Vorremmo riconoscere gli sforzi del Dr. Mary-Lou Vallano, Dipartimento di Neuroscienze e Fisiologia, per i suoi preziosi contributi nel scrivere per questo manoscritto.

1. Chirurgia per Rat Oggetto SAH iniettato con 0,15 ml di sangue autologo arteriosa <ol><li> Il ratto è anestetizzato con 0,1 mg / kg di ketamina / TMA cocktail roditore e lasciata riposare per 5 min. </li><li> Anestesia adeguata è confermato dalla riduzione di riflesso degli arti posteriori. </li><li> L&#39;utilizzo di un rasoio elettronico un collo al naso zona di capelli intorno al sub-occipitale è rasata. </li><li> L&#39;animale viene posto supino sul tavolo operatorio e la coda è deterso con Betadine per garantire una incisione sterile. </li><li> Un diritto da 1 centimetro linea mediana incisione è assunto l&#39;aspetto ventrale della coda </li><li> La dissezione è prorogata fino l&#39;arteria coda è identificato e isolato. </li><li> Utilizzando una sterile 26-gauge catetere, l&#39;arteria coda è cannulata e 0,15 ml di sangue arterioso viene aspirata in una siringa. </li><li> Una garza sterile viene avvolto intorno alla incisione per assicurare emostasi prima dell&#39;applicazione del vetbond per sigillare l&#39;incisione. </li><li> Tratto si è attivata prono sul tavolo e la zona rasata sul sub-occipitale è deterso con betadine. </li><li> Utilizzo di un accesso verticale incisione mediana si ottiene l&#39;cisterna magna. </li><li> Una volta identificato un ago calibro 25 è inserito nella cisterna magna e 0,15 ml di CSF viene prelevata con una siringa per evitare maggiori pressioni intracranici con iniezione di volume di sangue autologo. </li><li> Ora, il 0,15 ml di sangue estratto dall&#39;arteria coda viene iniettata lentamente in cisterna magna. </li><li> L&#39;ago rimane in posizione per 30 sec per garantire la coagulazione nello spazio subaracnoideo e poi estratto con cautela. </li><li> L&#39;emostasi è assicurata e l&#39;incisione viene chiusa con un dispositivo di pinzatura. </li><li> L&#39;animale è ora posizionato prono su una superficie di riscaldamento con una testa 20 ° verso il basso la posizione per 20 min per permettere al sangue di coagularsi nelle cisterne intorno dell&#39;arteria basilare. </li><li> Passi 1,1-1,15 si ripetono nel corso del secondo intervento 48 ore di distanza. </li&gt; </ol> 2. Chirurgia per Rat Oggetto SAH iniettato con 0,15 ml di soluzione salina <ol><li> Il ratto è anestetizzato con 0,1 mg / kg di ketamina / TMA cocktail roditore e lasciata riposare per 5 min. </li><li> Anestesia adeguata è confermato dalla riduzione di riflesso degli arti posteriori. </li><li> L&#39;utilizzo di un rasoio elettronico un collo al naso zona di capelli intorno al sub-occipitale è rasata. </li><li> L&#39;animale viene posto supino sul tavolo operatorio e la coda è deterso con Betadine per garantire una incisione sterile. </li><li> Un diritto da 1 centimetro linea mediana incisione è assunto l&#39;aspetto ventrale della coda </li><li> La dissezione è prorogata fino l&#39;arteria coda è identificato e isolato. </li><li> Utilizzando una sterile 26-gauge catetere, l&#39;arteria coda è cannulata e 0,15 ml di sangue arterioso viene aspirata in una siringa. </li><li> Una garza sterile viene avvolto intorno alla incisione per assicurare emostasi prima dell&#39;applicazione del vetbond per sigillare l&#39;incisione. </li><li> Il rat è attivata prona sul tavolo e la zona rasata sul sub-occipitale è verniciato con betadine. </li><li> Utilizzo di un accesso verticale incisione mediana si ottiene l&#39;cisterna magna. </li><li> Una volta identificato un ago calibro 25 è inserito nella cisterna magna e 0,15 ml di CSF viene prelevata con una siringa ed il campione viene memorizzato. </li><li> Ora, il 0,15 ml di soluzione salina normale (37 ° C) viene iniettata lentamente in cisterna magna. </li><li> L&#39;ago viene lasciato in sede per 30 sec e estratto con cautela. </li><li> L&#39;emostasi è assicurata e l&#39;incisione viene chiusa con un dispositivo di pinzatura. </li><li> L&#39;animale è ora posizionato prono su una superficie di riscaldamento con una testa 20 ° verso il basso la posizione per 20 min. </li><li> Passi 2,1-2,15 si ripetono nel corso del secondo intervento 48 ore di distanza. </li></ol> 3. Rat Sacrificio <ol><li> Il giorno 5 dopo il secondo intervento chirurgico, i ratti vengono sacrificati mediante perfusione cardiaca. </li><li> Il ratto è dato una dose letale (00,2 ml / kg) di Fatal Plus (Vortech PHARMACEUTICALS LTD., Dearborn, MI) </li><li> Con una incisione mediana verticale, la cavità addominale viene raggiunta e il peritoneo è aperto. </li><li> Una toracotomia anteriore viene eseguita e il cuore è esposto. </li><li> Utilizzando un 26-gauge catetere collegato ad una soluzione di tampone fosfato (PBS pH 7,4 ea 37 °), l&#39;animale viene svuotato di sangue e viene poi perfusi con paraformaldeide al 4%. </li><li> Dopo garantendo un&#39;adeguata perfusione, la perfusione viene arrestata e il ratto viene portata al tavolo decapitazione. </li><li> Dopo decapitazione, un rongeur osso viene utilizzato per rimuovere il cranio per la rimozione del cervello. </li><li> Il cervello e del tronco cerebrale sono accuratamente estratta dalla volta cranica e posto in una soluzione di paraformaldeide 4% e conservata a 4 ° C per 48 ore. </li></ol> 4. Creazione di sezioni per valutare vasospasmo <ol><li> Il cervello di ratto che è stato immerso in 30% saccarosio per 4 giorni è portato a thcriostato e per il sezionamento. </li><li> Una volta crioprotetti, 12 sezioni di m sono creati utilizzando il criostato, con l&#39;anteriore arteria cerebellare inferiore (AICA) come punto di partenza per garantire la coerenza tra soggetti. </li><li> 20 sezioni vengono creati per ogni animale, che termina a livello dell&#39;arteria cerebellare superiore (SCA). </li><li> Le sezioni sono posti su un vetrino e valutati per vasospasmo con metodologia istologica </li></ol>

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M., Guarino, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cortical+blood+flow+and+cerebral+perfusion+pressure+in+a+new+noncraniotomy+model+of+subarachnoid+hemorrhage+in+the+rat.">Cortical blood flow and cerebral perfusion pressure in a new noncraniotomy model of subarachnoid hemorrhage in the rat.</a> Stroke. 26, 1086-1091 (1995).</li><li>Cheng, G., Wei, L., Zhi-Dan, S., Shi-Guang, Z., Xiang-Zhen, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Atorvastatin+ameliorates+cerebral+vasospasm+and+early+brain+injury+after+subarachnoid+hemorrhage+and+inhibits+caspase-dependent+apoptosis+pathway.">Atorvastatin ameliorates cerebral vasospasm and early brain injury after subarachnoid hemorrhage and inhibits caspase-dependent apoptosis pathway.</a> BMC Neurosci. 10, 7-17 (2009).</li><li>Jackowski, A., Crockard, A., Burnstock, G., Russell, R. 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Y., Sagher, O., Keep, R., Hua, Y., Xi, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+experimental+rat+models+of+early+brain+injury+after+subarachnoid+hemorrhage.">Comparison of experimental rat models of early brain injury after subarachnoid hemorrhage.</a> Neurosurgery. 65 (2), 331-343 (2009).</li><li>Megyesi, J. F., Vollrath, B., Cook, D. A., Findlay, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+animal+models+of+cerebral+vasospasm:+a+review.">In vivo animal models of cerebral vasospasm: a review.</a> Neurosurgery. 46, 448-460 (2000).</li><li>Prunell, G. F., Mathiesen, T., Diemer, N. H., Svendgaard, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+subarachnoid+hemorrhage:+Subarachnoid+blood+volume,+mortality+rate,+neuronal+death,+cerebral+blood+flow,+and+perfusion+pressure+in+three+different+rat+models.">Experimental subarachnoid hemorrhage: Subarachnoid blood volume, mortality rate, neuronal death, cerebral blood flow, and perfusion pressure in three different rat models.</a> Neurosurgery. 52, 165-176 (2003).</li><li>Prunell, G. F., Mathiesen, T., Svendgaard, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+subarachnoid+hemorrhage:+Cerebral+blood+flow+and+brain+metabolism+during+the+acute+phase+in+three+different+models+in+the+rat.">Experimental subarachnoid hemorrhage: Cerebral blood flow and brain metabolism during the acute phase in three different models in the rat.</a> Neurosurgery. 54, 426-436 (2004).</li><li>Ryba, M. S., Gordon-Krajcer, W., Walski, M., Chalimoniuk, M., Chrapusta, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hydroxylamine+attenuates+the+effects+of+simulated+subarachnoid+hemorrhage:+implication+for+the+role+of+oxidative+stress+in+cerebral+vasospasm.">Hydroxylamine attenuates the effects of simulated subarachnoid hemorrhage: implication for the role of oxidative stress in cerebral vasospasm.</a> Neurol. Res. 31, 195-199 (1999).</li><li>Satoh, M., Parent, A. D., Zhang, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibitory+effect+with+antisense+mitogen-activated+protein+kinase+oligodeoxynucleotide+against+cerebral+vasospasm+in+rats.">Inhibitory effect with antisense mitogen-activated protein kinase oligodeoxynucleotide against cerebral vasospasm in rats.</a> Stroke. 33, 775-781 (2002).</li><li>Suzuki, H., Kanamaru, K., Tsunoda, H., Inada, H., Kuroki, M., Sun, H., Waga, S., Tanaka, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heme+oxygenase-1+gene+induction+as+an+intrinsic+regulation+against+delayed+cerebral+vasospasm+in+rats.">Heme oxygenase-1 gene induction as an intrinsic regulation against delayed cerebral vasospasm in rats.</a> J. Clin. Invest. 104, 59-66 (1999).</li><li>Swift, D. M., Solomon, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subarachnoid+hemorrhage+fails+to+produce+vasculopathy+or+chronic+blood+flow+changes+in+rats.">Subarachnoid hemorrhage fails to produce vasculopathy or chronic blood flow changes in rats.</a> Stroke. 19, 878-882 (1988).</li><li>Vatter, H., Weidauer, S., Konczalla, J., Dettmann, E., Zimmermann, M., Raabe, A., Preibisch, C., Zanella, F., Seifert, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Time+course+in+the+development+of+cerebral+vasospasm+after+experimental+subarachnoid+hemorrhage:+clinical+and+neuroradiological+assessment+of+the+rat+double+hemorrhage+model.">Time course in the development of cerebral vasospasm after experimental subarachnoid hemorrhage: clinical and neuroradiological assessment of the rat double hemorrhage model.</a> Neurosurgery. 58, 1190-1197 (2006).</li><li>Veelken, J. A., Laing, R. J., Jakubowski, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Sheffield+model+of+subarachnoid+hemorrhage+in+rats.">The Sheffield model of subarachnoid hemorrhage in rats.</a> Stroke. 26, 1279-1283 (1995).</li><li>Zubkov, A. Y., Nanda, A., Zhang, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Signal+transduction+pathways+in+cerebral+vasospasm.">Signal transduction pathways in cerebral vasospasm.</a> Pathophysiology. 9, 47-61 (2003).</li></ol>

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Primati, con una composizione più simile genetica e caratteristiche anatomiche per l&#39;umano, più strettamente imitare gli eventi di ritardo vasospasmo cerebrale e possono più facilmente subire non invasiva per immagini (risonanza magnetica e angiografia) per monitorare i cambiamenti delle arterie, che roditori 8. Tuttavia, i modelli di primati sono un costo proibitivo e associato con cura più complessa e le questioni etiche, di modelli animali di piccole dimensioni. Piccoli animali SAH modelli che sono...

<table border="1"><tbody><tr><td> Nome di attrezzature / reagente </td><td> Azienda </td><td> Numero di catalogo </td></tr><tr><td> Ratti maschi SD (250-300 g) </td><td> Taconic </td><td> SD-M </td></tr><tr><td> 26 g Cateteri </td><td> Webster </td><td> 8416683 </td></tr><tr><td> 25 G Aghi </td><td> Bufalo </td><td> 305122 </td></tr><tr><td> 1 cc Siringhe </td><td> Negozi centrali </td><td> 54245 </td></tr><tr><td> La ketamina / Xylazina cocktail </td><td> Animal Care (SUNY) * </td><td> - </td></tr><tr><td> Betadine </td><td> Negozi centrali </td><td> 51458 </td></tr><tr><td> Saccarosio </td><td> Sigma </td><td> S9378-1kg </td></tr><tr><td> Paraformaldeide </td><td> Sigma </td><td> P6148-500G </td></tr><tr><td> Soluzione tampone fosfato </td><td> Pescatore </td><td> BP-399-4 </td></tr><tr><td> Tavolo operatorio </td><td> Harvard </td><td> PY2 72-2590 </td></tr><tr><td> Ottobre composto (crioprotezione) </td><td> VWR </td><td> 25608-930 </td></tr><tr><td> SuperFrost diapositive </td><td> Pescatore </td><td> 12-550-15 </td></tr></tbody></table> * Sintetizzato presso il Dipartimento di cura degli animali da laboratorio, SUNY Upstate Medical University. Aggiungere 1 cc [100 mg / ml] di DBU a 10 ml [100 mg / ml] di Ketamina.

a low mortality rat model to assess delayed cerebral vasospasm after experimental subarachnoid hemorrhage

Obiettivo: caratterizzare e stabilire un modello riproducibile che dimostra in ritardo vasospasmo cerebrale dopo emorragia subaracnoidea aneurismatica (SAH) nei ratti, al fine di individuare gli eventi scatenanti, i cambiamenti fisiopatologici e target potenziali per il trattamento. Metodi: Ventotto maschi Sprague-Dawley (250-300 g) sono stati arbitrariamente assegnati ad uno dei due gruppi - SAH o controllo salino. Emorragia subaracnoidea ratto nel gruppo SAH (n = 15) è stata indotta mediante doppia iniezione di sangue autologo, 48 hr a parte, nella cisterna magna. Analogamente, soluzione salina normale (n = 13) è stato iniettato nella cisterna magna del gruppo di controllo salina. Ratti sono stati sacrificati al giorno cinque dopo l&#39;iniezione e sangue secondo i cervelli sono stati conservati per l&#39;analisi istologica. Il grado di vasospasmo è stata misurata utilizzando sezioni dell&#39;arteria basilare, misurando l&#39;area in sezione trasversale interna luminale utilizzando NIH Image-J software. Il significato eratestato con Tukey / Kramer &#39;s l&#39;analisi statistica. Risultati: Dopo l&#39;analisi delle sezioni istologiche, basilare superficie luminale dell&#39;arteria sezione trasversale erano più piccole in SAH rispetto al gruppo salina, coerente con vasospasmo cerebrale nel primo gruppo. Nel gruppo SAH, zona interna dell&#39;arteria basilare (0,056 micron ± 3) erano significativamente più piccole da vasospasmo cinque giorni dopo l&#39;iniezione di sangue secondo (sette giorni dopo l&#39;iniezione di sangue iniziale), rispetto al gruppo di controllo salina con area interna (0,069 ± 3, p = 0,004). Non ci sono stati casi di mortalità da vasospasmo cerebrale. Conclusione: Il doppio ratto induce una lieve emorragia subaracnoidea, capaci di mantenersi, vasospasmo dell&#39;arteria basilare che può essere utilizzato per studiare i meccanismi fisiopatologici di vasospasmo cerebrale in un modello animale di piccole dimensioni. Un tasso di mortalità basso e accettabile è un criterio significativo per essere soddisfatti per un ideale modello animale SAH in modo che i meccanismi di vasospasmo può essere elucidated 7, 8. Ulteriori modifiche del modello può essere fatto per regolare la maggiore gravità del vasospasmo e gli esami neurologici.

Entro i protocolli descritti sopra, ci sono diversi passaggi che riteniamo richiedono una migliore caratterizzazione del modello di quello che è stato descritto precedentemente in letteratura. Qui ci concentriamo sui passi che sono essenziali al fine di ottenere una bassa mortalità riproducibile cerebrale modello animale piccolo vasospasmo ed evitare potenziali problemi associati a questo modello se non effettuate correttamente. 1. Il sangue autologo Draw dal Arteria Tail: <p class="jove_...

Watch this Scientific Journal Video about A Low Mortality Rat Model to Assess Delayed Cerebral Vasospasm After Experimental Subarachnoid Hemorrhage at JoVE.com

A Low Mortality Rat Model to Assess Delayed Cerebral Vasospasm After Experimental Subarachnoid Hemorrhage

Emorragia subaracnoidea aneurismatica (SAH) è il sanguinamento che si verifica nello spazio subaracnoideo quando una rottura dell&#39;aneurisma. Mentre la morbilità e la mortalità di questo evento è stato su un declino a causa di migliori approcci terapeutici, il rischio di vasospasmo dopo emorragia subaracnoidea continua ad essere la stessa che era diversi anni fa. L&#39;importanza della creazione di un modello animale completo e riproducibile per identificare gli eventi scatenanti di vasospasmo cerebrale è stato al centro di ricerche sin dal primo utilizzo di ratti in un modello sperimentale di vasospasmo nel 1979 da Barry<em&gt; Et al.</em&gt; I primi lavori nei ratti ha dimostrato che una singola iniezione di sangue autologo nella cisterna magna portato a acuta (entro pochi minuti), ma non in ritardo vasospasmo cerebrale<sup&gt; 3, 6, 14</sup&gt;. Qui caratterizzare una bassa mortalità modello di ratto SAH che si traduce in riproducibili vasospasmo ritardato.

Un basso Rat modello di mortalità per valutare ritardata vasospasmo cerebrale dopo emorragia subaracnoidea sperimentale

Objective: To characterize and establish a reproducible model that demonstrates delayed cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH) ...

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14.2K Views.  SUNY Upstate Medical University. Emorragia subaracnoidea aneurismatica (SAH) è il sanguinamento che si verifica nello spazio subaracnoideo quando una rottura dell'aneurisma. Mentre la morbilità e la mortalità di questo evento è stato su un declino a causa di migliori approcci terapeutici, il rischio di vasospasmo dopo emorragia subaracnoidea continua ad essere la stessa che era diversi anni fa. L'importanza della creazione di un modello animale completo e riproducibile per identificare gli ev...

Video: A Low Mortality Rat Model to Assess Delayed Cerebral Vasospasm After Experimental Subarachnoid Hemorrhage

Modeling Stroke in Mice - Middle Cerebral Artery Occlusion with the Filament Model

Stroke is among the most frequent causes of death and adult disability, especially in highly developed countries. However, treatment options to date are very limited. To meet the need for novel therapeutic approaches, experimental stroke research frequently employs rodent models of focal cerebral ischaemia. Most researchers use permanent or transient occlusion of the middle cerebral artery (MCA) in mice or rats.
Proximal occlusion of the middle cerebral artery (MCA) via the intraluminal suture technique (so called filament or suture model) is probably the most frequently used model in experimental stroke research. The intraluminal MCAO model offers the advantage of inducing reproducible transient or permanent ischaemia of the MCA territory in a relatively non-invasive manner. Intraluminal approaches interrupt the blood flow of the entire territory of this artery. Filament occlusion thus arrests flow proximal to the lenticulo-striate arteries, which supply the basal ganglia. Filament occlusion of the MCA results in reproducible lesions in the cortex and striatum and can be either permanent or transient. In contrast, models inducing distal (to the branching of the lenticulo-striate arteries) MCA occlusion typically spare the striatum and primarily involve the neocortex. In addition these models do require craniectomy. In the model demonstrated in this article, a silicon coated filament is introduced into the common carotid artery and advanced along the internal carotid artery into the Circle of Willis, where it blocks the origin of the middle cerebral artery. In patients, occlusions of the middle cerebral artery are among the most common causes of ischaemic stroke. Since varying ischemic intervals can be chosen freely in this model depending on the time point of reperfusion, ischaemic lesions with varying degrees of severity can be produced. Reperfusion by removal of the occluding filament at least partially models the restoration of blood flow after spontaneous or therapeutic (tPA) lysis of a thromboembolic clot in humans.
In this video we will present the basic technique as well as the major pitfalls and confounders which may limit the predictive value of this model.

Filamentous occlusion of the Middle cerebral artery is a common model for studying ischemic stroke in mice.

Corsa di modellazione in Mouse - Medio occlusione dell&#39;arteria cerebrale con il Modello Filament

Stroke is among the most frequent causes of death and adult disability, especially in highly developed countries. However, treatment options to date are ...

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Medicina

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

The number of patients with end-stage renal disease, and the number of kidney allograft recipients continuously increases. Episodes of acute cellular allograft rejection (AR) are a negative prognostic factor for long-term allograft survival, and its timely diagnosis is crucial for allograft function 1. At present, AR can only be definitely diagnosed by core-needle biopsy, which, as an invasive method, bares significant risk of graft injury or even loss. Moreover, biopsies are not feasible in patients taking anticoagulant drugs and the limited sampling site of this technique may result in false negative results if the AR is focal or patchy. As a consequence, this gave rise to an ongoing search for new AR detection methods, which often has to be done in animals including the use of various transplantation models.
Since the early 60s rat renal transplantation is a well-established experimental method for the examination and analysis of AR 2. We herein present in addition small animal positron emission tomography (PET) using 18F-fluorodeoxyglucose (FDG) to assess AR in an allogeneic uninephrectomized rat renal transplantation model and propose graft FDG-PET imaging as a new option for a non-invasive, specific and early diagnosis of AR also for the human situation 3. Further, this method can be applied for follow-up to improve monitoring of transplant rejection 4.

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

We herein present a rat renal transplantation model to non-invasively assess acute allograft rejection using positron emission tomography with 18F-fluorodeoxyglucose.

Imaging non invasivo di acuta Alloinnesto rigetto dopo trapianto renale Rat Utilizzo<sup&gt; 18</sup&gt; F-FDG PET

The number of patients with end-stage renal  disease, and the number of kidney allograft recipients continuously increases.  Episodes of acute cellular ...

Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as studies on graft preservation and ischemia-reperfusion injury 1,2, immunological responses 3,4, hemodynamics 5,6, and small-for-size syndrome 7. The rat OLT is among the most difficult animal models in experimental surgery and demands advanced microsurgical skills that take a long time to learn. Consequently, the use of this model has been limited. Since the reliability and reproducibility of results are key components of the experiments in which such complex animal models are used, it is essential for surgeons who are involved in rat OLT to be trained in well-standardized and sophisticated procedures for this model.
While various techniques and modifications of OLT in rats have been reported 8 since the first model was described by Lee et al. 9 in 1973, the elimination of the hepatic arterial reconstruction 10 and the introduction of the cuff anastomosis technique by Kamada et al. 11 were a major advancement in this model, because they simplified the reconstruction procedures to a great degree. In the model by Kamada et al., the hepatic rearterialization was also eliminated. Since rats could survive without hepatic arterial flow after liver transplantation, there was considerable controversy over the value of hepatic arterialization. However, the physiological superiority of the arterialized model has been increasingly acknowledged, especially in terms of preserving the bile duct system 8,12 and the liver integrity 8,13,14.
In this article, we present detailed surgical procedures for a rat model of OLT with hepatic arterial reconstruction using a 50% partial graft after ex vivo liver resection. The reconstruction procedures for each vessel and the bile duct are performed by the following methods: a 7-0 polypropylene continuous suture for the supra- and infrahepatic vena cava; a cuff technique for the portal vein; and a stent technique for the hepatic artery and the bile duct.

Orthotopic liver transplantation in rats is an indispensable experimental model for biomedical research. Here we present our surgical procedures for orthotopic rat liver transplantation with hepatic arterial reconstruction using a 50% partial graft.

Procedure chirurgiche per un modello di Rat parziale trapianto di fegato con la ricostruzione arteriosa epatica

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

Optimized System for Cerebral Perfusion Monitoring in the Rat Stroke Model of Intraluminal Middle Cerebral Artery Occlusion

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal models of acute ischemic stroke allows to confirm successful arterial occlusion and exclude subarachnoid hemorrhage. Cerebral perfusion monitoring can also be used to study intracranial collateral circulation, which is emerging as a powerful determinant of stroke outcome and a possible therapeutic target. Despite a recognized role of Laser Doppler perfusion monitoring as part of the current guidelines for experimental cerebral ischemia, a number of technical difficulties exist that limit its widespread use. One of the major issues is obtaining a secure and prolonged attachment of a deep-penetration Laser Doppler probe to the animal skull. In this video, we show our optimized system for cerebral perfusion monitoring during transient middle cerebral artery occlusion by intraluminal filament in the rat. We developed in-house a simple method to obtain a custom made holder for twin-fibre (deep-penetration) Laser Doppler probes, which allow multi-site monitoring if needed. A continuous and prolonged monitoring of cerebral perfusion could easily be obtained over the intact skull.

Cerebral perfusion monitoring has been demonstrated to improve accuracy in ischemic stroke models. Technical difficulties often limit the use of this essential tool for cerebrovascular research. In this video, an optimized system is shown to obtain a single or multi-site hemodynamic monitoring during intraluminal middle cerebral artery occlusion in rats.

Sistema ottimizzato per il monitoraggio perfusione cerebrale nel modello Rat Stroke of intraluminale occlusione dell&#39;arteria cerebrale media

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal ...

A Murine Model of Subarachnoid Hemorrhage

In this video publication a standardized mouse model of subarachnoid hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of Willis perforation (CWp) and proven by intracranial pressure (ICP) monitoring. Thereby a homogenous blood distribution in subarachnoid spaces surrounding the arterial circulation and cerebellar fissures is achieved. Animal physiology is maintained by intubation, mechanical ventilation, and continuous on-line monitoring of various physiological and cardiovascular parameters: body temperature, systemic blood pressure, heart rate, and hemoglobin saturation. Thereby the cerebral perfusion pressure can be tightly monitored resulting in a less variable volume of extravasated blood. This allows a better standardization of endovascular filament perforation in mice and makes the whole model highly reproducible. Thus it is readily available for pharmacological and pathophysiological studies in wild type and genetically altered mice.

A standardized mouse model of subarachnoid hemorrhage by intraluminal Circle of Willis perforation is described. Vessel perforation and subarachnoid bleeding are monitored by intracranial pressure monitoring. In addition various vital parameters are recorded and controlled to maintain physiologic conditions.

Un modello murino di emorragia subaracnoidea

In this video publication a standardized mouse model of subarachnoid  hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of  Willis ...

The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and controllable animal models that simulate both conditions are presently uncommon. Therefore, new models are needed in order to mimic human pathophysiological conditions resulting from SAH.
This report describes the technical nuances of a rabbit blood-shunt SAH model that enables control of intracerebral pressure (ICP). An extracorporeal shunt is placed between the arterial system and the subarachnoid space, which enables examiner-independent SAH in a closed cranium. Step-by-step procedural instructions and necessary equipment are described, as well as technical considerations to produce the model with minimal mortality and morbidity. Important details required for successful surgical creation of this robust, simple and consistent ICP-controlled SAH rabbit model are described.

The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures &#8212; subclavian artery cannulation and transcutaneous cisterna magna puncture, which enables close mimicking of human pathophysiological conditions after SAH. We present step-by-step instructions and discuss key surgical points for successful experimental SAH creation.

Il Coniglio Blood-shunt modello per lo studio di acuta e sequele tardive di emorragia subaracnoidea: Aspetti tecnici

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and ...

Leprdb Mouse Model of Type 2 Diabetes: Pancreatic Islet Isolation and Live-cell 2-Photon Imaging Of Intact Islets

Type 2 diabetes is a chronic disease affecting 382 million people in 2013, and is expected to rise to 592 million by 2035 1. During the past 2 decades, the role of beta-cell dysfunction in type 2 diabetes has been clearly established 2. Research progress has required methods for the isolation of pancreatic islets. The protocol of the islet isolation presented here shares many common steps with protocols from other groups, with some modifications to improve the yield and quality of isolated islets from both the wild type and diabetic Leprdb (db/db) mice. A live-cell 2-photon imaging method is then presented that can be used to investigate the control of insulin secretion within islets.

Leprdb Mouse Model of Type 2 Diabetes: Pancreatic Islet Isolation and Live-cell 2-Photon Imaging Of Intact Islets

We present here a protocol for the isolation of islets from the mouse model of type 2 diabetes, Leprdb and details of a live-cell assay for measurement of insulin secretion from intact islets that utilizes 2 photon microscopy.

LEPR<sup&gt; Db</sup&gt; Mouse Modello di diabete di tipo 2: pancreas isolamento Islet e live-cella 2-Photon Imaging di isolotti Intact

Type 2 diabetes is a chronic disease affecting 382 million people in 2013, and is expected to rise to 592 million by 2035 1. During the past 2 decades, ...

Integrated Compensatory Responses in a Human Model of Hemorrhage

Hemorrhage is the leading cause of trauma-related deaths, partly because early diagnosis of the severity of blood loss is difficult. Assessment of hemorrhaging patients is difficult because current clinical tools provide measures of vital signs that remain stable during the early stages of bleeding due to compensatory mechanisms. Consequently, there is a need to understand and measure the total integration of mechanisms that compensate for reduced circulating blood volume and how they change during ongoing progressive hemorrhage. The body&#39;s reserve to compensate for reduced circulating blood volume is called the &#39;compensatory reserve&#39;. The compensatory reserve can be accurately evaluated with real-time measurements of changes in the features of the arterial waveform measured with the use of a high-powered computer. Lower Body Negative Pressure (LBNP) has been shown to simulate many of the physiological responses in humans associated with hemorrhage, and is used to study the compensatory response to hemorrhage. The purpose of this study is to demonstrate how compensatory reserve is assessed during progressive reductions in central blood volume with LBNP as a simulation of hemorrhage.

The purpose of this protocol is to demonstrate the techniques for measuring compensatory responses to reduced central blood volume using lower body negative pressure as a noninvasive experimental model of human hemorrhage which can be used to quantify the total integration of compensatory mechanisms to blood volume deficit in humans.

Le risposte compensative integrati in un modello umano di emorragia

Hemorrhage is the leading cause of trauma-related deaths, partly because early diagnosis of the severity of blood loss is difficult. Assessment of ...

Transplantation of Adipose Tissue-Derived Stem Cell Sheet to Reduce Leakage After Partial Colectomy in A Rat Model

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of clinical efficacy, they are until now imperfect solutions. Stem cell therapy using ASC sheets could provide a solution to this problem. ASCs are considered as promising candidates for promoting tissue healing because of their trophic and immunomodulatory properties. Here, we provide methods to produce high-density ASC sheets, that are transplanted onto a colorectal anastomosis in a rat model to reduce the leakage. ASCs formed cell sheets in thermo-responsive culture dishes that could be easily detached. On the day of the transplantation, a partial colectomy with a 5-suture colorectal anastomosis was performed. Animals were immediately transplanted with 1 ASC sheet per rat. ASC sheets adhered spontaneously to the anastomosis without any glue, suture, or any biomaterial. Animal groups were sacrificed 3 and 7 days postoperatively. Compared to transplanted animals, the incidence of anastomotic abscesses and leakage was higher in control animals. In our model, the transplantation of ASC sheets after colorectal anastomosis was successful and associated with a lower leakage rate.

The preparation and transplantation of adipose tissue derived stem cell (ASC) sheets onto insufficiently sutured colorectal anastomoses in a rat model is presented. This novel application shows that ASC sheets can reduce colorectal anastomosis leakage.

Trapianto di cellule staminali di tessuto adiposo-derivato foglio per ridurre le perdite dopo la colectomia parziale in un modello del ratto

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of ...

Implantation of hiPSC-derived Cardiac-muscle Patches after Myocardial Injury in a Guinea Pig Model

Due to the limited regeneration capacity of the heart in adult mammals, myocardial infarction results in an irreversible loss of cardiomyocytes. This loss of relevant amounts of heart muscle mass can lead to the heart failure. Besides heart transplantation, there is no curative treatment option for the end-stage heart failure. In times of organ donor shortage, organ independent treatment modalities are needed. Left-ventricular assist devices are a promising therapy option, however, especially as destination therapy, limited by its side-effects like stroke, infections and bleedings. In recent years, several cardiac repair strategies including stem cell injection, cardiac progenitors or myocardial tissue engineering have been investigated. Recent improvements in cell biology allow for the differentiation of large amounts of cardiomyocytes derived from human induced pluripotent stem cells (iPSC). One of the cardiac repair strategies currently under evaluation is to transplant artificial heart tissue. Engineered heart tissue (EHT) is a three-dimensional in vitro created cardiomyocyte network, with functional properties of native heart tissue. We have created EHT-patches from hiPSC derived cardiomyocytes. Here we present a protocol for the induction of left ventricular myocardial cryoinjury in a guinea pig, followed by implantation of hiPSC derived EHT on the left ventricular wall.

Here we present a protocol for the induction of left ventricular cryoinjury followed by the implantation of a cardiac muscle patch, derived from human iPS-cell cardiomyocytes in a guinea pig model.

Impianto di hiPSC-derivato del muscolo cardiaco patch dopo lesione del miocardio in un modello della cavia

Due to the limited regeneration capacity of the heart in adult mammals, myocardial infarction results in an irreversible loss of cardiomyocytes. This loss ...