Marco Bueter e Florian Seyfried sono stati sostenuti dalla Deutsche Forschungsgemeinschaft (DFG). Thomas A Lutz è stato sostenuto dal Fondo nazionale svizzero per la ricerca (FNS). Marco Bueter e Thomas Lutz A ulteriore beneficiare di un finanziamento dal National Institute of Health (NIH) e dal Centro Zurigo Integrativa Fisiologia Umana (ZIHP). Carel Le Roux W è stata sostenuta da un Dipartimento di Salute premio scienziato medico. Imperial College di Londra riceve il sostegno del regime di finanziamento NIHR Biomedical Research Centre.

1. Cura preoperatoria <ol><li> Togliere gli alimenti dal ratto durante la notte prima dell&#39;intervento. </li><li> Indurre l&#39;anestesia in camera con il 4-5% del flusso isoflurano e O2 di 2 l / min. </li><li> Shave addome da sterno al bacino con rasoio elettrico. </li><li> Mettere ratto anestetizzato in posizione supina sul pad riscaldamento isotermico. </li><li> Applicare una pomata oculare (Vitagel) prima di mettere il muso dei ratti in dell&#39;ogiva. </li><li> Mantenere l&#39;anestesia con isoflurano concentrazione del 2-3% e il flusso O2 di 2 l / min. </li><li> Disinfettare la cute con Betadine-Solution. </li><li> Confermare la profondità dell&#39;anestesia, con un pizzico pinze tra le dita della zampa posteriore. </li><li> Somministrare 5,7 mg / kg per via intraperitoneale Enrofloxacin come profilassi antibiotica perioperatoria, e 1 mg / kg Flunixin per l&#39;analgesia. </li></ol> 2. La laparotomia mediana <ol><li> Eseguire un&#39;incisione mediana con bisturi a partire proprio sotto processo xifoideo (lama n ° 10). </li><li> Mobilitare la pelle circonferenzialmente da Sottolinying muscoli addominali con le forbici Metzenbaum. </li><li> Aprire cavità addominale. </li><li> Installare divaricatori per facilitare l&#39;esposizione ottimale del campo operatorio. </li></ol> 3. Biliopancreatica e arti Alimentare <ol><li> Identificare dove duodeno o del digiuno prossimale passa sotto il colon. </li><li> Transetto del piccolo intestino di circa 10 cm aborally da qui e legare entrambe le estremità del budello (PDS 5-0). </li><li> Posizionare moncone prossimale delle due estremità in quadrante superiore sinistro dell&#39;addome come sarà successivamente usate per formare la parte biliopancreatica di Roux-en-Y ricostruzione. </li><li> Posizionare moncone distale delle due estremità in quadrante superiore destro dell&#39;addome come sarà successivamente usate per formare la parte alimentare di Roux-en-Y ricostruzione. </li></ol> 4. Jejuno-digiunostomia <ol><li> Identificare con valvola ileocoecal cieco e l&#39;ileo. </li><li> Seguire ileo per via orale per circa 25 cm. Il Jejuno-digiunostomia verrà inserito qui come punto di partenzadel canale comune di Roux-en-Y ricostruzione. </li><li> Recuperare arto biliopancreatica dal quadrante superiore sinistro dell&#39;addome e posizionarla vicino al canale comune dove si prevede di eseguire Jejuno-digiunostomia. </li><li> Biliopancreatica arto sicura e canale comune con ritenzione punto (PDS 6-0). </li><li> Incidere entrambi i cicli più di circa 10 mm con micro con le forbici. </li><li> Creare Jejuno-digiunostomia eseguendo da lato a lato anastomosi con suture interrotte PDS (6-0). </li><li> Primo lato dorsale e completa poi il lato ventrale della anastomosi. </li></ol> 5. Tasca gastrica <ol><li> Identificare giunzione gastro-esofagea. </li><li> Mobilitare questa area dissezione gastro-epatiche e gastro-splenica legamenti con le forbici Metzenbaum. </li><li> Spostare a sinistra dell&#39;arteria gastrica e fibre vagali di sinistra para-esofagea fascio lateralmente per evitare sanguinamenti maggiori e danni del nervo vagale quando piccola tasca gastrica è stato creato. </li><li> Esporre gastro-esofageo junction mettendo un batuffolo di cotone retro-oesophageally. </li><li> Coagulare piccoli vasi dello stomaco frontale mediante dispositivo di cauterizzazione disponibile in commercio - anche per prevenire emorragie. </li><li> Transetto stomaco circa 5 mm al di sotto gastro-esofageo giunzione creando tasca gastrica di una dimensione non superiore al 2-3% della dimensione originale utilizzando stomaco delicati, forbici curve. </li><li> Chiudi gastrico residuo (PDS 5-0). </li></ol> 6. Gastro-digiunostomia <ol><li> Recuperare arto alimentare dal quadrante superiore destro dell&#39;addome e posizionarla vicino al tasca gastrica. </li><li> Creare Gastro-digiunostomia eseguendo end-to-side anastomosi (PDS 7-0). </li><li> Prima lato posteriore completa e poi il lato anteriore di anastomosi. </li></ol> 7. Chiusura addominale <ol><li> Ridurre l&#39;anestesia, riducendo la concentrazione isoflurano a 1,5%. </li><li> Primo strato muscolare della parete addominale con suture continue PDS (4-0). </li><li> Somministrare 100 pl di 0,3 mg / mlsoluzione di buprenorfina per via sottocutanea per analgesia. </li><li> Inoltre ridurre la concentrazione di isoflurano fino a 1%. </li><li> Chiudere la pelle con punti staccati Vicryl (4-0). </li></ol> 8. Cura postoperatoria <ol><li> Fermare isoflurano e continuare con O2. </li><li> Somministrare 5 ml di soluzione salina calda per la sostituzione del fluido in tre depositi sottocutanei. </li><li> Rat posizione sotto la luce rossa fino al recupero completo. </li><li> Ritorna il ratto alla gabbia a casa. </li></ol> 9. Risultati rappresentativi Animali e abitazioni Ratti maschi Wistar (Harlan Laboratories Inc., Prugnolo, UK;. Elevage Janvier, Le-Genest-St Isola, Francia) di peso compreso tra 350 e 500 g sono stati alloggiati individualmente sotto un h 12/12 h ciclo luce-buio a temperatura ambiente di 21 ± 2 ° C. Acqua e standard di chow erano disponibili ad libitum, salvo diversa indicazione. Tutti gli esperimenti sono stati eseguiti sotto una licenza rilasciata dalla Casa Office, UK (PL70-6669) o approvati dal veterinario del Cantone di Zurigo, Svizzera. Tutti i ratti hanno dato una settimana di ambientamento prima di essere randomizzati a bypass gastrico o sham-operation. Dopo l&#39;intervento chirurgico, i ratti hanno ricevuto una dieta liquida per 3 giorni prima l&#39;accesso alla normalità Chow è stato reinstallato. Peso corporeo I dati del nostro modello bypass gastrico ratto sono coerenti con i precedenti risultati che la chirurgia di bypass gastrico è efficace per ridurre il peso corporeo e in particolare per mantenere la perdita di peso corporeo (Figura 2). Media pre-operatoria di peso corporeo di ratti utilizzati per il bypass gastrico e sham-operazioni è stata simile (sham: 433,4 ± 8,3 g vs bypass: 420,7 ± 8,4 g, p = 0,28). Cinque giorni dopo l&#39;intervento sham comandi azionati pesato significativamente più rispetto ai ratti di bypass gastrico (sham: 422,2 ± 8,3 g vs bypass: 374,7 ± 7,6 g, p &lt;0,001). Il giorno postoperatorio 60, la differenza del peso corporeo era quasi 170 g (sham: 533,2 ± 8,1 g vs bypass: 366,2 ± 10,8 g, p &lt;0,001). Assunzione di cibo L&#39;assunzione di cibo andamento simile come peso corporeo e ridotta nei ratti bypass gastrico rispetto al sham azionati ad libitum ratti alimentati. Figura 3 mostra il consumo medio giornaliera cibo per entrambi i gruppi (postoperatoria giorni 1-60). L&#39;assunzione di cibo giornaliera è stata costantemente inferiore dopo bypass gastrico (sham: 29,9 ± 0,2 g vs bypass: 25,7 ± 0,3 g, p &lt;0,001). Gut ormoni Sangue da tutti i ratti sono stati raccolti il giorno del termine dello studio 8,16. Gli animali avevano ad libitum l&#39;accesso al cibo la sera prima e sono stati decapitati all&#39;inizio del ciclo di luce giorno post-operatorio 60. Il sangue è stato ottenuto, immediatamente centrifugati a 3000 rpm per 10 minuti a 4 ° C, e conservato a -20 ° C fino a quando i campioni sono stati analizzati in duplicato in un unico passaggio. PYY immunoreattività simile era meavati con un saggio radioimmunologico specifico e sensibile, che misura, sia la lunghezza (PYY1-36) e il frammento (PYY3-36). GLP-1 è stata misurata mediante stabilito in-house radioimmunologici 17,18. Le differenze di assunzione di cibo può essere in parte spiegato da un aumento dei livelli plasmatici postprandiali di peptide YY (PYY) e il peptide glucagone-simile 1 (GLP-1) come ratti di bypass gastrico hanno mostrato livelli significativamente più elevati per PYY (Sham: 26 ± 2 pmol / L vs . bypass: 141 ± 14 pmol / L, p &lt;0,001) e GLP-1 (sham: 40 ± 5 pmol / L vs bypass: 215 ± 23 pmol / L, p &lt;0,001; Figura 4). <img alt="Figura 1" src="/files/ftp_upload/3940/3940fig1.jpg" /> Figura 1. Anatomia di bypass gastrico. Schema di anatomia piccolo intestino prima (A) e dopo (B) un&#39;operazione di bypass gastrico. Le diverse sfumature di rosso rappresentano approssimativamente corrispondenti segmenti del piccolo intestino con il rosso che rappresenta il mezzo di foregut(Esofago, stomaco, duodeno e digiuno prossimale), la luce rossa che rappresenta il midgut (digiuno prossimale e media, ileo prossimale) ed il rosso scuro che rappresenta la hindgut (ileo, cieco). <img alt="Figura 2" src="/files/ftp_upload/3940/3940fig2.jpg" /> Figura 2. . Perdita di peso corporeo dopo chirurgia di bypass gastrico nei ratti cambio del peso del corpo per un gruppo rappresentativo di ratti dopo bypass gastrico (-) (n = 52) e sham-operati ratti (-) (n = 52) durante un periodo di osservazione 60 giorni. I dati sono stati raccolti da pubblicazioni precedenti 6,8-10 e sono mostrati come valori medi ± SEM (*** = p &lt;0,001). <img alt="Figura 3" src="/files/ftp_upload/3940/3940fig3.jpg" /> Figura 3. Assunzione di cibo media dopo l&#39;intervento chirurgico di bypass gastrico nei ratti. Media giornaliera di alimenti di un gruppo rappresentativo di ratti dopo bypass gastrico (nero, n = 52) e sham-operati ratti (bianco, n = 52) per tutta laperiodo post-operatorio di 60 giorni. I dati sono stati raccolti da pubblicazioni precedenti 6,8-10 e sono mostrati come valori medi ± SEM. <img alt="Figura 3" src="/files/ftp_upload/3940/3940fig3.jpg" /> Figura 4. Postprandiale PYY e GLP-1 i livelli sierici dopo l&#39;intervento chirurgico di bypass gastrico post-prandiale nei ratti. PYY e il livello di GLP-1 nel siero per i ratti di bypass gastrico (nero, n = 18) e sham-operati ratti (bianco, n = 22). I dati sono stati raccolti da pubblicazioni precedenti 8,16 e sono mostrati come valori medi ± SEM.

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E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deoxycholate+is+an+important+releaser+of+peptide+YY+and+enteroglucagon+from+the+human+colon.">Deoxycholate is an important releaser of peptide YY and enteroglucagon from the human colon.</a> Gut. 34, 1219-1224 (1993).</li><li>Nakatani, H., Kasama, K., Oshiro, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Serum+bile+acid+along+with+plasma+incretins+and+serum+high-molecular+weight+adiponectin+levels+are+increased+after+bariatric+surgery.">Serum bile acid along with plasma incretins and serum high-molecular weight adiponectin levels are increased after bariatric surgery.</a> Metabolism. 58, 1400-1407 (2009).</li><li>Ashrafian, H., le Roux, C. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metabolic+surgery+and+gut+hormones+-+a+review+of+bariatric+entero-humoral+modulation.">Metabolic surgery and gut hormones - a review of bariatric entero-humoral modulation.</a> Physiol. Behav. 97, 620-631 (2009).</li><li>Choban, P. S., Flancbaum, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+Roux+limb+lengths+on+outcome+after+Roux-en-Y+gastric+bypass:+a+prospective,+randomized+clinical+trial.+Obes.">The effect of Roux limb lengths on outcome after Roux-en-Y gastric bypass: a prospective, randomized clinical trial. Obes.</a> Surg. 12, 540-545 (2002).</li><li>Gleysteen, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Five-year+outcome+with+gastric+bypass:+Roux+limb+length+makes+a+difference.">Five-year outcome with gastric bypass: Roux limb length makes a difference.</a> Surg. Obes. Relat. Dis. 5, 242-247 (2009).</li><li>Lee, S., Sahagian, K. G., Schriver, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relationship+between+varying+Roux+limb+lengths+and+weight+loss+in+gastric+bypass.">Relationship between varying Roux limb lengths and weight loss in gastric bypass.</a> Curr. Surg. 63, 259-263 (2006).</li><li>Hayes, M. R., Kanoski, S. E., De Jonghe, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+common+hepatic+branch+of+the+vagus+is+not+required+to+mediate+the+glycemic+and+food+intake+suppressive+effects+of+glucagon-like-peptide-1.">The common hepatic branch of the vagus is not required to mediate the glycemic and food intake suppressive effects of glucagon-like-peptide-1.</a> Am. J. Physiol. Regul. Integr. Comp. Physiol. 301, R1479-R1485 (2011).</li></ol>

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La Roux-en-Y bypass gastrico procedura per l&#39;uomo è stato descritto da Mason nel 1967 e modificato nella sua forma attuale Torres nel 1983 19. Attualmente, la procedura consiste in una piccola tasca gastrica e il bypass dell&#39;intestino prossimale di piccole dimensioni. Una illustrazione schematica dell&#39;anatomia pre-e post-operatoria è riportato in figura 1. Il bypass gastrico nell&#39;uomo induce e mantiene la perdita di peso corporeo di circa il 15-3...

<table border="1"><tbody><tr><td> Nome generico </td><td> Marca </td><td> Azienda </td><td> Numero di catalogo </td></tr><tr><td> Enrofloxacina </td><td> Baytril 2,5% </td><td> Provet AG </td><td> 1036 </td></tr><tr><td> Flunixin </td><td> Finadyne </td><td> Graeub </td><td> 908040 </td></tr><tr><td> Buprenorfina </td><td> Temgesic </td><td> Reckitt Benckiser </td><td> 138976 </td></tr><tr><td> Isoflurano </td><td> IsoFlo </td><td> Graeub </td><td> 902035 </td></tr><tr><td> Vitamina A </td><td> Vitagel </td><td> Bausch &amp; Lomb </td><td> 690 </td></tr><tr><td> Soluzione di iodio </td><td> Betadine </td><td> Mundipharma </td><td> 111141 </td></tr><tr><td> NaCl 0,9% </td><td> NaCl 0,9% </td><td> B. Braun </td><td> 534534 </td></tr></tbody></table> Tabella 1. Droga. <table border="1"><tbody><tr><td> Nome </td><td> Dimensione </td><td> Azienda </td><td> Numero di catalogo </td></tr><tr><td> PDS II </td><td> 4-0 </td><td> Ethicon </td><td> Z924H </td></tr><tr><td> PDS II </td><td> 5-0 </td><td> Ethicon </td><td> Z925H </td></tr><tr><td> PDS II </td><td> 6-0 </td><td> Ethicon </td><td> PUU2971E </td></tr><tr><td> PDS II </td><td> 7-0 </td><td> Ethicon </td><td> Z1370E </td></tr><tr><td> Vicryl </td><td> 4-0 </td><td> Ethicon </td><td> V451H </td></tr></tbody></table> Tabella 2. Punti di sutura. <table border="1"><tbody><tr><td> Nome </td><td> Azienda </td><td> Numero di catalogo </td></tr><tr><td> Scalpel maniglia No. 3 </td><td> Aesculap </td><td> BB073R </td></tr><tr><td> Lame di bisturi No. 10 </td><td> Swann-Morton </td><td> 0301 </td></tr><tr><td> Porta aghi </td><td> Aesculap </td><td> BM124R </td></tr><tr><td> Tessuto pinza </td><td> Aesculap </td><td> BD555R </td></tr><tr><td> Forbici Metzenbaum, diritto </td><td> Aesculap </td><td> BC022R </td></tr><tr><td> Metzenbaum forbici, curvo </td><td> Aesculap </td><td> BC023R </td></tr><tr><td> Forbici curve delicate, </td><td> Aesculap </td><td> BC061R </td></tr><tr><td> Pinza arteria, curvo </td><td> Aesculap </td><td> BH109R </td></tr><tr><td> Pinze arterie, curve, denti 1x2 </td><td> Aesculap </td><td> BH121R </td></tr><tr><td> Probe, doppio attacco </td><td> Aesculap </td><td> BN113R </td></tr><tr><td> Porta aghi Micro </td><td> Aesculap </td><td> FM 541R </td></tr><tr><td> Micro pinza </td><td> Aesculap </td><td> FM571R </td></tr><tr><td> Micro forbici </td><td> AESCulap </td><td> FM470R </td></tr><tr><td> Occhio cautery monouso </td><td> John Weiss internazionale </td><td> 0111122 </td></tr><tr><td> Cotton fioc </td><td> Hartmann AG </td><td> 9679369 </td></tr></tbody></table> Tabella 3. Attrezzature chirurgiche.

roux-en-y gastric bypass operation in rats

Attualmente, la terapia più efficace per il trattamento dell&#39;obesità patologica per indurre significativi e mantenuto la perdita di peso corporeo con un beneficio sulla mortalità provata è di 1,2 chirurgia bariatrica. Di conseguenza, si è registrato un costante aumento del numero di operazioni effettuate obesi in tutto il mondo negli ultimi anni, con la Roux-en-Y bypass gastrico (bypass gastrico) è la più comunemente eseguita funzionamento 3. In questo contesto, è importante comprendere i meccanismi fisiologici con cui bypass gastrico induce e mantiene la perdita di peso corporeo. Questi meccanismi non sono ancora pienamente compreso, ma possono includere la fame ridotti e una maggiore sazietà 4,5, l&#39;aumento della spesa di energia 6,7, la preferenza per il cibo modificato ad alto contenuto di grassi e zuccheri 8,9, il sale e il trattamento dell&#39;acqua alterata del rene 10 come così come alterazioni della flora intestinale 11. Tali cambiamenti osservati dopo bypass gastrico può almeno in parte derivano dal modo in cuila chirurgia altera l&#39;ambiente ormonale bypass gastrico perché aumenta il rilascio postprandiale di peptide-YY (PYY) e glucagone-simile-peptide-1 (GLP-1), ormoni che vengono rilasciati dall&#39;intestino in presenza di sostanze nutritive e che riducono mangiare 12. Negli ultimi due decenni numerosi studi utilizzando ratti sono stati eseguiti per investigare ulteriormente cambiamenti fisiologici dopo bypass gastrico. Il bypass gastrico modello di ratto ha dimostrato di essere uno strumento prezioso, non sperimentale, almeno per quanto imita da vicino il profilo temporale e l&#39;entità della perdita di peso umano, ma anche permette ai ricercatori di controllare e manipolare critici fattori anatomici e fisiologici, compreso l&#39;uso di controlli adeguati. Di conseguenza, esiste una vasta gamma di modelli di ratto di bypass gastrici disponibili in letteratura recensione altrove in dettaglio 13-15. La descrizione della tecnica chirurgica esatta di questi modelli varia ampiamente e differisce per esempio in termini di dimensioni sacchetto, artolunghezze, e la conservazione del nervo vagale. Se riferito, i tassi di mortalità sembrano variare da 0 a 35% 15. Inoltre, la chirurgia è stata effettuata quasi esclusivamente in ratti maschi di ceppi differenti ed età. Diete pre-e post-operatorio varia anche in modo significativo. Variazioni tecniche e sperimentali in modelli di bypass gastrico di ratto pubblicato complicare il confronto e l&#39;identificazione dei potenziali meccanismi fisiologici coinvolti nel bypass gastrico. Non vi è alcuna chiara evidenza che uno di questi modelli è superiore, ma c&#39;è un bisogno emergente per la standardizzazione delle procedure per ottenere dati omogenei e comparabili. Questo articolo si propone quindi di riassumere e discutere i dettagli tecnici e sperimentale del nostro modello validato e pubblicato rat bypass gastrico.

Watch this Scientific Journal Video about Roux-en-Y Gastric Bypass Operation in Rats at JoVE.com

Roux-en-Y Gastric Bypass Operation in Rats

Numerosi studi che utilizzano modelli di ratto di bypass gastrico sono stati recentemente condotti per scoprire i meccanismi fisiologici alla base di Roux-en-Y operazioni di bypass gastrico. Questo articolo mira a dimostrare e discutere i dettagli tecnici e sperimentale del nostro modello di ratto pubblicato bypass gastrico per capire vantaggi e limiti di questo strumento sperimentale.

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 ...

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JoVE Journal

Medicine

23.4K Views.  University Hospital Zürich. Numerosi studi che utilizzano modelli di ratto di bypass gastrico sono stati recentemente condotti per scoprire i meccanismi fisiologici alla base di Roux-en-Y operazioni di bypass gastrico. Questo articolo mira a dimostrare e discutere i dettagli tecnici e sperimentale del nostro modello di ratto pubblicato bypass gastrico per capire vantaggi e limiti di questo strumento sperimentale.

Video: Roux-en-Y Gastric Bypass Operation in Rats

Acute Myocardial Infarction in Rats

With heart failure leading the cause of death in the USA (Hunt), biomedical research is fundamental to advance medical treatments for cardiovascular diseases. Animal models that mimic human cardiac disease, such as myocardial infarction (MI) and ischemia-reperfusion (IR) that induces heart failure as well as pressure-overload (transverse aortic constriction) that induces cardiac hypertrophy and heart failure (Goldman and Tarnavski), are useful models to study cardiovascular disease. In particular, myocardial ischemia (MI) is a leading cause for cardiovascular morbidity and mortality despite controlling certain risk factors such as arteriosclerosis and treatments via surgical intervention (Thygesen). Furthermore, an acute loss of the myocardium following myocardial ischemia (MI) results in increased loading conditions that induces ventricular remodeling of the infarcted border zone and the remote non-infarcted myocardium. Myocyte apoptosis, necrosis and the resultant increased hemodynamic load activate multiple biochemical intracellular signaling that initiates LV dilatation, hypertrophy, ventricular shape distortion, and collagen scar formation. This pathological remodeling and failure to normalize the increased wall stresses results in progressive dilatation, recruitment of the border zone myocardium into the scar, and eventually deterioration in myocardial contractile function (i.e. heart failure). The progression of LV dysfunction and heart failure in rats is similar to that observed in patients who sustain a large myocardial infarction, survive and subsequently develops heart failure (Goldman). The acute myocardial infarction (AMI) model in rats has been used to mimic human cardiovascular disease; specifically used to study cardiac signaling mechanisms associated with heart failure as well as to assess the contribution of therapeutic strategies for the treatment of heart failure. The method described in this report is the rat model of acute myocardial infarction (AMI). This model is also referred to as an acute ischemic cardiomyopathy or ischemia followed by reperfusion (IR); which is induced by an acute 30-minute period of ischemia by ligation of the left anterior descending artery (LAD) followed by reperfusion of the tissue by releasing the LAD ligation (Vasilyev and McConnell). This protocol will focus on assessment of the infarct size and the area-at-risk (AAR) by Evan's blue dye and triphenyl tetrazolium chloride (TTC) following 4-hours of reperfusion; additional comments toward the evaluation of cardiac function and remodeling by modifying the duration of reperfusion, is also presented. Overall, this AMI rat animal model is useful for studying the consequence of a myocardial infarction on cardiac pathophysiological and physiological function.

The rat model of acute myocardial infarction (AMI) is useful to study the consequence of a MI on cardiac pathophysiological and physiological function.

Infarto acuto del miocardio in topi

With heart failure leading the cause of death in the USA (Hunt), biomedical research is fundamental to advance medical treatments for cardiovascular ...

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Medicina

Orthotopic Liver Transplantation in Rats

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat liver transplantation in 1979 by Kamada et al.3, this model has remained the gold standard despite various proposed alternative techniques4. Nevertheless, its broader use is limited by its steep learning curve5.
In this video paper, we show a simple and easy-to-establish revision of Kamada's two-cuff technique. The suprahepatic vena cava anastomosis is performed manually with a running suture, and the vena porta and infrahepatic vena cava anastomoses are performed utilizing a quick-linker cuff system6. Manufacturing the quick-linker kit is shown in a separate video paper.

We present an easy-to-establish revision of the classical two-cuff technique for orthotopic liver transplantation in rat.

Trapianto di fegato in ratti

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat ...

Intraoperative Gastroscopy for Tumor Localization in Laparoscopic Surgery for Gastric Adenocarcinoma

Determining resection margins for gastric cancer, which are not exposed to the serosal surface of the stomach, is the most important procedure during totally laparoscopic gastrectomy (TLG). The aim of this protocol is to introduce a procedure for intraoperative gastroscopy, in order to directly mark tumors during TLG for gastric cancer in the middle third of the stomach. Patients who were diagnosed with adenocarcinoma in the middle third of the stomach were enrolled in this case series. Before surgery, additional gastroscopy for tumor localization is not performed. Under general anesthesia, laparoscopic mobilization of the stomach is performed first. After the first portion of the duodenum is mobilized from the pancreas and clamped, the surgeon moves to the other side for the gastroscopic procedure. On the insertion of a gastroscope through the oral cavity into the stomach, 2 - 3 cc of indigo carmine is administered via an endoscopic injector into the gastric muscle layer at the proximal margin of the stomach. The location of stained serosa in the laparoscopic view is used to guide distal subtotal gastrectomy, however, total gastrectomy is performed if the tumor is too close to the esophagogastric junction. A specimen is sampled after distal gastrectomy to confirm sufficient length from resection margin to tumor before reconstruction. In our case series, all patients had tumor-free margins and required no additional resection. There was no morbidity related to the gastroscopic procedure, and the time required for the procedure has gradually decreased to about five minutes. Intraoperative gastroscopy for tumor localization is an accurate and tolerated method for gastric cancer patients undergoing totally laparoscopic distal gastrectomy.

In early gastric cancer, the aim of surgery is to precisely remove the distal stomach including the primary tumor. To do this, accurate localization of the tumor is crucial, especially in totally laparoscopic surgery. This protocol describes a procedure for intraoperative gastroscopy in totally laparoscopic subtotal gastrectomy.

Gastroscopia intraoperatoria per tumore Localizzazione in chirurgia laparoscopica per adenocarcinoma gastrico

Determining resection margins for gastric cancer, which are not exposed to the serosal surface of the stomach, is the most important procedure during ...

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. ...

Techniques of Sleeve Gastrectomy and Modified Roux-en-Y Gastric Bypass in Mice

Obesity is a major public health issue, with a prevalence of 4 to 28% for men and 6.2 to 36.5% for women in Europe (from 2003 to 2008). Morbid obesity is frequently associated with metabolic complications, such as type 2 diabetes, hypertension, and dyslipidemia, reducing life expectancy and quality. In the absence of any effective noninvasive treatments, bariatric surgery is a valuable therapeutic option for patients with morbid obesity (body mass index (BMI) &#62;40 kg/m2), leading to long-term, sustained weight loss and improvements in metabolic complications. However, the underlying cellular and molecular mechanisms sustaining the beneficial effects of bariatric surgery are not yet fully understood. Due to the numerous genetically-modified strains available, the mouse model is the most convenient animal model to explore the molecular mechanisms behind the pleiotropic beneficial effects of bariatric surgeries. Here, we detailed the optimized healthcare methods and surgical protocols in mice for the two most widely-used bariatric surgeries: the sleeve gastrectomy and the modified Roux-en-Y gastric bypass. Deciphering the molecular mechanisms underlying the therapeutic effects of bariatric surgeries offers the promise of identifying new therapeutics targets.

Bariatric surgery is the most efficient way to reduce body weight and the deadly metabolic complications (diabetes, obesity, and dyslipidemia) frequently associated with morbid obesity. Mouse models of bariatric surgery represent a unique asset for deciphering molecular mechanisms behind the beneficial effects of these surgeries on diabetes, hypertension, and dyslipidemia.

Tecniche di sleeve gastrectomy e modificata Roux-en-Y bypass gastrico nei topi

Obesity is a major public health issue, with a prevalence of 4 to 28% for men and 6.2 to 36.5% for women in Europe (from 2003 to 2008). Morbid obesity is ...

Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization and for minimizing organ damage resulting from prolonged extracorporal circulation. The goal of this paper was to demonstrate a fully functional and clinically relevant model of cardiopulmonary bypass in a mouse. We report on the device design, perfusion circuit optimization, and microsurgical techniques. This model is an acute model, which is not compatible with survival due to the need for multiple blood drawings. Because of the range of tools available for mice (e.g., markers, knockouts, etc.), this model will facilitate investigation into the molecular mechanisms of organ damage and the effect of cardiopulmonary bypass in relation to other comorbidities.

This paper describes how to perform cardiopulmonary bypass in mice. This novel model will facilitate the investigation of the molecular mechanisms involved in organ damage.

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 ...

One-anastomosis Gastric Bypass (OAGB) in Rats

The goal of this protocol is to set up a preclinical model of bariatric surgery and, more specifically, OAGB in obese rats. Based on this preclinical model, longitudinal studies can be carried out to provide an improved understanding of the mechanisms underlying the outcomes seen after bariatric surgery in humans. For this purpose, rats are operated on through a laparotomy under general anesthesia with isoflurane. First, the surgeon creates a long and tubular gastric pouch: after greater curve and hiatal dissection, the nonglandular stomach is stapled and removed. Then, the remaining stomach is also stapled in order to create a gastric tube and exclude the antrum of the stomach. After that, the surgeon performs a single end-to-side gastrojejunostomy 35 cm from the duodenojejunal angle. This limb length has been chosen in order to reproduce the same ratio between the biliopancreatic limb (BPL) and common limb (CL) length as in human bariatric surgery. The operation ends by aponeurotic and cutaneous closure. The early postoperative management consists of subcutaneous hydration, an intramuscular prophylactic antibiotic injection, a parietal injection of xylocaine, the administration of painkillers, and a progressive reintroduction of diet.

One-anastomosis Gastric Bypass OAGB in Rats

This protocol is for the performance of one-anastomosis gastric bypass (OAGB) on rat. The operator performs a long and tubular-stapled gastric pouch followed by hand-sewn anastomosis. For this model, the operator reproduces the same ratio between biliopancreatic and common limb in humans; therefore, the biliopancreatic limb measures 35 cm.

Esclusione gastrica di un-anastomosi (OAGB) in ratti

The goal of this protocol is to set up a preclinical model of bariatric surgery and, more specifically, OAGB in obese rats. Based on this preclinical ...

Preparation of Decellularized Kidney Scaffolds in Rats

Tissue engineering is a cutting-edge discipline in biomedicine. Cell culture techniques can be applied for regeneration of functional tissues and organs to replace diseased or damaged organs. Scaffolds are needed to facilitate the generation of three-dimensional organs or tissues using differentiated stem cells in vivo. In this report, we describe a novel method for developing vascularized scaffolds using decellularized rat kidneys. Eight-week-old Sprague-Dawley rats were used in this study, and heparin was injected into the heart to facilitate flow into the renal vessels, allowing heparin to perfuse into the renal vessels. The abdominal cavity was opened, and the left kidney was collected. The collected kidneys were perfused for 9 h using detergents, such as Triton X-100 and sodium dodecyl sulfate, to decellularize the tissue. Decellularized kidney scaffolds were then gently washed with 1% penicillin/streptomycin and heparin to remove cellular debris and chemical residues. Transplantation of stem cells with the decellularized vascular scaffolds is expected to facilitate the generation of new organs. Thus, the vascularized scaffolds may provide a foundation for tissue engineering of organ grafts in the future.

This protocol introduces a method to develop a scaffold using decellularized rat kidneys. The protocol includes decellularization and recellularization processes to confirm bioavailability. Decellularization is performed using Triton X-100 and sodium dodecyl sulfate.

Preparazione di impalcature renali decellularizzate nei ratti

Tissue engineering is a cutting-edge discipline in biomedicine. Cell culture techniques can be applied for regeneration of functional tissues and organs ...

A Protocol for Roux-en-Y Gastric Bypass in Rats using Linear Staplers

Roux-en-Y gastric bypass (RYGB) is commonly performed for the treatment of severe obesity and type 2 diabetes. However, the mechanism of weight loss and metabolic changes are not well understood. Multiple factors are thought to play a role, including reduced caloric intake, decreased nutrient absorption, increased satiety, the release of satiety-promoting hormones, shifts in bile acid metabolism, and alterations in the gut microbiota.
The rat RYGB model presents an ideal framework to study these mechanisms. Prior work on mouse models have had high mortality rates, ranging from 17 to 52%, limiting their adoption. Rat models demonstrate more physiologic reserve to surgical stimulus and are technically easier to adopt as they allow for the use of surgical staplers. One challenge with surgical staplers, however, is that they often leave a large gastric pouch which is not representative of RYGB in humans. 
In this protocol, we present a RYGB protocol in rats that result in a small gastric pouch using surgical staplers. Utilizing two stapler fires which remove the forestomach of the rat, we obtain a smaller gastric pouch similar to that following a typical human RYGB. Surgical stapling also results in better hemostasis than sharp division. Additionally, the forestomach of the rat does not contain any glands and its removal should not alter the physiology of RYGB.
Weight loss and metabolic changes in the RYGB cohort were significant compared to the sham cohort, with significantly lower glucose tolerance at 14 weeks. Furthermore, this protocol has an excellent survival of 88.9% after RYGB. The skills described in this protocol can be acquired without previous microsurgical experience. Once mastered, this procedure will provide a reproducible tool for studying the mechanisms and effects of RYGB.

Roux-en-Y gastric bypass (RYGB) is performed to treat obesity and diabetes. However, the mechanisms underlying RYGB's efficacy are not fully understood, and studies are limited by technical difficulty leading to high mortality in animal models. This article provides instructions on how to perform RYGB in rats with high success rates.

Un protocollo per il bypass gastrico Roux-en-Y nei ratti che utilizza cucitrici lineari

Roux-en-Y gastric bypass (RYGB) is commonly performed for the treatment of severe obesity and type 2 diabetes. However, the mechanism of weight loss and ...

Robotic Central Pancreatectomy with Roux-en-Y Pancreaticojejunostomy

Central pancreatectomy is a parenchyma-sparing alternative to distal pancreatectomy in patients with a benign or low-grade malignant tumor in the body of the pancreas. The aim of central pancreatectomy is to prevent postoperative life-long endocrine and exocrine insufficiency. The downside of central pancreatectomy is the high rate of postoperative pancreatic fistula, which is the main reason that many surgeons do not routinely use central pancreatectomy in eligible patients. Most studies report open or laparoscopic central pancreatectomy with a pancreatico-gastrostomy anastomosis in adults. This is the first description of a standardized approach to robotic central pancreatectomy with Roux-en-Y pancreaticojejunostomy reconstruction in an adolescent (16-year-old boy) with a pseudopapillary tumor in the body of the pancreas. The operation time was 248 min with 20 mL of blood loss. The postoperative course was uneventful except for the short-term medical treatment for a grade B pancreatic fistula. Robotic central pancreatectomy can be safely applied in selected patients in experienced centers.

Robotic central pancreatectomy may be used in selected patients in experienced centers. This protocol presents all steps and the feasibility of a robotic central pancreatectomy with Roux-en-Y pancreaticojejunostomy in a 16-year-old adolescent patient.

Pancreatectomia centrale robotica con Roux-en-Y Pancreaticojejunostomy

Central pancreatectomy is a parenchyma-sparing alternative to distal pancreatectomy in patients with a benign or low-grade malignant tumor in the body of ...