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tutti gli esperimenti sugli animali sono stati eseguiti in conformità con la legge di protezione animale tedesco (TierSchG) e sono stati approvati dal comitato locale benessere degli animali (ufficio di stato della Sassonia inferiore per tutela del consumatore e sicurezza alimentare, protocollo TSA 14/1556). Il peso minimo del mouse è adatto per questo modello è di 25 g.
 1. preoperative preparazioni Nota: tutte le procedure vengono eseguite in condizioni di pulito, non sterile, con strumenti sterilizzati nell'autoclave.
<ol><li>Posto 50-60 8 cm lungo propilene hollow fibre in parallelo in un tubo e collegare con un adattatore a T su entrambi i lati. Riempire lo spazio tra le fibre cave e il ramo esterno dell'adattatore a T con colla.
	<ol><li>Consenti almeno 24 ore per la colla per indurire. Tagliare le fibre cave che si estende dal T-adattatore tramite un microtomo standard e accostare il tubo di silicone per i siti di collegamento.</li>
	</ol></li> <li>Inserire un ago di metallo 27 G (da un branule di 26G) in una cannula di poliuretano Fr 2. Utilizzare una lama chirurgica e micro-forbici al microscopio (ingrandimento X 8-12) per rendere le finestrature di tre o quattro di circa 0,15 cm entro il terzo distale della cannula per assicurare ottimale venoso flusso di ritorno.
	<ol><li>Rimuovi il filo quando completato. Utilizzare tamponi di cotone per dissezione smussa e retrazione delle strutture. Utilizzare tamponi di garza (5 x 5 cm 2) per assorbire il liquido in eccesso per prevenire la disidratazione del tessuto.</li>
	</ol></li> <li>Preparare il volume di riempimento. Per il completamento, aggiungere 30 UI eparina / mL soluzione e 2,5% v/v di una soluzione di 8,4% di NaHCO 3 per il buffer di adescamento. Conservare questa soluzione a 4 ° C fino a quando non usato.</li>
	<li>Riempire il circuito CPB (cioè, pompa, trapper aria, tubi e due cannule) con 850 µ l di soluzione di adescamento tramite cannula venosa. Una volta riempito, tenere il CPB in circolazione fino a quando l'animale è pronto per l'inserimento di una canula.</li>
</ol> 2. Anestesia degli animali <ol><li>per amministrare l'anestesia, metti l'animale in una camera di induzione inferiore 2,5% miscela di ossigeno/isoflurano v/v. Confermare la corretta anestesia valutando pedale ritiro reflex, reflex pizzico di coda e riflesso batter occhio. Applicare l'unguento veterinario degli occhi per evitare secchezza degli occhi.</li>
	<li>Metti l'animale su un tappetino riscaldante con funzione di regolazione di temperatura. Misurare la temperatura corporea con una sonda inserita per via rettale e collegato al sistema di acquisizione dati.</li>
	<li>Dopo anestesia completa è realizzata, intubare il orotracheally animale utilizzando una braunule di plastica di 20G, inserendolo oralmente e spingendolo nella trachea sotto controllo visivo. Iniziare la ventilazione meccanica utilizzando un vaporizzatore di isoflurane collegato ad un ventilatore del mouse. A seconda del peso dell'animale, regolare la ventilazione in modo che si ottiene un volume corrente di 250-350 µ l.</li>
	<li>Per assicurare la completa analgesia, iniettare 5 mg/kg peso corporeo degli animali carprofen per via sottocutanea. Si raccomanda che la concentrazione di isoflurane siano tenuti fra 1.3-2.5% in ossigeno. Isoflurano concentrazione può essere regolata manualmente a seconda della fase della procedura.</li>
</ol> 3. Procedure chirurgiche <ol><li>dopo l'anestesia completa e l'intubazione è raggiunto, eseguire un'incisione della pelle del midline al collo con forbici bene affilate, separare i muscoli in modo schietto ed esporre la vena giugulare di destra e sinistra dell'arteria carotica. Durante la preparazione, coagulare piccoli vasi con un veterinario coagulatore collegato alla pinza bipolare per garantire la perdita di anima minima.</li>
	<li>Dopo la preparazione dei vasi giugulari, cranialmente legare il segmento distale dell'arteria carotide comune sinistra alla sua biforcazione con punti di sutura seta 8-0.</li>
	<li>Collegare l'estremità distale di una cannula di 27 G alla tubazione di affluenza arteriosa utilizzando un connettore in silicone (ID di 0,5 mm, 1 mm di diametro), posto 8-0 sutura seta slip nodi a segmento prossimale dell'arteria e inserire la punta della cannula nell'arteria carotica.</li>
	<li>Dopo il corretto posizionamento della punta della cannula, avanzare la punta della cannula in modo che depone 3-4 mm prossimalmente al arco aortico. Difficoltà la punta della cannula fissando con nodi di seta 8-0.</li>
	<li>Uso microsurgical pinze e le forbici, eseguire la dissezione smussata e forte, esporre la vena giugulare di destra da smussato preparazione del tessuto lateralmente al muscolo sternocleidomastoideo, vicino alla clavicola e mettere un 8-0 seta nodo all'estremità distale e un 8-0 ciclo all'estremità prossimale.</li>
	<li>Dopo la legatura dell'estremità distale della vena giugulare, posizionare la punta della cannula venosa nella vena giugulare di destra e progressi verso l'atrio destro e fissarla con il nodo di seta. Per ottimo ritorno venoso, può essere necessario per far avanzare la punta venosa nel ventricolo destro.</li>
	<li>Una volta ottenuta la posizione corretta della cannula, svolgere eparinizzazione sistemica con l'aggiunta di 2,5 IU dell'eparina per grammo del peso corporeo degli animali tramite iniezione endovenosa diretta nella vena giugulare.</li>
	<li>Per il monitoraggio in tempo reale pressione invasiva, incannulare l'arteria femorale di sinistra. Esporre la zona inguinale e separare delicatamente l'arteria femorale comune dalla vena femorale e del nervo femorale sotto ingrandimento X 25.
	<ol><li>Legano la parte distale dell'arteria femorale. Temporaneamente occludere la parte prossimale con un cappio e praticare una piccola incisione sulla parete anteriore utilizzando micro-forcipe.</li>
		<li>In seguito, inserire una cannula di poliuretano Fr 1 nell'arteria femorale e fissarlo con una seta 10-0 suturare. Questa cannula viene utilizzata per estrarre i campioni di sangue per analisi di gas del sangue.</li>
	</ol></li> <li>Dopo posizionamento di successo delle cannule arteriose e venose, avviare l'esclusione cardiopolmonare di accendere la pompa. Il tempo da intubazione a partire del CPB è circa 45 min. iniziare ad usare una portata di 0,5 mL/min, a seconda delle misurazioni della pressione sistemica e aumentare il flusso sanguigno all'interno di 2 min a flusso pieno (4-6 mL/min) aumentando la velocità della pompa.</li>
	<li>Sotto monitoraggio completo, eseguire un sternotomy superiore utilizzando delle forbici affilate, a partire da manubrio e andando 5 mm in direzione caudale. Coagulare i bordi sternali immediatamente per evitare il sanguinamento. Esporre l'arco aortico tirando l'arteria carotica di destra in direzione craniale.</li>
	<li>Per un controllo ottimale, circonferenzialmente gratis arco aortico da timo e nel tessuto circostante per facilitare il bloccaggio. Luogo un ciclo di 8-0 seta intorno all'aorta ascendente. Per inserire il morsetto a croce per cardioplegia, tirare l'ansa seta in direzione craniale per esporre meglio l'aorta ascendente.</li>
</ol> 4. Esclusione cardiopolmonare e l'emogasanalisi <ol><li>per analisi di gas del sangue (BGA), utilizzare tubi capillari di vetro per raccogliere 60-90 µ l di sangue arterioso tramite catetere dell'arteria femorale.
	<ol><li>Utilizzare un piccolo morsetto del tubo di silicone e staccare il tubo dal catetere. Rilasciare la pressione lentamente sul morsetto per consentire il riempimento controllato del tubo capillare.</li>
		<li>Ricollegare il tubo di silicone al catetere. Inserire i tubi capillari nel sangue analizzatore di gas per misure presso i seguenti punti di tempo: 10 min dopo l'inizio di CPB con ventilazione (BGA1) dopo 40 min di CPB e 30 min di arresto respiratorio (BGA3) dopo 55 min di CPB, 45 min di respiratori e 20 min di arresto cardiaco (BGA4)</li></ol></li> <li>sotto flusso stabile di CPB, initia arresto respiratorio te arrestando ventilazione.</li>
	<li>Dopo il termine della ventilazione, impostare il riscaldamento pad a 28 º c e iniziare il raffreddamento topicamente l'animale alla temperatura corporea di 28 ° c entro i primi 20 min di arresto respiratorio utilizzando una garza imbevuta di soluzione salina fredda.</li>
	<li>Una volta ottenuta una temperatura corporea di 28 ° c e dopo un totale di 30 min di arresto respiratorio, amministrare 0,3 mL di soluzione di KCl 7,45% nel circuito del CPB per avviare cardioplegia.</li>
	<li>Per attraversare la pressione dell'aorta, tirare l'ansa di seta precedentemente posizionata (punto 3.10) in direzione craniale per una migliore esposizione e posizionare un morsetto di micro-serrefine sulla parte ascendente dell'aorta.</li>
	<li>Eseguire un totale di 60 min arresto respiratorio e 30 min di arresto cardiaco. Rimuovere il morsetto di micro-serrefine dall'aorta ascendente di avviare riperfusione del cuore. Contemporaneamente, ri-ventilare e caldo l'animale a 37 ° C.</li>
	<li>Dopo riperfusione è completato e l'animale ha raggiunto normothermia, terminare il CPB spegnendo la pompa e continuare a monitorare le misurazioni fisiologiche (ad es. temperatura corporea, ECG e pressione sanguigna invasiva) per 20 min.</li>
	<li>Alla fine dell'esperimento, prosciugare l'animale in anestesia completo (5% isoflurano) e raccogliere il sangue e gli organi per ulteriore analisi 14.</li>
</ol>

<ol>
	<li>Edmunds, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiopulmonary+Bypass+after+50+Years.">Cardiopulmonary Bypass after 50 Years.</a> N. Engl. J. Med. 351 (16), 1601-1603 (2004).</li><li>Goto, T., Maekawa, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cerebral+dysfunction+after+coronary+artery+bypass+surgery.">Cerebral dysfunction after coronary artery bypass surgery.</a> J. Anesth. 28 (2), 242-248 (2014).</li><li>Uysal, S., Reich, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurocognitive+outcomes+of+cardiac+surgery.">Neurocognitive outcomes of cardiac surgery.</a> J. Cardiothorac. Vasc. Anesth. 27 (5), 958-971 (2013).</li><li>Ballaux, P. K., Gourlay, T., Ratnatunga, C. P., Taylor, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+literature+review+of+cardiopulmonary+bypass+models+for+rats.">A literature review of cardiopulmonary bypass models for rats.</a> Perfusion. 14 (6), 411-417 (1999).</li><li>Jungwirth, B., de Lange, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+cardiopulmonary+bypass:+development,+applications,+and+impact.">Animal models of cardiopulmonary bypass: development, applications, and impact.</a> Semin. Cardiothorac. Vasc. Anesth. 14 (2), 136-140 (2010).</li><li>G&#252;nzinger, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+rat+model+of+cardiopulmonary+bypass+with+cardioplegic+arrest+and+hemodynamic+assessment+by+conductance+catheter+technique.">A rat model of cardiopulmonary bypass with cardioplegic arrest and hemodynamic assessment by conductance catheter technique.</a> Basic Res Cardiol. 102 (6), 508-517 (2007).</li><li>Waterbury, T., Clark, T. J., Niles, S., Farivar, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+model+of+cardiopulmonary+bypass+for+deep+hypothermic+circulatory+arrest.">Rat model of cardiopulmonary bypass for deep hypothermic circulatory arrest.</a> J. Thorac. Cardiovasc. Surg. 141 (6), 1549-1551 (2011).</li><li>Schnoering, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+newly+developed+miniaturized+heart-lung+machine-expression+of+inflammation+in+a+small+animal+model.">A newly developed miniaturized heart-lung machine-expression of inflammation in a small animal model.</a> Artif. Organs. 34 (11), 911-917 (2010).</li><li>Kim, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+responses+of+tissues+from+the+brain,+heart,+kidney,+and+liver+to+resuscitation+following+prolonged+cardiac+arrest+by+examining+mitochondrial+respiration+in+rats.">The responses of tissues from the brain, heart, kidney, and liver to resuscitation following prolonged cardiac arrest by examining mitochondrial respiration in rats.</a> Oxid. Med. Cell. Longev. 2016, (2016).</li><li>Shappell, S. B., Gurpinar, T., Lechago, J., Suki, W. N., Truong, L. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+obstructive+uropathy+in+severe+combined+immunodeficient+(SCID)+mice:+lymphocyte+infiltration+is+not+required+for+progressive+tubulointerstitial+injury.">Chronic obstructive uropathy in severe combined immunodeficient (SCID) mice: lymphocyte infiltration is not required for progressive tubulointerstitial injury.</a> J. Am. Soc. Nephrol. 9 (6), 1008-1017 (1998).</li><li>Majzoub, J. A., Muglia, L. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Knockout+mice.">Knockout mice.</a> N. Engl. J. Med. , 904-907 (1996).</li><li>Houser, S. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+Models+of+Heart+Failure+A+Scientific+Statement+From+the+American+Heart+Association.">Animal Models of Heart Failure A Scientific Statement From the American Heart Association.</a> Circ. Res. 111 (1), 131-150 (2012).</li><li>Russell, J. C., Proctor, S. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Small+animal+models+of+cardiovascular+disease:+tools+for+the+study+of+the+roles+of+metabolic+syndrome,+dyslipidemia,+and+atherosclerosis.">Small animal models of cardiovascular disease: tools for the study of the roles of metabolic syndrome, dyslipidemia, and atherosclerosis.</a> Cardiovasc. Pathol. 15 (6), 318-330 (2006).</li><li>Iurascu-Gagea, M., Craig, S., Suckow, M. A., Stevens, K. A., Wilson, R. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Euthanasia+and+necropsy.">Euthanasia and necropsy.</a> The laboratory rabbit, guinea pig, hamster, and other rodents. , 117-141 (2012).</li></ol>

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Abbiamo sviluppato un modello clinicamente rilevante perfettamente funzionante di CPB in un mouse. Con più di trenta ceppi di topi con malattie cardiovascolari, il nostro modello potrebbe essere un punto di partenza per lo sviluppo di nuovi protocolli prospettici correlati a CPB. Inoltre, a causa della pletora di reagenti specifici del mouse e topi knockout, questo modello non può solo sostituire l'attuale modello di ratto di CPB ma faciliterà la dissezione dei meccanismi molecolari coinvolti nel danno dell'organo CPB...

Fin dall'introduzione dell'esclusione cardiopolmonare (CPB) in clinica, esso ha svolto un ruolo essenziale in cardiochirurgia1. In cardiochirurgia moderna, prolungato tempo CPB è essenziale per eseguire vaste ricostruzioni aortiche e procedure unite. Anche se i progressi tecnologici sono stati enormi, l'uso della circolazione extracorporea è associato con intra - e postoperatorie sistemica e locale organo danni2,3.
Grandi modelli animali sono stati sviluppati per studiare il ruolo di CPB su processi fisiologici4,5. Anche se questi modelli hanno fornito alcune del CPB connesse complicazioni, sono estremamente costosi e strumenti molecolari (ad es., anticorpi) sono molto limitati. Un'alternativa più efficiente è stata sviluppata nei piccoli animali. Dal loro sviluppo, sono stati condotti diversi studi per ottimizzare un modello CPB in ratti e conigli5,6,7,8,9. Questi modelli costituiscono una buona base per le misurazioni dei processi patofisiologici malattia; Tuttavia, essi sono ancora insufficienti per indagare immunologia umorale e cellulare a causa della mancanza di anticorpi pertinenti e reagenti. Ciò altera il loro ruolo in questo campo di ricerca.
Recentemente abbiamo sviluppato un modello murino di CPB. A causa di una vasta gamma di reagenti specifici del mouse e topi modificati geneticamente, modelli murini sono in generale il modello di scelta per la ricerca fisiologica, molecolari ed immunologici10,11. Pertanto, il nostro modello faciliterà lo studio di CPB in relazione alle varie comorbidità come ci sono molti ceppi di topi disponibili con malattie clinicamente rilevante12,13. Di conseguenza, questo articolo descrive, in dettaglio, come eseguire CPB in topi. Ossigeno e i parametri emodinamici sono strettamente monitorati dopo l'arresto di profondità respiratoria e circolatoria.

<table border="0" cellpadding="0" cellspacing="0" width="1191">
	<tbody>
		<tr height="23">
			<td height="23" style="height:23px;width:297px;">Sterofundin</td>
			<td style="width:279px;">B.Braun Petzold GmbH</td>
			<td style="width:181px;">PZN:8609189</td>
			<td style="width:435px;">priming volume, 1:1 with Tetraspan</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">Tetraspan 6% HES Solution</td>
			<td style="width:279px;">B. Braun Melsungen AG</td>
			<td style="width:181px;">PZN: 05565416</td>
			<td>priming volume, 1:1 with Sterofundin</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">Heparin Natrium 25.000</td>
			<td style="width:279px;">Ratiopharm GmbH</td>
			<td style="width:181px;">PZN: 3029843</td>
			<td>2.5 IU per ml of priming solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">NaHCO3 8,4% Solution</td>
			<td style="width:279px;">B. Braun Melsungen AG</td>
			<td style="width:181px;">PZN: 1579775</td>
			<td>3% in priming solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">KCL 7,45 % Solution</td>
			<td style="width:279px;">B. Braun Melsungen AG</td>
			<td style="width:181px;">PZN: 2418577</td>
			<td>0.1 ml for cardioplegia</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">Carprofen</td>
			<td style="width:279px;">Zoetis Inc., USA</td>
			<td>PZN:00289615 08859153</td>
			<td>5 mg/kg/BW</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">1 Fr PU Catheter</td>
			<td style="width:279px;">Instechlabs INC., USA</td>
			<td style="width:181px;">C10PU-MCA1301</td>
			<td>carotid artery</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">2 Fr PU Catheter</td>
			<td style="width:279px;">Instechlabs INC., USA</td>
			<td style="width:181px;">C20PU-MJV1302</td>
			<td>jugular vein</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vasofix Safety catheter 20G</td>
			<td style="width:279px;">B.Braun Medical</td>
			<td style="width:181px;">4268113S-01</td>
			<td>orotracheal intubation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">8-0 Silk suture braided</td>
			<td>Ashaway Line &#38; Twine Mfg. Co., USA</td>
			<td style="width:181px;">75290</td>
			<td>ligature</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">Isoflurane</td>
			<td style="width:279px;">Piramal Critical Care Deutschland GmbH</td>
			<td style="width:181px;">PZN:9714675</td>
			<td>narcosis</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">CLINITUBES blood capillaries</td>
			<td style="width:279px;">Radiomed GmbH</td>
			<td style="width:181px;">51750132</td>
			<td>blood sampling 60 - 95 microliter</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Spring Scissors - 6mm Blades</td>
			<td>Fine Science Tools GmbH</td>
			<td style="width:181px;">15020-15</td>
			<td>instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Spring Scissors - 2mm Blades</td>
			<td>Fine Science Tools GmbH</td>
			<td style="width:181px;">15000-03</td>
			<td>instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Halsted-Mosquito Hemostat</td>
			<td>Fine Science Tools GmbH</td>
			<td style="width:181px;">13009-12</td>
			<td>instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dumont #55 Forceps</td>
			<td>Fine Science Tools GmbH</td>
			<td style="width:181px;">11295-51</td>
			<td>instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Castroviejo Micro Needle Holder - 9cm</td>
			<td>Fine Science Tools GmbH</td>
			<td style="width:181px;">12060-02</td>
			<td>instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micro Serrefines</td>
			<td>Fine Science Tools GmbH</td>
			<td>18555-01</td>
			<td>instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Bulldog Serrefine</td>
			<td>Fine Science Tools GmbH</td>
			<td style="width:181px;">18050-28</td>
			<td>instruments</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:297px;">MiniVent Ventilator for Mice (Model 845)</td>
			<td style="width:279px;">Harvard Apparatus</td>
			<td style="width:181px;">73-0044</td>
			<td>mechanical ventilation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">Isoflurane Vaporizer Drager 19.1</td>
			<td style="width:279px;">Dr&#228;gerwerk AG &#38; Co. KGaA</td>
			<td style="width:181px;"></td>
			<td>anesthesia 1.3 -2.5%</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PowerLab data acquisition device 4/35</td>
			<td>ADInstruments Ltd, New Zealand</td>
			<td style="width:181px;">PL3504</td>
			<td>invasive pressure, ECG, temperature</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ABL 800 Flex</td>
			<td>Radiometer GmbH</td>
			<td style="width:181px;"></td>
			<td>blood gas analysis</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:297px;">NMRI mice</td>
			<td style="width:279px;">Charles River Laboratories</td>
			<td style="width:181px;">Crl:NMRI(Han)</td>
			<td>male, 30-35 g, 12 weeks old, housed at least 1 week before the experiment</td>
		</tr>
	</tbody>
</table>

cardiopulmonary bypass in a mouse model: a novel approach

Come bypass cardiopolmonare prolungata diventa più essenziale durante gli interventi cardiaci, una crescente domanda clinica si pone per l'ottimizzazione della procedura e per minimizzare il danno d'organo derivanti da circolazione extracorporea prolungata. L'obiettivo di questa carta era di dimostrare un modello completamente funzionale e clinicamente rilevante dell'esclusione cardiopolmonare in un mouse. Segnaliamo il dispositivo progettazione, ottimizzazione del circuito di perfusione e le tecniche microsurgical. Questo modello è un modello acuto, che non è compatibile con la sopravvivenza a causa della necessità per più disegni di sangue. A causa della gamma di strumenti disponibili per i topi (ad es., marcatori, forature, ecc.), questo modello faciliterà la ricerca sui meccanismi molecolari del danno d'organo e l'effetto dell'esclusione cardiopolmonare in relazione agli altri comorbilità.

Questo protocollo descrive il circuito di perfusione, interventi chirurgici e il monitoraggio dei parametri fisiologici durante CPB di un mouse. Quando eseguita da un Microchirurgo adeguatamente qualificato, i risultati si ottengono costantemente e riproducibile.
Per mantenere la perfusione tissutale adeguata, la pressione arteriosa media è sempre tra 40 e 60 mmHg regolando il flusso di sangue CPB e aggiunta di volume supplemen...

Watch this Scientific Journal Video about Cardiopulmonary Bypass in a Mouse Model: A Novel Approach at JoVE.com

Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

Questo articolo descrive come eseguire il bypass cardiopolmonare in topi. Questo modello di romanzo faciliterà l'indagine dei meccanismi molecolari coinvolti nel danno dell'organo.

Esclusione cardiopolmonare in un modello murino: un nuovo approccio

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization ...

cardiopulmonary-bypass-in-a-mouse-model-a-novel-approach

Research

JoVE Journal

Medicine

10.8K Views.  Hannover Medical School. Questo articolo descrive come eseguire il bypass cardiopolmonare in topi. Questo modello di romanzo faciliterà l'indagine dei meccanismi molecolari coinvolti nel danno dell'organo.

Video: Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

Vascular Occlusion Training for Inclusion Body Myositis: A Novel Therapeutic Approach

Inclusion body myositis (IBM) is a rare idiopathic inflammatory myopathy. It is known to produces remarkable muscle weakness and to greatly compromise function and quality of life. Moreover, clinical practice suggests that, unlike other inflammatory myopathies, the majority of IBM patients are not responsive to treatment with immunosuppressive or immunomodulatory drugs to counteract disease progression1. Additionally, conventional resistance training programs have been proven ineffective in restoring muscle function and muscle mass in these patients2,3. Nevertheless, we have recently observed that restricting muscle blood flow using tourniquet cuffs in association with moderate intensity resistance training in an IBM patient produced a significant gain in muscle mass and function, along with substantial benefits in quality of life4. Thus, a new non-pharmacological approach for IBM patients has been proposed. Herein, we describe the details of a proposed protocol for vascular occlusion associated with a resistance training program for this population.

The present article presents the details pertaining to the application of resistance training associated to vascular occlusion in IBM patients.

Formazione occlusione vascolare per Miosite inclusione del corpo: un nuovo approccio terapeutico

Inclusion body myositis (IBM) is a rare idiopathic inflammatory myopathy. It is known to produces remarkable muscle weakness and to greatly compromise ...

Ricerca

Medicina

Bronchial Thermoplasty:  A Novel Therapeutic Approach to Severe Asthma

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled manner to reduce excessive airway smooth muscle. Reducing airway smooth muscle decreases the ability of the airways to constrict, thereby reducing the frequency of asthma attacks. Bronchial thermoplasty is delivered by the Alair System and is performed in three outpatient procedure visits, each scheduled approximately three weeks apart. The first procedure treats the airways of the right lower lobe, the second treats the airways of the left lower lobe and the third and final procedure treats the airways in both upper lobes. After all three procedures are performed the bronchial thermoplasty treatment is complete.
Bronchial thermoplasty is performed during bronchoscopy with the patient under moderate sedation. All accessible airways distal to the mainstem bronchi between 3 and 10 mm in diameter, with the exception of the right middle lobe, are treated under bronchoscopic visualization. Contiguous and non-overlapping activations of the device are used, moving from distal to proximal along the length of the airway, and systematically from airway to airway as described previously. Although conceptually straightforward, the actual execution of bronchial thermoplasty is quite intricate and procedural duration for the treatment of a single lobe is often substantially longer than encountered during routine bronchoscopy. As such, bronchial thermoplasty should be considered a complex interventional bronchoscopy and is intended for the experienced bronchoscopist. Optimal patient management is critical in any such complex and longer duration bronchoscopic procedure. This article discusses the importance of careful patient selection, patient preparation, patient management, procedure duration, postoperative care and follow-up to ensure that bronchial thermoplasty is performed safely.
Bronchial thermoplasty is expected to complement asthma maintenance medications by providing long-lasting asthma control and improving asthma-related quality of life of patients with severe asthma. In addition, bronchial thermoplasty has been demonstrated to reduce severe exacerbations (asthma attacks) emergency rooms visits for respiratory symptoms, and time lost from work, school and other daily activities due to asthma.

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled manner to reduce excessive airway smooth muscle. Reducing airway smooth muscle decreases the ability of the airways to constrict, thereby reducing the frequency of asthma attacks.

Termoplastica bronchiale: un nuovo approccio terapeutico per l&#39;asma grave

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled ...

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient management strategies. Orthotopic mouse models of oral cancer have been developed to facilitate the study of factors that impact invasion and serve as model system for evaluating anti-tumor therapeutics. In these systems, visualization of disseminated tumor cells within oral cavity tissues has typically been conducted by either conventional histology or with in vivo bioluminescent methods. A primary drawback of these techniques is the inherent inability to accurately visualize and quantify early tumor cell invasion arising from the primary site in three dimensions. Here we describe a protocol that combines an established model for squamous cell carcinoma of the tongue (SCOT) with two-photon imaging to allow multi-vectorial visualization of lingual tumor spread. The OSC-19 head and neck tumor cell line was stably engineered to express the F-actin binding peptide LifeAct fused to the mCherry fluorescent protein (LifeAct-mCherry). Fox1nu/nu mice injected with these cells reliably form tumors that allow the tongue to be visualized by ex-vivo application of two-photon microscopy. This technique allows for the orthotopic visualization of the tumor mass and locally invading cells in excised tongues without disruption of the regional tumor microenvironment. In addition, this system allows for the quantification of tumor cell invasion by calculating distances that invaded cells move from the primary tumor site. Overall this procedure provides an enhanced model system for analyzing factors that contribute to SCOT invasion and therapeutic treatments tailored to prevent local invasion and distant metastatic spread. This method also has the potential to be ultimately combined with other imaging modalities in an in vivo setting.

A comprehensive overview of the techniques involved in generating a mouse model of oral cancer and quantitative monitoring of tumor invasion within the tongue through multi-photon microscopy of labeled cells is presented. This system can serve as a useful platform for the molecular assessment and drug efficacy of anti-invasive compounds.

Multi-fotone Imaging di invasione delle cellule tumorali in un modello di mouse ortotopico di carcinoma orale a cellule squamose

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient ...

Normothermic Cardiac Arrest and Cardiopulmonary Resuscitation: A Mouse Model of Ischemia-Reperfusion Injury

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused by whole-body hypoperfusion.5,6 Successful reproduction of whole-body hypoperfusion in rodent models has been fraught with 
difficulty.7-9,9,10 Models which employ focal ischemia have repeatedly demonstrated results which do not translate to the clinical setting, and larger 
animal models which allow for whole body hypoperfusion lack access to the full toolset of genetic manipulation possible in the mouse.11,12 However, in recent 
years a mouse model of cardiac arrest and cardiopulmonary resuscitation has emerged which can be adapted to model AKI.13 This model reliably reproduces 
physiologic, functional, anatomic, and histologic outcomes seen in clinical AKI, is rapidly repeatable, and offers all of the significant advantages of a murine surgical 
model, including access to genetic manipulative techniques, low cost relative to large animals, and ease of use. Our group has developed extensive experience 
with use of this model to assess a number of organ-specific outcomes in AKI.14,15

A powerful model for perioperative and critical care related acute kidney injury is presented. Using whole body hypoperfusion induced by cardiac arrest it is possible to nearly replicate the histologic and functional changes of clinical AKI.

Normotermico arresto cardiaco e rianimazione cardiopolmonare: un modello murino di ischemia-riperfusione

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused ...

Roux-en-Y Gastric Bypass Operation in Rats

Currently, the most effective therapy for the treatment of morbid obesity to induce significant and maintained body weight loss with a proven mortality benefit is bariatric surgery1,2. Consequently, there has been a steady rise in the number of bariatric operations done worldwide in recent years with the Roux-en-Y gastric bypass (gastric bypass) being the most commonly performed operation3. Against this background, it is important to understand the physiological mechanisms by which gastric bypass induces and maintains body weight loss. These mechanisms are yet not fully understood, but may include reduced hunger and increased satiation4,5, increased energy expenditure6,7, altered preference for food high in fat and sugar8,9, altered salt and water handling of the kidney10 as well as alterations in gut microbiota11. Such changes seen after gastric bypass may at least partly stem from how the surgery alters the hormonal milieu because gastric bypass increases the postprandial release of peptide-YY (PYY) and glucagon-like-peptide-1 (GLP-1), hormones that are released by the gut in the presence of nutrients and that reduce eating12.
During the last two decades numerous studies using rats have been carried out to further investigate physiological changes after gastric bypass. The gastric bypass rat model has proven to be a valuable experimental tool not least as it closely mimics the time profile and magnitude of human weight loss, but also allows researchers to control and manipulate critical anatomic and physiologic factors including the use of appropriate controls. Consequently, there is a wide array of rat gastric bypass models available in the literature reviewed elsewhere in more detail 13-15. The description of the exact surgical technique of these models varies widely and differs e.g. in terms of pouch size, limb lengths, and the preservation of the vagal nerve. If reported, mortality rates seem to range from 0 to 35%15. Furthermore, surgery has been carried out almost exclusively in male rats of different strains and ages. Pre- and postoperative diets also varied significantly.
Technical and experimental variations in published gastric bypass rat models complicate the comparison and identification of potential physiological mechanisms involved in gastric bypass. There is no clear evidence that any of these models is superior, but there is an emerging need for standardization of the procedure to achieve consistent and comparable data. This article therefore aims to summarize and discuss technical and experimental details of our previously validated and published gastric bypass rat model.

Numerous studies using gastric bypass rat models have been recently conducted to uncover the underlying physiological mechanisms of Roux-en-Y gastric bypass operations. This article aims to demonstrate and discuss the technical and experimental details of our published gastric bypass rat model to understand advantages and limitations of this experimental tool.

Roux-en-Y operazione di bypass gastrico in Rats

Currently, the most effective therapy for the treatment of morbid obesity to induce significant and maintained body weight loss with a proven mortality ...

A Novel Surgical Approach for Intratracheal Administration of Bioactive Agents in a Fetal Mouse Model

Prenatal pulmonary delivery of cells, genes or pharmacologic agents could provide the basis for new therapeutic strategies for a variety of genetic and acquired diseases. Apart from congenital or inherited abnormalities with the requirement for long-term expression of the delivered gene, several non-inherited perinatal conditions, where short-term gene expression or pharmacological intervention is sufficient to achieve therapeutic effects, are considered as potential future indications for this kind of approach. Candidate diseases for the application of short-term prenatal therapy could be the transient neonatal deficiency of surfactant protein B causing neonatal respiratory distress syndrome1,2 or hyperoxic injuries of the neonatal lung3. Candidate diseases for permanent therapeutic correction are Cystic Fibrosis (CF)4, genetic variants of surfactant deficiencies5 and &alpha;1-antitrypsin deficiency6.
Generally, an important advantage of prenatal gene therapy is the ability to start therapeutic intervention early in development, at or even prior to clinical manifestations in the patient, thus preventing irreparable damage to the individual. In addition, fetal organs have an increased cell proliferation rate as compared to adult organs, which could allow a more efficient gene or stem cell transfer into the fetus. Furthermore, in utero gene delivery is performed when the individual's immune system is not completely mature. Therefore, transplantation of heterologous cells or supplementation of a non-functional or absent protein with a correct version should not cause immune sensitization to the cell, vector or transgene product, which has recently been proven to be the case with both cellular and genetic therapies7.
In the present study, we investigated the potential to directly target the fetal trachea in a mouse model. This procedure is in use in larger animal models such as rabbits and sheep8, and even in a clinical setting9, but has to date not been performed before in a mouse model. When studying the potential of fetal gene therapy for genetic diseases such as CF, the mouse model is very useful as a first proof-of-concept because of the wide availability of different transgenic mouse strains, the well documented embryogenesis and fetal development, less stringent ethical regulations, short gestation and the large litter size.
Different access routes have been described to target the fetal rodent lung, including intra-amniotic injection10-12, (ultrasound-guided) intrapulmonary injection13,14 and intravenous administration into the yolk sac vessels15,16 or umbilical vein17. Our novel surgical procedure enables researchers to inject the agent of choice directly into the fetal mouse trachea which allows for a more efficient delivery to the airways than existing techniques18.

We developed a novel surgical approach for intratracheal administration of bioactive agents into the mouse fetus. The delivery route is more efficient in targeting the fetal mouse lungs than the commonly used intra-amniotic injection. This procedure has to date not been described in a mouse model.

Un nuovo approccio chirurgico per somministrazione endotracheale di agenti bioattivi in ​​un modello murino fetale

Prenatal pulmonary delivery of cells, genes or pharmacologic agents could provide the basis for new therapeutic strategies for a variety of genetic and ...

Evaluation of a Novel Laser-assisted Coronary Anastomotic Connector - the Trinity Clip - in a Porcine Off-pump Bypass Model

To simplify and facilitate beating heart (i.e., off-pump), minimally invasive coronary artery bypass surgery, a new coronary anastomotic connector, the Trinity Clip, is developed based on the excimer laser-assisted nonocclusive anastomosis technique. The Trinity Clip connector enables simplified, sutureless, and nonocclusive connection of the graft to the coronary artery, and an excimer laser catheter laser-punches the opening of the anastomosis. Consequently, owing to the complete nonocclusive anastomosis construction, coronary conditioning (i.e., occluding or shunting) is not necessary, in contrast to the conventional anastomotic technique, hence simplifying the off-pump bypass procedure. Prior to clinical application in coronary artery bypass grafting, the safety and quality of this novel connector will be evaluated in a long-term experimental porcine off-pump coronary artery bypass (OPCAB) study. In this paper, we describe how to evaluate the coronary anastomosis in the porcine OPCAB model using various techniques to assess its quality. Representative results are summarized and visually demonstrated.

This paper describes a novel nonocclusive coronary anastomotic connector in a porcine off-pump coronary artery bypass (OPCAB) model. This easy-to-use coronary connector has intrinsic potential to facilitate minimally invasive OPCAB surgery.

Valutazione di un romanzo Laser-assistita coronarica anastomosi Connector - Clip Trinity - in un suino Off-pompa bypass Modello

To simplify and facilitate beating heart (i.e., off-pump), minimally invasive coronary artery bypass surgery, a new coronary anastomotic connector, the ...

A Novel Approach for the Administration of Medications and Fluids in Emergency Scenarios and Settings

The available routes of administration commonly used for medications and fluids in the acute care setting are generally limited to oral, intravenous, or intraosseous routes, but in many patients, particularly in the emergency or critical care settings, these routes are often unavailable or time-consuming to access. A novel device is now available that offers an easy route for administration of medications or fluids via rectal mucosal absorption (also referred to as proctoclysis in the case of fluid administration and subsequent absorption). Although originally intended for the palliative care market, the utility of this device in the emergency setting has recently been described. Specifically, reports of patients being treated for dehydration, alcohol withdrawal, vomiting, fever, myocardial infarction, hyperthyroidism, and cardiac arrest have shown success with administration of a wide variety of medications or fluids (including water, aspirin, lorazepam, ondansetron, acetaminophen, methimazole, and buspirone). Device placement is straightforward, and based on the observation of expected effects from the medication administrations, absorption is rapid. The rapidity of absorption kinetics are further demonstrated in a recent report of the measurement of phenobarbital pharmacokinetics. We describe here the placement and use of this device, and demonstrate methods of pharmacokinetic measurements of medications administered by this method.

This study presents a novel device that offers an easy route to quickly provide medications and fluids in patients with limited or difficult IV access. This small-diameter device is placed in the distal third of the rectum, allowing for ongoing medication and fluids administration.

Un nuovo approccio per la somministrazione di farmaci e fluidi in scenari di emergenza e impostazioni

The available routes of administration commonly used for medications and fluids in the acute care setting are generally limited to oral, intravenous, or ...

Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. Research models for bypass graft failure are essential for a better understanding of pathobiological and pathophysiological processes during graft patency loss. Large animal models, such as pigs or sheep, resemble human anatomical structures but require special facilities and equipment. This video describes a rat vein interposition model to investigate vein graft patency loss. Rats are inexpensive and easy to handle. Compared to mouse models, the convenient size of rats permits better operability and enables a sufficient amount of material to be obtained for further diverse analysis. In brief, the inferior epigastric vein of a donor rat is harvested and used to replace a segment of the femoral artery. Anastomosis is conducted via single stitches and sealed with fibrin glue. Graft patency can be monitored non-invasively using duplex sonography. Myointimal hyperplasia, which is the main cause for graft patency loss, develops progressively over time and can be calculated from histological cross sections.

This video demonstrates a model to study the development of myointimal hyperplasia after venous interposition surgery in rats.

Vena interposizione Modello: un modello adatto per studiare Bypass Graft pervietà

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. ...

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 ...