著者の謝辞があります。

すべての動物実験と動物愛護委員会 (低いザクセン州オフィス プロトコル食の安全と消費者保護のための承認されたドイツ動物保護法 (TierSchG) に準拠して行ったTSA 14/1556)。このモデルに適したマウスの最小限の重量が 25 g.
 1 術前準備 注: すべての手続は、きれいな、非無菌条件下でオートクレーブ楽器。。
<ol><li>場所 50 60 8 cm 長いプロピレン中空管の並行ファイバー両側 T-アダプターを接続してください。接着剤と中空繊維と T-アダプターの外部のブランチの間のスペースを埋める。
	<ol><li>許可少なくとも 24 h 硬化する接着剤。標準ミクロトームを使用して T-アダプターから広がっている中空繊維をカットし、シリコン チューブを接続サイトにプルします</li>。
	</ol></li> <li>(26 G branule) から 27 G の金属針を 2 Fr ポリウレタン カニューレに挿入します。外科ブレードを使用し、マイクロ剪刀、約 0.15 cm 各確保するためカニューレの遠位 3 分の 1 以内の 3 ~ 4 ピンホール最適な静脈を顕微鏡 (8-12 倍の倍率) で元に戻る流量。
	<ol><li>削除完了時のワイヤ。鈍的切離と構造の撤回に綿棒を使用します。組織の脱水を防ぐために余分な水分を吸収するガーゼ綿棒 (5 × 5 cm 2) を使用します</li>。
	</ol></li> <li>プライミング ボリュームを準備します。完了すると、30 mL ソリューション 2.5% v v/バッファリング NaHCO 3 の 8.4% 溶液をプライミングあたりヘパリン IU を追加します。4 ° C まで使用でこのソリューションを保存します</li>。
	<li>を埋める CPB 回路 (すなわち、ポンプ、空気のわな猟師、チューブ、両方カニューレ) 静脈カニューレを介して 850 μ L プライミング溶液で。一度いっぱい、動物穿刺の準備が整うまで、体外循環の循環を維持します</li>。
</ol> 2。動物の麻酔、麻酔を管理する誘導商工会議所 2.5% 未満に動物を配置する <ol><li>v/イソフルラン/酸素混合物。ペダル逃避反射、尾ピンチ反射、目の瞬目反射を評価することによって、適切な麻酔を確認します。目の乾燥を避けるために獣医の目軟膏を適用します</li>。
	<li>は、温度調節機能を持つ地球温暖化パッドに動物を配置します。プローブを直腸に挿入し、データ集録システムに接続されている体温を測定します</li>。
	<li>による麻酔が達成された後は、動物 orotracheally 20 G プラスチック製 braunule を使用して、口頭でそれを挿入してビジュアル制御の下で気管にプッシュを挿管します。マウスの人工呼吸器に接続されているイソフルラン気化器を使用して換気を開始します。動物の重量に応じて 250 350 μ L の一回換気量を達成、換気を調整する。</li>。
	完全無痛を保証するために <li>5 mg/kg の動物体重カルプロフェンを皮下注入します。イソフルラン麻酔濃度を酸素では 1.3 2.5% の間維持することをお勧めします。イソフルラン濃度は、手続の段階に応じて手動で調整すること</li>。
</ol> 3。手術 <ol><li>における麻酔・気管挿管が達成された後シャープ罰金ハサミ首に正中皮膚切開を実行、ぶっきらぼうな方法で筋肉を分離、右の内頚静脈と左頚動脈を公開します。準備中、最小限の失血を確保するためバイポーラ鉗子に接続されている獣医凝固装置を用いた血管を凝固します</li>。
	<li>頚静脈血管の準備の後 8-0 絹糸を使用してその分岐する左総頚動脈の遠位のセグメントを縛る頭側</li>。
	<li>シリコン コネクタ (ID 0.5 mm、1 mm 外径)、場所 8-0 絹糸スリップを使用して動脈流入管に 27 G カニューレの先端節動脈と内頚動脈にカニューレの先端を挿入の近位のセグメントに接続します</li>。
	<li>カニューレ先端部の適切な配置後、カニューレ先端が事前に、それは 3-4 mm 近位大動脈弓を産みます。8-0 絹の結び目を確保することでカニューレ先端を修正します</li>。
	<li>による顕微鏡下鉗子、はさみ、鈍とシャープな郭清を実行、胸鎖乳突筋の鎖骨の近くに、横に組織の鈍的準備によって右の内頚静脈を公開、遠位端で 8 0 8-0 絹結び目を配置近位端でループします</li>。
	<li>頚静脈の遠位端の血管を結紮後右頚静脈と右心房方向への進行状況に静脈カニューレの先端を置き、絹の結び目を使用してセキュリティで保護します。最適な静脈還流には右心室に入る静脈のヒントを事前に必要がある場合があります</li>。
	<li>正しいカニューレの位置を達成すると後で 2.5 頚静脈に直接静脈注射を介して動物の体重の 1 グラムあたりヘパリン IU を追加することによって全身パリン投与群を実行します</li>。
	<li>リアルタイム侵襲的圧監視、左大腿動脈を cannulate。鼠径部を公開し、25 倍の倍率の下で大腿神経大腿静脈から総大腿動脈でそっと分ける。
	<ol><li>Ligate 大腿動脈の遠位部。一時的にスリップ ノットの近位部分を閉塞し、マイクロ鉗子を用いた前壁に小切開を作る</li>。
		<li>はその後、大腿動脈に 1 Fr ポリウレタン カニューレを挿入、シルク 10-0 縫合糸で固定します。このカニューレは血液ガス分析血液サンプルを抽出するために使用します</li>。
	</ol></li> <li>動脈と静脈カニューレの成功後、ポンプを回して人工心肺を開始。挿管から体外循環の開始までの時間は約 45 分ポンプの速度を増やすことによって全身の圧力測定とは完全に流れ (4-6 mL/分) 2 分以内の増加血流によって、0.5 mL/分の流量を使用して起動します</li>。
	<li>完全監視下胸骨から始まって、尾側方向に 5 mm の鋭いはさみを使用して上部胸骨を実行します。出血を避けるためにすぐに胸骨端を凝固します。右頚動脈を頭蓋の方向に引いて大動脈弓を公開します</li>。
	<li>最適制御のため無料胸腺とクランプを容易にする周囲の組織から大動脈弓の円周。上行大動脈の周りに 8-0 シルク ループを配置します。心術のクロスのクランプを配置するプルより良い公開する頭蓋方向にシルクのループ上行大動脈</li>。
</ol> 4。人工心肺や血液ガス分析 <ol><li>血液ガス分析 (BGA)、大腿動脈カテーテル経由で 60-90 μ L 動脈血を収集するガラス毛細管を使用します。
	<ol><li>は、シリコーン チューブに小さなクランプを使用して、カテーテルからチューブを切り離します。キャピラリー チューブの制御充填できるようにクランプのゆっくりと圧力を解放します</li>。
		<li>では、カテーテルにシリコン チューブを接続し直します。キャピラリー チューブを挿入次の時点で測定用血液ガス分析器: (BGA1) の換気と体外循環の開始後 10 分 体外循環と呼吸の阻止 (BGA2) 15 分 25 分後 40 分の体外循環、呼吸の阻止 (BGA3) の 30 分後 CPB の 55 分、呼吸器、45 分、心不全 (BGA4) の 20 分後</li></ol></li> <li>安定した消保院流、コスモスイニシアテ ・換気を停止することによって呼吸停止します</li>。
	<li>換気が終了した後地球温暖化パッドを 28 ° C に設定して、冷たい生理食塩水に浸したガーゼを使用して呼吸停止の最初の 20 分以内の 28 ° C の体温に動物を局所的に冷却を開始します</li>。
	<li>28 ° C の体温が達成されれば、呼吸停止の 30 分の合計後、心術を開始する体外循環回路に 7.45 %kcl 溶液 0.3 mL を管理します</li>。
	<li>の大動脈のクランプをクロスより露出するため頭蓋方向に引いて、以前に配置したシルク ループ (ステップ 3.10)、大動脈の上行する部分にマイクロ serrefine クランプを配置します</li>。
	<li>は、合計 60 分の呼吸停止と心停止の 30 分を実行します。心の再灌流を開始する上行大動脈からマイクロ serrefine クランプを削除します。同時に再換気温かく、動物を 37 ° C</li>
	<li>再灌流が完了し、動物に常温療法、達した後ポンプをオフにする体外循環を終了して 20 分における生理的な測定 (例えば 体温、心電図、観血的血圧) の監視を継続</li>
	<li>実験の終わりには、exsanguinate 完全麻酔 (5% イソフルラン) 動物と血と 14 のさらなる分析のための器官を収集します</li>。
</ol>

<ol>
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著者が明らかに何もありません。

完全に機能している臨床的に関連するモデル CPB のマウスを開発しました。心血管疾患を持つマウスの以上 30 系統は、私たちモデルは CPB に関連する新しい将来のプロトコルの開発の開始点を使用できます。また、マウス固有試薬およびノックアウト マウスの茄多、ためこのモデルは CPB の現在のラットのモデルを置き換えることはできません、体外循環関連臓器の損傷に関与する分子メカ?...

心肺のバイパス (CPB) のクリニックへの導入、以来それは心臓手術1で重要な役割を果たしてきました。現代の心臓外科で長時間の体外循環時間豊富な大動脈再建と結合された手順を実行することが欠かせません。技術の進歩は、途方もないされていますが、体外循環の使用が関連付けられている術中と術後全身と地方機関損傷2,3。
大動物モデルは生理的過程4,5CPB の役割を調査するために開発されています。ただし、これらのモデルは、合併症、CPB のいくつかの洞察を提供している、彼らは非常に高価と分子ツール (抗体など) は非常に限られます。小動物でコスト効率的な代替物を開発しました。その開発以来複数の研究はラットおよびウサギ5,6,7,8,9で体外循環モデルを最適化するために行われています。これらのモデルは、疾患の病態生理学的プロセスの測定の良い基盤を提供します。しかし、彼らは関連抗体・試薬の不足のため携帯電話や体液の免疫学を調査するためにまだ十分ではありません。これは、研究のこの分野での役割を損ないます。
最近 CPB のマウス ・ モデルを開発しました。さまざまなマウス固有試薬、遺伝子組み換えマウスによるマウス モデルは一般的には生理学的、分子、および免疫学的研究10,11の選択のモデル.したがって、我々 のモデルは臨床的に関連する疾患12,13で利用できる多くのマウス系統があるので、さまざまな合併症に関連して体外循環の研究を促進します。したがって、本稿で、具体的には、マウスで体外循環を実行する方法。酸素と血行動態的パラメーターは、深い呼吸・循環の逮捕後密接に監視されます。

<table border="0" cellpadding="0" cellspacing="0" width="1191">
	<tbody>
		<tr height="23">
			<td height="23" style="height:23px;width:297px;">Sterofundin</td>
			<td style="width:279px;">B.Braun Petzold GmbH</td>
			<td style="width:181px;">PZN:8609189</td>
			<td style="width:435px;">priming volume, 1:1 with Tetraspan</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">Tetraspan 6% HES Solution</td>
			<td style="width:279px;">B. Braun Melsungen AG</td>
			<td style="width:181px;">PZN: 05565416</td>
			<td>priming volume, 1:1 with Sterofundin</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">Heparin Natrium 25.000</td>
			<td style="width:279px;">Ratiopharm GmbH</td>
			<td style="width:181px;">PZN: 3029843</td>
			<td>2.5 IU per ml of priming solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">NaHCO3 8,4% Solution</td>
			<td style="width:279px;">B. Braun Melsungen AG</td>
			<td style="width:181px;">PZN: 1579775</td>
			<td>3% in priming solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">KCL 7,45 % Solution</td>
			<td style="width:279px;">B. Braun Melsungen AG</td>
			<td style="width:181px;">PZN: 2418577</td>
			<td>0.1 ml for cardioplegia</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">Carprofen</td>
			<td style="width:279px;">Zoetis Inc., USA</td>
			<td>PZN:00289615 08859153</td>
			<td>5 mg/kg/BW</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">1 Fr PU Catheter</td>
			<td style="width:279px;">Instechlabs INC., USA</td>
			<td style="width:181px;">C10PU-MCA1301</td>
			<td>carotid artery</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">2 Fr PU Catheter</td>
			<td style="width:279px;">Instechlabs INC., USA</td>
			<td style="width:181px;">C20PU-MJV1302</td>
			<td>jugular vein</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vasofix Safety catheter 20G</td>
			<td style="width:279px;">B.Braun Medical</td>
			<td style="width:181px;">4268113S-01</td>
			<td>orotracheal intubation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">8-0 Silk suture braided</td>
			<td>Ashaway Line &#38; Twine Mfg. Co., USA</td>
			<td style="width:181px;">75290</td>
			<td>ligature</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">Isoflurane</td>
			<td style="width:279px;">Piramal Critical Care Deutschland GmbH</td>
			<td style="width:181px;">PZN:9714675</td>
			<td>narcosis</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">CLINITUBES blood capillaries</td>
			<td style="width:279px;">Radiomed GmbH</td>
			<td style="width:181px;">51750132</td>
			<td>blood sampling 60 - 95 microliter</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Spring Scissors - 6mm Blades</td>
			<td>Fine Science Tools GmbH</td>
			<td style="width:181px;">15020-15</td>
			<td>instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Spring Scissors - 2mm Blades</td>
			<td>Fine Science Tools GmbH</td>
			<td style="width:181px;">15000-03</td>
			<td>instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Halsted-Mosquito Hemostat</td>
			<td>Fine Science Tools GmbH</td>
			<td style="width:181px;">13009-12</td>
			<td>instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dumont #55 Forceps</td>
			<td>Fine Science Tools GmbH</td>
			<td style="width:181px;">11295-51</td>
			<td>instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Castroviejo Micro Needle Holder - 9cm</td>
			<td>Fine Science Tools GmbH</td>
			<td style="width:181px;">12060-02</td>
			<td>instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micro Serrefines</td>
			<td>Fine Science Tools GmbH</td>
			<td>18555-01</td>
			<td>instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Bulldog Serrefine</td>
			<td>Fine Science Tools GmbH</td>
			<td style="width:181px;">18050-28</td>
			<td>instruments</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:297px;">MiniVent Ventilator for Mice (Model 845)</td>
			<td style="width:279px;">Harvard Apparatus</td>
			<td style="width:181px;">73-0044</td>
			<td>mechanical ventilation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">Isoflurane Vaporizer Drager 19.1</td>
			<td style="width:279px;">Dr&#228;gerwerk AG &#38; Co. KGaA</td>
			<td style="width:181px;"></td>
			<td>anesthesia 1.3 -2.5%</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PowerLab data acquisition device 4/35</td>
			<td>ADInstruments Ltd, New Zealand</td>
			<td style="width:181px;">PL3504</td>
			<td>invasive pressure, ECG, temperature</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ABL 800 Flex</td>
			<td>Radiometer GmbH</td>
			<td style="width:181px;"></td>
			<td>blood gas analysis</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">NMRI mice</td>
			<td style="width:279px;">Charles River Laboratories</td>
			<td style="width:181px;">Crl:NMRI(Han)</td>
			<td>male, 30-35 g, 12 weeks old, housed at least 1 week before the experiment</td>
		</tr>
	</tbody>
</table>

cardiopulmonary bypass in a mouse model: a novel approach

心臓介入中に長時間心肺がますます不可欠になると手順の最適化のため、長時間体外循環に起因する臓器障害を最小限に抑えるため臨床需要の増加が発生します。このホワイト ペーパーの目的は、マウスで人工心肺の完全に機能と臨床的に関連するモデルを実証することでした。デバイス設計、灌流回路の最適化、および顕微鏡技術について報告する.このモデルは、複数の血の図面の必要性のための生存と互換性がない急性モデルです。マウス (例えばマーカー、ノックアウトのなど) に利用できるツールの範囲のための器官の損傷との関係において人工心肺の影響の分子機構に調査容易になりますこのモデル併存疾患。

このプロトコルを記述する灌流回路、外科的処置、およびマウスの CPB 中の生理的パラメーターの監視します。十分に熟練した微小血管によって実行されるとき、一貫性と再現性をもって結果が。
適切な組織灌流を維持するために平均動脈圧は常に体外循環血流を調整し、余分なボリュームを追加することによって 40 と 6...

Watch this Scientific Journal Video about Cardiopulmonary Bypass in a Mouse Model: A Novel Approach at JoVE.com

Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

本稿では、マウスで人工心肺を実行する方法について説明します。この小説のモデルは臓器の損傷に関与する分子メカニズムの調査を促進します。

マウス モデルにおける心肺バイパス: 新しいアプローチ

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization ...

cardiopulmonary-bypass-in-a-mouse-model-a-novel-approach

Research

JoVE Journal

Medicine

10.8K ビュー.  Hannover Medical School. 本稿では、マウスで人工心肺を実行する方法について説明します。この小説のモデルは臓器の損傷に関与する分子メカニズムの調査を促進します。

ビデオ: Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

Vascular Occlusion Training for Inclusion Body Myositis: A Novel Therapeutic Approach

Inclusion body myositis (IBM) is a rare idiopathic inflammatory myopathy. It is known to produces remarkable muscle weakness and to greatly compromise function and quality of life. Moreover, clinical practice suggests that, unlike other inflammatory myopathies, the majority of IBM patients are not responsive to treatment with immunosuppressive or immunomodulatory drugs to counteract disease progression1. Additionally, conventional resistance training programs have been proven ineffective in restoring muscle function and muscle mass in these patients2,3. Nevertheless, we have recently observed that restricting muscle blood flow using tourniquet cuffs in association with moderate intensity resistance training in an IBM patient produced a significant gain in muscle mass and function, along with substantial benefits in quality of life4. Thus, a new non-pharmacological approach for IBM patients has been proposed. Herein, we describe the details of a proposed protocol for vascular occlusion associated with a resistance training program for this population.

The present article presents the details pertaining to the application of resistance training associated to vascular occlusion in IBM patients.

封入体筋炎のための血管閉塞のトレーニング：新しい治療アプローチ

Inclusion body myositis (IBM) is a rare idiopathic inflammatory myopathy. It is known to produces remarkable muscle weakness and to greatly compromise ...

医学

Bronchial Thermoplasty:  A Novel Therapeutic Approach to Severe Asthma

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled manner to reduce excessive airway smooth muscle. Reducing airway smooth muscle decreases the ability of the airways to constrict, thereby reducing the frequency of asthma attacks. Bronchial thermoplasty is delivered by the Alair System and is performed in three outpatient procedure visits, each scheduled approximately three weeks apart. The first procedure treats the airways of the right lower lobe, the second treats the airways of the left lower lobe and the third and final procedure treats the airways in both upper lobes. After all three procedures are performed the bronchial thermoplasty treatment is complete.
Bronchial thermoplasty is performed during bronchoscopy with the patient under moderate sedation. All accessible airways distal to the mainstem bronchi between 3 and 10 mm in diameter, with the exception of the right middle lobe, are treated under bronchoscopic visualization. Contiguous and non-overlapping activations of the device are used, moving from distal to proximal along the length of the airway, and systematically from airway to airway as described previously. Although conceptually straightforward, the actual execution of bronchial thermoplasty is quite intricate and procedural duration for the treatment of a single lobe is often substantially longer than encountered during routine bronchoscopy. As such, bronchial thermoplasty should be considered a complex interventional bronchoscopy and is intended for the experienced bronchoscopist. Optimal patient management is critical in any such complex and longer duration bronchoscopic procedure. This article discusses the importance of careful patient selection, patient preparation, patient management, procedure duration, postoperative care and follow-up to ensure that bronchial thermoplasty is performed safely.
Bronchial thermoplasty is expected to complement asthma maintenance medications by providing long-lasting asthma control and improving asthma-related quality of life of patients with severe asthma. In addition, bronchial thermoplasty has been demonstrated to reduce severe exacerbations (asthma attacks) emergency rooms visits for respiratory symptoms, and time lost from work, school and other daily activities due to asthma.

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled manner to reduce excessive airway smooth muscle. Reducing airway smooth muscle decreases the ability of the airways to constrict, thereby reducing the frequency of asthma attacks.

気管支熱形成術：重篤な喘息の新しい治療アプローチ

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled ...

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient management strategies. Orthotopic mouse models of oral cancer have been developed to facilitate the study of factors that impact invasion and serve as model system for evaluating anti-tumor therapeutics. In these systems, visualization of disseminated tumor cells within oral cavity tissues has typically been conducted by either conventional histology or with in vivo bioluminescent methods. A primary drawback of these techniques is the inherent inability to accurately visualize and quantify early tumor cell invasion arising from the primary site in three dimensions. Here we describe a protocol that combines an established model for squamous cell carcinoma of the tongue (SCOT) with two-photon imaging to allow multi-vectorial visualization of lingual tumor spread. The OSC-19 head and neck tumor cell line was stably engineered to express the F-actin binding peptide LifeAct fused to the mCherry fluorescent protein (LifeAct-mCherry). Fox1nu/nu mice injected with these cells reliably form tumors that allow the tongue to be visualized by ex-vivo application of two-photon microscopy. This technique allows for the orthotopic visualization of the tumor mass and locally invading cells in excised tongues without disruption of the regional tumor microenvironment. In addition, this system allows for the quantification of tumor cell invasion by calculating distances that invaded cells move from the primary tumor site. Overall this procedure provides an enhanced model system for analyzing factors that contribute to SCOT invasion and therapeutic treatments tailored to prevent local invasion and distant metastatic spread. This method also has the potential to be ultimately combined with other imaging modalities in an in vivo setting.

A comprehensive overview of the techniques involved in generating a mouse model of oral cancer and quantitative monitoring of tumor invasion within the tongue through multi-photon microscopy of labeled cells is presented. This system can serve as a useful platform for the molecular assessment and drug efficacy of anti-invasive compounds.

口腔扁平上皮癌の同所性マウスモデルにおける腫瘍細胞の浸潤の多光子イメージング

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient ...

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 ...

Roux-en-Y Gastric Bypass Operation in Rats

Currently, the most effective therapy for the treatment of morbid obesity to induce significant and maintained body weight loss with a proven mortality benefit is bariatric surgery1,2. Consequently, there has been a steady rise in the number of bariatric operations done worldwide in recent years with the Roux-en-Y gastric bypass (gastric bypass) being the most commonly performed operation3. Against this background, it is important to understand the physiological mechanisms by which gastric bypass induces and maintains body weight loss. These mechanisms are yet not fully understood, but may include reduced hunger and increased satiation4,5, increased energy expenditure6,7, altered preference for food high in fat and sugar8,9, altered salt and water handling of the kidney10 as well as alterations in gut microbiota11. Such changes seen after gastric bypass may at least partly stem from how the surgery alters the hormonal milieu because gastric bypass increases the postprandial release of peptide-YY (PYY) and glucagon-like-peptide-1 (GLP-1), hormones that are released by the gut in the presence of nutrients and that reduce eating12.
During the last two decades numerous studies using rats have been carried out to further investigate physiological changes after gastric bypass. The gastric bypass rat model has proven to be a valuable experimental tool not least as it closely mimics the time profile and magnitude of human weight loss, but also allows researchers to control and manipulate critical anatomic and physiologic factors including the use of appropriate controls. Consequently, there is a wide array of rat gastric bypass models available in the literature reviewed elsewhere in more detail 13-15. The description of the exact surgical technique of these models varies widely and differs e.g. in terms of pouch size, limb lengths, and the preservation of the vagal nerve. If reported, mortality rates seem to range from 0 to 35%15. Furthermore, surgery has been carried out almost exclusively in male rats of different strains and ages. Pre- and postoperative diets also varied significantly.
Technical and experimental variations in published gastric bypass rat models complicate the comparison and identification of potential physiological mechanisms involved in gastric bypass. There is no clear evidence that any of these models is superior, but there is an emerging need for standardization of the procedure to achieve consistent and comparable data. This article therefore aims to summarize and discuss technical and experimental details of our previously validated and published gastric bypass rat model.

Numerous studies using gastric bypass rat models have been recently conducted to uncover the underlying physiological mechanisms of Roux-en-Y gastric bypass operations. This article aims to demonstrate and discuss the technical and experimental details of our published gastric bypass rat model to understand advantages and limitations of this experimental tool.

ラットにおけるルー-en-Y法胃バイパス手術

Currently, the most effective therapy for the treatment of morbid obesity to induce significant and maintained body weight loss with a proven mortality ...

A Novel Surgical Approach for Intratracheal Administration of Bioactive Agents in a Fetal Mouse Model

Prenatal pulmonary delivery of cells, genes or pharmacologic agents could provide the basis for new therapeutic strategies for a variety of genetic and acquired diseases. Apart from congenital or inherited abnormalities with the requirement for long-term expression of the delivered gene, several non-inherited perinatal conditions, where short-term gene expression or pharmacological intervention is sufficient to achieve therapeutic effects, are considered as potential future indications for this kind of approach. Candidate diseases for the application of short-term prenatal therapy could be the transient neonatal deficiency of surfactant protein B causing neonatal respiratory distress syndrome1,2 or hyperoxic injuries of the neonatal lung3. Candidate diseases for permanent therapeutic correction are Cystic Fibrosis (CF)4, genetic variants of surfactant deficiencies5 and &alpha;1-antitrypsin deficiency6.
Generally, an important advantage of prenatal gene therapy is the ability to start therapeutic intervention early in development, at or even prior to clinical manifestations in the patient, thus preventing irreparable damage to the individual. In addition, fetal organs have an increased cell proliferation rate as compared to adult organs, which could allow a more efficient gene or stem cell transfer into the fetus. Furthermore, in utero gene delivery is performed when the individual's immune system is not completely mature. Therefore, transplantation of heterologous cells or supplementation of a non-functional or absent protein with a correct version should not cause immune sensitization to the cell, vector or transgene product, which has recently been proven to be the case with both cellular and genetic therapies7.
In the present study, we investigated the potential to directly target the fetal trachea in a mouse model. This procedure is in use in larger animal models such as rabbits and sheep8, and even in a clinical setting9, but has to date not been performed before in a mouse model. When studying the potential of fetal gene therapy for genetic diseases such as CF, the mouse model is very useful as a first proof-of-concept because of the wide availability of different transgenic mouse strains, the well documented embryogenesis and fetal development, less stringent ethical regulations, short gestation and the large litter size.
Different access routes have been described to target the fetal rodent lung, including intra-amniotic injection10-12, (ultrasound-guided) intrapulmonary injection13,14 and intravenous administration into the yolk sac vessels15,16 or umbilical vein17. Our novel surgical procedure enables researchers to inject the agent of choice directly into the fetal mouse trachea which allows for a more efficient delivery to the airways than existing techniques18.

We developed a novel surgical approach for intratracheal administration of bioactive agents into the mouse fetus. The delivery route is more efficient in targeting the fetal mouse lungs than the commonly used intra-amniotic injection. This procedure has to date not been described in a mouse model.

胎児のマウスモデルにおける生物活性剤の気管内投与のための新しい手術アプローチ

Prenatal pulmonary delivery of cells, genes or pharmacologic agents could provide the basis for new therapeutic strategies for a variety of genetic and ...

Evaluation of a Novel Laser-assisted Coronary Anastomotic Connector - the Trinity Clip - in a Porcine Off-pump Bypass Model

To simplify and facilitate beating heart (i.e., off-pump), minimally invasive coronary artery bypass surgery, a new coronary anastomotic connector, the Trinity Clip, is developed based on the excimer laser-assisted nonocclusive anastomosis technique. The Trinity Clip connector enables simplified, sutureless, and nonocclusive connection of the graft to the coronary artery, and an excimer laser catheter laser-punches the opening of the anastomosis. Consequently, owing to the complete nonocclusive anastomosis construction, coronary conditioning (i.e., occluding or shunting) is not necessary, in contrast to the conventional anastomotic technique, hence simplifying the off-pump bypass procedure. Prior to clinical application in coronary artery bypass grafting, the safety and quality of this novel connector will be evaluated in a long-term experimental porcine off-pump coronary artery bypass (OPCAB) study. In this paper, we describe how to evaluate the coronary anastomosis in the porcine OPCAB model using various techniques to assess its quality. Representative results are summarized and visually demonstrated.

This paper describes a novel nonocclusive coronary anastomotic connector in a porcine off-pump coronary artery bypass (OPCAB) model. This easy-to-use coronary connector has intrinsic potential to facilitate minimally invasive OPCAB surgery.

ブタオフポンプバイパスモデルで - トリニティクリップ - 小説レーザ支援冠動脈吻合コネクタの評価

To simplify and facilitate beating heart (i.e., off-pump), minimally invasive coronary artery bypass surgery, a new coronary anastomotic connector, the ...

A Novel Approach for the Administration of Medications and Fluids in Emergency Scenarios and Settings

The available routes of administration commonly used for medications and fluids in the acute care setting are generally limited to oral, intravenous, or intraosseous routes, but in many patients, particularly in the emergency or critical care settings, these routes are often unavailable or time-consuming to access. A novel device is now available that offers an easy route for administration of medications or fluids via rectal mucosal absorption (also referred to as proctoclysis in the case of fluid administration and subsequent absorption). Although originally intended for the palliative care market, the utility of this device in the emergency setting has recently been described. Specifically, reports of patients being treated for dehydration, alcohol withdrawal, vomiting, fever, myocardial infarction, hyperthyroidism, and cardiac arrest have shown success with administration of a wide variety of medications or fluids (including water, aspirin, lorazepam, ondansetron, acetaminophen, methimazole, and buspirone). Device placement is straightforward, and based on the observation of expected effects from the medication administrations, absorption is rapid. The rapidity of absorption kinetics are further demonstrated in a recent report of the measurement of phenobarbital pharmacokinetics. We describe here the placement and use of this device, and demonstrate methods of pharmacokinetic measurements of medications administered by this method.

This study presents a novel device that offers an easy route to quickly provide medications and fluids in patients with limited or difficult IV access. This small-diameter device is placed in the distal third of the rectum, allowing for ongoing medication and fluids administration.

緊急シナリオと設定における医薬品および流体の投与のための新しいアプローチ

The available routes of administration commonly used for medications and fluids in the acute care setting are generally limited to oral, intravenous, or ...

Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. Research models for bypass graft failure are essential for a better understanding of pathobiological and pathophysiological processes during graft patency loss. Large animal models, such as pigs or sheep, resemble human anatomical structures but require special facilities and equipment. This video describes a rat vein interposition model to investigate vein graft patency loss. Rats are inexpensive and easy to handle. Compared to mouse models, the convenient size of rats permits better operability and enables a sufficient amount of material to be obtained for further diverse analysis. In brief, the inferior epigastric vein of a donor rat is harvested and used to replace a segment of the femoral artery. Anastomosis is conducted via single stitches and sealed with fibrin glue. Graft patency can be monitored non-invasively using duplex sonography. Myointimal hyperplasia, which is the main cause for graft patency loss, develops progressively over time and can be calculated from histological cross sections.

This video demonstrates a model to study the development of myointimal hyperplasia after venous interposition surgery in rats.

静脈介在モデル：バイパスグラフト開存を研究するための適切なモデル

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. ...

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 ...

マウス モデルにおける心肺バイパス: 新しいアプローチ

マウスモデルにおける心肺バイパス: 新しいアプローチ