Unterstützt durch NIH 1P01HL089407-01A1 (Lawrence, PI), Animal-Core A, NIH 1 K01 HL080962-01A2 (Myers, PI).

1. Maus Anästhesieverfahren <ol><li> Die Tiere sind aus ihrem Käfig genommen und in ein Betäubungsmittel präoperativen Induktion Kammer für Gas Induktion mit 2% Isofluran und 100% Sauerstoff mit einer Flussrate von 0,5 Liter pro Minute (Verdampfer gesteuert). </li><li> Mäuse sind in der Narkoseeinleitung Kammer sediert, entfernt von Kammer-und der ventralen Bauch ist mit elektrischen Haarschneider rasiert, gepflegt oder in der Kammer, wenn ein Haar Enthaarungsmittel verwendet wird (1 min). </li><li> Noch unter Sedierung, sie wogen, werden die Augen mit einer sterilen Augensalbe und dann in Rückenlage recumbancy an einem warmen Wasser zirkuliert Heizgerät gelegt unter Vollnarkose mit einer Nase Konus mit Isofluran und Sauerstoff geschmiert. An diesem Punkt der Sauerstoffmenge auf 0,2 Liter pro Minute reduziert. </li><li> Der Bauch ist mit Chlorhexidin-Lösung besprüht (Chlorhexidindiacetat schnell bacterialcidal und persistent) und wischte sich mit steriler Gaze dreimal oder bis zum OP-Gebiet ist sauber. </li></ol> 2. Maus Micro-chirurgischen und Recovery-Verfahren <ol><li> Ein ventralen Mittellinie Schnitt (2 cm) ist mit Iris Schere durch die Haut und Bauchwand Freilegung der Bauch gemacht. Ein steriler Kochsalzlösung getränkt 2x2 in Gaze wird verwendet, um den Darm des Tieres linken Seite widerspiegeln. Eine Maus restrainer ist in Ort ermöglicht die Visualisierung der unteren Hohlvene (IVC) setzen. </li><li> An diesem Punkt ist die Isofluran zu einer Wartung Niveau von 2% reduziert und der Sauerstoff bleibt bei 0,2 Liter pro Minute für den Rest des chirurgischen Eingriffs. </li><li> Stumpfe Präparation wird erleichtert, mit einem sterilen Tupfer Applikator und extra zarte Iris halbgebogen Gewebe Zange. Es muss im Umgang mit Maus Gewebe für sie äußerst fragil eingenommen werden. </li><li> Alle IVC Seitenäste, von der Nierenvenen der iliakalen Bifurkation, werden mit 7-0 nicht-reaktiven Prolene Fäden ligiert. Zurück Niederlassungen blieb Patent. </li><li> Bei der elektrolytischen IVC-Modell (EIM), eine 25G Edelstahl-Nadel, die an einer versilberten Kupferdraht (KY-30 bis 1-GRN, Electrospec, Dover, NJ) in der exponierten caudal IVC, (Abbildung 1) eingefügt und gegen die vordere Wand (Anode) positioniert. </li><li> Ein weiterer Draht wird subkutan implantiert Abschluss der Schaltung (Kathode). </li><li> Ein Strom von 250 μAmps über 15 Minuten wurde mit einem Grass S48 Rechtecksignal Stimulator und ein konstanter Strom-Einheit (Grass Technologies, ein Astro-Med, Inc., West Warwick, RI). Der Gleichstrom führt zur Bildung von toxischen Produkten der Elektrolyse, dass die Endothel-Oberfläche des IVC zur Förderung einer thrombogenen Umwelt und nachfolgende Thrombusbildung erodieren. </li><li> Nach 15 Minuten wird die Nadel entfernt und ein Wattestäbchen wird sanft in Kontakt mit der Umgebung, um die Blutung zu verhindern statt </li><li> In sham Tieren ist die Nadel in die IVC gelegt und dann entfernt, ohne Anwendung des Stroms. </li><li> Die Laparotomie Website wird in einem Zwei-Schicht verschlossen. Unser Labor verwendet 5-0 Vicryl, ein synthetisches Nahtmaterial, in einem kontinuierlichen Muster, um die Bauchdecke zu schließen. Adhesive Hautleim wird verwendet, um die Haut zu schließen. Es gibt keine externen Nähte für die Herausnahme. </li><li> Mäuse werden dann in einer individuellen Maus Käfig wieder, dicht postoperativ (45 Minuten bis 2 Stunden) unter einer Wärmelampe (min Abstand 24 cm entfernt von Käfig) beobachtet, dann wieder an ihren ursprünglichen Wohneinheiten. Schmerzmittel und entzündungshemmende Mittel können verwendet werden, wenn sie nicht mit Studienziel nicht stören. Wissenschaftliche Begründung für Schmerzmittel vorenthalten, Veterinär-und ICUCA Genehmigung erforderlich sind. Unser Labor hat festgestellt, dass Buprenorphin ,0.05-0 .1 mg / kg SQ ein ausgezeichnetes pre-und post-operative Analgetikum für Mäuse ist. Darüber hinaus kann Lidocain bei 4mg/kg (0,4 ml / kg einer 1% igen Lösung) in die Inzisionsstelle infundiert werden, wenn systemische Analgetika können experimentelle Ergebnisse zu verwechseln. </li><li> Wir in der Regel nicht erleben postoperativen Beeinträchtigungen durch DVT. Allerdings können negative post-operative Effekte sind spontane Anästhesie Tod, chirurgische Blutverlust und partielle Parese aufgrund Nerventrauma. Tiere, die nur langsam regenerieren können eingespeist Nutria Gel oder feuchten chow auf dem Boden des Käfigs, das Tier die Genesung zu erleichtern platziert. Tiere mit sever postoperative Komplikationen human eingeschläfert werden. </li><li> Am zugewiesen Zeitpunkt ist die Euthanasie menschlich in Übereinstimmung mit den Empfehlungen der American Veterinary Medical Association Richtlinien zur Euthanasie für Nager gesetzt durchgeführt. </li></ol> 3. Repräsentative Ergebnisse Die EIM ist ein Modell, Thrombus bildet in Gegenwart von kontinuierlichen Blutfluss. Die folgenden Abbildungen zeigen Parameter in diesem Roman Modell untersucht. <img src="/files/ftp_upload/2737/2737fig1.jpg" alt="Abbildung 1" /> Abbildung 1 definesthe anatomischen Bereich in dem ein 25G EdelstahlNadel, an einem versilberten Kupferdraht (KY-30 bis 1-GRN, Electrospec, Dover, NJ) eingesetzt wird. Beachten Sie, dass ein Lymphknoten fast vorhanden ist und hilft als ein Wahrzeichen der kaudalen IVC zu finden. <img src="/files/ftp_upload/2737/2737fig2.jpg" alt="Abbildung 2" /> Abbildung 2: Thrombus Gewicht von 2 Tagen nach EIM in verschiedene Stämme. Wie erwartet, war Thrombusgewichts niedriger PAI-1 KO-Mäuse und größere in Hyperkoagulabilität Delta CT-Mäuse (^ CT). <img src="/files/ftp_upload/2737/2737fig3.jpg" alt="Abbildung 3" /> Abbildung 3: Löslich P-Selektin, einem Hersteller für tiefe Venenthrombose, wurde in verschiedenen Stämmen gemessen. Die niedrigsten Werte wurden in PAI-1 KO Mäusen gefunden, während die höchsten in Hyperkoagulabilität delta CT-Mäusen gefunden wurden. <img src="/files/ftp_upload/2737/2737fig4.jpg" alt="Abbildung 4" /> Abbildung 4 Zusammenhang zwischen Thrombus Gewicht und löslichen P-Selektin wurde in verschiedene Stämme gefunden. Zu beachten ist, in die Gruppe, desto kürzer Thrombus Größe erzeugt, wurde das lösliche P-Selektin nachgewiesen dem niedrigsten (PAI-1 KO-Mäuse). In der Gruppe, der größte Thrombus Größe erzeugt, wurde das lösliche P-Selektin erkannt höchsten (Hyperkoagulabilität delta CT). <img src="/files/ftp_upload/2737/2737fig5.jpg" alt="Abbildung 5" /> Abbildung 5: Um die Anwesenheit von Blut fließen zu demonstrieren, Ultraschall wurde in Mäusen unterziehen EIM durchgeführt. Abbildung 5 zeigt die Ultraschall in einem Maus-2 Tage nach EIM durchgeführt. Dann wurde die Probe zu ernten. Beachten Sie die, dass der Thrombus Form Thrombusbildung schlage in Anwesenheit von Flow

<ol>
	<li>Diaz, J. A., Hawley, A. E., Alvarado, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thrombogenesis+with+continuous+blood+flow+in+the+inferior+vena+cava.+A+novel+mouse+model.">Thrombogenesis with continuous blood flow in the inferior vena cava. A novel mouse model.</a> Thromb Haemost. 104, 366-375 (2010).</li><li>Eitzman, D. T., Westrick, R. J., Nabel, E. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plasminogen+activator+inhibitor-1+and+vitronectin+promote+vascular+thrombosis+in+mice.">Plasminogen activator inhibitor-1 and vitronectin promote vascular thrombosis in mice.</a> Blood. 95, 577-580 (2000).</li><li>Myers, D., Farris, D., Hawley, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selectins+influence+thrombosis+in+a+mouse+model+of+experimental+deep+venous+thrombosis.">Selectins influence thrombosis in a mouse model of experimental deep venous thrombosis.</a> J Surg Res. 108, 212-221 (2002).</li><li>Myers, D. D., Hawley, A. E., Farris, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=P-selectin+and+leukocyte+microparticles+are+associated+with+venous+thrombogenesis.">P-selectin and leukocyte microparticles are associated with venous thrombogenesis.</a> J Vasc Surg. 38, 1075-1089 (2003).</li><li>Pierangeli, S. S., Barker, J. H., Stikovac, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+human+IgG+antiphospholipid+antibodies+on+an+in+vivo+thrombosis+model+in+mice.">Effect of human IgG antiphospholipid antibodies on an in vivo thrombosis model in mice.</a> Thromb Haemost. 71, 670-674 (1994).</li><li>Pierangeli, S. S., Liu, X. W., Barker, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+thrombosis+in+a+mouse+model+by+IgG,+IgM+and+IgA+immunoglobulins+from+patients+with+the+antiphospholipid+syndrome.">Induction of thrombosis in a mouse model by IgG, IgM and IgA immunoglobulins from patients with the antiphospholipid syndrome.</a> Thromb Haemost. 74, 1361-1367 (1995).</li></ol>

Die Maus ist eine hervorragende Recherche-Tool für in vivo-Experimente. So sind die Entwicklung von Mausmodellen, die genau imitieren Krankheiten oder pathologischen Bedingungen notwendig. Mehrere Venenthrombose Mausmodellen wurden verwendet, um DVT einschließlich Studie: photochemische (2), Stase (3-4) und mechanisches Trauma (5-6). Allerdings verwenden keines dieser Modelle eine interne endothelial Reiz Thrombus in Gegenwart von kontinuierlichen Blutfluss erzeugen. Die EIM, mit Elektrolyt-Stimulus in der IVC, fördert den venösen Thrombosen durch endotheliale Aktivierung in Gegenwart des Blutflusses (1). Es ist technisch einfach, leicht reproduzierbar und hat eine hohe Überlebensrate (99%). Trotz der Tatsache, dass ein elektrischer Strom verwendet wird, um die Elektrolyse zu schaffen, wird die Wärme nicht erzeugt (1). Die EIM-Modell eng simuliert klinische DVT, und dient daher als ein wertvolles Werkzeug für die Untersuchung der inneren Mechanismen der Thrombusbildung und Auflösung. Die EIM-Modell ist auch für die Weiterentwicklung neuer Therapieansätze gegen Thrombose Krankheit nützlich.

<table border="1"><tbody><tr><th> Name des Reagenzes </th><th> Firma </th><th> Katalog-Nummer </th><th> Kommentare (optional) </th></tr><tr><td> DMEM </td><td> Invitrogen </td><td> ABCD1234 </td><td></td></tr></tbody></table>

electrolytic inferior vena cava model (eim) of venous thrombosis

Tiermodelle dienen eine wichtige Rolle in tiefe Venenthrombose (DVT) Forschung, um Thrombenbildung Thrombus Auflösung zu untersuchen und mögliche therapeutische Verbindungen (1) zu testen. Neue Verbindungen in der Behandlung und Prävention von TVT ausgenutzt werden derzeit entwickelt. Die Auslieferung der potenziellen therapeutischen Antagonist Verbindungen an ein betroffenes thrombosierten Vene wurde problematisch. Im Rahmen der therapeutischen Anwendungen, ein Modell, das partielle Stasis nutzt und konsequent erzeugt Thromben innerhalb einer großen Vene wurde vor kurzem gegründet. Die elektrolytische Inferior Vena Cava Model (EIM) ist Mausmodell der DVT, dass Thrombusbildung in Gegenwart von kontinuierlichen Blutfluss ermöglicht. Dieses Modell erlaubt Therapeutika in Kontakt mit dem Thrombus in einer dynamischen Art und Weise sein, und ist empfindlicher als andere Modelle der DVT (1). Darüber hinaus dieses Thrombose-Modell eng simuliert klinische Situationen der Thrombusbildung und ist ideal, um venöse Endothelzellaktivierung, Leukozytenmigration, venösen Thrombosen, zu studieren und therapeutischen Anwendungen (1) zu testen. Die EIM-Modell ist technisch einfach, leicht reproduzierbar, schafft einheitliche Thromben Größen und ermöglicht einer großen Stichprobe (dh Thrombus und Venenwand), die für analytische Zwecke erforderlich ist.

Watch this Scientific Journal Video about Electrolytic Inferior Vena Cava Model EIM of Venous Thrombosis at JoVE.com

Electrolytic Inferior Vena Cava Model EIM of Venous Thrombosis

Die elektrolytische Induktion der endothelialen Aktivierung der inneren Oberfläche der Inferior Vena Cava Ergebnisse im venösen Typ Thrombusbildung durch eine endotheliale Aktivierung und teilweisen Blut-Stase, zwei Komponenten der Virchow-Trias.

Elektrolytische Inferior Vena Cava Model (EIM) von Venenthrombosen

Animal models serve a vital role in deep venous thrombosis (DVT) research in order to study thrombus formation, thrombus resolution and to test potential ...

electrolytic-inferior-vena-cava-model-eim-of-venous-thrombosis

Research

JoVE Journal

Medicine

Electrolytic Inferior Vena Cava Model (EIM) of Venous Thrombosis

17.0K Aufrufe.  University of Michigan. Die elektrolytische Induktion der endothelialen Aktivierung der inneren Oberfläche der Inferior Vena Cava Ergebnisse im venösen Typ Thrombusbildung durch eine endotheliale Aktivierung und teilweisen Blut-Stase, zwei Komponenten der Virchow-Trias.

Serie: Electrolytic Inferior Vena Cava Model EIM of Venous Thrombosis

Mouse Complete Stasis Model of Inferior Vena Cava Thrombosis

Venous thromboembolism (VTE) includes both deep vein thrombosis (DVT) and pulmonary embolism (PE). In the United States (U.S.), the high morbidity and mortality rates make VTE a serious health concern 1-2. After heart disease and stroke, VTE is the third most common vascular disease 3. In the U.S. alone, there is an estimated 900,000 people affected each year, with 300,000 deaths occurring annually 3. A reliable in vivo animal model to study the mechanisms of this disease is necessary.
The advantages of using the mouse complete stasis model of inferior vena cava thrombosis are several. The mouse model allows for the administration of very small volumes of limited availability test agents, reducing costs dramatically. Most promising is the potential for mice with gene knockouts that allow specific inflammatory and coagulation factor functions to be delineated. Current molecular assays allow for the quantitation of vein wall, thrombus, whole blood, and plasma for assays. However, a major concern involving this model is the operative size constraints and the friability of the vessels. Also, due to the small IVC sample weight (mean 0.005 grams) it is necessary to increase animal numbers for accurate statistical analysis for tissue, thrombus, and blood assays such as real-time polymerase chain reaction (RT-PCR), western blot, enzyme-linked immunosorbent (ELISA), zymography, vein wall and thrombus cellular analysis, and whole blood and plasma assays 4-8.
The major disadvantage with the stasis model is that the lack of blood flow inhibits the maximal effect of administered systemic therapeutic agents on the thrombus and vein wall.

The mouse complete stasis model of inferior vena cava thrombosis yields quantifiable amounts of vein wall tissue and thrombus. It has proven useful for evaluating interactions between the vein wall and the occlusive thrombus and in assessing the progression from acute to chronic inflammation.

Maus Komplette Stasis Model of Inferior Vena Cava Thrombose

Venous thromboembolism (VTE) includes both deep vein thrombosis (DVT) and pulmonary embolism (PE).  In the United States (U.S.), the high morbidity and ...

Forschung

Medizin

Implantation of Inferior Vena Cava Interposition Graft in Mouse Model

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. The long-term clinical results showed excellent patency rates, however, with significant incidence of stenosis. To investigate the cellular and molecular mechanisms of vascular neotissue formation and prevent stenosis development in tissue engineered vascular grafts (TEVGs), we developed a mouse model of the graft with approximately 1 mm internal diameter. First, the TEVGs were assembled from biodegradable tubular scaffolds fabricated from a polyglycolic acid nonwoven felt mesh coated with &#949;-caprolactone and L-lactide copolymer. The scaffolds were then placed in a lyophilizer, vacuumed for 24 hr, and stored in a desiccator until cell seeding. Second, bone marrow was collected from donor mice and mononuclear cells were isolated by density gradient centrifugation. Third, approximately one million cells were seeded on a scaffold and incubated O/N. Finally, the seeded scaffolds were then implanted as infrarenal vena cava interposition grafts in C57BL/6 mice. The implanted grafts demonstrated excellent patency (&#62;90%) without evidence of thromboembolic complications or aneurysmal formation. This murine model will aid us in understanding and quantifying the cellular and molecular mechanisms of neotissue formation in the TEVG.

To improve our knowledge of cellular and molecular neotissue formation, a murine model of the TEVG was recently developed. The grafts were implanted as infrarenal vena cava interposition grafts in C57BL/6 mice. This model achieves similar results to those achieved in our clinical investigation, but over a far shortened time-course.

Die Implantation von Vena Cava Interposition Graft in Maus-Modell

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. ...

Orthotopic Injection of Breast Cancer Cells into the Mammary Fat Pad of Mice to Study Tumor Growth.

Breast cancer growth can be studied in mice using a plethora of models. Genetic manipulation of breast cancer cells may provide insights into the functions of proteins involved in oncogenic progression or help to discover new tumor suppressors. In addition, injecting cancer cells into mice with different genotypes might provide a better understanding of the importance of the stromal compartment. Many models may be useful to investigate certain aspects of disease progression but do not recapitulate the entire cancerous process. In contrast, breast cancer cells engraftment to the mammary fat pad of mice better recapitulates the location of the disease and presence of the proper stromal compartment and therefore better mimics human cancerous disease. In this article, we describe how to implant breast cancer cells into mice orthotopically and explain how to collect tissues to analyse the tumor milieu and metastasis to distant organs. Using this model, many aspects (growth, angiogenesis, and metastasis) of cancer can be investigated simply by providing a proper environment for tumor cells to grow.

Cancer is a complex disease that is influenced by the tissue surrounding the tumor as well as local pro- and anti-inflammatory mediators. Therefore, orthotropic injection models, rather than subcutaneous models may be useful to study cancer progression in a manner that better mimics human pathology.

Orthotopische Injektion von Brustkrebszellen in das Brustfettpolster von Mäusen, das Tumorwachstum zu studieren.

Breast cancer growth can be studied in mice using a plethora of models. Genetic manipulation of breast cancer cells may provide insights into the ...

Patch Angioplasty in the Rat Aorta or Inferior Vena Cava

Pericardial patches are commonly used in vascular surgery to close vessels. To facilitate studies of the neointimal hyperplasia that forms on the patch, we developed a rat model of patch angioplasty that can be used in either a vein or an artery, creating a patch venoplasty or a patch arterioplasty, respectively. Technical aspects of this model are discussed. The infra-renal IVC or aorta are dissected and then clamped proximally and distally. A 3 mm venotomy or arteriotomy is performed in the infrarenal inferior vena cava or aorta of 6 to 8 week-old Wistar rats. A bovine pericardial patch (3 mm x 1.5 mm x 0.6 mm) is then used to close the site using a 10-0 nylon suture. Compared to arterial patches, venous patches show increased neointimal thickness on postoperative day 7. This novel model of pericardial patch angioplasty can be used to examine neointimal hyperplasia on vascular biomaterials, as well as to compare the differences between the arterial and venous environments.

We have established a model of pericardial patch angioplasty that can be used in either small-diameter veins or arteries. This model can be used to compare venous and arterial neointimal hyperplasia formation.

Patch-Angioplastie in der Ratte Aorta oder Vena Cava

Pericardial patches are commonly used in vascular surgery to close vessels. To facilitate studies of the neointimal hyperplasia that forms on the patch, ...

A Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep learning curve. The introduction of the cuff technique for anastomosis of the portal vein (PV) and infrahepatic vena cava (IHVC) has significantly simplified the transplant procedure in rats. However, due to the short anterior wall of the recipients' suprahepatic vena cava (SHVC), the cuff technique is very difficult to use for the reconstruction of the SHVC. Most researchers in this field still use the hand-suture technique for SHVC reconstruction, which makes it the bottleneck step in rat orthotopic liver transplantation. The magnetic anastomosis technique (i.e., magnamosis) is a method of connecting two vessels using the attractive force between two magnets. Our recent study has shown that the magnetic anastomosis technique is superior to the hand-suture technique for SHVC reconstruction in rats. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using the novel magnetic anastomosis technique. In this model, the reconstruction of the PV and IHVC was performed by the standard cuff technique, while the reconstruction of the bile duct (BD) was performed by a stent technique. The hepatic re-arterialization was not performed. The magnetic anastomosis technique made SHVC reconstruction much easier and significantly shortened the anphepatic phase. After a reasonable learning curve, even researchers without advanced microsurgical skills can produce reliable and reproducible results using this rat model of OLT.

The reconstruction of the suprahepatic vena cava (SHVC) remains a difficult step in rat orthotopic liver transplantation. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using a novel magnetic anastomosis technique.

Einem Rattenmodell der orthotopen Lebertransplantation mit einer neuartigen magnetischen Anastomose-Technik für Suprahepatic Vena Cava Wiederaufbau

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep ...

Ferric Chloride-induced Canine Carotid Artery Thrombosis: A Large Animal Model of Vascular Injury

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, we have translated a vascular injury model from a small animal into a large animal (canine), with slight modifications that can be used for pre-clinical screening of prophylactic and thrombolytic agents. In addition to the surgical methods, the modified protocol describes the step-by-step methods to assess carotid artery canalization by angiography, detailed instructions to process both the brain and carotid artery for histological analysis to verify carotid canalization and cerebral hemorrhage, and specific parameters to complete an assessment of downstream thromboembolic events by utilizing magnetic resonance imaging (MRI). In addition, specific procedural changes from the previously well-established small animal model necessary to translate into a large animal (canine) vascular injury are discussed.

Here, we present the modifications necessary to a well characterized and commonly used small animal ferric chloride-induced (FeCl3) carotid artery injury model for use in a large animal vascular injury model. The resulting model can be utilized for pre-clinical trial assessment of both prophylactic and thrombolytic pharmacological and mechanical interventions.

Eisenchlorid-induzierte eckzahn Halsschlagader Thrombosen: Eine große Tiermodell der Gefäßverletzung

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

Messung der isometrischen Kontraktilität der Maus mesenterialen Arterie mit Draht Myographie

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Image Acquisition Method for the Sonographic Assessment of the Inferior Vena Cava

Over the past several decades, clinicians have incorporated several applications of diagnostic point-of-care ultrasound (POCUS) into medical decision-making. Among the applications of POCUS, imaging the inferior vena cava (IVC) is practiced by a wide variety of specialties, such as nephrology, emergency medicine, internal medicine, critical care, anesthesiology, pulmonology, and cardiology. Although each specialty uses IVC data in slightly different ways, most medical specialties, at minimum, attempt to use IVC data to make predictions about intravascular volume status. While the relationship between IVC sonographic data and intravascular volume status is complex and highly context-dependent, all clinicians should collect the sonographic data in standardized ways to ensure repeatability. This paper describes standardized IVC image acquisition including patient positioning, transducer selection, probe placement, image optimization, and the pitfalls and limitations of IVC sonographic imaging.&#160;This paper also describes the commonly performed anterior IVC long-axis view and three other views of the IVC that can each provide helpful diagnostic information when the anterior long-axis view is difficult to obtain or interpret.

Point-of-care ultrasound evaluation of the inferior vena cava (IVC) is commonly utilized to identify, among other things, the volume status. Imaging should be performed systematically to ensure repeatability. This manuscript reviews the methods and pitfalls of sonographic IVC examination.

Bildaufnahmeverfahren zur sonographischen Beurteilung der Vena cava inferior

Over the past several decades, clinicians have incorporated several applications of diagnostic point-of-care ultrasound (POCUS) into medical ...

Point-Of-Care Ultrasound Screening for Proximal Lower Extremity Deep Venous Thrombosis

Acute lower extremity deep venous thrombosis (DVT) is a serious vascular disorder that requires accurate and early diagnosis to prevent life-threatening sequelae. While whole leg compression ultrasound with color and spectral Doppler is commonly performed in radiology and vascular labs, point-of-care ultrasound (POCUS) is becoming more common in the acute care setting. Providers appropriately trained in focused POCUS can perform a rapid bedside examination with high sensitivity and specificity in critically ill patients. This paper describes a simplified yet validated approach to POCUS by describing a three-zone protocol for lower extremity DVT POCUS image acquisition. The protocol explains the steps in obtaining vascular images at six compression points in the lower extremity. Beginning at the level of the proximal thigh and moving distally to the popliteal space, the protocol guides the user through each of the compression points in a stepwise manner: from the common femoral vein to the femoral and deep femoral vein bifurcation, and, finally, to the popliteal vein. Further, a visual aid is provided that may assist providers during real-time image acquisition. The goal in presenting this protocol is to help make proximal lower extremity DVT exams more accessible and efficient for POCUS users at the patient&#39;s bedside.

Traditionally, lower extremity deep venous thrombosis (DVT) is diagnosed by radiology-performed venous duplex ultrasound. Providers appropriately trained in focused point-of-care ultrasound (POCUS) can perform a rapid bedside examination with high sensitivity and specificity in critically ill patients. We describe the scanning technique for focused POCUS DVT lower extremity examination.

Point-of-Care-Ultraschall-Screening für proximale tiefe Venenthrombose der unteren Extremität

Acute lower extremity deep venous thrombosis (DVT) is a serious vascular disorder that requires accurate and early diagnosis to prevent life-threatening ...

Strategien für eine effiziente vaskuläre Anastomose bei der Lebertransplantation der Maus

Vascular Reconstruction with the Cuff Technique in Mouse Orthotopic Liver Transplantation

Mouse orthotopic liver transplantation is an effective methodology for investigating the underlying mechanisms of liver ischemia and reperfusion injury. However, the technical challenges pose a barrier to utilizing this valuable experimental model and passing on these skills to the next generation. The most challenging aspect of this procedure is vascular reconstruction, including the portal vein (PV), infrahepatic inferior vena cava (IHIVC), and suprahepatic inferior vena cava. The use of plastic cuffs, rather than sutures, allows for smoother PV and IHIVC reconstruction. Vessels are reconstructed by attaching a cuff made from an intravenous catheter to the tip of the graft vessel and interposing the cuff into the recipient vessel. The two most crucial aspects are properly visualizing the inner lumen of the vessel and avoiding the use of excessive force. Our aim is to provide a technical overview of vascular reconstructions using the cuff technique in recipient surgery. These technical tips for the cuff technique are expected to help microsurgeons facilitate vascular reconstruction and advance their research.

Autor im Rampenlicht: Erforschung der kalten Ischämie und der warmen Reperfusionsschädigung bei der Fettlebertransplantation

This protocol provides technical information for vessel reconstruction using the cuff technique in mouse orthotopic liver transplantation.

Gefäßrekonstruktion mit der Manschettentechnik bei der orthotopen Lebertransplantation der Maus

Mouse orthotopic liver transplantation is an effective methodology for investigating the underlying mechanisms of liver ischemia and reperfusion injury. ...