Name: electrolytic inferior vena cava model (eim) of venous thrombosis
Uploaded: 2011-07-12T13:00:00
Duration: 6 min 3 s
Description: Watch this Scientific Journal Video about Electrolytic Inferior Vena Cava Model EIM of Venous Thrombosis at JoVE.com

Ondersteund door NIH 1P01HL089407-01A1 (Lawrence, PI), Animal Core A, NIH een K01 HL080962-01A2 (Myers, PI).

1. Muis Anesthesie Procedures <ol><li> Dieren uit hun kooi en geplaatst in een verdovingsmiddel pre-operatieve inductie kamer voor gasinductie op 2% isofluraan en 100% zuurstof bij een debiet van 0,5 liter per minuut (vaporizer gecontroleerd). </li><li> Muizen worden verdoofd in de verdoving inductie kamer, verwijderd van kamer en het ventrale buik wordt geschoren met de tondeuse, of onderhouden in de kamer als een haar ontharingscrème wordt gebruikt (1 min). </li><li> Hoewel nog steeds onder sedatie, ze zijn gewogen, zijn de ogen gesmeerd met steriele oogzalf, en vervolgens geplaatst in dorsale recumbancy op een warm water circuleert apparatuur voor verwarming van onder algehele narcose met een neus met behulp van isofluraan en zuurstof. Op dit punt de zuurstof tarief wordt verlaagd tot 0,2 liter per minuut. </li><li> De buik is bespoten met chloorhexidine-oplossing (chloorhexidine diacetaat wordt snel bacterialcidal en persistent) en veegde met een steriel gaasje drie keer of totdat de operatiewond schoon is. </li></ol> 2. Muis Micro-chirurgische en recovery procedures <ol><li> Een ventrale middellijn incisie (2 cm) is gemaakt met iris schaar door de huid en de buikwand het blootstellen van de buik. Een steriele zoutoplossing gedrenkt 2x2 in gaas wordt gebruikt om de darmen naar links van het dier kant weer te geven. Een muis restrainer is opgezet zodat visualisatie van de vena cava inferior (IVC). </li><li> Op dit punt wordt de isofluraan teruggebracht tot een onderhouds niveau van 2% en de zuurstof percentage blijft op 0,2 liter per minuut voor de rest van de chirurgische procedure. </li><li> Stompe dissectie wordt vergemakkelijkt met een steriele applicator staafje en een extra delicate iris half gebogen weefsel tang. Voorzichtigheid is geboden bij het hanteren muisweefsels want ze zijn uiterst kwetsbaar. </li><li> Alle IVC zijtakken, van de nier-aders naar de iliacale bifurcatie, worden geligeerd met 7-0 niet-reactief prolene hechtingen. Terug takken bleven patent. </li><li> In de elektrolytische IVC model (EIM), een 25G roestvrij stalen naald, bevestigd aan een verzilverd koperdraad (KY-30-1-GRN, Electrospec, Dover, New Jersey) is ingebracht in de blootgestelde caudale IVC, (figuur 1) en geplaatst tegen de voorste muur (anode). </li><li> Een andere draad wordt geïmplanteerd subcutaan het invullen van het circuit (kathode). </li><li> Een stroom van 250 μAmps meer dan 15 minuten werd toegepast met behulp van een Grass S48 blokgolf stimulator en een constante stroom eenheid (Grass Technologies, een Astro-Med, Inc, West Warwick, RI). De directe stroom resulteert in de vorming van giftige producten van elektrolyse dat de endotheliale oppervlak van het IVC het bevorderen van een trombogene omgeving en de daaropvolgende trombusvorming eroderen. </li><li> Na 15 minuten, wordt de naald verwijderd en een wattenstaafje wordt voorzichtig in contact met het gebied om het bloeden te voorkomen </li><li> In schijnvertoning dieren, wordt de naald geplaatst in de IVC en vervolgens verwijderd, zonder toepassing van de huidige. </li><li> De laparotomie site is gesloten in een twee-lagen mode. Ons laboratorium maakt gebruik van 5-0 Vicryl, een synthetisch hechtdraad, in een doorlopend patroon af te sluiten van de buikwand. Adhesive huidlijm wordt gebruikt om de huid te sluiten. Er zijn geen externe hechtingen voor verwijdering. </li><li> Muizen worden dan teruggevonden in een individuele muizen kooi, op de voet post-operatief waargenomen (45 minuten tot 2 uur) onder een warmtelamp (min. afstand 24 centimeter uit de buurt van kooi), keerde daarna terug naar hun oorspronkelijke woningen. Pijnstillers en anti-inflammatoire middelen kunnen worden gebruikt als het niet interfereert met studie doelstelling. Wetenschappelijke redenen om pijnstillers te weigeren, veterinaire en ICUCA goedkeuring nodig zijn. Ons laboratorium heeft uitgewezen dat Buprenorfine ,0.05-0 0,1 mg / kg SQ is een uitstekende pre-en post-operatieve analgeticum voor muizen. Bovendien kan Lidocaïne op 4mg/kg (0,4 ml / kg van een 1% oplossing) worden toegediend in de incisie site wanneer systemische analgetica kan experimentele resultaten verwarren. </li><li> We meestal niet ervaren post-operatieve schadelijke effecten als gevolg van DVT. Kan echter negatieve post-operatieve effecten zijn spontane narcose dood, chirurgische bloedverlies, en gedeeltelijke parese als gevolg van zenuw trauma. Dieren die zijn traag om te herstellen kunnen worden ingevoerd nutria gel of vochtige chow op de vloer van de kooi om het dier terugwinning te vergemakkelijken. Dieren met verbreken post-operatieve complicaties op humane wijze worden gedood. </li><li> Op de toegewezen tijdstip, is euthanasie humaan uitgevoerd in overeenstemming met de aanbevelingen uiteengezet door de American Veterinary Medical Association richtsnoeren inzake Euthanasie voor knaagdieren in te stellen. </li></ol> 3. Representatieve resultaten Het EIM is een model dat trombus vormen in de aanwezigheid van continue bloedstroom. De volgende cijfers laten zien parameters onderzocht in deze roman model. <img src="/files/ftp_upload/2737/2737fig1.jpg" alt="Figuur 1" /> Figuur 1 definesthe anatomische gebied waar een 25G roestvrij staalnaald, bevestigd aan een verzilverd koperdraad (KY-30-1-GRN, Electrospec, Dover, New Jersey) is geplaatst. Merk op dat een lymfatische node is vrijwel aanwezig is en helpt als een mijlpaal op zoek naar de caudale IVC. <img src="/files/ftp_upload/2737/2737fig2.jpg" alt="Figuur 2" /> Figuur 2: trombus gewicht 2 dagen na EIM in verschillende stammen. Zoals verwacht, trombus gewicht was lager in de PAI-1 KO muizen en groter in hypercoagulatie Delta CT muizen (^ CT). <img src="/files/ftp_upload/2737/2737fig3.jpg" alt="Figuur 3" /> Figuur 3: Oplosbaar P-selectine, een maker voor diepe veneuze trombose, werd gemeten in verschillende stammen. De laagste niveaus werden gevonden in PAI-1 KO muizen terwijl de hoogste niveaus werden gevonden in hypercoagulatie delta CT muizen. <img src="/files/ftp_upload/2737/2737fig4.jpg" alt="Figuur 4" /> Figuur 4 Correlatie tussen trombus gewicht en oplosbaar P-selectine werd gevonden in verschillende stammen. Van de nota, in de groep die de kortere trombus grootte gegenereerd, de oplosbare P-selectine werd aangetoond was de laagste (PAI-1 KO muizen). In de groep die de grootste trombus grootte gegenereerd, de oplosbare P-selectine ontdekt was het hoogst (hypercoagulatie delta CT). <img src="/files/ftp_upload/2737/2737fig5.jpg" alt="Figuur 5" /> Figuur 5: Met het oog op de aanwezigheid van de bloedstroom te tonen, echografie werd uitgevoerd bij muizen ondergaan EIM. Figuur 5 toont de echografie uitgevoerd in een muis 2 dagen na EIM. Dan is de monster was oogst. Let op de dat de trombus vorm trombusvorming suggereren in aanwezigheid van stroom

<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>

De muis is een uitstekend research tool voor in vivo experimenten. Daarom is de ontwikkeling van muismodellen die nauw nabootsen ziekten of pathologische omstandigheden noodzakelijk zijn. Meerdere veneuze trombose muismodellen zijn gebruikt om DVT onderzoek, waaronder: fotochemische (2), stasis (3-4) en mechanische trauma (5-6). Echter, geen van deze modellen maken gebruik van een intern endotheel stimulans om een ​​thrombus in de aanwezigheid van continue bloedstroom te genereren. Het EIM, met behulp van elektrolytische stimulans binnen de IVC, bevordert de veneuze trombogenese door endotheel activatie in de aanwezigheid van de bloedstroom (1). Het is technisch eenvoudig, gemakkelijk reproduceerbaar en heeft een hoog overlevingspercentage (99%). Ondanks het feit dat een elektrische stroom wordt gebruikt om de elektrolyse te maken, wordt warmte niet gegenereerd (1). Het EIM-model simuleert nauw klinische DVT, en dus dient als een waardevol instrument voor het bestuderen van de intrinsieke mechanismen van trombusvorming en resolutie. Het EIM-model is ook nuttig voor de vooruitgang van de nieuwe therapeutische aanpak voor de bestrijding trombose ziekte.

<table border="1"><tbody><tr><th> Naam van het serum </th><th> Vennootschap </th><th> Catalogus nummer </th><th> Commentaar (optioneel) </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

Diermodellen dienen een belangrijke rol in diepe veneuze trombose (DVT) onderzoek met het oog op trombusvorming, trombus resolutie te bestuderen en om potentiële therapeutische stoffen (1) test. Nieuwe verbindingen te worden gebruikt in de behandeling en preventie van DVT worden momenteel ontwikkeld. De levering van potentiële therapeutische antagonist verbindingen aan een getroffen thrombosed ader is problematisch. In het kader van therapeutische toepassingen, een model dat gedeeltelijke stasis gebruikt en consequent genereert trombi binnen een belangrijke ader is onlangs vastgesteld. De elektrolytische vena cava inferior Model (EIM) is muismodel van DVT die trombusvorming in de aanwezigheid van continue bloedstroom mogelijk maakt. Dit model maakt therapeutische middelen om in contact met de trombus op een dynamische manier, en is gevoeliger dan de andere modellen van DVT (1). Daarnaast is deze trombose model simuleert nauw klinische situaties van trombusvorming en is ideaal om veneuze endotheel activatie, migratie van leukocyten, veneuze trombogenese, bestuderen en therapeutische toepassingen (1) test. Het EIM-model is technisch eenvoudig, gemakkelijk reproduceerbaar, creëert consistente trombi maten en zorgt voor een grote steekproef (dat wil zeggen trombus en ader muur) die nodig is voor analytische doeleinden.

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

De elektrolytische inductie van endotheel activatie aan het interne oppervlak van de vena cava inferior resulteert in een veneuze soort trombusvorming te wijten aan endotheel activatie en gedeeltelijke bloed stasis, twee componenten van de triade Virchow&#39;s.

Elektrolytische Inferior Vena Cava Model (EIM) van veneuze trombose

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

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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.

Muis Compleet Stasis Model van Inferior Vena Cava trombose

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

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.

Implantatie van de vena cava inferior Interpositie Graft in muismodel

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

Ferric Chloride-induced Thrombosis Mouse Model on Carotid Artery and Mesentery Vessel

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric chloride injury is now a well-established technique to rapidly and accurately induce the formation of thrombi in exposed veins or artery of small and large diameter. This model has played a key role in the study of the pathophysiology of thrombosis, in the discovery and validation of novel antithrombotic drugs and in the understanding of the mechanism of action of these new agents. Here, the implementation of this technique on a mesenteric vessel and carotid artery in mice is presented. The method describes how to label circulating leukocytes and platelets with a fluorescent dye and to observe, by intravital microscopy on the exposed mesentery, their accumulation at the injured vessel wall which leads to the formation of a thrombus. On the carotid artery, the occlusion caused by the clot formation is measured by monitoring the blood flow with a Doppler probe.

The FeCl3 induced thrombosis model in mice is described herein. A method to monitor thrombus growth by intravital microscopy observation on a mesenteric vessel and by blood flow measurement in the carotid artery is presented.

Ferric-Chloride geïnduceerde trombose Muis Model op halsslagader en Mesenterium Vessel

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric ...

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 het Rat Aorta of 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, ...

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.

Een Rat Model van Orthotopic met behulp van een roman magnetische Anastomosis techniek voor Suprahepatic Vena Cava wederopbouw levertransplantatie

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.

Canine halsslagader ferrichloride-geïnduceerde trombose: Een grote diermodel van vasculaire verwonding

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.

Isometrische Contractility meting van de muis mesenterische slagader met behulp van draad Myography

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

Murine Model of Central Venous Stenosis using Aortocaval Fistula with an Outflow Stenosis

Central venous stenosis is an important entity contributing to arteriovenous fistula (AVF) failure. A murine AVF model was modified to create a partial ligation of the inferior vena cava (IVC) in the outflow of the fistula, mimicking central venous stenosis. Technical aspects of this model are introduced. The aorta and IVC are exposed, following an abdominal incision. The infra-renal aorta and IVC are dissected for proximal clamping, and the distal aorta is exposed for puncture. The IVC at the midpoint between the left renal vein and the aortic bifurcation is carefully dissected to place an 8-0 suture beneath the IVC. After clamping the aorta and IVC, an AVF is created by puncturing the infra-renal aorta through both walls into the IVC with a 25 G needle, followed by ligating a 22 G intra-venous (IV) catheter and IVC together. The catheter is then removed, creating a reproducible venous stenosis without occlusion. The aorta and IVC are unclamped after confirming primary hemostasis. This novel model of central vein stenosis is easy to perform, reproducible, and will facilitate studies on AVF failure.

An aortocaval fistula was created by puncturing the murine infra-renal aorta through both walls into the inferior vena cava and was followed by creation of a stenosis in its outflow via partial ligation of the inferior vena cava. This reproducible model can be used to study central venous stenosis.

Murine model van centraal veneuze stenose met behulp van Aortocaval fistel met een uitstroom stenose

Central venous stenosis is an important entity contributing to arteriovenous fistula (AVF) failure. A murine AVF model was modified to create a partial ...

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.

Beeldacquisitiemethode voor de sonografische beoordeling van de inferieure vena cava

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 echografie screening voor proximale onderste extremiteit diepe veneuze trombose

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