Questo lavoro è stato sostenuto dalla Deutsche Forschungsgemeinschaft (SFB 656, Münster, in Germania, i progetti C7 e C6) e il IZKF Münster (Core unità SMAP). Gli autori sono grati a Truc Van Le, Anne Kanzog, Ute Neugebauer, Wiebke Gottschlich e Priebe romana per un&#39;eccellente assistenza tecnica e di Daniel Burkert e Sven Fatum per la produzione di radiofarmaci.

1. Donor Recovery Organ <ol><li> Impostare il microscopio stereotassica, record di peso del 8-10 settimane di età ratto (donatore e ricevente di peso corporeo dovrebbe corrispondere). </li><li> Anestetizzare il topo donatore (Lewis Brown Norway F1, LBN F1) con ossigeno / inalazione isoflurano (isoflurano 4% / 2 L / min di ossigeno). Mantenere l&#39;anestesia abbassando isoflurano al 2-2,5%. </li><li> Posizionare il topo anestetizzato sul pad chirurgia. Fissare le estremità del ratto con del nastro adesivo sul tampone e applicare pomata oftalmica (Bepanthen, Bayer) agli occhi del ratto. </li><li> Shave e disinfettare l&#39;addome del ratto e di eseguire una incisione mediana ventrale dal pube fino al confine caudale del fegato. Esporre il rene sinistro e sue navi spostando attentamente l&#39;intestino per il lato destro. Posizionare un foglio sottile di garza sul rene sinistro e inumidire con soluzione salina isotonica riscaldata per prevenire l&#39;essiccamento. </li><li> Sezionare con cura il tessuto adiposo con Dumont angolati Pinze Tip (Dumont SS-45 Pinza Inox Medical, Belle Strumenti Scienza, Cat.No. 11.203-25) che avvolge l&#39;uretere sinistro. Evitare il contatto diretto con l&#39;uretere, invece, mobilitare con sufficiente tessuto adiposo circostante. </li><li> Separare la vena renale sinistra dall&#39;arteria renale utilizzando Dumont angolati pinzetta Tip. Rimuovere tutti i grassi e tessuto connettivo da entrambe le navi e cauterizzare i vasi surrenali e testicoli. </li><li> Sezionare aorta e vena cava inferiore (IVC) sopra e sotto l&#39;incrocio con l&#39;arteria renale sinistra e la vena, rispettivamente, e cauterizzare tutte le arterie in uscita. </li><li> Bloccare l&#39;aorta surrenale con un morsetto microchirurgico (micro Serrefines, Belle Strumenti Scienza, art 18055-04) sopra l&#39;arteria renale sinistra, il più vicino possibile al dell&#39;arteria mesenterica superiore. Legare l&#39;aorta sottorenale con seta chirurgico (5-0, Vömel, arte no. 14739) di circa 5 mm sotto l&#39;arteria renale sinistra. </li><li> Legare la IVC sottorenale con seta chirurgico (5-0, Vömel), e bloccare la surrenale IVCcon un morsetto microchirurgico (micro Serrefines, Fine strumenti per le scienze, art 18055-03). </li><li> Tagliare la vena renale il più vicino possibile alla IVC con le forbici sottili (Iris Forbici - ToughCut dritte 11,5 centimetri, Fine strumenti per le scienze, art 14058-11). </li><li> Perfusione Lentamente il rene in situ con 2 ml di soluzione di perfusione HTK ghiacciata (CUSTODIOL HTK, Dr. Franz Köhler Chemie) con l&#39;inserimento di una cannula (Microlance 3 27G ¾, BD, art 302200) in aorta sottorenale. Confermare che i cambiamenti di colore renali (ora oliva colorato). </li><li> Prima, transezione dell&#39;aorta surrenale, poi l&#39;aorta sottorenale e infine dell&#39;uretere più vicino possibile alla vescica urinaria. </li><li> Rimuovere il rene e il suo apporto vascolare con l&#39;uretere e conservare in soluzione di perfusione HTK sul ghiaccio. </li><li> Euthanize il topo donatore rimuovendo i morsetti vascolari e l&#39;escissione consecutivo di cuore in anestesia. </li></ol> 2. Destinatario Preparazione e trapiantozione <ol><li> Impostare il microscopio stereotassica, peso record di ratto. </li><li> Anestetizzare il ratto destinatario (Lewis) utilizzando isoflurano 4%. Mantenere l&#39;anestesia abbassando isoflurano al 2-2,5%. </li><li> Posizionare il topo anestetizzato sul pad chirurgia. Fissare le estremità del ratto con del nastro adesivo sul tampone e applicare pomata oftalmica per gli occhi il ratto `s. Monitorare e controllare la temperatura corporea interna utilizzando un sensore di temperatura rettale e un pad di riscaldamento. Durante la chirurgia controllare ripetitivamente temperatura, la respirazione e la frequenza cardiaca dell&#39;animale. </li><li> Shave e disinfettare l&#39;addome ed eseguire una incisione mediana ventrale dal pube fino al confine caudale del fegato. Utilizzare un bisturi forcutting la pelle al fine di minimizzare il trauma pelle. Esporre il rene sinistro e sue navi spostando l&#39;intestino per il lato destro. </li><li> Rimuovere con cautela la capsula renale utilizzando Dumont angolati pinzetta TIP e tamponi di cotone. </li><li> Occludere sinistra dell&#39;arteria renale, vena e uretere vicino renaleilo con due legature con filo chirurgico (Mersilene 4-0, Ethicon, cat. n. EH6732H). Asportare il maggior parte del rene, lasciando solo l&#39;ilo renale. Pulire la superficie di taglio con tamponi di cotone e di verificare la presenza di sanguinamento. Aggiungi un&#39;altra legatura, se necessario. </li><li> Sezionare con cura senza mezzi termini attraverso il tessuto connettivo che copre l&#39;aorta sottorenale e vena cava sul sito caudale con tamponi di cotone. </li><li> Separare ogni nervo visibile dalla aorta sottorenale e vena cava usando Dumont angolati pinzetta Tip. </li><li> Sezionare attentamente il foglio di tessuto connettivo tra l&#39;aorta sottorenale e vena cava, formando due aperture di 2-4 mm di lunghezza: Il primo sotto la ramificazione dei vasi testicolari, e la seconda sopra la ramificazione dei vasi iliaci comuni. Cauterizzare le arterie e le vene iliolumbar in mezzo. </li><li> Disegnare un unico filo chirurgico (Mersilene 0;. Ethicon, cat no.EH6665E) in ciascuna delle due aperture. Utilizzando questi fili tirare il sottorenaleaorta e vena cava delicatamente il lato destro dell&#39;animale per accedere ai vasi ramificazione sul sito dorsale. Cauterize qualsiasi di questi vasi tra le due aperture e rimuovere i fili chirurgici successivi. </li><li> Avanti, occludere l&#39;aorta e vena cava inferiore per l&#39;applicazione in senso orario di quattro morsetti di microchirurgia (Micro Serrefines, Belle Strumenti Scienza, art 18055-04) che cominciano in apertura superiore con l&#39;aorta e termina con la vena cava in apertura superiore (Micro Serrefines , Belle Strumenti Scienza, art 18055-03). </li><li> In seguito con attenzione perforare il finale superiore della parte isolata dell&#39;aorta con un ago (Microlance 3 27G ¾, BD, art 302200), inserire la lama inferiore di una forbice fine primavera (Spring Vännäs Forbici - 3 mm Lamelle, raffinata Scienza Strumenti, cat.no 15.000-00) attraverso l&#39;apertura puntura ed eseguire una breve incisione longitudinale. Lavare l&#39;aorta isolata con ghiacciata soluzione di perfusione HTK per rimuovere il sangue residuo. </li><li> Allo stesso modo, con attenzione perforareil finale inferiore della parte isolata della vena cava con un ago, inserire la lama inferiore di una multa forbice molla attraverso l&#39;apertura puntura ed eseguire una incisione longitudinale che termina poco sotto l&#39;incisione aortica. Ancora una volta, lavare il vaso per rimuovere il sangue residuo. </li><li> Rimuovere il trapianto di rene dalla soluzione di storage. E posizionarla sotto la precedente posizione del rene sinistro, e assicurarsi che sia orientata correttamente. </li><li> Controllare l&#39;orientamento della arteria renale innesti e garantire che non ci sono colpi di scena. Quindi posizionare un foglio sottile di garza sopra l&#39;innesto ed inumidire con soluzione di perfusione HTK ghiacciata per raffreddarlo, e per prevenire l&#39;essiccamento. </li><li> Fissare l&#39;apertura della parte surrenale del pezzo aortica in cui l&#39;arteria renale dell&#39;innesto si apre con filo chirurgico (Ethilon 9-0, Ethicon, cat n 2809G..) - Prima al superiore, quindi all&#39;estremità inferiore di l&#39;incisione aortica. Chiudere l&#39;incisione in senso orario con una sutura continua con filo chirurgico(Ethilon 9-0), Dumont angolati pinzetta punta ed un porta aghi Castroviejo curvo (Belle Strumenti Scienza, cat. N. 12061-01). Successivamente, coprire la prima sutura con tessuto connettivo attraverso una seconda sutura continua antiorario. Ora, solo la parte inferiore del pezzo aortica dell&#39;innesto rimane da chiudere. Regolarmente raffreddare l&#39;innesto con ghiacciata soluzione di perfusione HTK. </li><li> Profumato l&#39;innesto con 1 ml di ghiacciata soluzione di perfusione HTK attraverso un ago smussato inserito nell&#39;estremità aperta del pezzo aortica innesti convalidare la tenuta e l&#39;integrità della sutura, così come la rimozione di sangue rimanente nel trapianto. Successivamente, legare l&#39;estremità aperta del pezzo aortica con seta chirurgica (5-0, Vömel). </li><li> Controllare l&#39;orientamento della vena renale innesti e garantire che non ci sono colpi di scena. </li><li> Fissare l&#39;apertura della vena dell&#39;innesto con filo chirurgico (Ethilon 9-0) - prima la tomaia, poi all&#39;estremità inferiore della incisione alla vena cava. Chiudere l&#39;incisionein senso orario con una sutura continua con filo chirurgico (Ethilon 9-0), Dumont angolati pinzetta punta ed un porta ago curvo Castroviejo. Regolarmente raffreddare l&#39;innesto con ghiacciata soluzione di perfusione HTK. </li><li> Antiorario rimuovere le fascette di microchirurgia, a cominciare dalla vena cava superiore morsetto. Dopo aver rimosso l&#39;ultimo morsetto aortica, stop attentamente emorragia finalmente mite con tamponi di cotone. Dopo pochi secondi l&#39;innesto diventa roseo-tinta, e contrazioni uretric dovrebbe apparire. </li><li> Per l&#39;inserimento del innesti dell&#39;uretere in vescica `s il destinatario rimuovere con attenzione una piccola zona della muscolatura nella parte superiore della vescica utilizzando un Vannas Primavera Forbice (Studente Vannas Primavera Scissor Diritto, fine Strumenti di scienze, Cat.No. 91.500-09 ). Quindi, rimuovere il tessuto circostante (per lo più grasso) da dell&#39;uretere innesto utilizzando Dumont angolati pinzetta Tip. Pertanto, afferrare con attenzione la punta dell&#39;uretere e delicatamente separare il circostante grasso, tessuto connettivo e vasi incorporati froM essa tirando in senso opposto. Fissare la punta uretric al termine di un filo chirurgico (Prolene 6-0, Ethicon, cat. No. 8697H) e perforare la parte superiore della vescica con l&#39;ago inserito precedentemente preparato. Tirare l&#39;uretere attraverso la vescica e uscirne alla base attraverso una seconda perforazione. </li><li> Fissare la precedenza dagli ureteri separato tessuto nella parte superiore della vescica con filo chirurgico (Ethilon 9-0). Tagliare la punta del uretere con il filo chirurgico annesso e tirare l&#39;uretere nella vescica spingendo delicatamente dal basso. Successivamente, chiudere la seconda puntura vescicale con filo chirurgico (Ethilon 9-0). </li><li> Chiudere la parete addominale e la pelle da due suture continue ma separate utilizzando filo di sutura chirurgico (Mersilene 0). Disinfettare la ferita con Pividon Iodio e amministrare buprenorfina (0,1 mg per kg / BW / bid) sc per tre giorni dopo l&#39;intervento chirurgico per controllare il dolore delle ferite. </li></ol> 3. Imaging Allograft Rifiuto<ol><li> Anestetizzare ratto non a digiuno con isoflurano 2-2,5%. </li><li> 30 MBq FDG (30 MBq FDG in 0,1 ml 0,9% NaCl) sono somministrati iv tramite cateterizzazione una vena della coda laterale utilizzando un catetere G 24 (Braun, Introcan, 4.252.500-01). In seguito, spurgare il catetere con 0,9 ml di soluzione di NaCl allo 0,9%. </li><li> Lasciare il topo in un dispositivo di immobilizzazione senza anestesia fino all&#39;inizio della scansione e idratare l&#39;animale da iniezione endovenosa di 1 ml di soluzione di NaCl allo 0,9% ogni ora. </li><li> Rat ri-anestetizzare con isoflurano 2% subito prima che la ricerca inizi. </li><li> Mettere ratto anestetizzato in una ad alta risoluzione da camera-based multi-filo animale PET fotocamera quadHIDAC (Oxford Positron Systems Ltd, Oxford, UK). La risoluzione spaziale della PET scanner è di 1,0 mm ed è costante su tutta FOV (diametro, 165 mm; lunghezza assiale di 280 mm). Monitoraggio e controllo della temperatura corporea durante la scansione utilizzando un sensore di temperatura rettale e l&#39;anestesia controllo con un ossimetro di impulso. </li><li> Avviare acquisizione dinamica di 60 min partenza 180 min dopo la FDG-iniezione per ridurre l&#39;accumulo di tracciante nei reni causati dalla escrezione renale di FDG. </li><li> Applicare 5 MBq di 18 F-fluoro iv ed eseguire un&#39;altra scansione PET immediatamente dopo l&#39;iniezione 18 F-fluoro per 60 min senza spostare la posizione del ratto nello scanner per l&#39;identificazione di parenchima renale e per il calcolo del 18 F-spazio 5. Ricostruire le immagini da entrambe le scansioni. Tracciare manualmente un volume di interesse (VOI) sulle immagini ricostruite 2 min dopo l&#39;iniezione di 18 F-fluoruro (perfusione-fase) intorno corteccia renale e trasferire il VOI alle immagini FDG. Escludere attentamente la pelvi renale da VOI. Ricostruire le immagini da entrambe le scansioni e tracciare manualmente un volume di interesse (VOI) intorno corteccia renale. Dati list-mode sono stati ricostruiti in immagini con una dimensione voxel di 0,4 × 0,4 × 0,4 millimetri 3. Escludere attentamente la pelvi renale da VOI. </li><li> Calcola FDG per il rapporto of conteggi totali e il volume e calcolare la percentuale della dose iniettata (% ID). Abbiamo usato MATLAB (versione R2011b, MathWorks, Natick, MA, USA) e di imaging tomografico (TIM) Versione 2.8 per l&#39;analisi delle immagini. </li></ol>

<ol>
	<li>Wu, O., Levy, A. R., Briggs, A., Lewis, G., Jardine, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+rejection+and+chronic+nephropathy:+a+systematic+review+of+the+literature.">Acute rejection and chronic nephropathy: a systematic review of the literature.</a> Transplantation. 87, 1330-1339 (2009).</li><li>Miller, B. F., Gonzales, E., Wilchins, L. J., Nathan, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kidney+transplantation+in+the+rat.">Kidney transplantation in the rat.</a> Nature. 194, 309-310 (1962).</li><li>Reuter, S., Schn&#246;ckel, U., Schr&#246;ter, R., Schober, O., Pavenst&#228;dt, H., Sch&#228;fers, M., Gabri&#235;ls, G., Schlatter, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-invasive+imaging+of+acute+renal+allograft+rejection+in+rats+using+small+animal+F-FDG-PET.">Non-invasive imaging of acute renal allograft rejection in rats using small animal F-FDG-PET.</a> PLoS. One. 4, e5296 (2009).</li><li>Reuter, S., Schn&#246;ckel, U., Edemir, B., Schr&#246;ter, R., Kentrup, D., Pavenst&#228;dt, H., Schober, O., Schlatter, E., Gabri&#235;ls, G., Sch&#228;fers, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Potential+of+noninvasive+serial+assessment+of+acute+renal+allograft+rejection+by+18F-FDG+PET+to+monitor+treatment+efficiency.">Potential of noninvasive serial assessment of acute renal allograft rejection by 18F-FDG PET to monitor treatment efficiency.</a> J. Nucl. Med. 51, 1644-1652 (2010).</li><li>Schn&#246;ckel, U., Reuter, S., Stegger, L., Schlatter, E., Sch&#228;fers, K. P., Hermann, S., Schober, O., Gabri&#235;ls, G., Sch&#228;fers, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+18F-fluoride+small+animal+PET+to+noninvasively+assess+renal+function+in+rats.">Dynamic 18F-fluoride small animal PET to noninvasively assess renal function in rats.</a> Eur. J. Nucl. Med. Mol. Imaging. 35, 2267-2274 (2008).</li><li>Grabner, A., Schn&#246;ckel, U., Kentrup, D., Sch&#228;fers, M., Reuter, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strategies+for+Non-Invasive+Molecular+Imaging+of+Acute+Allograft+Rejection+by+Gamma+Scintigraphy+and+Positron+Emission+Tomography.">Strategies for Non-Invasive Molecular Imaging of Acute Allograft Rejection by Gamma Scintigraphy and Positron Emission Tomography.</a> Curr. Radiopharm. 4, 10-23 (2011).</li><li>Racusen, L. C., Solez, K., Colvin, R. B., Bonsib, S. M., Castro, M. C., Cavallo, T., Croker, B. P., Demetris, A. J., Drachenberg, C. B., Fogo, A. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Banff+97+working+classification+of+renal+allograft+pathology.">The Banff 97 working classification of renal allograft pathology.</a> Kidney Int. 55, 713-723 (1999).</li><li>Edemir, B., Reuter, S., Borgulya, R., Schr&#246;ter, R., Neugebauer, U., Gabri&#235;ls, G., Schlatter, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+rejection+modulates+gene+expression+in+the+collecting+duct.">Acute rejection modulates gene expression in the collecting duct.</a> J. Am. Soc. Nephrol. 19, 538-546 (2008).</li><li>Velic, A., Gabri&#235;ls, G., Hirsch, J. R., Schr&#246;ter, R., Edemir, B., Paasche, S., Schlatter, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+rejection+after+rat+renal+transplantation+leads+to+downregulation+of+Na++and+water+channels+in+the+collecting+duct.">Acute rejection after rat renal transplantation leads to downregulation of Na+ and water channels in the collecting duct.</a> Am. J. Transplant. 5, 1276-1285 (2005).</li><li>Reuter, S., Velic, A., Edemir, B., Schr&#246;ter, R., Pavenst&#228;dt, H., Gabri&#235;ls, G., Bleich, M., Schlatter, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protective+role+of+NHE-3+inhibition+in+rat+renal+transplantation+undergoing+acute+rejection.">Protective role of NHE-3 inhibition in rat renal transplantation undergoing acute rejection.</a> Pflugers Arch. 456, 1075-1084 (2008).</li></ol>

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L&#39;imaging FDG-PET è una nuova opzione per la diagnosi di rigetto acuto. A causa della sua natura non invasiva e specifici, FDG-PET è vantaggi rispetto alla diagnostica classica da Core ago biopsia. In contrasto con la dimensione del campione limitata di una biopsia, analisi FDG-PET tutta innesto. Inoltre, si può applicare a pazienti in terapia anticoagulante, e si può effettuare misure di PET ripetutamente ad esempio per monitorare l&#39;efficienza di trattamento 4. Inoltre, abbiamo già dimo...

<table border="1">
 <tbody>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Equipment</td>
 </tr>
 <tr>
 <td>Mathieu Needle Holder - 14 cm</td>
 <td>Fine Science Tools</td>
 <td>12010-14</td>
 <td></td>
 </tr>
 <tr>
 <td>Castroviejo Micro Needle Holder - 9 cm</td>
 <td>Fine Science Tools</td>
 <td>12061-01</td>
 <td></td>
 </tr>
 <tr>
 <td>Surgical Scissors - Sharp_Blunt</td>
 <td>Fine Science Tools</td>
 <td>14001-12</td>
 <td></td>
 </tr>
 <tr>
 <td>Iris Scissors - ToughCut Straight 11.5 cm</td>
 <td>Fine Science Tools</td>
 <td>14058-11</td>
 <td></td>
 </tr>
 <tr>
 <td>Student Vannas Spring Scissors </td>
 <td>Fine Science Tools</td>
 <td>91500-09</td>
 <td></td>
 </tr>
 <tr>
 <td>Vannas Spring Scissors - 3 mm Blades</td>
 <td>Fine Science Tools</td>
 <td>15000-00</td>
 <td></td>
 </tr>
 <tr>
 <td>Student Tissue Forceps - 1x2 Teeth 12 cm</td>
 <td>Fine Science Tools</td>
 <td>91121-12</td>
 <td></td>
 </tr>
 <tr>
 <td>Dumont SS-45 Forceps - Inox Medical</td>
 <td>Fine Science Tools</td>
 <td>11203-25</td>
 <td></td>
 </tr>
 <tr>
 <td>Micro-Serrefine Clip Applicator with Lock</td>
 <td>Fine Science Tools</td>
 <td>18056-14</td>
 <td></td>
 </tr>
 <tr>
 <td>Micro-Serrefine 6 mm x 1 mm</td>
 <td>Fine Science Tools</td>
 <td>18055-03</td>
 <td></td>
 </tr>
 <tr>
 <td>Micro-Serrefine 4 mm x 0.75 mm</td>
 <td>Fine Science Tools</td>
 <td>18055-04</td>
 <td></td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Reagent</td>
 </tr>
 <tr>
 <td>Isoflurane (e.g. Forene 100% v/v)</td>
 <td>Abott</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>cutane antiseptic (e.g. Octeniderm) </td>
 <td>Sch&uuml;lke</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Povidone Iodine (e.g. Betaisodona)</td>
 <td>Mundipharma</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>ophthalmic ointment (e.g. Bepanthen)</td>
 <td>Bayer</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Buprenorphin (e.g. Temgesic)</td>
 <td>RB Pharmaceuticals</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>HTK perfusion solution (e.g. CUSTODIOL HTK)</td>
 <td>Dr. Franz K&ouml;hler Chemie</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>surgical thread Mersilene 0</td>
 <td>Ethicon</td>
 <td>EH6665E</td>
 <td></td>
 </tr>
 <tr>
 <td>surgical thread Mersilene 4-0</td>
 <td>Ethicon</td>
 <td>EH6732H</td>
 <td></td>
 </tr>
 <tr>
 <td>surgical thread Prolene 6-0</td>
 <td>Ethicon</td>
 <td>8697H</td>
 <td></td>
 </tr>
 <tr>
 <td>surgical thread Ethilon 9-0</td>
 <td>Ethicon</td>
 <td>2809G</td>
 <td></td>
 </tr>
 <tr>
 <td>surgical silk 5-0</td>
 <td>V&ouml;mel</td>
 <td>14739</td>
 <td></td>
 </tr>
 <tr>
 <td>Canula (e.g. Microlance 3, 27G &frac34;)</td>
 <td>BD</td>
 <td>302200</td>
 <td></td>
 </tr>
 </tbody>
</table>

non-invasive imaging of acute allograft rejection after rat renal transplantation using 18f-fdg pet

Il numero di pazienti con malattia renale allo stadio terminale, e il numero di riceventi trapianto allogenico di rene aumenta continuamente. Gli episodi di rigetto acuto cellulare (AR) sono un fattore prognostico negativo per la sopravvivenza del trapianto a lungo termine, e la sua diagnosi tempestiva è fondamentale per la funzionalità dell&#39;organo trapiantato 1. Allo stato attuale, AR può essere definitivamente diagnosticata solo mediante biopsia core-ago, che, come un metodo invasivo, scopre significativo rischio di lesioni o addirittura la perdita dell&#39;innesto. Inoltre, le biopsie non sono fattibili in pazienti che assumono farmaci anticoagulanti e il sito di campionamento limitato di questa tecnica può portare a risultati falsi negativi se l&#39;AR è focale o irregolare. Di conseguenza, questo ha dato luogo a una continua ricerca di nuovi metodi di rilevazione AR, che spesso deve essere fatto in animali, compreso l&#39;uso di vari modelli di trapianto. Dall&#39;inizio degli anni &#39;60 il trapianto renale ratto è un metodo sperimentale consolidata per l&#39;esazione e l&#39;analisi di AR 2. Noi qui presenti oltre piccola tomografia a emissione di positroni animali (PET) con 18 F-fluorodeossiglucosio (FDG) per valutare AR in un ratto uninephrectomized renale modello di trapianto allogenico e proporre innesto di imaging FDG-PET come una nuova opzione per un non-invasiva, specifica e la diagnosi precoce di AR anche per la situazione umana 3. Inoltre, questo metodo può essere applicato per il follow-up per migliorare il monitoraggio del rigetto del trapianto 4.

Istologia Durante AR leucociti, cioè soprattutto linfociti T sono reclutati nel trapianto, mentre la gravità del rigetto viene riflessa dal grado di infiammazione. Nel periodico-Acid-Schiff (PAS) colorazione raffigurato qui (figura 1), il trapianto renale mostra significativi segni istologici di AR, ovvero glomerulite, tubulitis, endothelialitis e infiltrazione innesto (figura 1, ATX Pod4) (POD = giorno post-operatorio) mentre segnali di rigetto sono...

Watch this Scientific Journal Video about Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET at JoVE.com

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

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

Siamo qui presentiamo un modello di trapianto di rene di ratto per valutare in modo non invasivo rigetto acuto utilizzando la tomografia ad emissione di positroni con 18F-fluorodeossiglucosio.

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 ...

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

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16.9K Views.  University of Münster. Siamo qui presentiamo un modello di trapianto di rene di ratto per valutare in modo non invasivo rigetto acuto utilizzando la tomografia ad emissione di positroni con 18F-fluorodeossiglucosio.

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

Non-invasive Optical Measurement of Cerebral Metabolism and Hemodynamics in Infants

Perinatal brain injury remains a significant cause of infant mortality and morbidity, but there is not yet an effective bedside tool that can accurately screen for brain injury, monitor injury evolution, or assess response to therapy. The energy used by neurons is derived largely from tissue oxidative metabolism, and neural hyperactivity and cell death are reflected by corresponding changes in cerebral oxygen metabolism (CMRO2). Thus, measures of CMRO2 are reflective of neuronal viability and provide critical diagnostic information, making CMRO2 an ideal target for bedside measurement of brain health.
Brain-imaging techniques such as positron emission tomography (PET) and single-photon emission computed tomography (SPECT) yield measures of cerebral glucose and oxygen metabolism, but these techniques require the administration of radionucleotides, so they are used in only the most acute cases.
Continuous-wave near-infrared spectroscopy (CWNIRS) provides non-invasive and non-ionizing radiation measures of hemoglobin oxygen saturation (SO2) as a surrogate for cerebral oxygen consumption. However, SO2 is less than ideal as a surrogate for cerebral oxygen metabolism as it is influenced by both oxygen delivery and consumption. Furthermore, measurements of SO2 are not sensitive enough to detect brain injury hours after the insult 1,2, because oxygen consumption and delivery reach equilibrium after acute transients 3. We investigated the possibility of using more sophisticated NIRS optical methods to quantify cerebral oxygen metabolism at the bedside in healthy and brain-injured newborns. More specifically, we combined the frequency-domain NIRS (FDNIRS) measure of SO2 with the diffuse correlation spectroscopy (DCS) measure of blood flow index (CBFi) to yield an index of CMRO2 (CMRO2i) 4,5.
With the combined FDNIRS/DCS system we are able to quantify cerebral metabolism and hemodynamics. This represents an improvement over CWNIRS for detecting brain health, brain development, and response to therapy in neonates. Moreover, this method adheres to all neonatal intensive care unit (NICU) policies on infection control and institutional policies on laser safety. Future work will seek to integrate the two instruments to reduce acquisition time at the bedside and to implement real-time feedback on data quality to reduce the rate of data rejection.

We combined frequency-domain near-infrared spectroscopy measures of cerebral hemoglobin oxygenation with diffuse correlation spectroscopy measures of cerebral blood flow index to estimate an index of oxygen metabolism. We tested the utility of this measure as a bedside screening tool to evaluate the health and development of the newborn brain.

Non invasiva di misurazione ottica del metabolismo cerebrale ed emodinamica nei neonati

Perinatal brain injury remains a significant cause of infant mortality  and morbidity, but there is not yet an effective bedside tool that can  accurately ...

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Transplantation into the Anterior Chamber of the Eye for Longitudinal, Non-invasive In vivo Imaging with Single-cell Resolution in Real-time

Intravital imaging has emerged as an indispensable tool in biological research. In the process, many imaging techniques have been developed to study different biological processes in animals non-invasively. However, a major technical limitation in existing intravital imaging modalities is the inability to combine non-invasive, longitudinal imaging with single-cell resolution capabilities. We show here how transplantation into the anterior chamber of the eye circumvents such significant limitation offering a versatile experimental platform that enables non-invasive, longitudinal imaging with cellular resolution in vivo. We demonstrate the transplantation procedure in the mouse and provide representative results using a model with clinical relevance, namely pancreatic islet transplantation. In addition to enabling direct visualization in a variety of tissues transplanted into the anterior chamber of the eye, this approach provides a platform to screen drugs by performing long-term follow up and monitoring in target tissues. Because of its versatility, tissue/cell transplantation into the anterior chamber of the eye not only benefits transplantation therapies, it extends to other in vivo applications to study physiological and pathophysiological processes such as signal transduction and cancer or autoimmune disease development.

Transplantation into the Anterior Chamber of the Eye for Longitudinal, Non-invasive In vivo Imaging with Single-cell Resolution in Real-time

A new approach combining intraocular transplantation and confocal microscopy enables longitudinal, non-invasive real-time imaging with single-cell resolution within grafted tissues in vivo. We demonstrate how to transplant pancreatic islets into the anterior chamber of the mouse eye.

Trapianto nella camera anteriore dell&#39;occhio per longitudinale, non invasiva<em&gt; In vivo</em&gt; Imaging con Single-cella di risoluzione in tempo reale

Intravital imaging has emerged as an indispensable tool in biological research.  In the process, many imaging techniques have been developed to study ...

Non-invasive Imaging and Analysis of Cerebral Ischemia in Living Rats Using Positron Emission Tomography with 18F-FDG

Stroke is the third leading cause of death among Americans 65 years of age or older1. The quality of life for patients who suffer from a stroke fails to return to normal in a large majority of patients2, which is mainly due to current lack of clinical treatment for acute stroke. This necessitates understanding the physiological effects of cerebral ischemia on brain tissue over time and is a major area of active research. Towards this end, experimental progress has been made using rats as a preclinical model for stroke, particularly, using non-invasive methods such as 18F-fluorodeoxyglucose (FDG) coupled with Positron Emission Tomography (PET) imaging3,10,17. Here we present a strategy for inducing cerebral ischemia in rats by middle cerebral artery occlusion (MCAO) that mimics focal cerebral ischemia in humans, and imaging its effects over 24 hr using FDG-PET coupled with X-ray computed tomography (CT) with an Albira PET-CT instrument. A VOI template atlas was subsequently fused to the cerebral rat data to enable a unbiased analysis of the brain and its sub-regions4. In addition, a method for 3D visualization of the FDG-PET-CT time course is presented. In summary, we present a detailed protocol for initiating, quantifying, and visualizing an induced ischemic stroke event in a living Sprague-Dawley rat in three dimensions using FDG-PET.

Non-invasive Imaging and Analysis of Cerebral Ischemia in Living Rats Using Positron Emission Tomography with 18F-FDG

Brain damage resulting from cerebral ischemia may be non-invasively imaged and studied in rats using pre-clinical positron emission tomography coupled with the injectable radioactive probe, 18F-fluorodeoxyglucose. Further, the use of modern software tools that include volume of interest (VOI) brain templates dramatically increase the quantitative information gleaned from these studies.

Imaging non invasiva e analisi di ischemia cerebrale in ratti Living Utilizzando Positron Emission Tomography con<sup&gt; 18</sup&gt; F-FDG

Stroke is the third leading cause of death among Americans 65 years of age or older1. The quality of life for patients who suffer from a stroke fails to ...

Murine Model for Non-invasive Imaging to Detect and Monitor Ovarian Cancer Recurrence

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of care consisting of surgical debulking and combination chemotherapy consisting of platinum and taxane compounds, almost 90% of patients recur within a few years. In these patients the development of chemoresistant disease limits the efficacy of currently available chemotherapy agents and therefore contributes to the high mortality. To discover novel therapy options that can target recurrent disease, appropriate animal models that closely mimic the clinical profile of patients with recurrent ovarian cancer are required. The challenge in monitoring intra-peritoneal (i.p.) disease limits the use of i.p. models and thus most xenografts are established subcutaneously. We have developed a sensitive optical imaging platform that allows the detection and anatomical location of i.p. tumor mass. The platform includes the use of optical reporters that extend from the visible light range to near infrared, which in combination with 2-dimensional X-ray co-registration can provide anatomical location of molecular signals. Detection is significantly improved by the use of a rotation system that drives the animal to multiple angular positions for 360 degree imaging, allowing the identification of tumors that are not visible in single orientation. This platform provides a unique model to non-invasively monitor tumor growth and evaluate the efficacy of new therapies for the prevention or treatment of recurrent ovarian cancer.

We describe a non-invasive animal imaging platform that allows the detection, quantification, and monitoring of ovarian cancer growth and recurrence. This intra-peritoneal xenograft model mimics the clinical profile of patients with ovarian cancer.

Modello murino per l&#39;imaging non invasivo per rilevare e monitorare Ovarian Cancer Ricorrenza

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of ...

Mouse Kidney Transplantation: Models of Allograft Rejection

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely replicates both the technical and pathological processes that occur in human renal transplantation. Although mouse models of allogeneic rejection in organs other than the kidney exist, and are more technically feasible, there is evidence that different organs elicit disparate rejection modes and dynamics, for instance the time course of rejection in cardiac and renal allograft differs significantly in certain strain combinations. This model is an attractive tool for many reasons despite its technical challenges. As inbred mouse strain haplotypes are well characterized it is possible to choose donor and recipient combinations to model acute allograft rejection by transplanting across MHC class I and II loci. Conversely by transplanting between strains with similar haplotypes a chronic process can be elicited were the allograft kidney develops interstitial fibrosis and tubular atrophy. We have modified the surgical technique to reduce operating time and improve ease of surgery, however a learning curve still needs to be overcome in order to faithfully replicate the model. This study will provide key points in the surgical procedure and aid the process of establishing this technique.

Here, we present a protocol to study the immunology of rejection. The surgical model presented reports a short operating time and a concise technique. Depending on the donor-recipient strain combination, the transplanted kidney may develop acute cellular rejection or chronic allograft damage, defined by interstitial fibrosis and tubular atrophy.

Mouse trapianto renale: Modelli di Allograft Rifiuto

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely ...

Determining Glucose Metabolism Kinetics Using 18F-FDG Micro-PET/CT

This paper describes the use of 18F-FDG and micro-PET/CT imaging to determine in vivo glucose metabolism kinetics in mice (and is transferable to rats). Impaired uptake and metabolism of glucose in multiple organ systems due to insulin resistance is a hallmark of type 2 diabetes. The ability of this technique to extract an image-derived input function from the vena cava using an iterative deconvolution method eliminates the requirement of the collection of arterial blood samples. Fitting of tissue and vena cava time activity curves to a two-tissue, three compartment model permits the estimation of kinetic micro-parameters related to the 18F-FDG uptake from the plasma to the intracellular space, the rate of transport from intracellular space to plasma and the rate of 18F-FDG phosphorylation. This methodology allows for multiple measures of glucose uptake and metabolism kinetics in the context of longitudinal studies and also provides insights into the efficacy of therapeutic interventions.

Determining Glucose Metabolism Kinetics Using 18F-FDG Micro-PET/CT

This study describes a protocol that uses 18F-FDG and positron emission tomography/computed tomography (PET/CT) imaging, together with kinetic modelling, to quantify the in vivo, real-time uptake of 18F-FDG into tissues.

Determinazione metabolismo del glucosio Cinetica Utilizzando<sup&gt; 18</sup&gt; F-FDG micro-PET / TC

This paper describes the use of 18F-FDG and micro-PET/CT imaging to determine in vivo glucose metabolism kinetics in mice (and is transferable to rats). ...

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 ...

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

In endothermic animals, brown adipose tissue (BAT) is activated to produce heat for defending body temperature in response to cold. BAT&#39;s ability to expend energy has made it a potential target for novel therapies to ameliorate obesity and associated metabolic disorders in humans. Though this tissue has been well studied in small animals, BAT&#39;s thermogenic capacity in humans remains largely unknown due to the difficulties of measuring its volume, activity, and distribution. Identifying and quantifying active human BAT is commonly performed using 18F-Fluorodeoxyglucose (18F-FDG) positron emission tomography and computed tomography (PET/CT) scans following cold-exposure or pharmacological activation. Here we describe a detailed image-analysis approach to quantify total-body human BAT from 18F-FDG PET/CT scans using an open-source software. We demonstrate the drawing of user-specified regions of interest to identify metabolically active adipose tissue while avoiding common non-BAT tissues, to measure BAT volume and activity, and to further characterize its anatomical distribution. Although this rigorous approach is time-consuming, we believe it will ultimately provide a foundation to develop future automated BAT quantification algorithms.

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

Using free, open-source software, we have developed an analytical approach to quantify total and regional brown adipose tissue (BAT) volume and metabolic activity of BAT using 18F-FDG PET/CT.

Tutto il corpo e quantificazione regionale dell'essere umano attivo marrone tessuto adiposo con 18F-FDG PET/CT

In endothermic animals, brown adipose tissue (BAT) is activated to produce heat for defending body temperature in response to cold. BAT&#39;s ability to ...

Semi-quantitative Assessment Using [18F]FDG Tracer in Patients with Severe Brain Injury

Patients with severe traumatic brain injury (sTBI) have difficulty knowing whether they are accurately expressing their thoughts and emotions because of disorders of consciousness, disrupted higher brain function, and verbal disturbances. As a consequence of an insufficient ability to communicate, objective evaluations are needed from family members, medical staff, and caregivers. One such evaluation is the assessment of functioning brain areas. Recently, multimodal brain imaging has been used to explore the function of damaged brain areas. [18F]-fluorodeoxyglucose positron emission tomography-computed tomography ([18F]FDG-PET/CT) is a successful tool for examining brain function. However, the assessment of brain glucose metabolism based on [18F]FDG-PET/CT is not standardized and depends on several varying parameters, as well as the patient&#39;s condition. Here, we describe a series of semiquantitative assessment protocols for a region-of-interest (ROI) image analysis using self-produced [18F]FDG tracers in patients with sTBI. The protocol focuses on screening the participants, preparing the [18F]FDG tracer in the hot lab, scheduling the acquisition of [18F]FDG-PET/CT brain images, and measuring glucose metabolism using the ROI analysis from a targeted brain area.

Semi-quantitative Assessment Using [18F]FDG Tracer in Patients with Severe Brain Injury

[18F]-fluorodeoxyglucose (FDG) positron emission tomography-computed tomography is useful for studying glucose metabolism related to brain function. Here, we present a protocol for an [18F]FDG tracer set-up and semiquantitative assessment of the region-of-interest analysis for targeted brain areas associated with clinical manifestations in patients with severe traumatic brain injury.

Valutazione semi-quantitativa utilizzando [18F] FDG Tracer in pazienti con trauma cranico severo

Patients with severe traumatic brain injury (sTBI) have difficulty knowing whether they are accurately expressing their thoughts and emotions because of ...

Optical Clearing and Imaging of Immunolabeled Kidney Tissue

Optical clearing techniques render tissue transparent by equilibrating the refractive index throughout a sample for subsequent three-dimensional (3-D) imaging. They have received great attention in all research areas for the potential to analyze microscopic multicellular structures that extend over macroscopic distances. Given that kidney tubules, vasculature, nerves, and glomeruli extend in many directions, which have been only partially captured by traditional two-dimensional techniques so far, tissue clearing also opened up many new areas of kidney research. The list of optical clearing methods is rapidly growing, but it remains difficult for beginners in this field to choose the best method for a given research question. Provided here is a simple method that combines antibody labeling of thick mouse kidney slices; optical clearing with cheap, non-toxic and ready-to-use chemical ethyl cinnamate; and confocal imaging. This protocol describes how to perfuse kidneys and use an antigen-retrieval step to increase antibody- binding without requiring specialized equipment. Its application is presented in imaging different multicellular structures within the kidney, and how to troubleshoot poor antibody penetration into tissue is addressed. We also discuss the potential difficulties of imaging endogenous fluorophores and acquiring very large samples and how to overcome them. This simple protocol provides an easy-to-setup and comprehensive tool to study tissue in three dimensions.

The combination of antibody labeling, optical clearing, and advanced light microscopy allows three-dimensional analysis of complete structures or organs. Described here is a simple method to combine immunolabeling of thick kidney slices, optical clearing with ethyl cinnamate, and confocal imaging that enables visualization and quantification of three-dimensional kidney structures.

Cancellazione ottica e imaging del tessuto renale immunoetichettato

Optical clearing techniques render tissue transparent by equilibrating the refractive index throughout a sample for subsequent three-dimensional (3-D) ...