This work was supported by the National Institutes of Health (HL114669). CM is a recipient of a postdoctoral research fellowship grant awarded by the Myocarditis Foundation, NJ.

DICHIARAZIONE ETICA: Tutte le procedure di polizia sono state condotte in conformità con le linee guida per la cura e l&#39;uso di animali da laboratorio e approvati dalla University of Nebraska-Lincoln, Lincoln, NE. 1. Induzione di autoimmune sperimentale miocardite <ol><li> Preparare la soluzione sciogliendo peptide MyHC-α 334-352 in 1x PBS ad una concentrazione finale di 2 ml mg/1.5. </li><li> Preparare la tossina della pertosse (PT) aggiungendo 1 ml di PBS sterile 1x in una fiala contenente 50 mg di PT liofilizzate per ottenere una concentrazione magazzino di 50 ng / ml. Prendere 20 ml di brodo in una provetta sterile da 1,5 ml e aggiungere 980 ml di sterile 1x PBS per ottenere una concentrazione di lavoro di 1 ng / ml. </li><li> Preparare adiuvante completo di Freund (CFA) con l&#39;aggiunta di 40 mg di Mycobacterium tuberculosis H37Rv (MTB) estratto a 10 ml di CFA per ottenere una concentrazione finale5 mg / ml. </li><li> Preparare l&#39;emulsione peptide-CFA. NOTA: Per l&#39;induzione EAM, l&#39;emulsione peptide-CFA è stato somministrato a 150 ml contenenti 100 mg di MyHC-α 334-352 per animale. Ad esempio, per immunizzare topi dieci usano 1,5 ml di emulsione peptide-CFA contenente 1 mg di MyHC-α 334-352. <ol><li> Per preparare 1,5 ml di emulsione, un&#39;aliquota 750 microlitri di MyHC-α 334-352 soluzione di peptide in una provetta da 1,5 ml, e CFA integrati con MTB in un&#39;altra provetta da 1,5 ml. Usando una siringa luer-lok 3 ml, aspirare la soluzione peptide prima, seguita dalla estratto CFA / MTB. </li><li> Collegare la siringa al rubinetto a 3 vie e collegare l&#39;altra uscita del rubinetto ad un vuoto siringa 3 ml. Regolare la pervietà del rubinetto in modo che la miscela peptidica-CFA scorre da una siringa all&#39;altra con ragionevolmente buona resistenza. </li><li> Mescolare spingendo il contenuto da una siringa all&#39;altra ripetutamente per ~ 1 min e poi posizionare l&#39;intero gruppo sul ghiaccio per ~ 3 min. Ripetere questaprocedura un minimo di 10x. </li><li> Determinare la consistenza dell&#39;emulsione ponendo attenzione una piccola goccia su ancora acqua in un becher da 100 ml in vetro. La gocciolina non dovrebbe disperdere in acqua. Se è così, continuare a mescolare fino ad ottenere la consistenza desiderata. </li><li> Trasferire il contenuto di emulsione da 3 ml siringhe in 1 ml siringhe Luer-Lok sostituendo una delle due siringhe da 3 ml collegati al rubinetto con la siringa da 1 ml, e inserire un ago 27 G ½ alla siringa 1 ml. </li></ol></li><li> Iniettare 150 ml di emulsione peptide-CFA in dosi per via sottocutanea divisi nelle due regioni inguinali di sei-otto settimane di età femminile A / J topi (~ 75 ml ciascuna). </li><li> Somministrare 100 ml di sospensione PT (100 ng) per via intraperitoneale a ciascun animale al giorno 0 e il giorno 2 post-immunizzazione. </li><li> Ripetere la procedura immunizzazione al giorno 7 somministrando 150 ml di emulsione peptide-CFA in dosi parziali sottocutanea in entrambi i lati di tegli sterno (~ 75 ml ciascuna). Preparare questa emulsione fresca come sopra. Poi, il giorno 21, sottoporre gli animali a immagini MRM, vedere il punto 3. </li></ol> 2. Manipolazione di animali <ol><li> Posizionare ciascun topo in una camera di induzione anestetica contenente 2% isoflurano miscela di aria con una piastra elettrica posta sotto per mantenere il calore, e trasferire l&#39;animale a un titolare animale appositamente progettato (Figura 1). </li><li> Immobilizzare l&#39;animale in posizione prona sul supporto animale così il muso si inserisce il cono per mantenere l&#39;anestesia (Figura 1). Fissare la testa del topo con un morso-bar attaccato ai denti anteriori del mouse. </li><li> Accendere il riscaldamento soffio d&#39;aria con il suo tubo di scarico inserita nel foro verticale dello scanner per mantenere la temperatura corporea degli animali durante l&#39;esperimento. </li><li> Mantenere l&#39;anestesia a 0,5 al 2% isoflurano con un flusso di 2 ml / min durante l&#39;intera sessione di imaging. Confermareanestesia con il metodo pinch punta, previsto nessun movimento. </li><li> Impostare un sensore pneumatico cuscino, coda di topo / caviglia cardiofrequenzimetro a fibra ottica ossimetria, e la sonda di temperatura rettale per controllare la respirazione, il battito cardiaco e la temperatura corporea, rispettivamente (Figura 1). Nota: gating cardiaco viene eseguita mediante ossimetria Pulse, che permette il monitoraggio non invasivo della saturazione arteriosa di ossigeno nel sangue. Il sensore pulsossimetria deve essere attaccato alla caviglia sinistra, e il piede deve essere fissato con un anello di filo e nastro per mantenere la caviglia nel sensore. Imaging MRM si ottiene gating la respirazione e segnali cardiaci senza l&#39;uso di mezzi di contrasto. </li></ol> 3. Acquisizione Immagine <ol><li> Dopo aver preparato la (Figura 1) animale, posizionare il mouse al centro dello scanner MRM con cuore posizionato al centro del campo di vista (FOV), dove il campo magnetico è massima omogeneità. NOTA: A wide-bore(89 mm) 9.4 T magnete verticale-foro dotato di gradienti triplice asse di 100 g / cm e 4 cm radiofrequenza (RF) bobina di imaging deve essere utilizzato per ottenere ad alta risoluzione tridimensionale (3D) immagini. Nota: Assicurati di forniture non-magnetici quando si utilizza uno scanner MRI. </li><li> Eseguire l&#39;interfaccia di imaging e selezionare &quot;Nuovo studio&quot; dal menu &quot;Opzioni di studio&quot;. Digitare &quot;mtune&quot; al bar dei comandi ed eseguirlo per tirare su il &quot;Tune GUI&quot;. Quindi selezionare &quot;Start Probe Tune&quot; e fare clic sul pulsante &quot;Autoscale&quot;. Il Tune GUI cambierà per mostrare il RFsignal. Utilizzare la melodia / coi remoti manopole alla fine della bobina per sintonizzare la bobina RF alla frequenza di risonanza protonica (400 MHz). Sulla scheda &quot;Start&quot; vai alla pagina &quot;Scansione preliminare&quot; per eseguire la calibrazione della frequenza e della potenza cliccando sui relativi pulsanti. Premi il pulsante XYZ (rapida) sul &#39;&quot;scheda di&quot; Shim studio&#39; linguetta per tirare su la pagina di spessoramento.Vai alla pagina di spessore, selezionare tutte le iterazioni, e premere il pulsante shim per eseguire l&#39;auto spessoramento. </li><li> Selezionare una sequenza esploratore dalla scheda &quot;Studio&quot; dell&#39;interfaccia di imaging per localizzare il cuore del mouse. Sulla scheda &quot;Acquire&quot; cambiare il FOV a 35 mm 2 e mantenere le impostazioni predefinite della macchina. Fare clic su Start per eseguire la sequenza; regolare la posizione del supporto animale se il cuore non è al centro del FOV, sintonizzare la bobina RF e avere nuovamente le immagini scout. </li><li> Fare clic sul pulsante &quot;GEMS&quot; sequenza sulla scheda &quot;Studio&quot;, quindi inserire i parametri di acquisizione sulla scheda &quot;Acquire&quot; corrispondente ottenere due piani ortogonali lungo l&#39;asse corto e lungo l&#39;asse del cuore 20,21. NOTA: I parametri tipici di acquisizione per la scansione gradient-echo sono: spessore di strato, 1 mm; tempo di ripetizione (TR), 200 msec; tempo di eco (TE), 2.67 msec (minimo); flip angle, 25 °; dimensione della matrice inplane, 128 x 128; FOV, 22 millimetri2; numero di acquisizioni, 4; e un tempo approssimativo di acquisizione di 1 min e 30 sec. </li><li> Sulla scheda &quot;Adv&quot;, selezionare la sequenza &quot;CINE&quot; per raccogliere i asse corto cine immagini RM ossimetria-gated impulsi al fine di misurare i parametri anatomici e funzionali LV. Regolare la posizione e l&#39;angolo delle fette di imaging basata sulla vista asse lungo del cuore con il mouse hovering e trascinamento. Inserire i seguenti parametri di imaging nella scheda &quot;Acquisisci&quot; per ottenere il gradiente echo sequenza cine: TR, 500 msec; TE, 5 msec; FOV 22 mm 2; matrice di acquisizione, 256 x 256; spessore di strato, 1 mm; numero di acquisizioni, 8; numero di fotogrammi, 6; e un tempo di acquisizione di ~ 17 min. Fare clic su Start per avviare l&#39;acquisizione. </li><li> Convertire le immagini acquisite in formato DICOM utilizzando la scheda &quot;I / O&quot; del software di imaging e trasferire i file corrispondenti al centro dati per l&#39;elaborazione. </li><li> Al termine della procedura di imaging, tuttiow i topi di recuperare dall&#39;anestesia all&#39;interno del filtro-top gabbie. Non lasciare incustoditi i topi fino a che riprendano conoscenza sufficiente a mantenere decubito sternale e conservarle per ulteriori studi, se necessario. </li></ol> 4. Analisi dei dati del Cardiac Magnetic Resonance Imaging Microscopy <ol><li> Utilizzare il software di settore per analizzare i parametri anatomici e funzionali del LV 19. Con il caricamento delle immagini cine formato DICOM nel software utilizzando il &quot;Image File Open (s)&quot; sottomenu del menu &quot;File&quot;. Sulla GUI selezionare &#39;MRGE&#39; come la tecnica di imaging, &#39;Cine&#39; come il tipo di immagine e &#39;asse corto medio-ventricolare&#39; come piano di vista dell&#39;immagine. </li><li> Specificare le cornici di tempo da utilizzare per end-diastole e sistole-end attraverso il &quot;Set temporale corrente per porre fine diastole&quot; e &quot;Set temporale corrente per porre fine sistole&quot;, rispettivamente sottomenu del menu &quot;Modifica&quot;. </li><li>In primo luogo fate clic sul pulsante di comando &quot;LV&quot; e poi il &quot;EPI&quot; pulsanti di comando sul pannello in basso a destra &quot;ENDO&quot; e per disegnare manualmente il endocardio ventricolo sinistro ed epicardio, rispettivamente. Rimuovere i muscoli papillari premendo il pulsante di comando corrispondente, allo scopo di aumentare la precisione dei calcoli. </li><li> Leggere i parametri quantificati LV come volume diastolico, il volume sistolica, gittata sistolica e frazione di eiezione sul pannello in alto a destra. Fare clic sul pulsante di comando &quot;Varie&quot; e poi il pulsante di comando &quot;Misura pinza&quot; per misurare i parametri LV quali lo spessore della parete, e il diametro ventricolare 22. </li><li> Fare clic sul sottomenu &quot;Salva Sia Immagine Pile e segmentazione&quot; del menu &quot;File&quot; per salvare le immagini, comprese le segmentazioni, in formato &#39;. Mat&#39; per rielaborare le immagini, se necessario. </li></ol>

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No conflicts of interests were declared.

Questo studio descrive la procedura MRM e la sua utilità come strumento non invasivo per verificare anomalie cardiache in topi affetti da miocardite autoimmune. Poiché le caratteristiche istologiche di EAM assomigliano miocardite postinfettiva degli esseri umani, modelli murini sono comunemente impiegati per delineare i meccanismi immunitari di lesioni miocardiche 23-25. Tuttavia, gli animali affetti da miocardite appaiono clinicamente normali, e la diagnosi è fatta sulla base di istologia al termine degli...

L&#39;insufficienza cardiaca è la causa principale delle morti, e miocardite è una causa predominante di insufficienza cardiaca nei giovani adolescenti 1. La maggior parte dei pazienti affetti da miocardite rimangono asintomatici e la malattia viene spontaneamente risolti 2. Tuttavia, il 10-20% dei soggetti colpiti in grado di sviluppare malattie croniche, portando a cardiomiopatia dilatativa (DCM) 3. Vari modelli animali sono stati sviluppati per studiare la patogenesi immunitaria di miocardite. La malattia può essere indotta in miocardite-suscettibile A / J e topi Balb / c immunizzando gli animali con miosina cardiaca catena pesante (MyHC)-α o dei suoi frammenti peptidici immunodominanti o infettando con agenti patogeni come virus Coxsackie B3 4-9. Questo studio coinvolge MyHC-α miocardite 334-352 indotta in A / J topi. Nonostante mostrando infiltrazioni del miocardio, gli animali miocardite colpite appaiono clinicamente normali; diagnosi si basa sulla valutazione istologica di cuori per l&#39;infiammazione 7 bisnd ecocardiografia 10. Microscopia a risonanza magnetica (MRM) è un metodo comunemente utilizzato per ottenere imaging cardiovascolare ad alta risoluzione piani tridimensionali, che consenta di valutare dettagli funzionali al livello dei vasi sanguigni minute (fino a 10 micron di diametro), ma questo livello di potere risolvente è non realizzabili con la routine risonanza magnetica (MRI) procedura di scansione, in cui, la risoluzione viene generalmente ottenuta 1 mm 11-14. MRM offre un vantaggio in quanto permette l&#39;acquisizione di immagini ad alta risoluzione e anche per ricavare i parametri di prestazione nei momenti iniziali della malattia processo 14. Clinicamente, l&#39;imaging MRM è stato ampiamente applicato per studiare i parametri funzionali del malato cuore, polmoni o cervello 15-17. In questo studio, è mostrato l&#39;uso di una tecnica MRM come strumento non invasivo per verificare anomalie cardiache in A / J topi affetti da miocardite autoimmune. Specificamente, tegli Imaging MRM permette la quantificazione dei parametri funzionali quali ventricolare sinistra (LV) volume telediastolico e frazione di eiezione (EF) con ragionevole precisione 18. Le definizioni dei rispettivi parametri sono: fine LV volume telediastolico, volume di sangue nel ventricolo sinistro a fine ciclo diastolica e frazione di eiezione, stroke volume / volume telediastolico. L&#39;analisi dei dati viene effettuata utilizzando il software liberamente disponibile segmento sviluppato per l&#39;elaborazione immagini cardiovascolari DICOM acquisiti da scanner a risonanza magnetica 19. I dati hanno rivelato un aumento dello spessore della parete LV negli animali myocarditic, corrispondente ad una diminuzione del volume telediastolico LV, gittata sistolica e frazione di eiezione, rispetto a questi parametri funzionali in topi sani.

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		<tr height="25">
			<td height="25" style="height:25px;width:509px;">Myhc-a 334-352 (DSAFDVLSFTAEEKAGVYK)</td>
			<td style="width:364px;">Neopeptide, Cambridge, MA</td>
			<td style="width:145px;"></td>
			<td style="width:167px;">Store at 4&#959;C</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">CFA</td>
			<td style="width:364px;">Sigma Aldrich, St Louis, MO</td>
			<td style="width:145px;">5881</td>
			<td>Store at 4&#959;C</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">MTB&#160; H37Rv extract&#160;</td>
			<td>Difco Laboratories, Detroit, MI</td>
			<td>231141</td>
			<td>Store at 4&#959;C</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">PT</td>
			<td>List Biologicals Laboratories, Campbell, CA</td>
			<td>181</td>
			<td>Store at 4&#959;C</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">1x PBS&#160;</td>
			<td>Corning, Manassas, VA</td>
			<td>21-040-CV</td>
			<td>Store at 4&#959;C</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isoflurane&#160;</td>
			<td>Piramal Healthcare, Mumbai, India</td>
			<td>NDC66794-013-25</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Female A/J mice&#160;</td>
			<td>Jackson Laboratories, Bar Harbor, ME</td>
			<td>646</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Leur-lok sterile 1 ml syrringe</td>
			<td>BD, Franklin Lakes, NJ</td>
			<td>309628</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Leur-lok sterile 3 ml syrringe</td>
			<td>BD, Franklin Lakes, NJ</td>
			<td>309657</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sterile needle, 18 G</td>
			<td>BD, Franklin Lakes, NJ</td>
			<td>305195</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sterile needle, 27 1/2 G</td>
			<td>BD, Franklin Lakes, NJ</td>
			<td>305109</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">3-way stopcock&#160;</td>
			<td>Smiths Medical ASD, Inc. Dublin, OH</td>
			<td>MX5311L</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Kerlix gauze bandage rolls&#160;</td>
			<td>Covidien, Mansfield, MA</td>
			<td>6720</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Kimwipes</td>
			<td>Kimberly-Clark Professional, Roswell, GA</td>
			<td>34155</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Protouch Stockinette&#160;</td>
			<td>Medline Industries, Mundelein, IL</td>
			<td>30-1001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sterile surgical scissors and forceps</td>
			<td style="width:364px;">INOX tool Corporation</td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micro oven</td>
			<td style="width:364px;">GE Healthcare,&#160;</td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ThermiPAQ hot and cold therapy system&#160;</td>
			<td>Theramics Corporation, Springfield, IL</td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Reptile heating lamp&#160;</td>
			<td>Energy Savers Unlimited, Inc. Carson, CA</td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">3M Transpore tapes&#160;</td>
			<td>Target Corporation, MN</td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Up and Up Polymyxin B sulfate/Bacitracin/Neomycin sulfate antibiotic ointment</td>
			<td>Target Corporation, MN</td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">North Safety DeciDamp-2PVC foam ear plugs</td>
			<td style="width:364px;">North Safety Products, Smithfield, RI</td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cotton tipped applicator, 6&#8217;&#8217; wooden stem&#160;</td>
			<td>Jorgensen Laboratories, Inc. Loveland, CO</td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Anesthesia induction chamber&#160;</td>
			<td>Summit Anesthesia Solutions, Ann Harbor, MI</td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Summit Anesthesia Support system for regulating flow of anesthesia&#160;</td>
			<td>Summit Anesthesia Solutions, Ann Harbor, MI</td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Specially designed animal holder</td>
			<td>Agilent Technologies, Santa Clara, CA</td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Bickford&#160;Omnicon F/Air&#160;anesthesia gas filter unit&#160;</td>
			<td>A.M. Bickford, Inc. Wales Center, NY</td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pulse-oximeter module, MR compatible small animal monitoring and gating system&#160;</td>
			<td>Small Animal Instruments, Inc. Stony Brook, NY</td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Oxygen cylinder&#160;</td>
			<td>Matheson-Tri Gas, North-Central Zone, Lincoln, NE</td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Gas regulator&#160;</td>
			<td>Western Medica, West Lake, OH</td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Signal breaking module, MR compatible small animal monitoring and gating system</td>
			<td>Small Animal Instruments, Inc. Stony Brook, NY</td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">9.4 T (400 MHZ) 89 mm vertical core bore MR scanner&#160;</td>
			<td>Agilent Technologies, Santa Clara, CA</td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">4-cm millipede micro-imaging RF coil&#160;</td>
			<td>Agilent Technologies, Santa Clara, CA</td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">SAM PC monitor&#160;</td>
			<td>Small Animal Instruments, Inc. Stony Brook, NY</td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Quantitative Medical Image analysis software&#160;</td>
			<td colspan="3">http://segment.heiberg.se;&#160; Segment v1.8 R1430,&#160; Medviso, Oresunds region, Sweden</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Matlab software&#160;</td>
			<td>The Mathworks, Inc.&#160; Natick, MA</td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Computer-Unix operating system</td>
			<td style="width:364px;"></td>
			<td style="width:145px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

noninvasive assessment of cardiac abnormalities in experimental autoimmune myocarditis by magnetic resonance microscopy imaging in the mouse

La miocardite è una infiammazione del miocardio, ma solo circa il 10% di questi spettacoli manifestazioni cliniche interessate della malattia. Per studiare gli eventi del sistema immunitario di lesioni del miocardio, vari modelli murini di miocardite sono stati ampiamente utilizzati. Questo studio ha coinvolto sperimentale miocardite autoimmune (EAM) indotto con miosina cardiaca catena pesante (MyHC)-α 334-352 in A / J topi; gli animali colpiti sviluppano miocardite linfocitica ma senza segni clinici evidenti. In questo modello, l&#39;utilità di microscopia a risonanza magnetica (MRM) come modalità non invasiva per determinare le cardiaci cambiamenti strutturali e funzionali in animali immunizzati con MyHC-α 334-352 è mostrato. EAM e topi sani sono stati ripresi con un 9.4 T (400 MHz) 89 millimetri scanner foro centrale verticale dotato di una sonda di imaging a radiofrequenza 4 centimetri millepiedi e 100 g / cm gradienti triple assi. Immagini cardiache sono state acquisite da animali anestetizzati utilizzando una sequenza di impulsi cine-based gradient-echo, e l&#39;animaLS sono stati monitorati dalla respirazione e pulsossimetria. L&#39;analisi ha rivelato un aumento dello spessore della parete ventricolare in topi EAM, con una corrispondente diminuzione del diametro interno dei ventricoli, se confrontato con topi sani. I dati suggeriscono che i cambiamenti morfologici e funzionali nei cuori infiammati possono essere non invasiva monitorato da MRM di animali vivi. In conclusione, MRM offre un vantaggio di valutare la progressione e regressione delle lesioni miocardiche in malattie causate da agenti infettivi, così come la risposta alle terapie.

In questa relazione, viene mostrato l&#39;utilità della tecnica MRM come una modalità non invasiva per determinare i cambiamenti strutturali e funzionali nei cuori di animali affetti da EAM. La miocardite è stata indotta in A / J topi immunizzando gli animali con MyHC-α 334-352 in CFA 7, e gli animali sono stati sottoposti ad esperimenti MRM al giorno 21 post-vaccinazione. La rappresentazione MRM è stata effettuata su animali vivi in ​​anestesia isoflurano a 9.4 T (400 MHz per i protoni), utilizzando...

Watch this Scientific Journal Video about Noninvasive Assessment of Cardiac Abnormalities in Experimental Autoimmune Myocarditis by Magnetic Resonance Microscopy Imaging in the Mouse at JoVE.com

Noninvasive Assessment of Cardiac Abnormalities in Experimental Autoimmune Myocarditis by Magnetic Resonance Microscopy Imaging in the Mouse

This study demonstrates the successful establishment of magnetic resonance microscopy imaging as a non-invasive tool to assess the cardiac abnormalities in mice affected with autoimmune myocarditis. The data indicate that the technique can be used to monitor the disease-progression in live animals.

La valutazione non invasiva della Cardiac Anomalie in Experimental Autoimmune miocardite da Risonanza Magnetica Imaging Microscopy nel mouse

Myocarditis is an inflammation of the myocardium, but only ~10% of those affected show clinical manifestations of the disease. To study the immune events ...

noninvasive-assessment-cardiac-abnormalities-experimental-autoimmune

Research

JoVE Journal

Medicine

9.8K Views.  University of Nebraska-Lincoln. This study demonstrates the successful establishment of magnetic resonance microscopy imaging as a non-invasive tool to assess the cardiac abnormalities in mice affected with autoimmune myocarditis. The data indicate that the technique can be used to monitor the disease-progression in live animals.

Video: Noninvasive Assessment of Cardiac Abnormalities in Experimental Autoimmune Myocarditis by Magnetic Resonance Microscopy Imaging in the Mouse

Magnetic Resonance Derived Myocardial Strain Assessment Using Feature Tracking

Purpose: An accurate and practical method to measure parameters like strain in myocardial tissue is of great clinical value, since it has been shown, that strain is a more sensitive and earlier marker for contractile dysfunction than the frequently used parameter EF. Current technologies for CMR are time consuming and difficult to implement in clinical practice. Feature tracking is a technology that can lead to more automization and robustness of quantitative analysis of medical images with less time consumption than comparable methods.
Methods: An automatic or manual input in a single phase serves as an initialization from which the system starts to track the displacement of individual patterns representing anatomical structures over time. The specialty of this method is that the images do not need to be manipulated in any way beforehand like e.g. tagging of CMR images.
Results: The method is very well suited for tracking muscular tissue and with this allowing quantitative elaboration of myocardium and also blood flow.
Conclusions: This new method offers a robust and time saving procedure to quantify myocardial tissue and blood with displacement, velocity and deformation parameters on regular sequences of CMR imaging. It therefore can be implemented in clinical practice.

An accurate and practical method to measure parameters like strain in myocardial tissue is of great clinical value, since it has been shown, that strain is a more sensitive and earlier marker for contractile dysfunction than the frequently used parameter EF.

Risonanza Magnetica Derivato valutazione di deformazione miocardica Utilizzo del rilevamento delle caratteristiche

Purpose: An accurate and practical method to measure parameters like strain in myocardial tissue is of great clinical value, since it has been shown, that ...

Ricerca

Medicina

Multi-modal Imaging of Angiogenesis in a Nude Rat Model of Breast Cancer Bone Metastasis Using Magnetic Resonance Imaging, Volumetric Computed Tomography and Ultrasound

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell proliferation in the bone marrow cavity as well as for interaction of tumor and bone cells resulting in local bone destruction. Our aim was to develop a model of experimental bone metastasis that allows in vivo assessment of angiogenesis in skeletal lesions using non-invasive imaging techniques.
For this purpose, we injected 105 MDA-MB-231 human breast cancer cells into the superficial epigastric artery, which precludes the growth of metastases in body areas other than the respective hind leg1. Following 25-30 days after tumor cell inoculation, site-specific bone metastases develop, restricted to the distal femur, proximal tibia and proximal fibula1. Morphological and functional aspects of angiogenesis can be investigated longitudinally in bone metastases using magnetic resonance imaging (MRI), volumetric computed tomography (VCT) and ultrasound (US).
MRI displays morphologic information on the soft tissue part of bone metastases that is initially confined to the bone marrow cavity and subsequently exceeds cortical bone while progressing. Using dynamic contrast-enhanced MRI (DCE-MRI) functional data including regional blood volume, perfusion and vessel permeability can be obtained and quantified2-4. Bone destruction is captured in high resolution using morphological VCT imaging. Complementary to MRI findings, osteolytic lesions can be located adjacent to sites of intramedullary tumor growth. After contrast agent application, VCT angiography reveals the macrovessel architecture in bone metastases in high resolution, and DCE-VCT enables insight in the microcirculation of these lesions5,6. US is applicable to assess morphological and functional features from skeletal lesions due to local osteolysis of cortical bone. Using B-mode and Doppler techniques, structure and perfusion of the soft tissue metastases can be evaluated, respectively. DCE-US allows for real-time imaging of vascularization in bone metastases after injection of microbubbles7.
In conclusion, in a model of site-specific breast cancer bone metastases multi-modal imaging techniques including MRI, VCT and US offer complementary information on morphology and functional parameters of angiogenesis in these skeletal lesions.

In the pathogenesis of bone metastasis, angiogenesis is a crucial process and therefore represents a target for imaging and therapy. Here, we present a rat model of site-specific breast cancer bone metastasis and describe strategies to non-invasively image angiogenesis in vivo using magnetic resonance imaging, volumetric computed tomography and ultrasound.

Multi-modal Imaging di angiogenesi in un modello di ratto Nudo di metastasi ossee cancro al seno utilizzando la risonanza magnetica, tomografia computerizzata volumetrica e Ultrasuoni

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell ...

Diffusion Tensor Magnetic Resonance Imaging in the Analysis of Neurodegenerative Diseases

Diffusion tensor imaging (DTI) techniques provide information on the microstructural processes of the cerebral white matter (WM) in vivo. The present applications are designed to investigate differences of WM involvement patterns in different brain diseases, especially neurodegenerative disorders, by use of different DTI analyses in comparison with matched controls. 	
DTI data analysis is performed in a variate fashion, i.e. voxelwise comparison of regional diffusion direction-based metrics such as fractional anisotropy (FA), together with fiber tracking (FT) accompanied by tractwise fractional anisotropy statistics (TFAS) at the group level in order to identify differences in FA along WM structures, aiming at the definition of regional patterns of WM alterations at the group level. Transformation into a stereotaxic standard space is a prerequisite for group studies and requires thorough data processing to preserve directional inter-dependencies. The present applications show optimized technical approaches for this preservation of quantitative and directional information during spatial normalization in data analyses at the group level. On this basis, FT techniques can be applied to group averaged data in order to quantify metrics information as defined by FT. Additionally, application of DTI methods, i.e. differences in FA-maps after stereotaxic alignment, in a longitudinal analysis at an individual subject basis reveal information about the progression of neurological disorders. Further quality improvement of DTI based results can be obtained during preprocessing by application of a controlled elimination of gradient directions with high noise levels.	
In summary, DTI is used to define a distinct WM pathoanatomy of different brain diseases by the combination of whole brain-based and tract-based DTI analysis.

Diffusion tensor imaging (DTI) basically serves as an MRI-based tool to identify in vivo the microstructure of the brain and pathological processes due to neurological disorders within the cerebral white matter. DTI-based analyses allow for application to brain diseases both at the group level and in single subject data.

Diffusion Tensor Imaging a risonanza magnetica in Analisi delle Malattie Neurodegenerative

Diffusion tensor imaging (DTI) techniques provide information on the microstructural processes of the cerebral white matter (WM) in vivo. The present ...

Implanting Glass Spinal Cord Windows in Adult Mice with Experimental Autoimmune Encephalomyelitis

Experimental autoimmune encephalomyelitis (EAE) in adult rodents is the standard experimental model for studying autonomic demyelinating diseases such as multiple sclerosis. Here we present a low-cost and reproducible glass window implantation protocol that is suitable for intravital microscopy and studying the dynamics of spinal cord cytoarchitecture with subcellular resolution in live adult mice with EAE. Briefly, we surgically expose the vertebrae T12-L2 and construct a chamber around the exposed vertebrae using a combination of cyanoacrylate and dental cement. A laminectomy is performed from T13 to L1, and a thin layer of transparent silicone elastomer is applied to the dorsal surface of the exposed spinal cord. A modified glass cover slip is implanted over the exposed spinal cord taking care that the glass does not directly contact the spinal cord. To reduce the infiltration of inflammatory cells between the window and spinal cord, anti-inflammatory treatment is administered every 2 days (as recommended by ethics committee) for the first 10 days after implantation. EAE is induced only 2-3 weeks after the cessation of anti-inflammatory treatment.
Using this approach we successfully induced EAE in 87% of animals with implanted windows and, using Thy1-CFP-23 mice (blue axons in dorsal spinal cord), quantified axonal loss throughout EAE progression. Taken together, this protocol may be useful for studying the recruitment of various cell populations as well as their interaction dynamics, with subcellular resolution and for extended periods of time. This intravital imaging modality represents a valuable tool for developing therapeutic strategies to treat autoimmune demyelinating diseases such as EAE.

We describe a method for implanting and maintaining glass windows over the exposed spinal cords of adult mice for studying experimental autoimmune encephalomyelitis. These windows provide chronic optical access to the spinal cord for monitoring cell dynamics at the subcellular level in live animals using two-photon microscopy.

Experimental autoimmune encephalomyelitis (EAE) in adult rodents is the standard experimental model for studying autonomic demyelinating diseases such as ...

Simultaneous PET/MRI Imaging During Mouse Cerebral Hypoxia-ischemia

Dynamic changes in tissue water diffusion and glucose metabolism occur during and after hypoxia in cerebral hypoxia-ischemia reflecting a bioenergetics disturbance in affected cells. Diffusion weighted magnetic resonance imaging (MRI) identifies regions that are damaged, potentially irreversibly, by hypoxia-ischemia. Alterations in glucose utilization in the affected tissue may be detectable by positron emission tomography (PET) imaging of 2-deoxy-2-(18F)fluoro-&#7429;-glucose ([18F]FDG) uptake. Due to the rapid and variable nature of injury in this animal model, acquisition of both modes of data must be performed simultaneously in order to meaningfully correlate PET and MRI data. In addition, inter-animal variability in the hypoxic-ischemic injury due to vascular differences limits the ability to analyze multi-modal data and observe changes to a group-wise approach if data is not acquired simultaneously in individual subjects. The method presented here allows one to acquire both diffusion-weighted MRI and [18F]FDG uptake data in the same animal before, during, and after the hypoxic challenge in order to interrogate immediate physiological changes.

The method presented here uses simultaneous positron emission tomography and magnetic resonance imaging. In the cerebral hypoxia-ischemia model, dynamic changes in diffusion and glucose metabolism occur during and after injury. The evolving and irreproducible damage in this model necessitates simultaneous acquisition if meaningful multi-modal imaging data are to be acquired.

Simultanea Imaging PET / MRI Durante mouse cerebrale ipossia-ischemia

Dynamic changes in tissue water diffusion and glucose metabolism occur during and after hypoxia in cerebral hypoxia-ischemia reflecting a bioenergetics ...

Assessment of Cardiac Morphological and Functional Changes in Mouse Model of Transverse Aortic Constriction by Echocardiographic Imaging

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable noninvasive method is essential to assess real-time cardiac morphological and functional changes in animal models of heart disease. Transthoracic echocardiography represents an important tool for noninvasive assessment of cardiac structure and function2. Here we used a high-resolution ultrasound imaging system to monitor myocardial remodeling and heart failure progression over time in a mouse model of TAC. B-mode, M-mode, and Doppler imaging were used to precisely assess cardiac hypertrophy, ventricular dilatation, and functional deterioration in mice following TAC. Color and pulse wave (PW) Doppler imaging was used to noninvasively measure pressure gradient across the aortic constriction created by TAC and to assess transmitral blood flow in mice. Thus transthoracic echocardiographic imaging provides comprehensive noninvasive measurements of cardiac dimensions and function in mouse models of heart disease.

The goal of this protocol is to noninvasively assess cardiac structural and functional changes in a mouse model of heart disease created by transverse aortic constriction, using B- and M-mode echocardiography and color/pulse wave Doppler imaging.

Valutazione del Cardiac morfologiche e funzionali Variazioni modello murino di trasversale aortica costrizione da parte ecocardiografica Imaging

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable ...

Quantitative Visualization of Leukocyte Infiltrate in a Murine Model of Fulminant Myocarditis by Light Sheet Microscopy

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently labelled structures in large biological specimens, and is down to cellular resolution. Meanwhile, the constant refinement of underlying protocols and the enhanced availability of specialized commercial systems enable us to investigate the microstructure of whole mouse organs and even allow for the characterization of cellular behavior in various live-cell imaging approaches. Here, we describe a protocol for the spatial whole-mount visualization and quantification of the CD45+ leukocyte population in inflamed mouse hearts. The method employs a transgenic mouse strain (CD11c.DTR)that has recently been shown to serve as a robust, inducible model for the study of the development of fulminant fatal myocarditis, characterized by lethal cardiac arrhythmias. This protocol includes myocarditis induction, intravital antibody-mediated cell staining, organ preparation, and LSFM with subsequent computer-assisted image post-processing. Although presented as a highly-adapted method for our particular scientific question, the protocol represents the blueprint of an easily adjustable system that can also target completely different fluorescent structures in other organs and even in other species.

Here, we describe a light-sheet microscopy approach to visualize the cardiac CD45+ leukocyte infiltrate in a murine model of aseptic fulminant myocarditis, which is induced by the intratracheal diphtheria toxin treatment of CD11c.DTR mice.

Visualizzazione quantitativa di leucociti infiltrati in un modello di murina della miocardite fulminante mediante microscopia a foglio chiaro

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently ...

Cardiac Magnetic Resonance Imaging at 7 Tesla

CMR at an ultra-high field (magnetic field strength B0&#160;&#8805; 7 Tesla) benefits from the signal-to-noise ratio (SNR) advantage inherent at higher magnetic field strengths and potentially provides improved signal contrast and spatial resolution. While promising results have been achieved, ultra-high field CMR is challenging due to energy deposition constraints and physical phenomena such as transmission field non-uniformities and magnetic field inhomogeneities. In addition, the magneto-hydrodynamic effect renders the synchronization of the data acquisition with the cardiac motion difficult. The challenges are currently addressed by explorations into novel magnetic resonance technology. If all impediments can be overcome, ultra-high field CMR may generate new opportunities for functional CMR, myocardial tissue characterization, microstructure imaging or metabolic imaging. Recognizing this potential, we show that multi-channel radio frequency (RF) coil technology tailored for CMR at 7 Tesla together with higher order B0 shimming and a backup signal for cardiac triggering facilitates high fidelity functional CMR. With the proposed setup, cardiac chamber quantification can be accomplished in examination times similar to those achieved at lower field strengths. To share this experience and to support the dissemination of this expertise, this work describes our setup and protocol tailored for functional CMR at 7 Tesla.

The sensitivity gain inherent to ultrahigh field magnetic resonance holds promise for high spatial resolution imaging of the heart. Here, we describe a protocol customized for functional cardiovascular magnetic resonance (CMR) at 7 Tesla using an advanced multi-channel radio-frequency coil, magnetic field shimming and a triggering concept.

Risonanza magnetica cardiaca a 7 Tesla

CMR at an ultra-high field (magnetic field strength B0&#160;&#8805; 7 Tesla) benefits from the signal-to-noise ratio (SNR) advantage inherent at higher ...

Phase Contrast Magnetic Resonance Imaging in the Rat Common Carotid Artery

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous studies have shown that abnormal blood flow may be associated with systemic vascular risk. Thus, PC-MRI can facilitate the translation of data obtained from animal models of cardiovascular diseases to pertinent clinical investigations. In this report, we describe the procedure for measuring blood flow in the common carotid artery (CCA) of rats using cine-gated PC-MRI and discuss relevant analysis methods. This procedure can be performed in a live, anesthetized animal and does not require euthanasia after the procedure. The proposed scanning parameters yield repeatable measurements for blood flow, indicating excellent reproducibility of the results. The PC-MRI procedure described in this article can be used for pharmacological testing, pathophysiological assessment, and cerebral hemodynamics evaluation.

The overall goal of this procedure is to measure the blood flow in the rat common carotid artery by using noninvasive phase contrast magnetic resonance imaging.

Fase di contrasto risonanza magnetica nell'arteria carotica comune ratto

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous ...

Quantification of Levator Ani Hiatus Enlargement by Magnetic Resonance Imaging in Males and Females with Pelvic Organ Prolapse

Here we present a protocol to examine the levator ani hiatus in males and females with pelvic organ prolapse, during the Valsalva maneuver and while evacuating acoustic gel, using a horizontally oriented 1.5 T magnetic resonance (MR) scanner. On midsagittal images, the vertical distance of pelvic organs is measured in millimeters relative to the hymen plane (female) and to the lower border of the symphysis pubis (male), preceded by - (above) or + ( below) signs. On axial images, the levator ani area is calculated in square centimeters with a free-hand tracing method from three key images, passing through the midsymphysis (level I), tangential to the lower border of the symphysis (level II), and at the maximal anterior rectal wall bulging (level III). Areas at rest and strained are compared to find evidence of a percentage of increase. The purpose is to provide objective evidence of the maximal extent of pelvic organs descent and hiatus enlargement without the interference of foreign objects or the examiner&#39;s proximity, so as to overcome the limitations of pelvic examination and transperineal sonography (i.e., subjectivity and sex-related limitations [female only]).

Here we present a protocol to standardize the measurement of the levator ani hiatus size&#160;by magnetic resonance imaging. The purpose is to extract biomechanical inferences from image analysis by comparing resting and straining values in patients of both sexes with pelvic prolapse, using consistent anatomical bony landmarks.

Quantificazione di Levator Ani iato allargamento mediante risonanza magnetica in maschi e femmine con prolasso degli organi pelvici

Here we present a protocol to examine the levator ani hiatus in males and females with pelvic organ prolapse, during the Valsalva maneuver and while ...