Name: long-term catheterization of the intestinal lymph trunk and collection of lymph in neonatal pigs
Uploaded: 2016-03-05T14:00:00.000+00:00
Duration: 6 min 25 s
Description: Watch this Scientific Journal Video about Long-Term Catheterization of the Intestinal Lymph Trunk and Collection of Lymph in Neonatal Pigs at JoVE.com

The work was supported in part by funding from Alberta Livestock and Meat Agency and Natural Science and Research Council Discovery grant to S. D. Proctor.

ビデオと原稿の両方で説明した実験動物でのすべての手順は、制度的動物実験委員会によって承認され、動物ケアのカナダ人評議会によって設定されたガイドラインに従いました。 1.外科麻酔や新生児ブタの外科的準備<ol><li>アザペロン（0.3ミリグラム/キログラム）、塩酸ケタミン（10mg / kgの）、デクスメデトミジン（15μgの/キログラム）：分離控室では、25キロの含有筋肉内鎮静麻酔薬カクテルと首の付け根の近くに豚をpremedicate。 注：強化された術中疼痛管理のためのプレ麻酔薬カクテルにブプレノルフィン（0.005から0.02ミリグラム/キログラム）を追加します。 </li><li>フェイスマスクを使用して（500ミリリットル、千ミリリットル/分のO 2で四から五パーセントイソフルラン）吸入イソフルラン吸入ガスと豚を麻酔。獣医の喉頭スコープ（17〜25センチメートル長いストレート刃）を使用して、声帯を視覚化し、ボーカルへの局所10％のリドカインの去勢を適用コー​​ド。リドカインスプレーは声帯の痙攣および気道閉塞の可能性を低減するために、挿管に30-60秒前に声帯に連絡することを許可します。 </li><li>声帯の間（5.0〜7.0ミリメートル内径（ID））カフ付き気管内チューブを渡すことで豚を挿管し、イソフルランガスで麻酔を維持（1,000〜2,000 mlの0.5から2.0パーセントイソフルラン/分O 2）が閉じを使用して、手術全体の麻酔システムを再呼吸回路。顎トーンによって麻酔のレベル、およびペダルと眼瞼反射応答の両方を評価します。期限切れの麻酔ガスを掃気し、手術室の外に排出されます。 </li><li> 70％イソプロピルアルコールリンスに続いて2.0％クロルヘキシジン手術用スクラブ溶液で耳の外面を清掃してください。手術中に、20 G静脈カテーテルを用いて、静脈内輸液（5〜10ミリリットル/ kg /時間乳酸リンゲル液）を提供するために、耳静脈をカテーテルを挿入。メッドの舌の粘膜表面にパルスオキシメータを確保iCalのテープは、心拍数および末梢血酸素飽和度（SpO 2）を監視します。 </li><li>背臥位で麻酔をかけた豚を置き、恥骨の腹側面に半ば胸部尾側から腹側腹部を剃ります。 2交互に2.0％クロルヘキシジン手術用スクラブし、滅菌水洗浄でこのエリアを清掃してください。 </li><li>手術室に麻酔豚を移し、70％イソプロピルアルコールリンスの最終的な手術用スクラブを適用し、それを乾燥させ、その後、動物を羽織ります。 </li><li>体温を監視するために、直腸内に、直腸温度プローブ約2〜4センチ挿入します。外科手術中に正常な体温（38-40°C）を維持するために、水の循環加熱パッド上豚を置きます。 </li><li>腹部の周りの上層象限パターンに配置された4タオルのカーテンで豚を羽織ります。約5℃、腹部の外側面に沿って第2のドレープ、剣状突起全体で最初のドレープを配置腹部正中線にメートルの横。骨盤の回腸稜と第四ドレープ全体で三ドレープを配置し、（反対側が）二ドレープのように、腹部の外側面腹部正中線に横約5cmに沿って配置されています。 </li><li>根底にあるタオルドレープの上に、手術部位へのアクセスを可能にするスリット開口と、大きなテーブルのドレープを置き、豚、全体の手術台をカバーしています。最終的なドレープは、大きなテーブルのドレープの上に配置された使い捨てsteriドレープです。 </li></ol>腸リンパ本幹の2腹部手術やカテーテル<ol><li>根本的な腹筋を公開するために手術用メスの刃で20センチの皮膚切開を行います。壁側腹膜を露出させるためにモノポーラ電気メス（20ワットの設定）で腹部の筋肉層を切開。腹部内臓やリンパ管にアクセスするためにMetzenbaumのはさみで頭頂腹膜20cmの直線セグメントを開きます。 </li><li&gt;手術中は開いた腹腔を維持するために外科的切開の頭蓋側面でリトラクタを配置します。 </li><li>全体の外科的処置のための暖かい（37°C）滅菌生理食塩水ですべての組織を湿ら。静かに腹腔から結腸、盲腸、回腸および空腸を含む小腸の大部分を持ち上げ、上腹部、肝臓やリンパ管にアクセスするには、豚の左脇腹にそれを体外に出します。優しく腸をサポートするためにスリングを形成するために追加のタオルドレープとの位置に体外に腸を固定します。 </li><li>リンパ管の位置を確認し、それが約4センチメートル右腎静脈、6センチメートル尾-medial-大静脈職長の腹側および膵臓22,36,37の近くに右肝葉の内臓の側面の下の頭蓋-内側を産みます。門脈36,37の右腹側セグメントに並置された半透明の構造としてリンパ管を識別します。 </li><li>リンパヴェスを切り離しエル優しくQチップアプリケーターで付着した組織を離れてからかうことにより、周囲の筋膜から。容器の外側の側面は、周囲の組織から分離されると、微鈍鉗子をひっくり返して容器の下の「トンネル」の開口部を作成します。 </li><li>細かいピンセットでリンパ管の下に3 2-0絹縫合糸を渡します。 、閉塞拡張とリンパで容器を埋めるために最初に最も尾側の縫合糸を結紮。意図的にリンパ管にカテーテルを固定する（4センチ）比較的長いこの縫合糸の端を残します。他の二つの縫合糸を配置し、互いから1.0センチメートルを分離し、保護された尾縫合糸に頭蓋約1.0〜1.5センチメートルです。血管内にカテーテルを速く確保を可能にするために &#39;一緩い結紮ネクタイ&#39;でこれら2本の縫合糸を残します。 注：（2頭蓋縫合の）ほとんどの尾側結紮縫合糸と中間縫合糸の間に位置するリンパ管のセグメントは、カテーテル挿入のためのサイトです。 suコマンドについて必要に応じて、トゥーレ材料は、2-0ポリグラクチン縫合糸は、2-0絹縫合糸の代わりにすることができます。 </li><li>アイリスハサミで容器に小さな穴をカットし、微鈍鉗子で血管を拡張。容器内に傾斜した端部に特殊なカテーテルチューブ（4.06外径（OD）X 2.31ミリメートルのID）の約1.0〜1.5センチメートルを挿入し、所定の位置にカテーテルを固定するための2つの頭蓋縫合糸を結びます。容器にカテーテルを固定するために尾縫合糸の長い縫合糸の端を使用してください。 </li><li>温かい生理食塩水のおびただしい量の体外に腸を洗浄し、静かに腸の正しい解剖学的な位置決めを確実に腹腔に戻します。 </li><li>左ミッド脇腹（腹側paralumbar窩5〜10センチメートル）でカテーテルを体外に出します。メスで皮膚切開を行い、カテーテルの具象化するための開口部を作成するために、皮膚表面に腹腔からトロカールを通過します。腹部CAVからカテーテルを体外に出すために、大きなケリー鉗子を使用しますトロカールの貫通開口性。 </li><li>ラウンド（テーパー）針で2-0ポリグラクチン縫合糸の単純連続縫合パターンで壁側腹膜を閉じます。ラウンド針に2-0ポリグラクチン縫合糸でシンプルな結節縫合パターンで腹部の筋肉層を閉じます。 </li><li>切断針に2-0ポリグラクチン縫合糸で表皮下のパターンで皮膚を閉じます。巾着縫合パターンおよび切断針上の2-0ナイロン縫合糸で皮膚に体外カテーテルを固定します。 </li><li>まだその配置を容易にし、リカバリ時に豚のストレスを軽減するために麻酔しながら、豚に特化したジャケットを置きます。 </li></ol> 3.手術後の回復とリンパコレクション<ol><li>前吸入麻酔の中止へ約10分で、即時術後の鎮痛を提供するために、筋肉内にbupenorphine（0.1ミリグラム/キログラム）を管理。 bupenorphineを続行（0.1ミリグラム/キログラム）24〜48時間を維持するために12時間ごとに術後の鎮痛。 </li><li>手術後の合併症のために7日間の期間ごとに8-12時間を豚を監視します。 </li><li>エチレンジアミン四酢酸（EDTA）でコーティングし、抗生物質を補充した500 mlのポリプロピレン製の洗瓶でリンパ液を集めます。ペニシリン（6000 IU）、ストレプトマイシン（6 mg）をアムホテリシンB（3 mg）を7日間ごとに12時間。 </li></ol>リポタンパク質ApoB48、トリグリセリド、コレステロールと総タンパクの4定量リンパから収集<ol><li> 4℃で5分間、1800×gでリンパサンプルを遠心します。上清を収集し、トリグリセリド、コレステロールおよび総タンパク質の定量化のためにそれを使用しています。 </li><li>希釈していないサンプル、サンプル希釈1：20蒸留水と1希釈した最終サンプル：蒸留水で100 3サンプルに上清を分割します。 </li><li>市販のキットを用いて、コレステロールレベルを測定するために希釈されていない上澄みを使用してください。 </li><li> 1時20分と午前1時10分を使用します0は、それぞれ、市販のキットおよびビシンコニン酸総タンパク質アッセイを用いてトリグリセリドおよび総タンパク質レベルを測定するため、サンプルを希釈しました。 </li></ol>リンパ38から収集したリポタンパク質ApoB48 5.定量化、 <ol><li>適応免疫ウエスタンブロッティング法38でリポタンパクApoB48の濃度を決定します。 3〜8％トリス - 酢酸塩、ドデシル硫酸ナトリウムポリアクリルアミドゲル電気泳動（SDS-PAGE）で区切り、総リンパ。 </li><li>ポリフッ化ビニリデン膜（0.45μmの）に分離したタンパク質を移し、アポB（1：4000）にヤギポリクローナル抗体でそれらをインキュベートし、その後、抗ヤギ二次抗体とそれをバインドします。 </li><li>精製されたげっ歯類ApoB48タンパク質標準との線形濃度測定の比較を用い​​て、化学発光とリポ蛋白ApoB48を定量化します。 </li></ol>

The authors have nothing to disclose.

腸リンパを収集する様々な実験動物モデルにおいて、脂質8-12に関与する機構と薬物13-15代謝、癌転移16,17、細胞輸送及び免疫機能18-26を調査するための優れた方法です。実際に、長期間にわたって、末梢（求心性）と中央（遠心性と大きなトランク船）リンパを大量に収穫する能力は、免疫調節剤を18-22と課題以下の細胞集団で発生する時間的な変化を...

リンパ系は、生理の​​understudied領域です。リンパカテーテル法の前臨床モデルは、異なる動物種1-8で発生し、脂質8-12および薬物代謝13-15、実験的治療17と癌転移16に関与するメカニズムを調査するために製薬産業や研究機関によって使用され、免疫機能18 -26。この研究は、リポタンパク質代謝の成分を測定するために、国内のブタモデルにおける腸管リンパトランクカテーテルの使用を探ります。リポタンパク質代謝を産生および分泌カイロミクロンの、ならびに関連する脂質および総蛋白質の変化に関与しています。一般的に使用される齧歯類モデルと人間の間のような脂質代謝の主要な違いがあるように、これらは私を、脂質を研究するためのより多くの比較可能な情報を提供することができる腸のリンパ節を収集するために豚のモデルを採用し、重要な考慮すべき事項であります人々 27-31でtabolism。 いくつかの外科技術は、大規模な動物種における腸管リンパを収集するために使用されている：頭蓋肩のアプローチ（ すなわち、胸管カテーテル法）5、横上部脇腹のアプローチ32-34、および腹側正中線またはparamedianアプローチ22,35。このビデオでは、具体的に腸のリンパトランクのカテーテル挿入のための腹側正中外科的アプローチを使用して、ブタにおいて外科的手順を説明します。慎重な手術法は、長期間にわたって大リンパの量とその成分を収集するためにリンパカテーテル法のこの方法が可能になります。 この技術は、様々な生理学的機能を調べる多くの分野への応用の無数を開きます。アプリケーションが含まれる可能性があり、これらに限定されないが、全身リポタンパク質と脂質代謝、免疫学、腫瘍の起源及び転移、腸機能、およびdevelopm耳鼻咽喉科や腸の炎症性疾患の進行。

<table><tbody><tr><td>Miller laryngoscope blade</td><td>Welch Allyn</td><td>68044</td><td>182 mm length</td></tr><tr><td>Surgivet advisor: Vital signs monitor</td><td>Surgivet</td><td>V9203</td><td></td></tr><tr><td>Rectal temperature probe</td><td>Surgivet</td><td>V3417</td><td></td></tr><tr><td>Mono-polar electrosurgery generator</td><td>Valley Lab</td><td></td><td></td></tr><tr><td>Metzenbaum scissors</td><td>Fine Science</td><td>14518-18</td><td></td></tr><tr><td>Tuffier retractor</td><td>Stevens</td><td>162-11-676</td><td></td></tr><tr><td>Mosquito forceps</td><td>Stevens</td><td>162-7-10</td><td></td></tr><tr><td>Kelly forceps-curved (14cm)</td><td>Stevens</td><td>162-7-38</td><td></td></tr><tr><td>Allis tissue forceps</td><td>Stevens</td><td>162-7-38</td><td></td></tr><tr><td>Forceps dressing-eye (10.2cm)</td><td>Stevens</td><td>162-18-780</td><td></td></tr><tr><td>Forceps dressing-Adison (12.1cm)</td><td>Stevens</td><td>162-17-2510</td><td></td></tr><tr><td>Needle Drivers</td><td>Stevens</td><td>162-V98-42</td><td></td></tr><tr><td>Iris scissors</td><td>Fine science</td><td>14058-11</td><td></td></tr><tr><td>Circulating water pump</td><td>Jorvet</td><td>J-783X</td><td></td></tr><tr><td>Maxitherm-Vinyl blanket</td><td>Jorvet</td><td>J-784C</td><td></td></tr><tr><td>Q tip applicators</td><td>Fisher Scientific</td><td>22-037-960</td><td></td></tr><tr><td>Catheterization&amp;nbsp; tubing (4.06 OD X 2.31 ID)</td><td>Braintree Scientific Inc.</td><td>MRE-160</td><td>Micro-Renethane implantation tubing</td></tr><tr><td>2-0 silk suture</td><td>Ethicon</td><td>LA556</td><td></td></tr><tr><td>2-0 polyglactin suture</td><td>Ethicon</td><td>J443H</td><td>2-0 vicryl</td></tr><tr><td>Large animal jacket</td><td>Lomir Biomedical Inc.</td><td>SSJ2YC</td><td></td></tr><tr><td>Polypropylene wash bottles</td><td>Fisher Scientific</td><td>03-409-22C</td><td>500 ml</td></tr><tr><td>Penicillin-Streptomycin</td><td>Sigma Aldrich</td><td>D4333</td><td></td></tr><tr><td>EDTA</td><td>Sigma Aldrich</td><td>60-00-4</td><td></td></tr><tr><td>Amphotericin B</td><td>Sigma Aldrich</td><td>A2411</td><td></td></tr><tr><td>Azaperone</td><td>Elanco Animal Health</td><td></td><td>Stresnil</td></tr><tr><td>Dexmedetomidine hydrochloride</td><td>Zoetis</td><td>6295</td><td>Dexdomitor</td></tr><tr><td>Isoflurane</td><td>Abbott Animal&amp;nbsp; Health</td><td>05260-5</td><td>IsoFlo</td></tr><tr><td>Ketamine hydrochloride</td><td>Zoetis</td><td>2626</td><td>Ketaset</td></tr><tr><td>Bupenorphine hydrochloride</td><td>Champion Alstoe Animal Health</td><td></td><td>DIN:02347510</td></tr><tr><td>6 mm Endotracheal tube</td><td>Jorvet</td><td>J-165d</td><td></td></tr><tr><td>10% Lidocaine spray</td><td>AstraZeneca</td><td></td><td>DIN:02003767</td></tr><tr><td>4 % Chlorhexidine surgical scrub</td><td>Partnar Animal Health</td><td>PCH-011</td><td>Diluted: 2.0% solution</td></tr><tr><td>3M Surgical steri- drape</td><td>3M Health Care</td><td>1040</td><td></td></tr><tr><td>SDS page gel</td><td>Invitrogen</td><td>EA0375BOX</td><td>3-8 % tris acetate</td></tr><tr><td>Polyvinylidene fluoride membrane</td><td>Millipore</td><td>IPVH00010</td><td>0.45 &amp;mu;m pore size</td></tr><tr><td>ApoB antibody&amp;nbsp;</td><td>EMD Millipore</td><td>AB742</td><td>1:4000 dilution</td></tr><tr><td>Donkey anti-goat IgG-HRP</td><td>Santa Cruz Biotechnology</td><td>Sc-2304</td><td></td></tr><tr><td>ECL Prime Western Blotting Reagent</td><td>GE Healthcare LifeSciences</td><td>RPN2232&amp;nbsp;&amp;nbsp;&amp;nbsp;</td><td></td></tr><tr><td>Triglyceride Kit</td><td>Wako Pure Chemicals</td><td>998-40391/994-40491</td><td></td></tr><tr><td>Total Cholesterol Kit</td><td>Wako Pure Chemicals</td><td>439-17501</td><td></td></tr><tr><td>Total Protein</td><td>&amp;nbsp;Pierce&amp;nbsp;</td><td>23225</td><td>Bicinchoninic Acid Assay</td></tr></tbody></table>

long-term catheterization of the intestinal lymph trunk and collection of lymph in neonatal pigs

Catheterization of the intestinal lymph trunk in neonatal pigs is a technique allowing for the long-term collection of large quantities of intestinal (central) efferent lymph. Importantly, the collection of central lymph from the intestine enables researchers to study both the mechanisms and lipid constitutes associated with lipid metabolism, intestinal inflammation and cancer metastasis, as well as cells involved in immune function and immunosurveillance. A ventral mid-line surgical approach permits excellent surgical exposure to the cranial abdomen and relatively easy access to the intestinal lymph trunk vessel that lies near the pancreas and the right ventral segment of the portal vein underneath the visceral aspect of the right liver lobe. The vessel is meticulously dissected and released from the surrounding fascia and then dilated with sutures allowing for insertion and subsequent securing of the catheter into the vessel. The catheter is exteriorized and approximately 1 L/24 hr of lymph is collected over a 7 day period. While this technique enables the collection of large quantities of central lymph over an extended period of time, the success depends on careful surgical dissection, tissue handling and close attention to proper surgical technique. This is particularly important with surgeries in young animals as the lymph vessels can easily tear, potentially leading to surgical and experimental failure. The video demonstrates an excellent surgical technique for the collection of intestinal lymph.

<html>新生児の豚の腸リンパ本幹のリンパカテーテル法は、7日間にわたって中央リンパの約1 L / 24時間の収集を可能にします。この実験で収集さリンパは、脂質代謝、すなわち総リンパタンパク質、ApoB48リポタンパク質、トリグリセリド、総タンパク質、およびコレステロールの成分を含有していました。表1のハイライト3匹のブタのプールされたリンパサンプルから?...

Watch this Scientific Journal Video about Long-Term Catheterization of the Intestinal Lymph Trunk and Collection of Lymph in Neonatal Pigs at JoVE.com

Long-Term Catheterization of the Intestinal Lymph Trunk and Collection of Lymph in Neonatal Pigs

We present a surgical procedure to catheterize the intestinal lymph trunk in neonatal pigs to collect large quantities of lipid metabolism components from efferent lymph.

新生豚のリンパ節の腸リンパトランクやコレクションの長期カテーテル法

Catheterization of the intestinal lymph trunk in neonatal pigs is a technique allowing for the long-term collection of large quantities of intestinal ...

long-term-catheterization-intestinal-lymph-trunk-collection-lymph

Research

JoVE Journal

Medicine

8.7K ビュー.  University of Alberta. We present a surgical procedure to catheterize the intestinal lymph trunk in neonatal pigs to collect large quantities of lipid metabolism components from efferent lymph.

ビデオ: Long-Term Catheterization of the Intestinal Lymph Trunk and Collection of Lymph in Neonatal Pigs

Multispectral Real-time Fluorescence Imaging for Intraoperative Detection of the Sentinel Lymph Node in Gynecologic Oncology

The prognosis in virtually all solid tumors depends on the presence or absence of lymph node metastases.1-3 Surgical treatment most often combines radical excision of the tumor with a full lymphadenectomy in the drainage area of the tumor. However, removal of lymph nodes is associated with increased morbidity due to infection, wound breakdown and lymphedema.4,5 As an alternative, the sentinel lymph node procedure (SLN) was developed several decades ago to detect the first draining lymph node from the tumor.6 In case of lymphogenic dissemination, the SLN is the first lymph node that is affected (Figure 1). Hence, if the SLN does not contain metastases, downstream lymph nodes will also be free from tumor metastases and need not to be removed. The SLN procedure is part of the treatment for many tumor types, like breast cancer and melanoma, but also for cancer of the vulva and cervix.7 The current standard methodology for SLN-detection is by peritumoral injection of radiocolloid one day prior to surgery, and a colored dye intraoperatively. Disadvantages of the procedure in cervical and vulvar cancer are multiple injections in the genital area, leading to increased psychological distress for the patient, and the use of radioactive colloid.
Multispectral fluorescence imaging is an emerging imaging modality that can be applied intraoperatively without the need for injection of radiocolloid. For intraoperative fluorescence imaging, two components are needed: a fluorescent agent and a quantitative optical system for intraoperative imaging. As a fluorophore we have used indocyanine green (ICG). ICG has been used for many decades to assess cardiac function, cerebral perfusion and liver perfusion.8 It is an inert drug with a safe pharmaco-biological profile. When excited at around 750 nm, it emits light in the near-infrared spectrum around 800 nm. A custom-made multispectral fluorescence imaging camera system was used.9.
The aim of this video article is to demonstrate the detection of the SLN using intraoperative fluorescence imaging in patients with cervical and vulvar cancer. Fluorescence imaging is used in conjunction with the standard procedure, consisting of radiocolloid and a blue dye. In the future, intraoperative fluorescence imaging might replace the current method and is also easily transferable to other indications like breast cancer and melanoma.

Fluorescence imaging is a promising innovative modality for image-guided surgery in surgical oncology. In this video we describe the technical procedure for detection of the sentinel lymph node using fluorescence imaging as showcased in gynecologic oncologicy. A multispectral fluorescence camera system, together with the fluorescent agent indocyanine green, is applied.

婦人科腫瘍におけるセンチネルリンパ節の術中検出のためのマルチスペクトルのリアルタイム蛍光イメージング

The prognosis in virtually all solid tumors depends on the presence or absence of lymph node metastases.1-3 Surgical treatment most often combines radical ...

医学

A Swine Model of Neonatal Asphyxia

Annually more than 1 million neonates die worldwide as related to asphyxia. Asphyxiated neonates commonly have multi-organ failure including hypotension, perfusion deficit, hypoxic-ischemic encephalopathy, pulmonary hypertension, vasculopathic enterocolitis, renal failure and thrombo-embolic complications. Animal models are developed to help us understand the patho-physiology and pharmacology of neonatal asphyxia. In comparison to rodents and newborn lambs, the newborn piglet has been proven to be a valuable model. The newborn piglet has several advantages including similar development as that of 36-38 weeks human fetus with comparable body systems, large body size (&tilde;1.5-2 kg at birth) that allows the instrumentation and monitoring of the animal and controls the confounding variables of hypoxia and hemodynamic derangements.
We here describe an experimental protocol to simulate neonatal asphyxia and allow us to examine the systemic and regional hemodynamic changes during the asphyxiating and reoxygenation process as well as the respective effects of interventions. Further, the model has the advantage of studying multi-organ failure or dysfunction simultaneously and the interaction with various body systems. The experimental model is a non-survival procedure that involves the surgical instrumentation of newborn piglets (1-3 day-old and 1.5-2.5 kg weight, mixed breed) to allow the establishment of mechanical ventilation, vascular (arterial and central venous) access and the placement of catheters and flow probes (Transonic Inc.) for the continuously monitoring of intra-vascular pressure and blood flow across different arteries including main pulmonary, common carotid, superior mesenteric and left renal arteries. Using these surgically instrumented piglets, after stabilization for 30-60 minutes as defined by Z&lt;10% variation in hemodynamic parameters and normal blood gases, we commence an experimental protocol of severe hypoxemia which is induced via normocapnic alveolar hypoxia. The piglet is ventilated with 10-15% oxygen by increasing the inhaled concentration of nitrogen gas for 2h, aiming for arterial oxygen saturations of 30-40%. This degree of hypoxemia will produce clinical asphyxia with severe metabolic acidosis, systemic hypotension and cardiogenic shock with hypoperfusion to vital organs. The hypoxia is followed by reoxygenation with 100% oxygen for 0.5h and then 21% oxygen for 3.5h. Pharmacologic interventions can be introduced in due course and their effects investigated in a blinded, block-randomized fashion.

Large animal models have good translational values in the examination of physiology and pharmacology of neonatal asphyxia. Using newborn piglets, we develop an experimental protocol to simulate neonatal asphyxia which has advantages of studying the systemic and regional hemodynamics, oxygen transport with biochemical and pathologic pathways and correlations.

新生児仮死の新型モデル

Annually more than 1 million neonates die worldwide as related to asphyxia. Asphyxiated neonates commonly have multi-organ failure including hypotension, ...

Lavage-induced Surfactant Depletion in Pigs As a Model of the Acute Respiratory Distress Syndrome (ARDS)

Various animal models of lung injury exist to study the complex pathomechanisms of human acute respiratory distress syndrome (ARDS) and evaluate future therapies. Severe lung injury with a reproducible deterioration of pulmonary gas exchange and hemodynamics can be induced in anesthetized pigs using repeated lung lavages with warmed 0.9% saline (50 ml/kg body weight). Including standard respiratory and hemodynamic monitoring with clinically applied devices in this model allows the evaluation of novel therapeutic strategies (drugs, modern ventilators, extracorporeal membrane oxygenators, ECMO), and bridges the gap between bench and bedside. Furthermore, induction of lung injury with lung lavages does not require the injection of pathogens/endotoxins that impact on measurements of pro- and anti-inflammatory cytokines. A disadvantage of the model is the high recruitability of atelectatic lung tissue. Standardization of the model helps to avoid pitfalls, to ensure comparability between experiments, and to reduce the number of animals needed.

Lavage-induced Surfactant Depletion in Pigs As a Model of the Acute Respiratory Distress Syndrome ARDS

Repeated pulmonary lavages in anesthetized pigs induce lung injury resembling major aspects of human acute respiratory distress syndrome (ARDS). For this purpose the lungs are repeatedly lavaged with 0.9% saline at 37 &#176;C. The goal of the protocol is a reproducible mitigation of gas exchange and hemodynamics for research in ARDS.

急性呼吸窮迫症候群（ARDS）のモデルとして、ブタにおける洗浄によって誘発される界面活性剤枯渇

Various animal models of lung injury exist to study the complex pathomechanisms of human acute respiratory distress syndrome (ARDS) and evaluate future ...

Surgical Placement of Catheters for Long-term Cardiovascular Exercise Testing in Swine

This protocol describes the surgical procedure to chronically instrument swine and the procedure to exercise swine on a motor-driven treadmill. Early cardiopulmonary dysfunction is difficult to diagnose, particularly in animal models, as cardiopulmonary function is often measured invasively, requiring anesthesia. As many anesthetic agents are cardiodepressive, subtle changes in cardiovascular function may be masked. In contrast, chronic instrumentation allows for measurement of cardiopulmonary function in the awake state, so that measurements can be obtained under quiet resting conditions, without the effects of anesthesia and acute surgical trauma. Furthermore, when animals are properly trained, measurements can also be obtained during graded treadmill exercise.
Flow probes are placed around the aorta or pulmonary artery for measurement of cardiac output and around the left anterior descending coronary artery for measurement of coronary blood flow. Fluid-filled catheters are implanted in the aorta, pulmonary artery, left atrium, left ventricle and right ventricle for pressure measurement and blood sampling. In addition, a 20 G&#160;catheter is positioned in the anterior interventricular vein to allow coronary venous blood sampling.
After a week of recovery, swine are placed on a motor-driven treadmill, the catheters are connected to pressure and flow meters, and swine are subjected to a five-stage progressive exercise protocol, with each stage lasting 3 min. Hemodynamic signals are continuously recorded and blood samples are taken during the last 30 sec of each exercise stage.
The major advantage of studying chronically instrumented animals is that it allows serial assessment of cardiopulmonary function, not only at rest but also during physical stress such as exercise. Moreover, cardiopulmonary function can be assessed repeatedly during disease development and during chronic treatment, thereby increasing statistical power and hence limiting the number of animals required for a study.

Here we present a protocol to assess cardiopulmonary function in awake swine, at rest and during graded treadmill exercise. Chronic instrumentation allows for repeated hemodynamic measurements uninfluenced by cardiodepressive anesthetic agents.

豚の長期心血管運動テストのためのカテーテルの外科的配置

This protocol describes the surgical procedure to chronically instrument swine and the procedure to exercise swine on a motor-driven treadmill. Early ...

Direct Cannula Implantation in the Cisterna Magna of Pigs

The glymphatic system is a waste clearance system in the brain that relies on the flow of cerebrospinal fluid (CSF) in astrocyte-bound perivascular spaces and has been implicated in the clearance of neurotoxic peptides such as amyloid-beta. Impaired glymphatic function exacerbates disease pathology in animal models of neurodegenerative diseases, such as Alzheimer's, which highlights the importance of understanding this clearance system. The glymphatic system is often studied by cisterna magna cannulations (CMc), where tracers are delivered directly into the cerebrospinal fluid (CSF). Most studies, however, have been carried out in rodents. Here, we demonstrate an adaptation of the CMc technique in pigs. Using CMc in pigs, the glymphatic system can be studied at a high optical resolution in gyrencephalic brains and in doing so bridges the knowledge gap between rodent and human glymphatics.

This article presents a step-by-step protocol for the direct cannula implantation in the cisterna magna of pigs.

豚のシスターナマグナにおける直接カニューレ移植

The glymphatic system is a waste clearance system in the brain that relies on the flow of cerebrospinal fluid (CSF) in astrocyte-bound perivascular spaces ...

神経科学

Microscopic Observation of Lymphocyte Dynamics in Rat Peyer's Patches

Na&#239;ve lymphocytes recirculate from the blood to the lymphoid tissues under physiological condition and it is commonly recognized as an important phenomenon in the gut immunity. The stroma of secondary lymphoid organs, such as Peyer&#8217;s patches (PPs) or mesenteric lymph nodes, are where na&#239;ve lymphocytes sense antigens. Na&#239;ve lymphocytes circulate through the bloodstream to reach high endothelial venules, the portal of entry into PPs. Some immunomodulators are estimated to influence lymphocyte migration, but the precise evaluation of microcirculation dynamics is very difficult, and establishing a method to observe lymphocyte migration in vivo can contribute to the clarification of the precise mechanisms. We refined the method of collecting lymphocytes from the lymph duct and observing the detailed dynamics of gut-tropic lymphocytes in rat PPs. We chose confocal laser scanning microscopy to observe rat PPs in vivo and recorded it using time-lapse photography. We can now obtain clear images that can contribute to the analysis of lymphocyte dynamics.

Here, we describe a precise method for collecting thoracic duct lymphocytes and observing the migration of gut-tropic lymphocytes in rat Peyer&#8217;s patches for 3 hours using time-lapse photography. This technique can clarify how the dynamics of lymphocytes are affected under inflammatory conditions.

ラット・ペイアーのパッチにおけるリンパ球ダイナミクスの顕微鏡観察

Na&#239;ve lymphocytes recirculate from the blood to the lymphoid tissues under physiological condition and it is commonly recognized as an important ...

Intubation, Central Venous Catheter, and Arterial Line Placement in Swine for Translational Research in Abdominal Transplantation Surgery

Translational surgical research models in swine are crucial for developing safe preclinical protocols. However, the success of the experimental surgeries does not solely rely on the research team's surgical skills; perioperative care and management procedures, like intubation, central venous line, and arterial line placement, are necessary and of the utmost importance for favorable experiment results. As it is uncommon for research teams to have anesthesiologists or any other staff other than the surgical team, the surgical team involved in translational research must acquire and/or develop the skills to perform the perioperative care. The purpose of this paper is to show the techniques of intubation, central venous catheter, and arterial line placement used and perfected at the Toronto Organ Preservation Laboratory over the last 10 years, to be used as a reference for future researchers joining either this team or any other lab performing translational research protocols in swine and/or abdominal transplantation.

To obtain the best possible results, surgeons performing translational research experiments should be proficient at intubation and vascular line placement. This paper describes the techniques used by the Toronto Organ Preservation Laboratory to perform these procedures.

Translational surgical research models in swine are crucial for developing safe preclinical protocols. However, the success of the experimental surgeries ...

豚の長期血管アクセスのためのヒックマンカテーテル

Hickman Catheter Use for Long-Term Vascular Access in a Preclinical Swine Model

Central venous catheters (CVCs) are invaluable devices in large animal research as they facilitate a wide range of medical applications, including blood monitoring and reliable intravenous fluid and drug administration. Specifically, the tunneled multi-lumen Hickman catheter (HC) is commonly used in swine models due to its lower extrication and complication rates. Despite fewer complications relative to other CVCs, HC-related morbidity presents a significant challenge, as it can significantly delay or otherwise negatively impact ongoing studies. The proper insertion and maintenance of HCs is paramount in preventing these complications, but there is no consensus on best practices. The purpose of this protocol is to comprehensively describe an approach for the insertion and maintenance of a tunneled HC in swine that mitigates HC-related complications and morbidity. The use of these techniques in &gt;100 swine has resulted in complication-free patent lines up to 8 months and no catheter-related mortality or infection of the ventral surgical site. This protocol offers a method to optimize the lifespan of the HC and guidance for approaching issues during use.

著者スポットライト:前臨床豚モデルにおける長期血管アクセスのためのヒックマンカテーテルの使用  

A reliable and reproducible approach for the insertion and maintenance of a tunneled Hickman catheter for long-term vascular access in swine is described. Placement of a central venous catheter allows for convenient daily sampling of whole blood from awake animals and intravenous administration of medication and fluids.

ヒックマンカテーテルによる前臨床豚モデルにおける長期血管アクセスのための使用

Central venous catheters (CVCs) are invaluable devices in large animal research as they facilitate a wide range of medical applications, including blood ...

新生豚のリンパ節の腸リンパトランクやコレクションの長期カテーテル法

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婦人科腫瘍におけるセンチネルリンパ節の術中検出のためのマルチスペクトルのリアルタイム蛍光イメージング

新生児仮死の新型モデル

急性呼吸窮迫症候群（ARDS）のモデルとして、ブタにおける洗浄によって誘発される界面活性剤枯渇

豚の長期心血管運動テストのためのカテーテルの外科的配置

豚のシスターナマグナにおける直接カニューレ移植

ラット・ペイアーのパッチにおけるリンパ球ダイナミクスの顕微鏡観察

Intubation, Central Venous Catheter, and Arterial Line Placement in Swine for Translational Research in Abdominal Transplantation Surgery

ヒックマンカテーテルによる前臨床豚モデルにおける長期血管アクセスのための使用

ヒックマンカテーテルによる前臨床豚モデルにおける長期血管アクセスのための使用

ヒックマンカテーテルによる前臨床豚モデルにおける長期血管アクセスのための使用