プロジェクト番号NVKP_16-1–2016-0017(「ナショナルハートプログラム」)は、NVKP_16資金提供スキームの下で資金提供されたハンガリー国立研究開発イノベーション基金からの支援を受けて実施されました。この研究は、センメルワイス大学の治療法開発およびバイオイメージングテーマ別プログラムの枠組みの中で、ハンガリーのイノベーション技術省のテーマ別エクセレンスプログラム(2020-4.1.1.-TKP2020)によって資金提供されました。

プロトコルは、施設の人間の研究倫理委員会のガイドラインに従い、臨床例の患者は研究に書面によるインフォームドコンセントを与えました。
1. 技術要件
<ol><li>3Dの取得と解析には、適切なソフトウェアとハードウェアを使用してください。心エコー検査装置のECGケーブルを使用します。さらに、以下に説明する完全な3D取得プロトコルには必須です。</li>
	<li>3D取得には、3D心エコープローブと3D互換超音波装置を使用してください。3D RV体積解析には、専用のソフトウェアを使用してください。</li>
</ol>2. 買収
<ol><li>ほとんどの場合、頂端ビューを使用してRVの3D取得を実行します。LVに焦点を当てたビューとは対照的に、別の患者のポジショニングが推奨されます。正しい頂端ビュー上の1つの肋間空間に切り替えることによって実質的により良い画質を達成できる場合、この短縮されたビューは、より良い3D画質を可能にし得る。縮み率は、3D 解析中に修正できます。<ol><li>左外側褥瘡位(左腕を頭の上に伸ばして左側に横たわっている患者)が推奨される標準的な根尖心エコー取得と比較して、トランスデューサーのより横方向の位置を可能にするために、患者をわずかに後ろに傾けてもらいます。</li>
		<li>RVのみを含む画像深度を選択します。 不必要に奥行きが大きいと、RV体積分析に関する有益な効果がないため、取得フレームレートが低下する可能性があります。</li>
	</ol></li>
	<li>2D心エコー画像から正しいRVフォーカスビューを確認します。このビューからでもRVの自由壁の視覚化が不十分な場合、期待される3D画質はさらなる分析に最適ではありません。</li>
	<li>4Dボタンを使用してライブ 3D イメージングに切り替え、RVビューをさらに修正することができます。</li>
	<li>3Dライブモードは見た目に非常に美しいかもしれませんが、 3D ビューには 12スライス モードを使用して、関心領域の三葉機画像と自由に変更できる9つの断面面を表示します。切断面の回転と正しい位置により、RVフリーウォール全体(流出路と先端セグメントを含む)の可視性を確認します。</li>
	<li>セクターの左傾化(タッチスクリーンの2ページ目)を使用して画像をさらに調整し、RVの視覚化を改善します。</li>
	<li>RV体積解析には、 マルチ ビートモードと シングルビート モードの2つの3D取得モードを使用します。ただし、すべての患者でこれらのアプローチの両方を使用しますが、場合によっては(例:.、特定の不整脈、患者の重度の呼吸困難)、後者のみが実行可能な場合があります。.</li>
	<li>シングルビートモードを使用して、画質とフレームレートのトレードオフを実現します。最適な画像深度、幅、フレームレート(タッチスクリーンの下部パネル)を選択し、それ以上のアクションなしでRVの3Dループを取得します。この方法は大多数の患者で実行可能です。ただし、マルチビートアプローチと比較して、一般的に画質とフレームレートが低くなります。<ol><li>平均心拍数(60〜70 /分)の場合、適切なRV分析のために、フレームレートの下限を16フレーム/秒に保ちます。ただし、頻脈が存在する場合は、さらに高いフレームレートをお勧めします。</li>
	</ol></li>
	<li>マルチビートモードを使用して、タッチスクリーンで選択できる所定の数の心臓サイクルから取得した3Dループを再構築します(2、3、4、6 ビート モードを使用できます)。シングルビート取得とは対照的に、一般的により良い画質とフレームレートが期待されます。ただし、比較的一定の心周期の長さと、必須の息止め操作による患者のコンプライアンスが必要です。操作は、いわゆるステッチアーティファクトを回避するために不可欠です:取得した3Dボリュームが一緒にステッチされると、不均等な心周期の長さおよび/または呼吸による動きがこの現象を引き起こす可能性があります。<ol><li>プローブの正しい位置と機械の設定(「シングルビート」モードと同様)の後、患者に深呼吸をして保持するように依頼します。この場合、拡張肺は通常画像全体を覆います。</li>
		<li>患者に、厳密にガイダンスを付けてゆっくりと息を吐くように依頼します。肺の収縮と並行して、RVは再び見えるようになります。</li>
		<li>RV全体(自由壁と中隔)が再び現れたら、この状態で息を止めるように患者に依頼します。</li>
		<li>画面上で マルチビート をクリックして取得を開始すると、指定された量の心臓サイクル中に3Dループが蓄積されます。</li>
		<li>取得の準備ができたら(RV全体が視覚化されます)、患者に再び自由に呼吸するように依頼します。</li>
		<li>取得したループをチェックして、ステッチングやドロップアウトアーティファクトがないかどうかを確認します。</li>
	</ol></li>
</ol>3. 4D RV解析
<ol><li>専用ソフトウェアを使用して、RVの3D体積解析を実行します。患者ライブラリからRVに焦点を当てた3Dループを選択した後、ボリュームフォルダにある測定ウィンドウからソフトウェアを開きます。</li>
	<li>ソフトウェアを開いた後、RVを4つの事前定義された切断面に向けます。<ol><li>左上下の長軸平面の三尖弁の中心に2つのマーカー(TVセンター)を配置します。回転ツールを使用して、画像の長軸をRVの実際の長軸に調整します。右上の参照画像は、正しい向きがどのように表示されるかを示しています。</li>
		<li>右上のパネルで、回転によって短軸画像を正しい位置に揃えます。前の手順と同様に、参照画像もこのプロセスに役立ちます。</li>
	</ol></li>
	<li>完了したら、[ ランドマークの設定 ] をクリックして解析の次のステップに進みます。2つの画像にランドマークを設定します。<ol><li>左側で、自由壁(TVフリーウォール)とセプタム(TVセプタム)の三尖弁輪と、以前に配置された頂端4室ビューのRV頂点をマークします。</li>
		<li>右側で、RVの後方(LV / RV後方)と前方挿入点(LV / RV前方)とRV自由壁(RV自由壁)を設定します。前のウィンドウと同様に、右上隅の参照画像は、正しいセットアップに関するヘルプです。すべてのランドマークを設定した後、ソフトウェアは自動的に次のウィンドウ(レビュー)にジャンプします。</li>
	</ol></li>
	<li>このウィンドウ(レビュー)で、必要に応じて、心周期全体を通して心内膜境界の自動検出を確認し、手動で修正します。デフォルトでは、9つのパネルを見ることができます:左側に3つの移動ループ(1つの長軸と2つの短軸)、中央に同じ画像の拡張末期フレーム、右側に収縮期末期のフレーム。<ol><li>誤った追跡の場合は、心内膜の境界(緑色の線)、追跡された境界をクリックして自由に修正します。短軸の回転ツールを使用して、RVの全周に沿って追跡する画像を確認します。 右側のパネルで[ ペンサイズ ]を選択して、補正の大きさを調整します。トラッキングが正しいと思われる場合は、同じパネルの[ 結果 ]をクリックします。</li>
	</ol></li>
	<li>最後のセクションでは、右上(ワークシートパネル )で最終的な3D体積データとその他の計算パラメータを確認します。RV容量と排出率に加えて、ソフトウェアは、線形(中軸、基礎、長軸)直径などの2Dパラメータ、および事前定義された頂端4チャンバービューから得られたFACおよびTAPSE値も表示します。ソフトウェアはまた、RVの長軸と短軸(左側)、RVの3Dライブモデル(中央上)、およびチャンバーの体積-時間曲線(右下)も表示します。<ol><li>追跡をさらに調整する必要がある場合は、右側のパネルでクリックすることで、以前のすべての手順を修正できます。トラッキングと3Dパラメータが有効であると考えられる場合は、同じパネルの[承認して終了]をクリックして結果を保存します。</li>
	</ol></li>
</ol>

<ol>
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RVの3D分析は、日常の心臓病学の実践における重要なステップを表しています。以前は無視されていた心腔の形態と機能への関心の高まりと並行して、これらの新しいソリューションは、心臓の右側に関する臨床的に意味のある情報を提供します。3D取得には、2D心エコー画像とは著しく異なるいくつかの側面がありますが、重要なポイントに特別な注意を払い、徹底したプロトコルを使用す?...

心エコー検査は、1950年代の最初の臨床応用から長い道のりを歩んできました1。最初の1次元超音波プローブは、チャンバーの壁と内腔の単純な直線直径を提供するように設計されました。しかし、それらは間違いなく心臓血管イメージングのマイルストーンを表しています。2次元(2D)超音波イメージングの開発は、形態と機能のはるかに正確な定量化を提供することによるもう一つの大きなステップであり、今でも日常の臨床診療における標準的な方法であると考えられています。それにもかかわらず、2D心エコー検査ベースの評価は依然として技術の主要な制限を伴います:いくつかの断層平面からの所与のチャンバーのイメージングは、3次元(3D)構造の形態および機能を適切に特徴付けない。この問題は、右心室(RV)の場合にさらに顕著である:比較的単純な弾丸型の左心室(LV)と比較して、RVは、線形直径または領域3を使用して適切に定量化できない複雑な形状2を有する。これらの広く知られている事実にもかかわらず、RVの形態および機能は通常、臨床診療においてそのような単純なパラメータによって測定される。
何十年もの間、RVは左のRVと比較して流通においてはるかに重要ではないと考えられていました。いくつかの画期的な論文は、RVの形状と機能の強力な予後的役割を示すこの立場を打ち負かしました4,5,6,7。多くの研究により、比較的単純な従来のパラメータを使用してもRV測定の増分値が実証されており、潜在的に意味のある臨床的価値を持つチャンバーをより正確に定量することの重要性と必要性が浮き彫りになっています。
3D心エコー検査は、心腔の2D評価のいくつかの制限を克服します。LVの場合も、体積の測定や幾何学的仮定のない機能パラメータは高い関心事になる可能性がありますが、RV8の評価では特に重要になる可能性があります。3D由来のRV体積および駆出率(EF)は、様々な心血管状態において有意な予後値を有することが示されている9、10。
現在、いくつかのベンダーは、ゴールドスタンダードの心臓磁気共鳴(MR)測定に対して検証された結果を備えた3D RV評価用の半自動ソリューションを提供しています11,12。3D評価の技術的要件は、今日の最先端の心臓血管イメージング部門の重要な部分であり、まもなくすべての心エコー検査ラボの一般的な機器の一部になると予想されています。3D取得と後処理に関する適切な専門知識があれば、3D RV分析を標準検査プロトコルに簡単に実装できます。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>3V-D/4V-D/4Vc-D</td>
			<td>General Electric</td>
			<td>n.a.</td>
			<td>ultrasound probe</td>
		</tr>
		<tr>
			<td>4D Auto RVQ</td>
			<td>General Electric</td>
			<td>n.a.</td>
			<td>software for analysis</td>
		</tr>
		<tr>
			<td>E9/E95</td>
			<td>General Electric</td>
			<td>n.a.</td>
			<td>ultrasound machine</td>
		</tr>
		<tr>
			<td>EchoPac v203</td>
			<td>General Electric</td>
			<td>n.a.</td>
			<td>software for analysis</td>
		</tr>
	</tbody>
</table>

morphological and functional assessment of the right ventricle using 3d echocardiography

伝統的に、心臓の右側は循環において小さな役割を果たすと考えられていました。しかし、ますます多くのデータが、右心室(RV)機能がさまざまな心血管障害において強力な診断力と予後力を持っていることを示唆しています。その複雑な形態と機能のために、従来の2次元心エコー検査によるRVの評価は限られています:日常の臨床診療は通常、単純な線形寸法と機能的測定に依存しています。3次元(3D)心エコー検査は、幾何学的仮定のないRVの体積定量を提供することにより、これらの制限を克服しました。ここでは、市販の主要なソフトウェアを使用してRVの3D心エコーデータを取得および分析するためのステップバイステップガイドを提供します。3D RVボリュームと排出率を定量化します。いくつかの技術的側面は、RVの取得と分析の品質を向上させるのにも役立つ可能性があり、実用的な方法で提示します。この方法の現在の機会と制限要因を確認し、現在の臨床診療における3D RV評価の潜在的なアプリケーションも強調します。

RVの3D分析は、さまざまな心血管疾患で実行可能です。症例1は、正常な心室容積と機能を持つ健康なボランティアです(図1)。症例2は僧帽弁修復後の患者で,従来の2D評価の相反する結果の典型例であり,TAPSEは著しく低下しているが,RV機能障害の徴候は見られず,正常な3D RV EFによりRVの全体的な収縮機能が維持されていることが確認された(<strong class...

Watch this Scientific Journal Video about Morphological and Functional Assessment of the Right Ventricle Using 3D Echocardiography at JoVE.com

Morphological and Functional Assessment of the Right Ventricle Using 3D Echocardiography

ここでは、主にこの技術の実現可能性を最大化する実用的な側面に焦点を当てて、右心室の3D体積評価のための段階的な取得および分析プロトコルを提供します。

3次元心エコー検査を用いた右心室の形態学的・機能的評価

Traditionally, it was believed that the right side of the heart has a minor role in circulation; however, more and more data suggest that right ...

morphological-functional-assessment-right-ventricle-using-3d

Research

JoVE Journal

Medicine

2.7K ビュー.  Semmelweis University Heart and Vascular Center. ここでは、主にこの技術の実現可能性を最大化する実用的な側面に焦点を当てて、右心室の3D体積評価のための段階的な取得および分析プロトコルを提供します。

ビデオ: Morphological and Functional Assessment of the Right Ventricle Using 3D Echocardiography

Transthoracic Echocardiography in Mice

In recent years, murine models have become the primary avenue for studying the molecular mechanisms of cardiac dysfunction resulting from changes in gene expression. Transgenic and gene targeting methods can be used to generate mice with altered cardiac size and function,1-3 and as a result, in vivo techniques are needed to evaluate their cardiac phenotype. Transthoracic echocardiography, pulse wave Doppler (PWD), and tissue Doppler imaging (TDI) can be used to provide dimensional measurements of the mouse heart and to quantify the degree of cardiac systolic and diastolic performance. Two-dimensional imaging is used to detect abnormal anatomy or movements of the left ventricle, whereas M-mode echo is used for quantification of cardiac dimensions and contractility.4,5 In addition, PWD is used to quantify localized velocity of turbulent flow,6 whereas TDI is used to measure the velocity of myocardial motion.7 Thus, transthoracic echocardiography offers a comprehensive method for the noninvasive evaluation of cardiac function in mice.

Transthoracic echocardiography offers a noninvasive method for the evaluation of cardiac function in mice. A combination of ultrasound and Doppler imaging modalities can be used to obtain dimensional measurements of the heart and intracardiac blood flow, which together provide an assessment of cardiac systolic and diastolic performance.

マウスにおける経胸壁心エコー

In recent years, murine models have become the primary avenue for studying the molecular mechanisms of cardiac dysfunction resulting from changes in gene ...

医学

Echocardiographic Assessment of the Right Heart in Mice

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential therapies. Given the expense and time involved in creating animal models of disease, it is critical that researchers have tools to accurately assess phenotypic expression of disease. Right ventricular dysfunction is the major manifestation of pulmonary hypertension. Echocardiography is the mainstay of the noninvasive assessment of right ventricular function in rodent models and has the advantage of clear translation to humans in whom the same tool is used. Published echocardiography protocols in murine models of PAH are lacking.
In this article, we describe a protocol for assessing RV and pulmonary vascular function in a mouse model of PAH with a dominant negative BMPRII mutation; however, this protocol is applicable to any diseases affecting the pulmonary vasculature or right heart. We provide a detailed description of animal preparation, image acquisition and hemodynamic calculation of stroke volume, cardiac output and an estimate of pulmonary artery pressure.

This article provides a protocol for the echocardiographic assessment of right ventricular size and pulmonary hypertension in mice. Applications include phenotype determination and serial assessment in transgenic and toxin-induced mouse models of cardiomyopathy and pulmonary vascular disease.

マウスの右心の心エコー評価

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential ...

Assessment of Right Ventricular Structure and Function in Mouse Model of Pulmonary Artery Constriction by Transthoracic Echocardiography

Emerging clinical data support the notion that RV dysfunction is critical to the pathogenesis of cardiovascular disease and heart failure1-3. Moreover, the RV is significantly affected in pulmonary diseases such as pulmonary artery hypertension (PAH). In addition, the RV is remarkably sensitive to cardiac pathologies, including left ventricular (LV) dysfunction, valvular disease or RV infarction4. To understand the role of RV in the pathogenesis of cardiac diseases, a reliable and noninvasive method to access the RV structurally and functionally is essential.
A noninvasive trans-thoracic echocardiography (TTE) based methodology was established and validated for monitoring dynamic changes in RV structure and function in adult mice. To impose RV stress, we employed a surgical model of pulmonary artery constriction (PAC) and measured the RV response over a 7-day period using a high-frequency ultrasound microimaging system. Sham operated mice were used as controls. Images were acquired in lightly anesthetized mice at baseline (before surgery), day 0 (immediately post-surgery), day 3, and day 7 (post-surgery). Data was analyzed offline using software.
Several acoustic windows (B, M, and Color Doppler modes), which can be consistently obtained in mice, allowed for reliable and reproducible measurement of RV structure (including RV wall thickness, end-diastolic&#160;and end-systolic dimensions), and function (fractional area change, fractional shortening, PA peak velocity, and peak pressure gradient) in normal mice and following PAC.
Using this method, the pressure-gradient resulting from PAC was accurately measured in real-time using Color Doppler mode and was comparable to direct pressure measurements performed with a Millar high-fidelity microtip catheter. Taken together, these data demonstrate that RV measurements obtained from various complimentary views using echocardiography are reliable, reproducible and can provide insights regarding RV structure and function. This method will enable a better understanding of the role of RV cardiac dysfunction.

Right ventricle (RV) dysfunction is critical to the pathogenesis of cardiovascular disease, yet limited methodologies are available for its evaluation. Recent advances in ultrasound imaging provide a noninvasive and accurate option for longitudinal RV study. Herein, we detail a step-by-step echocardiographic method using a murine model of RV pressure overload.

経胸壁心エコー検査による肺動脈狭窄のマウスモデルにおける右室の構造と機能の評価

Emerging clinical data support the notion that RV dysfunction is critical to the pathogenesis of cardiovascular disease and heart failure1-3. Moreover, ...

Myocardial Infarction and Functional Outcome Assessment in Pigs

Introduction of newly discovered cardiovascular therapeutics into first-in-man trials depends on a strictly regulated ethical and legal roadmap. One important prerequisite is a good understanding of all safety and efficacy aspects obtained in a large animal model that validly reflect the human scenario of myocardial infarction (MI). Pigs are widely used in this regard since their cardiac size, hemodynamics, and coronary anatomy are close to that of humans. Here, we present an effective protocol for using the porcine MI model using a closed-chest coronary balloon occlusion of the left anterior descending artery (LAD), followed by reperfusion. This approach is based on 90 min of myocardial ischemia, inducing large left ventricle infarction of the anterior, septal and inferoseptal walls. Furthermore, we present protocols for various measures of outcome that provide a wide range of information on the heart, such as cardiac systolic and diastolic function, hemodynamics, coronary flow velocity, microvascular resistance, and infarct size. This protocol can be easily tailored to meet study specific requirements for the validation of novel cardioregenerative biologics at different stages (i.e. directly after the acute ischemic insult, in the subacute setting or even in the chronic MI once scar formation has been completed). This model therefore provides a useful translational tool to study MI, subsequent adverse remodeling, and the potential of novel cardioregenerative agents.

This protocol describes the porcine myocardial infarction (MI) model using a 90 min closed-chest coronary balloon occlusion of the left anterior descending artery (LAD), followed by reperfusion. Furthermore, the protocol for several outcome parameters, such as cardiac function, hemodynamics, microvascular resistance, and infarct size, are also presented.

豚における心筋梗塞および機能的結果の評価

Introduction of newly discovered cardiovascular therapeutics into first-in-man trials depends on a strictly regulated ethical and legal roadmap. One ...

Chronic Thromboembolic Pulmonary Hypertension and Assessment of Right Ventricular Function in the Piglet

An original piglet model of Chronic Thromboembolic Pulmonary Hypertension (CTEPH) associated with chronic Right Ventricular (RV) dysfunction is described. Pulmonary Hypertension (PH) was induced in 3-week-old piglets by a progressive obstruction of the pulmonary vascular bed. A ligation of the left Pulmonary Artery (PA) was performed first through a mini-thoracotomy. Second, weekly embolizations of the right lower pulmonary lobe were done under fluoroscopic guidance with n-butyl-2-cyanoacrylate during 5 weeks. Mean Pulmonary Arterial Pressure (mPAP) measured by ritght heart catheterism, increased progressively, as well as Right Atrial pressure and Pulmonary Vascular Resistances (PVR) after 5 weeks compared to sham animals. Right Ventricular (RV) structural and functional remodeling were assessed by transthoracic echocardiography (RV diameters, RV wall thickness, RV systolic function). RV elastance and RV-pulmonary coupling were assessed by Pressure-Volume Loops (PVL) analysis with conductance method. Histologic study of the lung and the right ventricle were also performed. Molecular analyses on RV fresh tissues could be performed through repeated transcutaneous endomyocardial biopsies. Pulmonary microvascular disease in obstructed and unobstructed territories was studied from lung biopsies using molecular analyses and pathology. Furthermore, the reliability and the reproducibility was associated with a range of PH severity in animals. Most aspects of the human CTEPH disease were reproduced in this model, which allows new perspectives for the understanding of the underlying mechanisms (mitochondria, inflammation) and new therapeutic approaches (targeted, cellular or gene therapies) of the overloaded right ventricle but also pulmonary microvascular disease.

Chronic Thromboembolic Pulmonary Hypertension (CTEPH) and Right Ventricular (RV) dysfunction were induced in piglets by progressive obstruction of the pulmonary arteries. Consequences were remarkably similar to those observed in CTEPH patients. This animal model would be a very useful tool for pathophysiology and therapeutic experiments on CTEPH and RV failure.

慢性血栓塞栓性肺高血圧症とピグレットにおける右室機能の評価

An original piglet model of Chronic Thromboembolic Pulmonary Hypertension (CTEPH) associated with chronic Right Ventricular (RV) dysfunction is described. ...

Assessment of Cardiac Morphological and Functional Changes in Mouse Model of Transverse Aortic Constriction by Echocardiographic Imaging

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable noninvasive method is essential to assess real-time cardiac morphological and functional changes in animal models of heart disease. Transthoracic echocardiography represents an important tool for noninvasive assessment of cardiac structure and function2. Here we used a high-resolution ultrasound imaging system to monitor myocardial remodeling and heart failure progression over time in a mouse model of TAC. B-mode, M-mode, and Doppler imaging were used to precisely assess cardiac hypertrophy, ventricular dilatation, and functional deterioration in mice following TAC. Color and pulse wave (PW) Doppler imaging was used to noninvasively measure pressure gradient across the aortic constriction created by TAC and to assess transmitral blood flow in mice. Thus transthoracic echocardiographic imaging provides comprehensive noninvasive measurements of cardiac dimensions and function in mouse models of heart disease.

The goal of this protocol is to noninvasively assess cardiac structural and functional changes in a mouse model of heart disease created by transverse aortic constriction, using B- and M-mode echocardiography and color/pulse wave Doppler imaging.

心臓形態学的および心エコーイメージングによって横大動脈狭窄のマウスモデルにおける機能の変更の評価

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable ...

Transthoracic Speckle Tracking Echocardiography for the Quantitative Assessment of Left Ventricular Myocardial Deformation

The value of conventional echocardiography is limited by differences in inter-individual image interpretation and therefore largely dependent on the examiners&#39; expertise. Speckle tracking Echocardiography (STE) is a promising but technically challenging method that can be used to quantitatively assess regional and global systolic and diastolic myocardial performance. Myocardial strain and strain rate can be measured in all three dimensions&#160;&#8212; radial, circumferential, longitudinal &#8212; of myocardial deformation. Standard cross-sectional two-dimensional B-mode images are recorded and subsequently postprocessed by automated continuous frame-by-frame tracking and motion analysis of speckles within the myocardium. Images are recorded as digital loops and synchronized to a 3-lead EKG for timing purposes. Longitudinal deformation is assessed in the apical 4-, 3-, and 2-chamber views. Circumferential and radial deformation are measured in the parasternal short axis plane.
Optimal image quality and accurate tissue tracking are paramount for the correct determination of myocardial performance parameters. Utilizing transthoracic STE in a healthy volunteer, the present article is a detailed outline of the essential steps and potential pitfalls of quantitative echocardiographic myocardial deformation analysis.

Speckle tracking echocardiography is an emerging diagnostic imaging technique for the quantitative assessment of global and regional myocardial performance. Standard view echocardiographic motion images are recorded and deformation parameters are subsequently measured by automated continuous frame-by-frame tracking and motion analysis of speckles within the B-mode images of the myocardium.

左心室心筋変形の定量的評価のための経胸壁スペックル追跡心エコー検査

The value of conventional echocardiography is limited by differences in inter-individual image interpretation and therefore largely dependent on the ...

Assessment of the Anticoagulant and Anti-inflammatory Properties of Endothelial Cells Using 3D Cell Culture and Non-anticoagulated Whole Blood

In vivo, endothelial cells are crucial for the natural anticoagulation of circulating blood. Consequently, endothelial cell activation leads to blood coagulation. This phenomenon is observed in many clinical situations, like organ transplantation in the presence of pre-formed anti-donor antibodies, including xenotransplantation, as well as in ischemia/reperfusion injury. In order to reduce animal experimentation according to the 3R standards (reduction, replacement and refinement), in vitro models to study the effect of endothelial cell activation on blood coagulation would be highly desirable. However, common flatbed systems of endothelial cell culture provide a surface-to-volume ratio of 1 - 5 cm2 of endothelium per mL of blood, which is not sufficient for natural, endothelial-mediated anticoagulation. Culturing endothelial cells on microcarrier beads may increase the surface-to-volume ratio to 40 - 160 cm2/mL. This increased ratio is sufficient to ensure the &#34;natural&#34; anticoagulation of whole blood, so that the use of anticoagulants can be avoided. Here an in vitro microcarrier-based system is described to study the effects of genetic modification of porcine endothelial cells on coagulation of whole, non-anticoagulated human blood. In the described assay, primary porcine aortic endothelial cells, either wild type (WT) or transgenic for human CD46 and thrombomodulin, were grown on microcarrier beads and then exposed to freshly drawn non-anticoagulated human blood. This model allows for the measurement and quantification of cytokine release as well as activation markers of complement and coagulation in the blood plasma. In addition, imaging of activated endothelial cell and deposition of immunoglobulins, complement- and coagulation proteins on the endothelialized beads were performed by confocal microscopy. This assay can also be used to test drugs which are supposed to prevent endothelial cell activation and, thus, coagulation. On top of its potential to reduce the number of animals used for such investigations, the described assay is easy to perform and consistently reproducible.

We present an in vitro model which allows the study and analysis of coagulation in whole, non-anticoagulated blood. Anticoagulation in the system depends on the natural anticoagulation effect of healthy endothelial cells and endothelial cell activation will result in clotting.

3 D 細胞培養と非高血圧症全血を用いた血管内皮細胞の抗凝固および抗炎症作用の評価

In vivo, endothelial cells are crucial for the natural anticoagulation of circulating blood. Consequently, endothelial cell activation leads to blood ...

2D and 3D Echocardiography in the Axolotl (Ambystoma Mexicanum)

Cardiac malfunction as a result of ischemic heart disease is a major challenge, and regenerative therapies to the heart are in high demand. A few model species such as zebrafish and salamanders that are capable of intrinsic heart regeneration hold promise for future regenerative therapies for human patients. To evaluate the outcome of cardioregenerative experiments it is imperative that heart function can be monitored. The axolotl salamander (A. mexicanum) represents a well-established model species in regenerative biology attaining sizes that allows for evaluation of cardiac function. The purpose of this protocol is to establish methods to reproducibly measure cardiac function in the axolotl using echocardiography. The application of different anesthetics (benzocaine, MS-222, and propofol) is demonstrated, and the acquisition of two-dimensional (2D) echocardiographic data in both anesthetized and unanesthetized axolotls is described. 2D echocardiography of the three-dimensional (3D) heart can suffer from imprecision and subjectivity of measurements, and to alleviate this phenomenon a solid method, namely intra/inter-operator/observer analysis, to measure and minimize this bias is demonstrated. Finally, a method to acquire 3D echocardiographic data of the beating axolotl heart at a very high spatiotemporal resolution and with pronounced blood-to-tissue contrast is described. Overall, this protocol should provide the necessary methods to evaluate cardiac function and model anatomy, and flow dynamics in the axolotl using ultrasound imaging with applications in both regenerative biology and general physiological experiments.

2D and 3D Echocardiography in the Axolotl Ambystoma Mexicanum

Here we present echocardiography protocols for two-dimensional and three-dimensional image acquisition of the beating heart of the axolotl salamander (Ambystoma mexicanum), a model species in heart regeneration. These methods allow for longitudinal evaluation of cardiac function at a high spatiotemporal resolution.

ウーパールーパー (アホロートル メキシコマンネングサ) で 2 D および 3 D 心エコー検査

Cardiac malfunction as a result of ischemic heart disease is a major challenge, and regenerative therapies to the heart are in high demand. A few model ...

Invasive Hemodynamic Assessment for the Right Ventricular System and Hypoxia-Induced Pulmonary Arterial Hypertension in Mice

Pulmonary arterial hypertension (PAH) is a chronic and severe cardiopulmonary disorder. Mice are a popular animal model used to mimic this disease. However, the evaluation of right ventricular pressure (RVP) and pulmonary artery pressure (PAP) remains technically challenging in mice. RVP and PAP are more difficult to measure than left ventricular pressure because of the anatomical differences between the left and right heart systems. In this paper, we describe a stable right heart hemodynamic measurement method and its validation using healthy and PAH mice. This method is based on open-chest surgery and mechanical ventilation support. It is a complicated procedure compared to closed chest procedures. While a well-trained surgeon is required for this surgery, the advantage of this procedure is that it can generate both RVP and PAP parameters at the same time, so it is a preferable procedure for the evaluation of PAH models.

Here, we present a protocol to perform an invasive hemodynamic assessment of the right ventricle and pulmonary artery in mice using an open-chest surgery approach.

マウスにおける右心室系と低酸素誘発性肺動脈高血圧に対する侵襲的血行力学的評価

Pulmonary arterial hypertension (PAH) is a chronic and severe cardiopulmonary disorder. Mice are a popular animal model used to mimic this disease. ...