Supportato da NIH 1P01HL089407-01A1 (Lawrence, PI), Animal Nucleo A, NIH 1 K01 HL080962-01A2 (Myers, PI).

1. Topo Anestesia Procedure <ol><li> Gli animali vengono rimossi dal loro gabbia e messi in anestesia pre-operatoria camera di induzione per l&#39;induzione di gas al 2% isoflurano e il 100% di ossigeno ad un flusso di 0,5 litri al minuto (vaporizzatore controllato). </li><li> I topi sono sedato nella camera di induzione dell&#39;anestesia, rimosso dalla camera e l&#39;addome ventrale è rasata con cesoie elettriche, o mantenuti all&#39;interno della camera se un depilatorio capelli è usato (1 min). </li><li> Mentre ancora sotto sedazione, sono pesati, gli occhi sono lubrificati con pomata oftalmica sterile e, poi messa in recumbancy dorsale su un dispositivo di riscaldamento ad acqua calda in circolazione in anestesia generale con un cono con isoflurano e ossigeno. A questo punto il tasso di ossigeno si riduce a 0,2 litri al minuto. </li><li> L&#39;addome viene spruzzato con una soluzione di clorexidina (clorexidina diacetato è rapidamente bacterialcidal e persistente) e si asciugò con garza sterile o tre volte fino a quando il sito chirurgico è pulito. </li></ol> 2. Micro Mouse-chirurgici e procedure di ripristino <ol><li> Una incisione linea mediana ventrale (2 cm) è fatto con le forbici iris attraverso la pelle e la parete addominale esporre l&#39;addome. Una sterile 2x2 garze imbevute di soluzione salina viene utilizzato per riflettere l&#39;intestino alla sinistra dell&#39;animale. Un dispositivo di immobilizzazione mouse è messo in atto a consentire la visualizzazione della vena cava inferiore (IVC). </li><li> A questo punto, l&#39;isoflurano è ridotto ad un livello di manutenzione del 2% e il tasso di ossigeno resta a 0,2 litri al minuto per il resto della procedura chirurgica. </li><li> Dissezione Blunt è facilitato utilizzando un tampone sterile applicatore ed extra delicato tessuto dell&#39;iride pinze mezzo curvo. Bisogna fare attenzione nel maneggiare il mouse per tessuti sono estremamente fragili. </li><li> Tutti i rami laterali IVC, dalle vene renali alla biforcazione iliaca, sono legatura con 7-0 punti di sutura non reattivi prolene. Rami sono rimasti indietro di brevetto. </li><li> Nel elettrolitico IVC modello (EIM), un 25G in acciaio inossidabile ago, attaccato ad un filo di rame ricoperto d&#39;argento (KY-30-1-GRN, Electrospec, Dover, NJ) è inserito nel esposti caudale IVC, (Figura 1) e posizionato contro la parete anteriore (anodo). </li><li> Un altro filo è impiantato per via sottocutanea completare il circuito (catodo). </li><li> Una corrente di 250 μAmps di 15 minuti è stato applicato usando un Grass S48 stimolatore ad onda quadra e una unità di corrente costante (Tecnologie Erba, Un Astro-Med, Inc., West Warwick, RI). I risultati a corrente nella formazione di prodotti tossici di elettrolisi che erodono la superficie endoteliale della IVC promozione di un ambiente trombogenico e la successiva formazione di trombi. </li><li> Dopo 15 minuti, l&#39;ago viene rimosso e un batuffolo di cotone si tiene delicatamente in contatto con l&#39;area per prevenire le emorragie </li><li> Negli animali sham, l&#39;ago viene inserito nella IVC e poi rimosso, senza applicazione della corrente. </li><li> Il sito laparotomia è chiuso in due strati di moda. Il nostro laboratorio utilizza Vicryl 5-0, una sutura sintetica, in un modello continuo per chiudere la parete addominale. Colla pelle adesivo utilizzato per chiudere la pelle. Non ci sono suture esterne per la rimozione. </li><li> I topi sono poi recuperati in una gabbia di topo individuale, strettamente osservate post-operatoria (45 minuti a 2 ore) sotto una lampada riscaldante (distanza minima 24 cm di distanza dalla gabbia), poi tornati alle loro abitazioni originale. Farmaci antidolorifici e antinfiammatori può essere utilizzato se non interferisce con l&#39;obiettivo di studio. Giustificazione scientifica per trattenere farmaci per il dolore, veterinario, e l&#39;approvazione ICUCA sono necessari. Il nostro laboratorio ha scoperto che Buprenorfina ,0.05-0 0,1 mg / kg SQ è un analgesico ottimo pre-e post-operatoria per i topi. Inoltre, a 4mg/kg lidocaina (0,4 ml / kg di una soluzione all&#39;1%) può essere infusa nel sito di incisione, quando gli analgesici sistemici possono confondere i risultati sperimentali. </li><li> Noi in genere non si verificano effetti negativi post-operatoria a causa di TVP. Tuttavia, avversa post-operatoria effetti possono includere spontaneo morte sotto anestesia, perdita di sangue chirurgiche e paresi parziale a causa di un trauma nervoso. Gli animali che sono lenti per recuperare possono essere alimentati nutria gel o umidi chow posto sul pavimento della gabbia per facilitare il recupero dell&#39;animale. Gli animali con sever complicanze post-operatorie dovrebbero essere umanamente eutanasia. </li><li> Al punto di tempo assegnato, l&#39;eutanasia viene eseguita con umanità conformemente alle raccomandazioni stabilite dalla American Veterinary Medical Association Linee guida sull&#39;eutanasia per i roditori. </li></ol> 3. Rappresentante Risultati L&#39;EIM è un modello trombo che si forma in presenza di flusso di sangue continuo. Le figure seguenti mostrano i parametri indagati in questo modello di romanzo. <img src="/files/ftp_upload/2737/2737fig1.jpg" alt="Figura 1" /> Figura 1 zona definesthe anatomiche in cui un 25G in acciaio inoxago, attaccato ad un filo d&#39;argento di rame ricoperto (KY-30-1-GRN, Electrospec, Dover, NJ) è inserito. Si noti che un nodo linfatico è quasi presente e aiuta come punto di riferimento per trovare la caudale IVC. <img src="/files/ftp_upload/2737/2737fig2.jpg" alt="Figura 2" /> Figura 2: peso Trombo 2 giorni dopo EIM in diversi ceppi. Come previsto, il peso trombo era più bassa nel PAI-1 nei topi KO e più grande in ipercoagulabilità Delta CT topi (^ CT). <img src="/files/ftp_upload/2737/2737fig3.jpg" alt="Figura 3" /> Figura 3: P-selectina solubile, un creatore di trombosi venosa profonda, è stata misurata in diversi ceppi. I livelli più bassi sono stati trovati nel PAI-1 nei topi KO, mentre i più alti livelli sono stati trovati in ipercoagulabilità delta topi CT. <img src="/files/ftp_upload/2737/2737fig4.jpg" alt="Figura 4" /> Figura 4 Correlazione tra peso trombo e P-selectina solubile è stato trovato in diversi ceppi. Da segnalare, nel gruppo che ha generato la dimensione più corta del trombo, il P-selectina solubile è stata rilevata la più bassa (PAI-1 nei topi KO). Nel gruppo che ha generato la più grande dimensione del trombo, il P-selectina solubile è stata rilevata la più alta (ipercoagulabilità delta CT). <img src="/files/ftp_upload/2737/2737fig5.jpg" alt="Figura 5" /> Figura 5: Al fine di dimostrare la presenza di flusso di sangue, l&#39;ecografia è stata eseguita nei topi sottoposti a EIM. La Figura 5 mostra l&#39;ecografia eseguita in un mouse 2 giorni dopo EIM. Poi il campione è stato raccolto. Si noti che la forma del trombo suggeriscono la formazione di trombi in presenza di flusso

<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>

Nessun conflitto di interessi dichiarati.

Il mouse è uno strumento di ricerca di eccellenza per esperimenti in vivo. Così, lo sviluppo di modelli murini che assomigliano malattie o condizioni patologiche sono necessari. Diversi modelli murini trombosi venosa sono stati utilizzati per studiare TVP tra cui: fotochimico (2), stasi (3-4) e traumi meccanici (5-6). Tuttavia, nessuno di questi modelli utilizzano uno stimolo interno endoteliale per generare trombo in presenza di flusso di sangue continuo. L&#39;EIM, utilizzando stimolo elettrolitico all&#39;interno della IVC, promuove trombogenesi venosa mediante l&#39;attivazione endoteliale in presenza di flusso di sangue (1). È tecnicamente semplice, facilmente riproducibile ed ha un elevato tasso di sopravvivenza (99%). Nonostante il fatto che una corrente elettrica viene utilizzato per creare l&#39;elettrolisi, il calore non viene generato (1). Il modello simula EIM TVP strettamente clinico, e costituisce pertanto uno strumento prezioso per studiare i meccanismi intrinseci di formazione di trombi e risoluzione. Il modello EIM è utile anche per l&#39;avanzamento di nuovi approcci terapeutici contro il morbo di trombosi.

<table border="1"><tbody><tr><th> Nome del reagente </th><th> Azienda </th><th> Numero di catalogo </th><th> Commenti (opzionale) </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

I modelli animali hanno un ruolo vitale nella trombosi venosa profonda (TVP) ricerca per studiare la formazione di trombi, la risoluzione del trombo e per testare potenziali composti terapeutici (1). Nuove mescole da utilizzare nel trattamento e nella prevenzione della TVP sono attualmente in fase di sviluppo. La consegna dei potenziali composti antagonista terapeutico ad una vena interessata trombizzata è stato problematico. Nel contesto delle applicazioni terapeutiche, un modello che utilizza parziale stasi e genera costantemente trombi all&#39;interno di un filone importante è stato di recente istituzione. Il elettrolitica vena cava inferiore Model (EIM) è un modello murino di TVP che permette la formazione di trombi in presenza di flusso di sangue continuo. Questo modello consente agli agenti terapeutici di essere in contatto con il trombo in modo dinamico, ed è più sensibile rispetto ad altri modelli di trombosi venosa profonda (1). Inoltre, questo modello simula trombosi da vicino situazioni cliniche di formazione di trombi ed è ideale per studiare l&#39;attivazione delle cellule endoteliali venose, la migrazione dei leucociti, trombogenesi venoso, e per testare le applicazioni terapeutiche (1). Il modello di EIM è tecnicamente semplice, facilmente riproducibile, crea coerente dimensioni trombi e consente un ampio campione (cioè trombo e parete venosa) che è richiesto per scopi di analisi.

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

L&#39;induzione elettrolitica di attivazione endoteliale sulla superficie interna della vena cava risultati in formazione tipo trombo venoso a causa di attivazione endoteliale e parziale stasi del sangue, due componenti della triade di Virchow.

Elettrolitico vena cava inferiore Model (EIM) della Trombosi Venosa

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 Views.  University of Michigan. L'induzione elettrolitica di attivazione endoteliale sulla superficie interna della vena cava risultati in formazione tipo trombo venoso a causa di attivazione endoteliale e parziale stasi del sangue, due componenti della triade di Virchow.

Video: 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.

Modello del mouse completa stasi della vena cava inferiore Trombosi

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

Ricerca

Medicina

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.

L&#39;impianto di Vena cava inferiore Interposizione Graft in modello murino

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.

Iniezione ortotopico di cellule di carcinoma mammario nel mammaria Fat Pad di Mice per studiare la crescita del tumore.

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 Angioplastica nel ratto aorta o vena cava inferiore

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.

Un modello del ratto di orthotopic del trapianto del fegato usando una tecnica novella dell'anastomosi magnetico per la ricostruzione della Vena Cava Suprahepatic

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.

Trombosi dell'arteria carotica canini cloruro ferrico-indotta: Un grande modello animale di danno vascolare

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.

Misurazione di contrattilità isometrica dell'arteria mesenterica Mouse con filo Myography

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.

Metodo di acquisizione delle immagini per la valutazione ecografica della vena cava inferiore

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.

Screening ecografico point-of-care per trombosi venosa profonda degli arti inferiori prossimali

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

Strategie per un'efficiente anastomosi vascolare nel trapianto di fegato di topo

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.

Autore in primo piano: Esplorazione dell'ischemia fredda e del danno da riperfusione calda nel trapianto di fegato grasso

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

Ricostruzione vascolare con la tecnica della cuffia nel trapianto di fegato ortotopico di topo

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