Biz Dr. Corey Reynolds ve bu proje ve tesislerin tür erişim ile uzman yardımı için BCM Fare fenotipleme Core teşekkür etmek istiyorum. XHTW WM Keck Tıp Araştırma Vakfı Seçkin Genç Bilim İnsanı, ve aynı zamanda NIH / NHLBI hibe R01-HL089598 ve R01-HL091947 ve Müsküler Distrofi Birliği hibe # 69.238 tarafından desteklenmektedir. Bu çalışma aynı zamanda Heart CaMKII Sinyal Vakfı Leducq Alliance tarafından kısmen destekleniyor.

1. Görüntüleme Çalışmaları hazırlanması Fare <ol><li> Önce görüntüleme çalışmaları, indüksiyon odasında fare (2% izofluran 0,5 L / dk ile karıştırılarak% 100 O 2) uyutmak. Operatörün güvenliği için atık gaz filtre emin olun. Indüksiyon odasından çıkarın ve hayvan kürk yaka orta göğüs seviyesine traş, saç kesme makineleri kullanmak. Sonra epilasyon kremi ile kalan vücut saç kaldırmak. Bu preparat, farelerde potansiyel istenmeyen stres tepkileri en aza indirmek için bir gün önce görüntüleme çalışmaları yapılabilir. </li><li> Sklera kurutulması (kornea jel aplikatör ile iletişime önlemek) önlemek için her iki göze jel uygulayın duralube. Gömülü EKG ile bir ısıtma yastığı tepesinde yatar pozisyonda anestezi Farenizi vücut ısısını korumak için açar. </li><li> Işlem boyunca istikrarlı bir devlet sedasyon seviyesi (0.5 L / dk% 100 O 2 ile karışık izofluran% 1.0 -% 1.5) korumak için anestezi sistemine bağlı bir burun konisi içinde burunları yerleştirin . Sedasyon onaylamak için bir parmak veya kuyruk tutam gerçekleştirin. Gerekirse, anestezi düzeyi, hedef kalp hızı 450 ± 50 dakikada vuruş (bpm) elde etmek için ayarlanabilir. </li><li> Yavaşça sürekli izlemek ve ısıtma yastığı aracılığıyla vücut ısısını ayarlamak için bir rektal prob (yağlama sonra) yerleştirin. Dar bir aralık içinde vücut ısısını korumak için önemlidir (37,0 ° C ± 0.5 ° C), ısı ve kalp hızı hatta ılımlı değişiklikler farelerde kalp fonksiyonunu etkileyebilir. </li><li> Dört pençeleri ve teyp EKG elektrotlar, elektrot jeli uygulayın. </li></ol> 2. Kalp değerlendirilmesi Sistolik Fonksiyon <ol><li> Bu yordamı başlamak için öncelikle alan, kalp üzerinde, göğüste, önceden ısıtılmış ultrason jel bir katman uygulamak. Jel, yaklaşık vücut ısısı ısındı olmalıdır. Soğuk jel hacmi hızla vücut ısısı kaybı oranı ekleyebilirsiniz. Ultrasonik görüntüleme müdahale jel hava kabarcıkları kaçının. </li><li> Fareyi hafifçe yukarıya doğru hafif bir açı altında bir pozisyonda (baş yukarı) hareketsiz. Bir mikromanipülatör kullanarak, ultrason probu, prob ve kalp arasında bir 90 ° açı ile hareketsiz. </li><li> Kısa eksen ekokardiyografik ölçümler yapmaya başlamak için, toraks üzerine parasternally prob düşüktür. Bu sinyali deforme olarak, sternum üzerinde doğrudan ya da cilde prob koymaktan kaçının. </li><li> Öncelikle, parasternal kısa eksen boyunca bir görünüm elde etmek için iki boyutlu (2D) görüntüleme (&#39;B-mod&#39;) gerçekleştirecektir. Hayvanın kalp büyüklüğüne göre komple bir görüntü elde etmek için probun görüş alanı ayarlayın. </li><li> Bu yönde düzgün bir görüntü, sol ventrikül ve sağ ventrikül duvar hafif bir bölümü yer alacak. Ok (video gösterilir), anterolateral ve posteromedial papiller kaslar, LV arka ve ön duvarları, intraventriküler septum duvar ve sağ ventrikül duvar hafif bir kısmını gösterir. Bu ölçümler sırasında, potansiyel duvar hareket bozuklukları (örneğin, hipokinezi, akinasia, asenkroni) ya da anevrizmalar özellikle dikkat edin. </li><li> Sonra, kalp boyutları ve kontraktilite ince ölçümler elde etmek için, 1-boyutlu (1D) sağlayan M-mod ekokardiyografi, kullanacaktır. Mağaza sistolik sol ventrikül fonksiyon parametreleri sonraki değerlendirme için görüntüleri satın aldı. </li></ol> 3. Kalbin Diyastolik Fonksiyon değerlendirilmesi <ol><li> Parasternal kısa eksen hala izlerken, biz miyokard hareket hızını ölçmek için doku Doppler görüntüleme (TDI) gerçekleştirebilirsiniz. Radyal eksen değerlendirilmesi için bir arka sol ventrikül duvar ilgi bölge işaretlenir. Elde edilen dalga şekli diyastolik fonksiyonu değerlendirmek için dört zirveleri olacak: IVRT, E, A &#39;ve IZKS. S &#39;sistolik hızı temsil eder. Bu parametreler (Şekil 3B) temsilcisi sonuç bölümünde daha sonra açıklanacaktır. </li><li> Pulse-wave Doppler (PWD) küçük bir alan içinde belirli bir derinlikte miyokard dokusunda kan akış hızını ölçmek için kullanılabilir. Trans-mitral akım paterni görüntü için, Trendelenburg pozisyonuna hayvan geriye doğru eğin. Prob kalbin apeks ile ortogonal olduğu gibi, yukarı prob yatırın. </li><li> Şimdi 300-350 bpm, mitral kapak hareketi yavaş kalp hızı daha düşük izofluran düzeyini artırmak. Karşılaştırma çalışmaları için, hayvanlar arasında benzer kalp hızı sağlamak için önemlidir. </li><li> Vana genelinde debisini ölçmek için bir referans noktası olarak mitral kapak hareketleri gözünüzde canlandırın. Örnek hacmi, mitral kapak ucunda yerleştirin. </li><li> PWD kullanarak mitral kapak üzerinden akım paternleri ölçün. İki dalgaları karakteristik görülür, ventrikül pasif dolum (erken [D] dalga) temsil eden bir vetutarlı bir atriyal sistol (atriyal [A] dalga) ile aktif dolgulu. Sağlıklı bir kalp, E-dalga hızı, A dalga (Şekil 3C) biraz daha fazla. </li><li> Tüm ölçümler tamamlandıktan sonra, elektrot jeli (Standart gazlı bez ile hafifçe silerek) ve hayvan koltuk çıkarın. Dizginlenmemiş hayvan dik bir pozisyonda ısıtmalı EKG pad tepesinde kurtarmak ve anestezi kapatmak için izin verin. Fare zaman uyanır, kafes dönün. </li></ol> 4. Temsilci Transtorasik Ekokardiyografi Sonuçlar Noninvaziv ekokardiyografik çalışmada, sol ventrikül morfolojisi ve kardiyak hastalık fare modelleri fonksiyonu değerlendirmek için kullanılan olabilir, 4,5 Bu tekniğin yaygın olarak kullanılan ve intrakardiyak basınç-volüm halkası measurements.8 terminali prosedürler olarak sol ventrikül fonksiyonunu değerlendirmek için bir alternatif sağlar. veya ex vivo çalışma kalp modeli. transtorasik M-mode ekokardiyografi ile sol ventrikül odası ölçüleri 9 Değerlendirme anormal AG oda genişleme veya artmış duvar kalınlığı, kalp hastalığı çeşitli modeller tespiti için izin verir., Böylece nabız dalga Doppler ile birlikte 5, transtorasik ekokardiyografi, sistolik ve diyastolik kalp fonksiyonu farelerde non-invaziv değerlendirilmesi için kapsamlı bir cadde oluşturmaktadır. <img src="/files/ftp_upload/1738/1738fig1.jpg" alt="Şekil 1" /> Şekil 1. Ekokardiyografi ve Doppler Görüntüleme Vivo Micro-Görüntüleme Sistemi Vevo 770 Yüksek Çözünürlüklü Bakış (A) fizyolojik izleme ünitesi, EKG kurulu, ve RMV 30MHz tarama kafası ile VisualSonics entegre raylı sistem kullanır. (B) fare konumlandırılmış ve entegre EKG elektrotları (altın rengi) ile ısıtma kartı düzgün bir şekilde kısıtlama. EKG elektrotları içine dört bacağı bantlanmış vardır. (CD) nabız dalga Doppler için uygun açılarda gösteren fotoğraflar, diyastolik fonksiyon ölçümleri, protokolün 4. bölümde açıklandığı gibi elde etmek için kullanılır . Sistolik fonksiyon ölçümleri için prob yönlendirmesi video protokolü göstermek. <img src="/files/ftp_upload/1738/1738fig2.jpg" alt="Şekil 2" /> Şekil 2. Temsilci 2D Ekokardiyografi Görüntüler (B-mod) sol ventrikül anterior (KS) ve posterior (PW) duvarları, intraventriküler septum duvar (SW) ve lateral duvar (LW) gösteren (A) Para-sternal kısa eksen. Sol ventrikül lümen çapı, sol ventrikül iç çapı (LVID) olarak ölçülebilir. Yıldız posteromedial papiller kas gösterir. E / A zirve hızlarının belirlenmesi için mitral kapak üzerinde bir görüş alanı (B) Apikal dört odacık. MVAL, mitral kapak ön broşür; MVPL, mitral kapak posterior broşür, RV, sağ ventrikül; AG, sol ventrikül. <img src="/files/ftp_upload/1738/1738fig3.jpg" alt="Şekil 3" /> Şekil 3. Ekokardiyografi ve Doppler Veri Toplama ve Niceleme. Bu rakam, Şekil 2&#39;de sunulan 2 boyutlu ekokardiyografi görüntüleri ile ilgili elde edilen verilerin temsili görüntülerini içerir. (A) M-mod sistol sonu (ESD) ve diyastol sonu çapları (EDD) gösteren çizgiler ile izleme. LVAWD / S, sol ventrikül ön duvar kalınlığı (diyastol / sistol); LVPWD / S, sol ventrikül arka duvar kalınlığı (diyastol / sistol). (B) iz LV arka duvar doku Doppler görüntüleme Temsilcisi. IVRT, izovolümik gevşeme zamanı; IZKS, izovolümik kontraksiyon zamanı. E dalga erken diyastolik LV dolum sırasında mitral annulus hareket gelir ve A dalga geç LV dolum sırasında atriyal sistol kaynaklanır. S &#39;sistolik hız temsil eder. (C) mitral kapak broşür ipuçları nabız dalga Doppler kayıt, mitral erken diyastolik hız (E), diyastolik daralma geç diyastolik hız (A) ve E / A oranı elde edilebileceği hız desen sağlar . IVRT diyastolik fonksiyon ve dolum basınçları karakterize etmek için de yararlı bir değişkendir. ET ejeksiyon zamanı gösterir.

<ol>
	<li>Zhang, T. M. a. i. e. r., S, L., Dalton, N. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+deltaC+isoform+of+CaMKII+is+activated+in+cardiac+hypertrophy+and+induces+dilated+cardiomyopathy+and+heart+failure.">The deltaC isoform of CaMKII is activated in cardiac hypertrophy and induces dilated cardiomyopathy and heart failure.</a> Circ Res. 92 (8), 912-912 (2003).</li><li>Srivastava, D., Thomas, T., Lin, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+cardiac+mesodermal+and+neural+crest+development+by+the+bHLH+transcription+factor,+dHAND.">Regulation of cardiac mesodermal and neural crest development by the bHLH transcription factor, dHAND.</a> Nat Genet. 16 (2), 154-154 (1997).</li><li>Sood, S., Chelu, M. G., van Oort, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracellular+calcium+leak+due+to+FKBP12.6+deficiency+in+mice+facilitates+the+inducibility+of+atrial+fibrillation.">Intracellular calcium leak due to FKBP12.6 deficiency in mice facilitates the inducibility of atrial fibrillation.</a> Heart Rhythm. 5 (7), 1047-1047 (2008).</li><li>Gardin, J. M., Siri, F. M., Kitsis, R. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Echocardiographic+assessment+of+left+ventricular+mass+and+systolic+function+in+mice.">Echocardiographic assessment of left ventricular mass and systolic function in mice.</a> Circ Res. 76 (5), 907-907 (1995).</li><li>Tanaka, N., Dalton, N., Mao, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transthoracic+echocardiography+in+models+of+cardiac+disease+in+the+mouse.">Transthoracic echocardiography in models of cardiac disease in the mouse.</a> Circulation. 94 (5), 1109-1109 (1996).</li><li>Patten, R. D., Aronovitz, M. J., Bridgman, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+pulse+wave+and+color+flow+Doppler+echocardiography+in+mouse+models+of+human+disease.">Use of pulse wave and color flow Doppler echocardiography in mouse models of human disease.</a> J Am Soc Echocardiogr. 15 (7), 708-708 (2002).</li><li>Garcia-Fernandez, M. A., Azevedo, J., Moreno, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regional+Left+Ventricular+Diastolic+Dysfunction+Evaluated+by+Pulsed-Tissue+Doppler+Echocardiography.">Regional Left Ventricular Diastolic Dysfunction Evaluated by Pulsed-Tissue Doppler Echocardiography.</a> Echocardiography. 16 (5), 491-491 (1999).</li><li>Pacher, P., Nagayama, T., Mukhopadhyay, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+cardiac+function+using+pressure-volume+conductance+catheter+technique+in+mice+and+rats.">Measurement of cardiac function using pressure-volume conductance catheter technique in mice and rats.</a> Nat Protoc. 3 (9), 1422-1422 (2008).</li><li>Larsen, T. S., Belke, D. D., Sas, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Pflugers Arch. 437 (6), 979-979 (1999).</li><li>Roth, D. M., Swaney, J. S., Dalton, N. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+anesthesia+on+cardiac+function+during+echocardiography+in+mice.">Impact of anesthesia on cardiac function during echocardiography in mice.</a> Am J Physiol Heart Circ Physiol. 282 (6), H2134-H2134 (2002).</li></ol>

M-mod ekokardiyografi kullanarak farelerde farklı gruplar veya genotipler arasında karşılaştırma kolaylaştırmak için, farelerin bir kohort içinde anestezi düzeyi, 10 vücut ısısı, kalp atış hızı standartlaştırmak için önemlidir . PWD diyastolik fonksiyon değerlendirme için fare kalbin diyastolik özellikleri değerlendirmek için, kalp atım hızı düşürülmelidir. Hayvan hayati fonksiyonları yakından hemodinamik yetmezliği ve ölüm önlemek için dikkat edilmelidir. Öte yandan, M-model ekokardiyografi, kalp hızı dakikada 400 vuruş altına kullanılarak sistolik fonksiyon ölçümleri için diyastol sonu çapları artmış LV dolum nedeniyle artmasına neden olabilir.

* Gerekli yazılım Vevo yazılım da dahil olmak üzere B-Mode uygulaması, nabız dalga Doppler uygulama ve kardiyovasküler analiz araçları paketi.

transthoracic echocardiography in mice

Son yıllarda, fare modelleri, gen ekspresyonu değişiklikleri kaynaklanan kardiyak disfonksiyon moleküler mekanizmaları çalışmak için birincil cadde haline gelmiştir. Transgenik ve gen hedefleme yöntemleri değişmiş kalp boyutu ve fonksiyonu ile fareler üretmek için kullanılabilir<sup&gt; 1-3</sup&gt; Ve bir sonucu olarak,<em&gt; In vivo</em&gt; Teknikleri, kalp fenotip değerlendirmek için gerekli. Transtorasik ekokardiyografi, nabız dalga Doppler (PWD), ve doku Doppler görüntüleme (TDI), fare kalp boyutlu ölçümler sağlamak ve kardiyak sistolik ve diyastolik performans derecesini ölçmek için kullanılabilir. Ise M-mod eko kalp boyutları ve kontraktilite ölçümü için kullanılan iki boyutlu görüntüleme, anormal anatomi ya da sol ventrikül hareketleri algılamak için kullanılır.<sup&gt; 4,5</sup&gt; Buna ek olarak, PWD, türbülanslı akış yerelleştirilmiş hızını ölçmek için kullanılır.<sup&gt; 6</sup&gt; TDI ise miyokard hareket hızını ölçmek için kullanılır.<sup&gt; 7</sup&gt; Bu nedenle, transtorasik ekokardiyografi farelerde kalp fonksiyonu noninvaziv değerlendirilmesi için kapsamlı bir yöntem sunmaktadır.

Watch this Scientific Journal Video about Transthoracic Echocardiography in Mice at JoVE.com

Transthoracic Echocardiography in Mice

Transtorasik ekokardiyografide farelerde kalp fonksiyonunun değerlendirilmesi için noninvaziv bir yöntem sunmaktadır. Kalp ve intrakardiyak kan akımı, kalp sistolik ve diyastolik performans birlikte bir değerlendirme sağlamak boyutlu ölçümler elde etmek için kullanılan ultrason ve Doppler görüntüleme yöntemleri bir arada olabilir.

Fareler de Transtorasik Ekokardiyografi

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

Research

JoVE Journal

Medicine

61.3K Görüntüleme.  Baylor College of Medicine (BCM). Transtorasik ekokardiyografide farelerde kalp fonksiyonunun değerlendirilmesi için noninvaziv bir yöntem sunmaktadır. Kalp ve intrakardiyak kan akımı, kalp sistolik ve diyastolik performans birlikte bir değerlendirme sağlamak boyutlu ölçümler elde etmek için kullanılan ultrason ve Doppler görüntüleme yöntemleri bir arada olabilir.

Video: Transthoracic Echocardiography in Mice

Murine Echocardiography and Ultrasound Imaging

Rodent models of cardiac pathophysiology represent a valuable research tool to investigate mechanism of disease as well as test new therapeutics.1 Echocardiography provides a powerful, non-invasive tool to serially assess cardiac morphometry and function in a living animal.2 However, using this technique on mice poses unique challenges owing to the small size and rapid heart rate of these animals.3 Until recently, few ultrasound systems were capable of performing quality echocardiography on mice, and those generally lacked the image resolution and frame rate necessary to obtain truly quantitative measurements. Newly released systems such as the VisualSonics Vevo2100 provide new tools for researchers to carefully and non-invasively investigate cardiac function in mice. This system generates high resolution images and provides analysis capabilities similar to those used with human patients. Although color Doppler has been available for over 30 years in humans, this valuable technology has only recently been possible in rodent ultrasound.4,5 Color Doppler has broad applications for echocardiography, including the ability to quickly assess flow directionality in vessels and through valves, and to rapidly identify valve regurgitation. Strain analysis is a critical advance that is utilized to quantitatively measure regional myocardial function.6 This technique has the potential to detect changes in pathology, or resolution of pathology, earlier than conventional techniques. Coupled with the addition of three-dimensional image reconstruction, volumetric assessment of whole-organs is possible, including visualization and assessment of cardiac and vascular structures. Murine-compatible contrast imaging can also allow for volumetric measurements and tissue perfusion assessment.

This video demonstrates use of a rail-mounted high-frequency ultrasound probe to perform echocardiography on an anesthetized mouse. The methods describe both conventional two-dimensional and M-mode measurements of cardiac function in addition to newer, more powerful tools such as color Doppler, strain analysis, as well as general and targeted contrast imaging.

Murin Ekokardiyografi ve Ultrasonik Görüntüleme

Rodent models of cardiac pathophysiology represent a valuable research tool to investigate mechanism of disease as well as test new therapeutics.1  ...

Tıp

Murine Fetal Echocardiography

Transgenic mice displaying abnormalities in cardiac development and function represent a powerful tool for the understanding the molecular mechanisms underlying both normal cardiovascular function and the pathophysiological basis of human cardiovascular disease. Fetal and perinatal death is a common feature when studying genetic alterations affecting cardiac development 1-3. In order to study the role of genetic or pharmacologic alterations in the early development of cardiac function, ultrasound imaging of the live fetus has become an important tool for early recognition of abnormalities and longitudinal follow-up. Noninvasive ultrasound imaging is an ideal method for detecting and studying congenital malformations and the impact on cardiac function prior to death 4. It allows early recognition of abnormalities in the living fetus and the progression of disease can be followed in utero with longitudinal studies 5,6. Until recently, imaging of fetal mouse hearts frequently involved invasive methods. The fetus had to be sacrificed to perform magnetic resonance microscopy and electron microscopy or surgically delivered for transillumination microscopy. An application of high-frequency probes with conventional 2-D and pulsed-wave Doppler imaging has been shown to provide measurements of cardiac contraction and heart rates during embryonic development with databases of normal developmental changes now available 6-10. M-mode imaging further provides important functional data, although, the proper imaging planes are often difficult to obtain. High-frequency ultrasound imaging of the fetus has improved 2-D resolution and can provide excellent information on the early development of cardiac structures 11.

Fetal and perinatal death is a common feature when studying genetic alterations affecting cardiac development. High-frequency ultrasound imaging has improved 2-D resolution and can provide excellent information on early cardiac development and is an ideal method to detect the impact on cardiac structure and function prior to death.

Mürin Fetal Ekokardiyografi

Transgenic mice displaying abnormalities in  cardiac development and function represent a powerful tool for the understanding  the molecular mechanisms ...

In utero Measurement of Heart Rate in Mouse by Noninvasive M-mode Echocardiography

Congenital heart disease (CHD) is the most frequent noninfectious cause of death at birth. The incidence of CHD ranges from 4 to 50/1,000 births (Disease and injury regional estimates, World Health Organization, 2004). Surgeries that often compromise the quality of life are required to correct heart defects, reminding us of the importance of finding the causes of CHD. Mutant mouse models and live imaging technology have become essential tools to study the etiology of this disease. Although advanced methods allow live imaging of abnormal hearts in embryos, the physiological and hemodynamic states of the latter are often compromised due to surgical and/or lengthy procedures. Noninvasive ultrasound imaging, however, can be used without surgically exposing the embryos, thereby maintaining their physiology. Herein, we use simple M-mode ultrasound to assess heart rates of embryos at E18.5 in utero. The detection of abnormal heart rates is indeed a good indicator of dysfunction of the heart and thus constitutes a first step in the identification of developmental defects that may lead to heart failure.

In utero Measurement of Heart Rate in Mouse by Noninvasive M-mode Echocardiography

Ultra-high frequency ultrasound is a powerful live imaging tool to examine cardiac abnormalities in small animals. Its noninvasiveness allows maintaining the physiologic state of embryos. Herein, we show the use of M-mode ultrasound to measure the heart rates of embryos at E18.5 in utero.

Noninvaziv M-mod ekokardiyografi ile Fare Kalp Puan utero Ölçümünde

Congenital heart disease (CHD) is the most frequent noninfectious cause of death at birth. The incidence of CHD ranges from 4 to 50/1,000 births (Disease ...

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.

Transtorasik Ekokardiyografi ile Pulmoner Arter Konstrüksiyon Fare Modeli sağ ventrikül yapı ve fonksiyonlarının değerlendirilmesi

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

Ultrasound-guided Transthoracic Intramyocardial Injection in Mice

Murine models of cardiovascular disease are important for investigating pathophysiological mechanisms and exploring potential regenerative therapies. Experiments involving myocardial injection are currently performed by direct surgical access through a thoracotomy. While convenient when performed at the time of another experimental manipulation such as coronary artery ligation, the need for an invasive procedure for intramyocardial delivery limits potential experimental designs. With ever improving ultrasound resolution and advanced noninvasive imaging modalities, it is now feasible to routinely perform ultrasound-guided, percutaneous intramyocardial injection. This modality efficiently and reliably delivers agents to a targeted region of myocardium. Advantages of this technique include the avoidance of surgical morbidity, the facility to target regions of myocardium selectively under ultrasound guidance, and the opportunity to deliver injectate to the myocardium at multiple, predetermined time intervals. With practiced technique, complications from intramyocardial injection are rare, and mice quickly return to normal activity on recovery from anesthetic. Following the steps outlined in this protocol, the operator with basic echocardiography experience can quickly become competent in this versatile, minimally invasive technique.

Echocardiography-guided percutaneous intramyocardial injection represents an efficient, reliable, and targetable modality for the delivery of gene transfer agents or cells into the murine heart. Following the steps outlined in this protocol, the operator can quickly become competent in this versatile, minimally invasive technique.

Farelerde ultrason eşliğinde transtorasik intramiyokardiyal Enjeksiyon

Murine models of cardiovascular disease are important for investigating pathophysiological mechanisms and exploring potential regenerative therapies. ...

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.

Sol Ventriküler Miyokard Deformasyon Kantitatif Değerlendirme Transtorasik Speckle İzleme Ekokardiyografi

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

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.

Amfibi (Ambystoma Mexicanum) 2D ve 3D ekokardiyografinin yeri

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 ...

Electromagnetic Navigation Transthoracic Nodule Localization for Minimally Invasive Thoracic Surgery

The increased use of chest computed tomography (CT) has led to an increased detection of pulmonary nodules requiring diagnostic evaluation and/or excision. Many of these nodules are identified and excised via minimally invasive thoracic surgery; however, subcentimeter and subsolid nodules are frequently difficult to identify intra-operatively. This can be mitigated by the use of electromagnetic transthoracic needle localization. This protocol delineates the step-by-step process of electromagnetic localization from the pre-operative period to the postoperative period and is an adaptation of the electromagnetically guided percutaneous biopsy previously described by Arias et al. Pre-operative steps include obtaining a same day CT followed by the generation of a three-dimensional virtual map of the lung. From this map, the target lesion(s) and an entry site are chosen. In the operating room, the virtual reconstruction of the lung is then calibrated with the patient and the electromagnetic navigation platform. The patient is then sedated, intubated, and placed in the lateral decubitus position. Using a sterile technique and visualization from multiple views, the needle is inserted into the chest wall at the prechosen skin entry site and driven down to the target lesion. Dye is then injected into the lesion and, then, continuously during needle withdrawal, creating a tract for visualization intra-operatively. This method has many potential benefits when compared to the CT-guided localization, including a decreased radiation exposure and decreased time between the dye injection and the surgery. Dye diffusion from the pathway occurs over time, thereby limiting intra-operative nodule identification. By decreasing the time to surgery, there is a decrease in wait time for the patient, and less time for dye diffusion to occur, resulting in an improvement in nodule localization. When compared to electromagnetic bronchoscopy, airway architecture is no longer a limitation as the target nodule is accessed via a transparenchymal approach. Details of this procedure are described in a step-by-step fashion.

Presented here is a protocol for lung nodule localization using dye marking via electromagnetically navigated transthoracic needle access. The technique described here can be accomplished in the peri-operative period to optimize nodule localization and to successful resection when performing minimally invasive thoracic surgery.

Minimal İnvaziv Göğüs Cerrahisinde Elektromanyetik Navigasyon Transtorasik Nodül Lokalizasyonu

The increased use of chest computed tomography (CT) has led to an increased detection of pulmonary nodules requiring diagnostic evaluation and/or ...

Transthoracic Echocardiographic Examination in the Rabbit Model

Large animal models such as the rabbit are valuable for translational preclinical research. Rabbits have a similar cardiac electrophysiology compared to that of humans and that of other large animal models such as dogs and pigs. However, the rabbit model has the additional advantage of lower maintenance costs compared to other large animal models. The longitudinal evaluation of cardiac function using echocardiography, when appropriately implemented, is a useful methodology for preclinical assessment of novel therapies for heart failure with reduced ejection fraction (e.g. cardiac regeneration). The correct use of this non-invasive tool requires the implementation of a standardized examination protocol following international guidelines. Here we describe, step by step, a detailed protocol supervised by veterinary cardiologists for performing echocardiography in the rabbit model, and demonstrate how to correctly obtain the different echocardiographic views and imaging planes, as well as the different imaging modes available in a clinical echocardiography system routinely used in human and veterinary patients.

Here we describe, step by step, a detailed protocol for performing echocardiography in the rabbit model. We show how to correctly obtain the different echocardiographic views and imaging planes, as well as the different imaging modes available in a clinical echocardiography system routinely used in human and veterinary patients.

Tavşan modelinde transtortik ekokardiyografi sınavı

Large animal models such as the rabbit are valuable for translational preclinical research. Rabbits have a similar cardiac electrophysiology compared to ...

Transthoracic Echocardiography to Assess Post-Resuscitation Left Ventricular Dysfunction After Acute Myocardial Infarction and Cardiac Arrest in Pigs

One of the main causes of out-of-hospital cardiac arrest is acute myocardial infarction (AMI). After successful resuscitation from cardiac arrest, approximately 70% of patients die before hospital discharge due to post-resuscitation myocardial and cerebral dysfunction. In experimental models, myocardial dysfunction after cardiac arrest, characterized by an impairment in both left ventricular (LV) systolic and diastolic function, has been described as reversible but very little data are available in cardiac arrest models associated with AMI in pigs. Transthoracic echocardiography is the first-line diagnostic test for the assessment of myocardial dysfunction, structural changes and/or AMI extension. In this pig model of ischemic cardiac arrest, echocardiography was done at baseline and 2-4 and 96 hours after resuscitation. In the acute phase, the examinations are done in anesthetized, mechanically ventilated pigs (weight 39.8 &#177; 0.6 kg) and ECG is recorded continuously. Mono- and bi-dimensional, Doppler and tissue Doppler recordings are acquired. Aortic and left atrium diameter, end-systolic and end-diastolic left ventricular wall thicknesses, end-diastolic and end-systolic diameters and shortening fraction (SF) are measured. Apical 2-, 3-, 4-, and 5-chamber views are acquired, LV volumes and ejection fraction are calculated. Segmental wall motion analysis is done to detect the localization and estimate the extent of myocardial infarction. Pulsed Wave Doppler echocardiography is used to record trans-mitral flow velocities from a 4-apical chamber view and trans-aortic flow from a 5-chamber view to calculate LV cardiac output (CO) and stroke volume (SV). Tissue Doppler Imaging (TDI) of LV lateral and septal mitral anulus is recorded (TDI septal and lateral s&#39;, e&#39;, a&#39; velocities). All the recordings and measurements are done according to the recommendations of the American and European Societies of Echocardiography Guidelines.

Transthoracic echocardiography is the first-line diagnostic test for post-resuscitation left ventricular dysfunction and structural changes in a pig model of cardiac arrest.

Domuzlarda Akut Miyokard Enfarktüsü ve Kardiyak Arrest Sonrası Resüsitasyon Sonrası Sol Ventrikül Disfonksiyonunu Değerlendirmek İçin Transtorasik Ekokardiyografi