This project is supported in part by grant KKF-15-1.

HINWEIS: Die intrakoronare Acetylcholin-Tests wurde von der lokalen Ethikkommission genehmigt worden ist und das Protokoll nach den Richtlinien unserer Institution für die menschliche Forschung. 1. Herstellung der Acetylcholin Solutions (siehe Materialien Table) <ol><li> Mischen Sie die 20 mg Acetylcholin mit 2 ml Lösungsmittel mit dem Paket zur Verfügung gestellt. </li><li> Fügen Sie die 2 ml ACH-Lösung zu 98 ml NaCl 0,9%. Dies entspricht einer Dosis von 0,2 mg / ml und ist Stammlösung 1. In 9 ml Stammlösung von 1 bis 91 ml NaCl 0,9% genannt, Dies entspricht einer Dosis von 18 ug / ml und ist Stammlösung 2 genannt. </li><li> Bereiten Sie 3 Perfusion Spritzen (à 50 ml), die als &quot;hoch&quot;, &quot;mittel&quot; und &quot;niedrig&quot;. Füllen Sie den Perfusionsspritze 1 mit &quot;hoch&quot; mit 40 ml Stammlösung 2. </li><li> Füllen Sie den Perfusionsspritze 2 (mit der Bezeichnung &quot;Medium&quot;) mit 8 ml der Perfusionsspritze 1 (hoch) und fügen Sie 32 ml NaCl 0,9%. Dies entspricht einerDosis von 3,6 &amp; mgr; g / ml. </li><li> Füllen Sie den Perfusionsspritze 3 (mit der Bezeichnung &quot;low&quot;) mit 4 ml der Perfusionsspritze 2 (mittel) und fügen Sie 36 ml NaCl 0,9%. Dies entspricht einer Dosis von 0,36 ug / ml. </li></ol> 2. Herstellung der Spritzen für die intrakoronare Injektion von Acetylcholin <ol><li> Bereiten 5 Spritzen für die intrakoronare Injektion (4 Spritzen von 5 ml [# 1, # 2, # 3, # 4] und 1 Spritze von 10 ml). </li><li> Füllen Sie das 5-ml-Spritze # 1 mit 6 ml der Perfusionsspritze Bezeichnung &quot;low&quot; (siehe Schritt 1.5). Dies entspricht einer Dosis von etwa 2 &amp; mgr; g. </li><li> Füllen Sie das 5-ml-Spritze # 2 mit 6 ml der Perfusionsspritze mit &quot;mittel&quot;. Dies entspricht einer Dosis von etwa 20 ug. </li><li> Füllen Sie das 5-ml-Spritze # 3 mit 5,5 ml der Perfusionsspritze mit &quot;hoch&quot;. Dies entspricht einer Dosis von etwa 100 ug. </li><li> Füllen Sie die 10-ml-Spritze mit 11 ml von der Perfusionsspritze gekennzeichnet. 4; high &quot;Dies entspricht einer Dosis von etwa 200 ug. </li><li> Füllen Sie das 5-ml-Spritze # 4 mit 4,5 ml der Perfusionsspritze mit &quot;hoch&quot;. Dies entspricht einer Dosis von etwa 80 &amp; mgr; g. Setzen Sie diese Spritze beiseite und es nur für die Beurteilung der rechten Koronararterie verwenden. </li></ol> 3. Diagnostische Koronarangiographie <ol><li> Injizieren Lokalanästhesie entweder in der Nähe der rechten radialen Arterie (üblicherweise 2 ml Mepivacain) oder in der Nähe der rechten Femoralarterie (üblicherweise 15 ml Mepivacain). </li><li> Bestätigen Sie den Erfolg der lokalen Anästhesie ist durch die narkotisiert-Haut mit der Nadel stechen und Befragung des Patienten, wenn die Schmerz noch vorhanden ist. </li><li> Die Punktion der Arterie nach der Seldinger - Technik 13 mit einer Kanüle, dann stecken Sie den Draht durch die Kanüle und entfernen. Legen Sie die Hülle (in der Regel 5F) über den Draht. Führen Sie Koronarangiographie unter sterilen Bedingungen. </li><li> Voraus, um den Draht thrau die Hülle an die aufsteigende Aorta und legen Sie den diagnostischen Katheter über die Aortenklappe. Dann entfernen Sie den Draht und verbinden Sie den Katheter mit dem Kontrast Spritze. HINWEIS: Das Kontrastmittel enthält Iomeprol, Trometamol, Salzsäure und Wasser. </li><li> Platzieren Sie den diagnostischen Katheter für die linke Koronararterie in der linken Hauptstamm durch leichtes Ziehen und den Katheter drehen. Bestätigen korrekte Positionierung des Katheters durch Einspritzen von 2 ml Kontrastmittel. </li><li> Führen Sie Koronarangiographie nach der Judkins - Technik 14 mit manuellen Injektionen von etwa 10 ml Kontrast der Koronararterien in verschiedenen Ansichten zu visualisieren. HINWEIS: In der Regel LAO 40 ° und RAO 35 ° sind für die rechte Koronararterie und LAO 45 ° verwendet / CRAN 25 °, 30 ° RAO / CRAN 30 ° und 20 ° RAO / caud 30 ° für die linke Koronararterie verwendet. </li><li> Starten Sie Acetylcholin Prüfung nach Ausschluss jeglicher relevanten epikardialen Stenose (≥50%) Durch visuelle Beurteilung. </li></ol> 4. Die intrakoronare Injektion von Acetylcholin <ol><li> Injizieren 6 ml der 2 ug Spritze # 1 in die linke Koronararterie. Injizieren Sie dies innerhalb von 20 Sekunden mit kontinuierlicher Überwachung des EKG und die Symptome des Patienten (zB Schmerzen in der Brust, Atemnot, Schwindel). Führen Sie Koronarangiographie der linken Koronararterie (in der Regel eine 20 RAO ° / caud 30 ° Projektion ist am besten Projektion) nach der Injektion der 6 ml. </li><li> manuell über die Kontrast Spritze in den Katheter Führen durch Koronarangiographie 10 ml Kontrast injiziert wird. Die 12-Kanal-EKG sollte nach jeder Dosis Acetylcholin aufgezeichnet und ausgedruckt werden. Eine Pause von 1 min sollte zwischen jeder Dosis liegen. </li><li> Injizieren 6 ml der 20 &amp; mgr; g Spritze # 2 in die linke Koronararterie. Injizieren dieser innerhalb von 20 sec mit kontinuierlicher Überwachung des EKG und die Symptome des Patienten. Führen Sie Koronarangiographie der linken Koronararterie nach der Injektionder 6 ml wie oben erwähnt. </li><li> Injizieren 5,5 ml der 100 ug Spritze # 3 in die linke Koronararterie. Injizieren dieser innerhalb von 20 sec mit kontinuierlicher Überwachung des EKG und die Symptome des Patienten. Führen Koronarangiographie der linken Koronararterie nach der Injektion von 5,5 ml wie oben erwähnt. HINWEIS: Die meisten Patienten einige Symptome berichten, EKG-Veränderungen oder epikardialen Vasokonstriktion bei dieser Dosis zeigen. Manchmal muss die Geschwindigkeit der manuellen Injektion verlangsamt werden. Wie unten verschiedene Protokolle für ACH-Test genannt werden, sind in Verwendung mit einer anderen Geschwindigkeit der Einspritzung. Eine langsamere Injektion über einen Zeitraum von 3 min zur Injektion innerhalb von 20 sec im Vergleich kann auch möglich sein. </li><li> Wenn kein Krampf (dh&gt; 90% Vasokonstriktion im Vergleich zum entspannten Zustand des Schiffes nach 200 ug nitroglyercine Injektion) bei einer Dosis von 100 ug tritt weiterhin mit der 200 ug (10 ml Spritze) ACH - Dosis. Injizieren Sie den 11 ml innerhalb von 20 Sekunden mit Dauerüberwachungsing des EKG und die Symptome des Patienten. </li><li> Führen Sie Koronarangiographie der linken Koronararterie nach der Injektion der 11 ml, wie oben (Abschnitt 3) erwähnt. Hinweis: Obwohl die 200 ug Dosis bei vielen Patienten angewendet worden sei erwähnt, dass geeignete Dosen von intrakoronaren Acetylcholin bleiben vollständig definiert werden. Einige Autoren schlagen vor, dass 100 ug sollte die maximale Dosis für die linke Koronararterie sein. HINWEIS: Oft tritt Bradykardie und die Geschwindigkeit der Injektion muss verlangsamt werden. </li><li> Injizieren 80 &amp; mgr; g ACH (4,5 ml, Spritze # 4) in die rechte Koronararterie, wenn keine Auffälligkeiten während des Testens der linken Koronararterie zu sehen sind. Injizieren dieser innerhalb von 20 sec mit kontinuierlicher Überwachung des EKG und die Symptome des Patienten. Oft tritt Bradykardie und die Geschwindigkeit der Injektion muss verlangsamt werden längere Bradykardie und / oder Asystolie zu verhindern. </li><li> Führen Koronarangiographie der rechten Koronararterienach der Injektion der 4,5 ml (in der Regel eine 40 LAO ° / 0 ° Projektion ist am besten), wie oben (Abschnitt 3) erwähnt. </li><li> Injizieren intrakoronare Nitroglyzerin mit einer Dosis von 200 ug in jeder getesteten Arterie nach dem Test oder wenn schwere Symptome (dh schwere Schmerzen in der Brust oder schwere Atemnot), ischämischen EKG - Veränderungen oder epikardialen Krampf auftritt. </li><li> Bild der Behälter wie in Schritt 4.2 nach einer Minute genannten Reversion von Krampf zu dokumentieren. </li></ol>

<ol>
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The authors have nothing to disclose.

Es ist möglich, die Acetylcholin-Test in der täglichen klinischen Routine im Katheterlabor zu implementieren. Neben der Herstellung der ACH-Lösungen gibt es einige technische Probleme, die gelöst werden müssen, bevor der Test einschließlich strahlendurchlässige EKG-Start führt zur kontinuierlichen 12-Kanal-EKG-Registrierung. Dies ist wesentlich, um transiente ischämische EKG-Veränderungen während des Tests zu ermitteln. Darüber hinaus ist es wichtig zu wissen, daß die ACH-Lösungen können nur für 2 Stunde...

Angina pectoris ist das Markenzeichen der koronaren Herzkrankheit und das Konzept eines epikardialen Stenose verursacht myokardiale Ischämie und belastungsinduzierter Angina ist seit vielen Jahren etabliert. viele Patienten mit Angina pectoris haben nicht die typische Trias von retrosternale Schmerzen, Beginn während des Trainings und Linderung von Nitroglyzerin oder Rest jedoch. Häufig berichten Patienten in Ruhe Angina pectoris oder einer Kombination aus körperlicher Anstrengung und Ruhe Angina sowie Atemnot bei Anstrengung als mögliche Angina-Äquivalent. Im Jahr 1959 war Prinzmetal- das erste , das Konzept der vorübergehenden Krampf der Koronararterien verursacht Angina in Ruhe mit ST-Strecken - Hebung auf dem Elektrokardiogramm (EKG) , aber mit erhaltenen Belastungsfähigkeit 1 assoziiert einzuführen. Diese Hypothese wurde später mit der Koronarangiographie bestätigt und es schien , dass Koronarspasmus 2 bei Patienten mit epikardialen Stenosen oder normalen Koronararterien vorliegen können. Im 1980ies intrakoronare Acetylcholin Provokationstestung (ACH-Test) für den Nachweis von Koronararterienspasmus wurde in Japan gegründet und anschließend das Interesse an der klinischen Forschung für Koronarspasmus 3 erhöht. Doch nach Einführung der perkutanen koronaren Intervention im Jahr 1977 und der ersten Stent-Implantation im Jahr 1986 das Interesse an der koronaren vasomotorischen Störungen deutlich zurückgegangen zumindest in Europa und den Vereinigten Staaten. Dies kann auch für die invasive Natur des ACH-Test (aufgrund der kurzen Halbwertszeit von ACH es kann nur in die Koronararterien für die Beurteilung der Koronar-Spasmus verabreicht werden) zurückzuführen sein. Viele Patienten mit Anzeichen und Symptome einer myokardialen Ischämie nicht über alle relevanten epikardialen Stenose auf Koronarangiographie 4,5,6 Dennoch. Bei solchen Patienten intrakoronare Acetylcholin Provokationstestung ist nützlich, um eine klinisch relevante koronare vasomotorischen Störung und das Institut eine geeignete medizinische Behandlung zu erkennen <sup&gt; 7. Acetylcholin ist ein Neurotransmitter im Parasympathikus. Es wirkt über nikotinerg sowie muskarinergen (mAChR) -Rezeptoren. Letztere sind wichtig für die vaskuläre Homöostase und Acetylcholin bindet an mAChR als nicht-selektiver Agonist. Aktivierung dieser Rezeptoren an der endothelialen Ebene führt zu Stickoxid vermittelte Vasodilatation wohingegen die Aktivierung von mAChR auf den vaskulären glatten Muskelzellen führt zu Vasokonstriktion 8. In Abhängigkeit von der Integrität des Endothels und der Reaktivität der glatten Muskelzellen der Nettoeffekt der intrakoronaren Acetylcholin Verabreichung Vasodilatation oder Vasokonstriktion. Die physiologische Reaktion der Koronararterien in Reaktion auf Acetylcholin in Menschen ist nicht vollständig verstanden, aber es wurde berichtet, dass eine Vasodilatation sowie Vasokonstriktion von bis zu 25% des Gefäßdurchmessers physiologischen sein kann als bei Patienten mit normalen Koronararterien gezeigt und keine Angina pec9 toris. Die intrakoronare Acetylcholin Provokationstestung wurde von der European Society of Cardiology Richtlinien 10 sowie die japanischen Circulation Society Richtlinien 11 für die Beurteilung der epikardialen und / oder mikrovaskulären Krampf bei Patienten mit Angina pectoris und ungehinderten Koronararterien empfohlen. Die intrakoronare Acetylcholin Provokation Tests haben in der täglichen klinischen Routine im Katheterlabor unserer Institution im Jahr 2003. Seit 2006 ist ein standardisiertes Protokoll wurde 12 gefolgt etabliert. ACH-Tests bei allen Patienten mit Anzeichen und Symptome einer myokardialen Ischämie noch keine relevanten epikardialen Stenose (&lt;50%) auf die koronare Angiographie durchgeführt. ACH-Test wird durchgeführt, unmittelbar nach der diagnostischen Koronarangiographie nach dem Protokoll nachfolgend beschrieben. Ergometrin ist ein weiteres Mittel, das für Spasmus Provokationstests mit einem anderen Wirkmechanismus verwendet wird. ausführlichInformationen über ergonovine Tests können an anderer Stelle 12.

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	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:528px;">Vial of 20 mg acetylcholine chloride powder and 1 Ampoule of 2 mL diluent</td>
			<td style="width:151px;">Bausch &#38; Lomb&#160;</td>
			<td style="width:165px;">NDC 24 208-539-20</td>
			<td style="width:80px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">3 x 100 ml NaCl 0.9 %</td>
			<td>BBraun</td>
			<td>3200950</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">3 x syringe 50 ml each</td>
			<td>BBraun</td>
			<td>4187903</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#160;1 x 2 ml syringe</td>
			<td>BBraun</td>
			<td>4606027V</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#160;1 x 10 ml syringe</td>
			<td>BBraun</td>
			<td>4606108V</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2 x cannula 20 G 70mm</td>
			<td>BBraun</td>
			<td>4665791</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#160;5 x 5 ml syringe</td>
			<td>BBraun</td>
			<td>4606051V</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Contrast agent Imeron 350 with a 10ml syringe for contrast injection</td>
			<td>Bracco Imaging</td>
			<td>30699.03.00</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Coronary angiography suite (AXIOM Artis MP eco )</td>
			<td>Siemens</td>
			<td>n/a</td>
			<td></td>
		</tr>
	</tbody>
</table>

intracoronary acetylcholine provocation testing for assessment of coronary vasomotor disorders

Intracoronary acetylcholine provocation testing (ACH-test) is an established method for assessment of epicardial coronary artery spasm in the catheterization laboratory which was introduced more than 30 years ago. Due to the short half-life of acetylcholine it can only be applied directly into the coronary arteries. Several studies have demonstrated the safety and clinical usefulness of this test. However, acetylcholine testing is only rarely applied in the U.S. or Europe. Nevertheless, it has been shown that 62% of Caucasian patients with stable angina and unobstructed coronary arteries on coronary angiography suffer from coronary vasomotor disorders that can be diagnosed with acetylcholine testing. In recent years it has been appreciated that the ACH-test not only assesses the presence of epicardial spasm but that it can also be useful for the detection of coronary microvascular spam. In such cases no epicardial spasm is seen after injection of acetylcholine but ischemic ECG shifts are present together with a reproduction of the patient's symptoms during the test. This article describes the experience with the ACH-test and its implementation in daily clinical routine.

Die Interpretation der Acetylcholin-Test basiert auf drei Kriterien. Zuerst wird der Patient während der gesamten Prüfung gefragt, ob oder nicht die Symptome auftreten. Häufig berichten die Patienten über eine Reproduktion ihrer üblichen Symptome wie Schmerzen in der Brust, Atemnot oder andere Symptome. Dies ist ein wichtiger Punkt für die Gesamtinterpretation des Tests. Zweitens ist ein 12-Kanal-EKG-Registrierung wird während des Tests mit einem besonderen Schwerpunkt auf ischäm...

Watch this Scientific Journal Video about Intracoronary Acetylcholine Provocation Testing for Assessment of Coronary Vasomotor Disorders at JoVE.com

Intracoronary Acetylcholine Provocation Testing for Assessment of Coronary Vasomotor Disorders

Intracoronary acetylcholine testing has been established for the assessment of epicardial coronary spasm more than 30 years ago. Recently, the focus has shifted towards the microcirculation and it has been shown that microvascular spasm can be detected using ACH-testing. This article describes the ACH-test and its implementation in daily routine.

Die intrakoronare Acetylcholin Provokationstestung für die Bewertung der koronaren Vasomotorische Disorders

Intracoronary acetylcholine provocation testing (ACH-test) is an established method for assessment of epicardial coronary artery spasm in the ...

intracoronary-acetylcholine-provocation-testing-for-assessment

Research

JoVE Journal

Medicine

15.5K Aufrufe.  Robert-Bosch-Krankenhaus. Intracoronary acetylcholine testing has been established for the assessment of epicardial coronary spasm more than 30 years ago. Recently, the focus has shifted towards the microcirculation and it has been shown that microvascular spasm can be detected using ACH-testing. This article describes the ACH-test and its implementation in daily routine.

Serie: Intracoronary Acetylcholine Provocation Testing for Assessment of Coronary Vasomotor Disorders

A Protocol for Comprehensive Assessment of Bulbar Dysfunction in Amyotrophic Lateral Sclerosis (ALS)

Improved methods for assessing bulbar impairment are necessary for expediting diagnosis of bulbar dysfunction in ALS, for predicting disease progression across speech subsystems, and for addressing the critical need for sensitive outcome measures for ongoing experimental treatment trials. To address this need, we are obtaining longitudinal profiles of bulbar impairment in 100 individuals based on a comprehensive instrumentation-based assessment that yield objective measures. Using instrumental approaches to quantify speech-related behaviors is very important in a field that has primarily relied on subjective, auditory-perceptual forms of speech assessment1. Our assessment protocol measures performance across all of the speech subsystems, which include respiratory, phonatory (laryngeal), resonatory (velopharyngeal), and articulatory. The articulatory subsystem is divided into the facial components (jaw and lip), and the tongue. Prior research has suggested that each speech subsystem responds differently to neurological diseases such as ALS. The current protocol is designed to test the performance of each speech subsystem as independently from other subsystems as possible. The speech subsystems are evaluated in the context of more global changes to speech performance. These speech system level variables include speaking rate and intelligibility of speech.
The protocol requires specialized instrumentation, and commercial and custom software. The respiratory, phonatory, and resonatory subsystems are evaluated using pressure-flow (aerodynamic) and acoustic methods. The articulatory subsystem is assessed using 3D motion tracking techniques. The objective measures that are used to quantify bulbar impairment have been well established in the speech literature and show sensitivity to changes in bulbar function with disease progression. The result of the assessment is a comprehensive, across-subsystem performance profile for each participant. The profile, when compared to the same measures obtained from healthy controls, is used for diagnostic purposes. Currently, we are testing the sensitivity and specificity of these measures for diagnosis of ALS and for predicting the rate of disease progression. In the long term, the more refined endophenotype of bulbar ALS derived from this work is expected to strengthen future efforts to identify the genetic loci of ALS and improve diagnostic and treatment specificity of the disease as a whole. The objective assessment that is demonstrated in this video may be used to assess a broad range of speech motor impairments, including those related to stroke, traumatic brain injury, multiple sclerosis, and Parkinson disease.

A Protocol for Comprehensive Assessment of Bulbar Dysfunction in Amyotrophic Lateral Sclerosis ALS

Objective assessments of the physiological mechanisms that support speech are needed to monitor disease onset and progression in persons with ALS and to quantify treatment effects in clinical trials. In this video, we present a comprehensive, instrumentation-based protocol for quantifying speech motor performance in clinical populations.

Ein Protokoll für eine umfassende Beurteilung der bulbären Dysfunction in Amyotrophe Lateralsklerose (ALS)

Improved methods for assessing bulbar impairment are necessary for expediting diagnosis of bulbar dysfunction in ALS, for predicting disease progression ...

Forschung

Medizin

Use of a Hanging Weight System for Coronary Artery Occlusion in Mice

Murine studies of acute injury are an area of intense investigation, as knockout mice for different genes are becoming increasingly available 1-38. Cardioprotection by ischemic preconditioning (IP) remains an area of intense investigation. To further elucidate its molecular basis, the use of knockout mouse studies is particularly important 7, 14, 30, 39. Despite the fact that previous studies have already successfully performed cardiac ischemia and reperfusion in mice, this model is technically very challenging. Particularly, visual identification of the coronary artery, placement of the suture around the vessel and coronary occlusion by tying off the vessel with a supported knot is technically difficult. In addition, re-opening the knot for intermittent reperfusion of the coronary artery during IP without causing surgical trauma adds additional challenge. Moreover, if the knot is not tied down strong enough, inadvertent reperfusion due to imperfect occlusion of the coronary may affect the results. In fact, this can easily occur due to the movement of the beating heart.
Based on potential problems associated with using a knotted coronary occlusion system, we adopted a previously published model of chronic cardiomyopathy based on a hanging weight system for intermittent coronary artery occlusion during IP 39. In fact, coronary artery occlusion can thus be achieved without having to occlude the coronary by a knot. Moreover, reperfusion of the vessel can be easily achieved by supporting the hanging weights which are in a remote localization from cardiac tissues.
We tested this system systematically, including variation of ischemia and reperfusion times, preconditioning regiments, body temperature and genetic backgrounds39. In addition to infarct staining, we tested cardiac troponin I (cTnI)
as a marker of myocardial infarction in this model. In fact, plasma levels of cTnI correlated with infarct sizes (R2=0.8). Finally, we could show in several studies that this technique yields highly reproducible infarct sizes during murine IP and myocardial infarction6, 8, 30, 40, 41. Therefore, this technique may be helpful for researchers who pursue molecular mechanisms involved in cardioprotection by IP using a genetic approach in mice with targeted gene deletion. Further studies on cardiac IP using transgenic mice may consider this technique.

A murine model for myocardial ischemia and ischemic preconditioning is an important tool study cardioprotective mechanisms in vivo. Here, we report an easy applicable in situ model for cardiac IP using a hanging-weight system for coronary artery occlusion.

Die Verwendung eines Hanging Weight System für Okklusion in Mäuse

Murine studies of acute injury are an area of intense investigation, as knockout mice for different genes are becoming increasingly available 1-38. ...

Modified Technique for Coronary Artery Ligation in Mice

Myocardial infarction (MI) is one of the most important causes of mortality in humans1-3. In order to improve morbidity and mortality in patients with MI we need better knowledge about pathophysiology of myocardial ischemia. This knowledge may be valuable to define new therapeutic targets for innovative cardiovascular therapies4. Experimental MI model in mice is an increasingly popular small-animal model in preclinical research in which MI is induced by means of permanent or temporary ligation of left coronary artery (LCA)5. In this video, we describe the step-by-step method of how to induce experimental MI in mice.
The animal is first anesthetized with 2% isoflurane. The unconscious mouse is then intubated and connected to a ventilator for artificial ventilation. The left chest is shaved and 1.5 cm incision along mid-axillary line is made in the skin. The left pectoralis major muscle is bluntly dissociated until the ribs are exposed. The muscle layers are pulled aside and fixed with an eyelid-retractor. After these preparations, left thoracotomy is performed between the third and fourth ribs in order to visualize the anterior surface of the heart and left lung. The proximal segment of LCA artery is then ligated with a 7-0 ethilon suture which typically induces an infarct size ~40% of left ventricle. At the end, the chest is closed and the animals receive postoperative analgesia (Temgesic, 0.3 mg/50 ml, ip). The animals are kept in a warm cage until spontaneous recovery.

The surgical procedure used to induce experimental myocardial infarction in mice begins with left thoracotomy between the third and the fourth ribs in order to visualize the anterior surface of the heart and left lung. The left coronary artery is ligated, the chest is closed and the mouse is allowed to recover spontaneously.

Geändert Technik für Coronary Artery Ligation in Mäuse

Myocardial infarction (MI) is one of the most important causes of mortality in humans1-3. In order to improve morbidity and mortality in patients with MI ...

Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment (RMCA) in Individuals with Chronic Spinal Cord Injury

During breathing, activation of respiratory muscles is coordinated by integrated input from the brain, brainstem, and spinal cord. When this coordination is disrupted by spinal cord injury (SCI), control of respiratory muscles innervated below the injury level is compromised1,2 leading to respiratory muscle dysfunction and pulmonary complications. These conditions are among the leading causes of death in patients with SCI3. Standard pulmonary function tests that assess respiratory motor function include spirometrical and maximum airway pressure outcomes: Forced Vital Capacity (FVC), Forced Expiratory Volume in one second (FEV1), Maximal Inspiratory Pressure (PImax) and Maximal Expiratory Pressure (PEmax)4,5. These values provide indirect measurements of respiratory muscle performance6. In clinical practice and research, a surface electromyography (sEMG) recorded from respiratory muscles can be used to assess respiratory motor function and help to diagnose neuromuscular pathology. However, variability in the sEMG amplitude inhibits efforts to develop objective and direct measures of respiratory motor function6. Based on a multi-muscle sEMG approach to characterize motor control of limb muscles7, known as the voluntary response index (VRI)8, we developed an analytical tool to characterize respiratory motor control directly from sEMG data recorded from multiple respiratory muscles during the voluntary respiratory tasks. We have termed this the Respiratory Motor Control Assessment (RMCA)9. This vector analysis method quantifies the amount and distribution of activity across muscles and presents it in the form of an index that relates the degree to which sEMG output within a test-subject resembles that from a group of healthy (non-injured) controls. The resulting index value has been shown to have high face validity, sensitivity and specificity9-11. We showed previously9 that the RMCA outcomes significantly correlate with levels of SCI and pulmonary function measures. We are presenting here the method to quantitatively compare post-spinal cord injury respiratory multi-muscle activation patterns to those of healthy individuals.

Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment RMCA in Individuals with Chronic Spinal Cord Injury

The purpose of this publication is to present our original work on a multi-muscle surface electromyographic approach to quantitatively characterize respiratory muscle activation patterns in individuals with chronic spinal cord injury using vector-based analysis.

Auswertung der Respiratory Muscle Activation Mit Respiratory Motor Control Assessment (RMCA) bei Personen mit chronischen Spinal Cord Injury

During breathing, activation of respiratory muscles is coordinated by integrated input from the brain, brainstem, and spinal cord. When this coordination ...

A Novel Application of Musculoskeletal Ultrasound Imaging

Ultrasound is an attractive modality for imaging muscle and tendon motion during dynamic tasks and can provide a complementary methodological approach for biomechanical studies in a clinical or laboratory setting. Towards this goal, methods for quantification of muscle kinematics from ultrasound imagery are being developed based on image processing. The temporal resolution of these methods is typically not sufficient for highly dynamic tasks, such as drop-landing. We propose a new approach that utilizes a Doppler method for quantifying muscle kinematics. We have developed a novel vector tissue Doppler imaging (vTDI) technique that can be used to measure musculoskeletal contraction velocity, strain and strain rate with sub-millisecond temporal resolution during dynamic activities using ultrasound. The goal of this preliminary study was to investigate the repeatability and potential applicability of the vTDI technique in measuring musculoskeletal velocities during a drop-landing task, in healthy subjects. The vTDI measurements can be performed concurrently with other biomechanical techniques, such as 3D motion capture for joint kinematics and kinetics, electromyography for timing of muscle activation and force plates for ground reaction force. Integration of these complementary techniques could lead to a better understanding of dynamic muscle function and dysfunction underlying the pathogenesis and pathophysiology of musculoskeletal disorders.

We describe a new ultrasound-based vector tissue Doppler imaging technique to measure muscle contraction velocity, strain and strain rate with sub-millisecond temporal resolution during dynamic activities. This approach provides complementary measurements of dynamic muscle function and could lead to a better understanding of mechanisms underlying musculoskeletal disorders.

A Novel Anwendung von Muskel-Skelett-Ultraschall-Bildgebung

Ultrasound is an attractive modality for imaging muscle and tendon  motion during dynamic tasks and can provide a complementary methodological  approach ...

Ultrasound Based Assessment of Coronary Artery Flow and Coronary Flow Reserve Using the Pressure Overload Model in Mice

Transthoracic Doppler echocardiography (TTDE) is a clinically useful, noninvasive tool for studying coronary artery flow velocity and coronary flow reserve (CFR) in humans. Reduced CFR is accompanied by marked intramyocardial and pericoronary fibrosis and is used as an indication of the severity of dysfunction. This study explores, step-by-step, the real-time changes measured in the coronary flow velocity, CFR and systolic to diastolic peak velocity (S/D) ratio in the setting of an aortic banding model in mice. By using a Doppler transthoracic imaging technique that yields reproducible and reliable data, the method assesses changes in flow in the septal coronary artery (SCA), for a period of over two weeks in mice, that previously either underwent aortic banding or thoracotomy. 
During imaging, hyperemia in all mice was induced by isoflurane, an anesthetic that increased coronary flow velocity when compared with resting flow. All images were acquired by a single imager. Two ratios, (1) CFR, the ratio between hyperemic and baseline flow velocities, and (2) systolic (S) to diastolic (D) flow were determined, using a proprietary software and by two independent observers. Importantly, the observed changes in coronary flow preceded LV dysfunction as evidenced by normal LV mass and fractional shortening (FS). 
The method was benchmarked against the current gold standard of coronary assessment, histopathology. The latter technique showed clear pathologic changes in the coronary artery in the form of peri-coronary fibrosis that correlated to the flow changes as assessed by echocardiography. 
The study underscores the value of using a non-invasive technique to monitor coronary circulation in mouse hearts. The method minimizes redundant use of research animals and demonstrates that advanced ultrasound-based indices, such as CFR and S/D ratios, can serve as viable diagnostic tools in a variety of investigational protocols including drug studies and the study of genetically modified strains.

Coronary flow reserve (CFR) is useful for assessment of myocardial oxygen demand and evaluation of cardiovascular risk. This study establishes a step-by-step transthoracic Doppler echocardiographic (TTDE) method for longitudinal monitoring of the changes in CFR, as measured from coronary artery in mice, under the experimental pressure overload of aortic banding.

Ultraschall Basierend Beurteilung der koronaren Herzfluss und koronare Flussreserve Mit der Drucküberlastung Modell an der Maus

Transthoracic Doppler echocardiography (TTDE) is a clinically useful, noninvasive tool for studying coronary artery flow velocity and coronary flow ...

'Boden Food Plate': Novel Interactive Web-based Method for the Assessment of Dietary Intake

Different methods can be used in research to assess dietary intake, many of which are still paper-based. Written estimated food diaries are often utilized in clinical trials, despite being a burden for both study participants and researchers. This method requires participant literacy, it is time consuming, labor intensive, and can easily lead to under-reporting. With advancements in technology, there is a growing interest in electronic diaries that automate the dietary assessment process. These are focused on improving accuracy, reducing both time and cost and providing users with a visual and more enjoyable experience. The methodology presented here aimed to validate the &#39;Boden Food Plate&#39;, a novel web-based platform for self-recording of food and drink items, compared to a traditional estimated food diary. The application was also rated on a satisfaction scale by study participants using a paper-based questionnaire. Sixty-seven participants completed the dietary measures on both the three-day electronic and paper food diaries. For the analysis, only dietary data completed at both study time points (baseline and week six) was utilized. Despite small mean differences between dietary data collection methods, Bland Altman analysis showed fairly wide 95% limits of agreement between the electronic platform and the written estimated food diary and there were few cases which did not fall within the 95% confidence intervals. Overall, participants found the electronic food diary to be more fun than the paper method and as easy to use as hard copy diaries. The new platform has potential as a self-recording tool for the collection of dietary data, particularly when utilized in clinical trial settings. However, further validation studies are needed to improve the validity of this novel electronic dietary data collection tool.

The &#39;Boden Food Plate&#39; is an electronic food diary designed to be an interactive and fun method to collect dietary intake using visual depictions. The purpose of this study was to validate the web-based application against a traditional three-day estimated food diary method.

"Boden Essen Teller": neue interaktive Web-basierte Methode für die Bewertung der Nahrungsaufnahme

Different methods can be used in research to assess dietary intake, many of which are still paper-based. Written estimated food diaries are often utilized ...

Coronary Progenitor Cells and Soluble Biomarkers in Cardiovascular Prognosis after Coronary Angioplasty

Major adverse cardiovascular events (MACEs) negatively impact the cardiovascular prognosis of patients undergoing coronary angioplasty due to coronary ischemic injury. The extent of coronary damage and the mechanisms of vascular repair are factors influencing the future development of MACEs. Intrinsic vascular features like the plaque characteristics and coronary artery complexity have demonstrated prognostic information for MACEs. However, the use of intracoronary circulating biomarkers has been postulated as a convenient method for the early identification and prognosis of MACEs, as they more closely reflect dynamic mechanisms involving coronary damage and repair. Determination of coronary circulating biomarkers during angioplasty, such as the number of subpopulations of mononuclear progenitor cells (MPCs) as well as the concentration of soluble molecules reflecting inflammation, cell adhesion, and repair, allows for assessment of future developments and the prognosis of MACEs 6 months post coronary angioplasty. This method is highlighted by its translational nature and better performance than peripheral blood circulating biomarkers regarding prediction of MACEs and its effect on the cardiovascular prognosis, which may be applied for risk stratification of patients with coronary artery disease undergoing angioplasty.

Development of major adverse cardiovascular events, which impact cardiovascular prognosis after coronary angioplasty, are influenced by the extent of coronary damage and vascular repair. The use of novel coronary cellular and soluble biomarkers, reactive to vascular damage and repair, are useful to predict the development of MACEs and prognosis.

Koronare Vorläuferzellen und lösliche Biomarker bei kardiovaskulärer Prognossie nach koronarer Angioplastie

Major adverse cardiovascular events (MACEs) negatively impact the cardiovascular prognosis of patients undergoing coronary angioplasty due to coronary ...

In Vitro Model of Coronary Angiogenesis

Here, we describe an in vitro culture assay to study coronary angiogenesis. Coronary vessels feed the heart muscle and are of clinical importance. Defects in these vessels represent severe health risks such as in atherosclerosis, which can lead to myocardial infarctions and heart failures in patients. Consequently, coronary artery disease is one of the leading causes of death worldwide. Despite its clinical importance, relatively little progress has been made on how to regenerate damaged coronary arteries. Nevertheless, recent progress has been made in understanding the cellular origin and differentiation pathways of coronary vessel development. The advent of tools and technologies that allow researchers to fluorescently label progenitor cells, follow their fate, and visualize progenies in vivo have been instrumental in understanding coronary vessel development. In vivo studies are valuable, but have limitations in terms of speed, accessibility, and flexibility in experimental design. Alternatively, accurate in vitro models of coronary angiogenesis can circumvent these limitations and allow researchers to interrogate important biological questions with speed and flexibility. The lack of appropriate in vitro model systems may have hindered the progress in understanding the cellular and molecular mechanisms of coronary vessel growth. Here, we describe an in vitro culture system to grow coronary vessels from the sinus venosus (SV) and endocardium (Endo), the two progenitor tissues from which many of the coronary vessels arise. We also confirmed that the cultures accurately recapitulate some of the known in vivo mechanisms. For instance, we show that the angiogenic sprouts in culture from SV downregulate COUP-TFII expression similar to what is observed in vivo. In addition, we show that VEGF-A, a well-known angiogenic factor in vivo, robustly stimulates angiogenesis from both the SV and Endo cultures. Collectively, we have devised an accurate in vitro culture model to study coronary angiogenesis.

In vitro models of coronary angiogenesis can be utilized for the discovery of the cellular and molecular mechanisms of coronary angiogenesis. In vitro explant cultures of sinus venosus and endocardium tissues show robust growth in response to VEGF-A and display a similar pattern of COUP-TFII expression as in vivo.

In Vitro Modell der koronaren Angiogenese

Here, we describe an in vitro culture assay to study coronary angiogenesis. Coronary vessels feed the heart muscle and are of clinical importance. Defects ...

Testing Acetylcholine Followed by Adenosine for Invasive Diagnosis of Coronary Vasomotor Disorders

More than 50% of patients with signs and symptoms of myocardial ischemia undergoing coronary angiography have unobstructed coronary arteries. Coronary vasomotor disorders (impaired vasodilatation and/or enhanced vasoconstriction/spasm) represent important functional causes for such a clinical presentation. Although impaired vasodilatation may be assessed with non-invasive techniques such as positron emission tomography or cardiac magnetic resonance imaging, there is currently no reliable non-invasive technique for the diagnosis of coronary spasm available. Thus, invasive diagnostic procedures (IDP) have been developed for the diagnosis of coronary vasomotor disorders including spasm testing as well as assessment of coronary vasodilatation. The identification of the underlying type of disorder (so called endotype) allows the initiation of targeted pharmacological treatments. Despite the fact that such an approach is recommended by the current European Society of Cardiology guidelines for the management of chronic coronary syndromes based on the CorMicA study, comparability of results as well as multicenter trials are currently hampered by major differences in institutional protocols for coronary functional testing. This article describes a comprehensive IDP protocol including intracoronary acetylcholine provocation testing for diagnosis of epicardial/microvascular spasm, followed by Doppler wire-based assessment of coronary flow reserve (CFR) and hyperemic microvascular resistance (HMR) in search of coronary vasodilatory impairment.

Coronary vasomotion disorders represent frequent functional causes of angina in patients with unobstructed coronaries. The underlying mechanism of angina (endotype) in these patients can be determined by a comprehensive invasive diagnostic procedure based on acetylcholine provocation testing followed by Doppler-derived assessment of the coronary flow reserve and microvascular resistance.

Testen von Acetylcholin gefolgt von Adenosin zur invasiven Diagnose von koronaren vasomotorischen Störungen

More than 50% of patients with signs and symptoms of myocardial ischemia undergoing coronary angiography have unobstructed coronary arteries. Coronary ...