo Model of Cell Proliferation

This protocol describes how to establish an enteroid culture system from neonatal mouse or premature human intestine as well as an efficient method to collect milk from mice.

母乳 Enteroids の増殖を促進する: 細胞増殖のEx Vivoモデル

Human small intestinal enteroids are derived from the crypts and when grown in a stem cell niche contain all of the epithelial cell types. The ability to ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

ワイヤー図法を使用してマウスの腸間膜動脈の等尺性収縮力測定

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

The Lower Body Positive Pressure Treadmill for Knee Osteoarthritis Rehabilitation

Here, based on a clinician&rsquo;s point-of-view, we propose a two-model lower body positive pressure (LBPP) protocol (walking and squatting models) in addition to a clinical, functional assessment methodology, including details for further encouragement of the development of non-drug surgical intervention strategies in knee osteoarthritis patients. However, we only present the effect of LBPP training in improvement of pain and knee function in one patient through three-dimensional gait analysis. The exact, long-term effects of this approach should be explored in future studies.

Here, based on a clinician&#8217;s point-of-view, we propose a two-model lower body positive pressure (LBPP) protocol (walking and squatting models) in addition to a clinical, functional assessment methodology, including details for further encouragement of the development of non-drug surgical intervention strategies in knee osteoarthritis patients. However, we only present the effect of LBPP training in improvement of pain and knee function in one patient through three-dimensional gait analysis. The exact, long-term effects of this approach should be explored in future studies.

膝変形性関節症リハビリテーションのための下半身ポジティブ圧力トレッドミル

Here, based on a clinician&rsquo;s point-of-view, we propose a two-model lower body positive pressure (LBPP) protocol (walking and squatting models) in ...

Noninvasive Electrocardiography in the Perinatal Mouse

Electrocardiography (ECG) has long been relied upon as an effective and reliable method of assessing cardiovascular (and cardiopulmonary) function in both human and animal models of disease. Individual heart rate, rhythm, and regularity, combined with quantitative parameters collected from ECG, serve to assess the integrity of the cardiac conduction system as well as the integrated physiology of the cardiac cycle. This article provides a comprehensive description of the methods and techniques used to perform a noninvasive ECG on perinatal and neonatal mouse pups as early as the first postnatal day, without requiring the use of anesthetics. This protocol was designed to directly address a need for a standardized and repeatable method for obtaining ECG in newborn mice. From a translational perspective, this protocol proves to be entirely effective for characterization of congenital cardiopulmonary defects generated using transgenic mouse lines, and particularly for analysis of defects causing lethality at or during the first postnatal days. This protocol also aims to directly address a gap in the scientific literature to characterize and provide normative data associated with maturation of the early postnatal cardiac conduction system. This method is not limited to a specific postnatal timepoint, but rather allows for ECG data collection in neonatal mouse pups from birth to postnatal day 10 (P10), a window that is of critical importance for modeling human diseases in vivo, with particular emphasis on congenital heart disease (CHD).

Here, we present a noninvasive electrocardiography (ECG) protocol, optimized for early postnatal mice, that does not require the use of anesthetics.

周産期マウスにおける非侵襲性心電図

Electrocardiography (ECG) has long been relied upon as an effective and reliable method of assessing cardiovascular (and cardiopulmonary) function in both ...

A Murine Model of Fetal Exposure to Maternal Inflammation to Study the Effects of Acute Chorioamnionitis on Newborn Intestinal Development

Chorioamnionitis is a common precipitant of preterm birth and is associated with many of the morbidities of prematurity, including necrotizing enterocolitis (NEC). However, a mechanistic link between these two conditions remains yet to be discovered. We have adopted a murine model of chorioamnionitis involving lipopolysaccharide (LPS)-induced fetal exposure to maternal inflammation (FEMI). This model of FEMI induces a sterile maternal, placental, and fetal inflammatory cascade, which is also present in many cases of clinical chorioamnionitis. Although models exist that utilize live bacteria and more accurately mimic the pathophysiology of an ascending infection resulting in chorioamnionitis, these methods may cause indirect effects on development of the immature intestinal tract and the associated developing microbiome. Using this protocol, we have demonstrated that LPS-induced FEMI results in a dose-dependent increase in pregnancy loss and preterm birth, as well as disruption of normal intestinal development in offspring. Further, we have demonstrated that FEMI significantly increases intestinal injury and serum cytokines in offspring, while simultaneously decreasing goblet and Paneth cells, both of which provide a first line of innate immunity against intestinal inflammation. Although a similar model of LPS-induced FEMI has been used to model the association between chorioamnionitis and subsequent abnormalities of the central nervous system, to our knowledge, this protocol is the first to attempt to elucidate a mechanistic link between chorioamnionitis and later perturbations in intestinal development as a potential link between chorioamnionitis and NEC.

We developed a model of chorioamnionitis to simulate fetal exposure to maternal inflammation (FEMI) without complications of live organisms to examine the effects of FEMI on development of the offspring&#8217;s intestinal tract. This allows for study of mechanistic causes for development of intestinal injury following chorioamnionitis.

新生児腸管の発達に及ぼす急性絨毛膜炎の影響を研究する母体炎症への胎児暴露のマウスモデル

Chorioamnionitis is a common precipitant of preterm birth and is associated with many of the morbidities of prematurity, including necrotizing ...

Sterile Pericarditis in Aachener Minipigs As a Model for Atrial Myopathy and Atrial Fibrillation

Atrial fibrillation (AF) is the most common arrhythmia caused by structural remodeling of the atria, also called atrial myopathy. Current therapies only target the electrical abnormalities and not the underlying atrial myopathy. For the development of novel therapies, a reproducible large animal model of atrial myopathy is necessary. This paper presents a model of sterile pericarditis-induced atrial myopathy in Aachener minipigs. Sterile pericarditis was induced by spraying sterile talcum and leaving a layer of sterile gauze over the atrial epicardial surface. This led to inflammation and fibrosis, two crucial components of the pathophysiology of atrial myopathy, making the atria susceptible to the induction of AF. Two pacemaker electrodes were positioned epicardially on each atrium and connected to two pacemakers from different manufacturers. This strategy allowed for repeated non-invasive atrial programmed stimulation to determine the inducibility of AF at specified time points after surgery. Different protocols to test AF inducibility were used. The advantages of this model are its clinical relevance, with AF inducibility and the rapid induction of inflammation and fibrosis-both present in atrial myopathy-and its reproducibility. The model will be useful in the development of novel therapies targeting atrial myopathy and AF.

We describe a sterile pericarditis model in minipigs to study atrial myopathy and atrial fibrillation (AF). We present surgical and anesthetic techniques, strategies for vascular access, and a protocol to study the inducibility of AF.

心房ミオパチーおよび心房細動のモデルとしてのAachenerミニブタにおける滅菌心膜炎

Atrial fibrillation (AF) is the most common arrhythmia caused by structural remodeling of the atria, also called atrial myopathy. Current therapies only ...

Effect of Hyaluronic Acid 35 kDa on an In Vitro Model of Preterm Small Intestinal Injury and Healing Using Enteroid-Derived Monolayers

In vitro scratch wound assays are commonly used to investigate the mechanisms and characteristics of epithelial healing in a variety of tissue types. Here, we describe a protocol to generate a two-dimensional (2D) monolayer from three-dimensional (3D) non-human primate enteroids derived from intestinal crypts of the terminal ileum. These enteroid-derived monolayers were then utilized in an in vitro scratch wound assay to test the ability of hyaluronan 35 kDa (HA35), a human milk HA mimic, to promote cell migration and proliferation along the epithelial wound edge. After the monolayers were grown to confluency, they were manually scratched and treated with HA35 (50 &#181;g/mL, 100 &#181;g/mL, 200 &#181;g/mL) or control (PBS). Cell migration and proliferation into the gap were imaged using a transmitted-light microscope equipped for live-cell imaging. Wound closure was quantified as percent wound healing using the Wound Healing Size Plugin in ImageJ.&#160;The scratch area and rate of cell migration and the percentage of wound closure were measured over 24 h. HA35 in vitro accelerates wound healing in small intestinal enteroid monolayers, likely through a combination of cell proliferation at the wound edge and migration to the wound area. These methods can potentially be used as a model to explore intestinal regeneration in the preterm human small intestine.

Effect of Hyaluronic Acid 35 kDa on an In Vitro Model of Preterm Small Intestinal Injury and Healing Using Enteroid-Derived Monolayers

This protocol describes a method to establish and perform a scratch wound assay on two-dimensional (2D) monolayers derived from three-dimensional (3D) enteroids isolated from non-human primate ileum.

エンテロイド由来単分子膜を用いた早産小腸障害および治癒の in vitro モデルに対するヒアルロン酸35 kDaの効果

In vitro scratch wound assays are commonly used to investigate the mechanisms and characteristics of epithelial healing in a variety of tissue types. ...

周産期に窒息ラム モデル: 新生児蘇生のためのモデル

周産期に窒息ラムモデル: 新生児蘇生のためのモデル