Este trabalho foi apoiado pelo Deutsche Forschungsgemeinschaft (SFB 656, Münster, Alemanha, projetos de C7 e C6) eo IZKF Münster (Core unidade SMAP). Os autores são gratos a Van Le Truc, Anne Kanzog, Ute Neugebauer, Wiebke Gottschlich e Roman Priebe para assistência técnica excelente e Daniel Burkert e Sven Fatum para a produção de radioisótopos.

1. Recuperação Organ Donor <ol><li> Configure o microscópio estereotáxica, peso registro da 8-10 semanas de idade rat (doador e receptor de peso corporal deve corresponder). </li><li> Anestesiar o rato doador (Lewis Brown Noruega F1, LBN F1) usando oxigênio / inalação de isoflurano (isoflurano 4% / 2 L / min de oxigênio). Manutenção da anestesia através da redução de 2-2,5% de isoflurano. </li><li> Colocar o rato anestesiado no bloco operatório. Fix extremidades do rato com fita adesiva sobre o bloco e aplicar pomada oftálmica (Bepanthen, Bayer) para os olhos do rato. </li><li> Barbear e desinfectar o abdómen do rato e de uma incisão na linha mediana ventral a partir do púbis para a borda caudal do fígado. Expor o rim esquerdo e seus navios movendo cuidadosamente o intestino para o lado direito. Depositar uma fina folha de gaze sobre o rim esquerdo e humedecer com solução salina isotônica aquecido para evitar a dessecação. </li><li> Dissecar cuidadosamente o tecido adiposo com Dumont Pinça ponta angular (SS-Dumont45 Pinça Inox Medical, Ferramentas Ciência Fine, Cat.No. 11203-25) que envolve o uréter esquerdo. Evite qualquer contato direto com o ureter, em vez disso, mobilizá-lo com suficiente circundante tecido adiposo. </li><li> Separe a veia renal esquerda da artéria renal Dumont usando uma pinça ponta angular. Remova toda a gordura e tecido conjuntivo de ambos os navios e cauterizar os vasos renais e testicular. </li><li> Dissecar aorta ea veia cava inferior (IVC), acima e abaixo da sua junção com a artéria e veia renal esquerda, respectivamente, e todas as artérias cauterizar saída. </li><li> Braçadeira supra-renal da aorta com uma braçadeira microcirúrgico (Micro Serrefines, ferramentas finas Science, N ° de Cat. 18055-04) acima da artéria renal esquerda, tão próximo quanto possível da artéria mesentérica superior. Ligadura aorta infrarenal utilizando seda cirúrgico (5-0, Vömel, nenhuma arte. 14739) a cerca de 5 mm abaixo da artéria renal esquerda. </li><li> Ligadura da VCI infra-renal com seda cirúrgica (5-0, Vömel), e prenda a VCI supra-renalcom uma pinça de microcirurgia (Micro Serrefines, Ferramentas Ciência Fine, Cat.No. 18.055-03). </li><li> Corte a veia renal tão perto quanto possível do IVC com uma tesoura fina (Iris Scissors - ToughCut reta 11,5 centímetros, Ferramentas Ciência Fine, Cat.No. 14.058-11). </li><li> Lentamente perfundir o rim in situ com solução de perfusão 2 ml gelada HTK (Custodiol HTK, Dr. Franz Köhler Chemie) inserindo uma cânula (Microlance 3 27G ¾, BD, Ref. ª 302200) na aorta infra-renal. Confirmar que as mudanças de cor renais (agora oliva cor). </li><li> Em primeiro lugar, cortar através de supra-renal da aorta, em seguida, a aorta infra-renal e, finalmente, o ureter tão próximo quanto possível da bexiga urinária. </li><li> Remover o rim e o seu fornecimento vascular juntamente com o ureter e armazenar em solução de perfusão HTK em gelo. </li><li> Eutanásia o rato dador por remoção dos grampos vasculares e excisão consecutivo do coração sob anestesia. </li></ol> 2. Preparação destinatário e transplanteção <ol><li> Configure o microscópio estereotáxica, peso registro do rato. </li><li> Anestesiar o rato destinatário (Lewis) utilizando isoflurano 4%. Manutenção da anestesia através da redução de 2-2,5% de isoflurano. </li><li> Colocar o rato anestesiado no bloco operatório. Fix extremidades do rato com fita adesiva sobre o bloco e aplicar pomada oftálmica para os olhos do rato `s. Monitor e controlo da temperatura do corpo do núcleo utilizando um sensor de temperatura rectal e uma almofada de aquecimento. Durante a cirurgia repetitivamente controlar a temperatura, a respiração e a frequência cardíaca dos animais. </li><li> Barbear e desinfectar o abdómen e uma incisão na linha mediana ventral a partir do púbis para a borda caudal do fígado. Usar um bisturi forcutting a pele, a fim de minimizar o trauma da pele. Expor o rim esquerdo e seus vasos movendo o intestino para o lado direito. </li><li> Remova cuidadosamente a cápsula renal usando uma pinça Dumont ponta angular e cotonetes. </li><li> Ocluir o lado esquerdo da artéria renal, veia e perto do ureter renalhilo com duas ligaduras usando fio cirúrgico (Mersilene 4-0, Ethicon, cat. nenhum. EH6732H). Impostos a maior parte do rim, deixando apenas o hilo renal. Limpe a superfície de corte com cotonetes e verificar se há sangramento. Adicionar outra ligadura se necessário. </li><li> Dissecar cuidadosamente sem rodeios através do tecido conjuntivo que cobre a aorta infra-renal e veia cava no site caudal com cotonetes. </li><li> Separar qualquer nervo visível a partir da aorta infra-renal e veia cava Dumont usando uma pinça ponta angular. </li><li> Dissecar cuidadosamente através da folha de tecido conjuntivo entre a aorta infra-renal e veia cava inferior, formando duas aberturas de 2-4 mm de comprimento: O primeiro abaixo da ramificação dos vasos testiculares, e a segunda uma acima da ramificação dos vasos ilíacos comuns. Iliolombar cauterizar as artérias e veias no meio. </li><li> Desenhe um único fio cirúrgico (Mersilene 0;. Ethicon, gato no.EH6665E) por cada uma das duas aberturas. Usando esses tópicos puxar a infra-aorta e veia cava suavemente para o lado direito do animal para ter acesso aos vasos ramificados no local dorsal. Cauterize qualquer destes navios entre as duas aberturas e remover os fios cirúrgicos posteriormente. </li><li> Em seguida, ocluir aorta e veia cava inferior através da aplicação dos ponteiros do relógio de quatro grampos microcirúrgicos (Micro Serrefines, ferramentas finas Science, N ° de Cat. 18055-04), com início na abertura superior com a aorta e terminando com a veia cava superior, na abertura (Micro Serrefines , Belas Tools ciência, Cat.No. 18.055-03). </li><li> Depois cuidadosamente furar o final superior da parte isolada da aorta com uma agulha (Microlance 3 27G ¾, BD, Ref. ª 302200), insira a lâmina inferior de uma tesoura de primavera (Vannas Primavera Scissors - 3 mm Lâminas, Fine Ciência Ferramentas, cat.no 15000-00), através da abertura de furos e realizar uma incisão longitudinal curto. Lave a aorta isolada com solução de perfusão HTK-gelada para remover qualquer vestígio de sangue restante. </li><li> Da mesma forma, cuidadosamente perfurantea parte inferior da parte final isolado da veia cava com uma agulha, inserir a lâmina inferior de uma mola de tesoura fina, através da abertura de punção e uma incisão longitudinal, terminando pouco abaixo da incisão da aorta. Mais uma vez, lave o navio para remover qualquer vestígio de sangue restante. </li><li> Remover o enxerto de rim a partir da solução de armazenamento. Coloque-o abaixo da posição anterior do rim esquerdo, e garantir que ele está na direção correta. </li><li> Verifique a orientação da artéria renal enxertos e garantir que não haja torções. Em seguida, coloque uma folha fina de gaze sobre o enxerto e umedecê-la com a solução de perfusão HTK gelada para resfriá-lo e evitar a dessecação. </li><li> Fixar a abertura da parte da supra-renal da aorta pedaço em que a artéria renal do enxerto se abre para com fio cirúrgico (9-0 Ethilon, Ethicon, o gato já 2809G..) - Em primeiro lugar no lado superior, em seguida, na extremidade inferior de a incisão da aorta. Fechar a incisão no sentido horário, com uma sutura contínua, usando fio cirúrgicoEthilon (9-0), Dumont fórceps ponta angular e um suporte de agulha Castroviejo curvo (Ferramentas Ciência Fine, cat. Nenhum. 12.061-01). Em seguida, cobrir a primeira sutura com tecido conjuntivo através de uma segunda sutura contínua anti-horário. Agora, apenas a parte inferior da peça de aorta do enxerto continua a ser fechado. Regularmente resfriar o enxerto com solução de perfusão HTK gelada. </li><li> Perfundir o enxerto com 1 ml de solução de perfusão HTK gelada através de uma agulha romba inserido na extremidade aberta da peça de aorta enxertos validar a integridade do aperto e da sutura, bem como a remoção do sangue remanescente no enxerto. Depois, ligadura da extremidade aberta da peça de aorta com seda cirúrgico (5-0, Vömel). </li><li> Verifique a orientação da veia renal enxertos e assegurar que não haja torções. </li><li> Fixar a abertura da veia de enxerto com fio cirúrgico (Ethilon 9-0) - pela primeira vez na parte superior, em seguida, na extremidade inferior da incisão na veia cava. Fechar a incisãono sentido horário, com uma sutura contínua, usando fio cirúrgico (Ethilon 9-0), Dumont fórceps ponta angular e um porta-agulha curvo Castroviejo. Regularmente resfriar o enxerto com solução de perfusão HTK gelada. </li><li> Sentido anti-horário remover os grampos de microcirurgia, começando com a braçadeira da veia cava superior. Após a remoção da última pinça aórtica, pare cuidadosamente sangramento eventualmente leve com cotonetes. Após alguns segundos, o enxerto vai virar róseo-matizado, e contrações uretric deve aparecer. </li><li> Para a inserção do ureter na bexiga enxertos `s do destinatário remova cuidadosamente um pequeno pedaço da musculatura, na parte superior da bexiga usando uma Primavera Scissor Vannas (Student Vannas Primavera tesoura reta, Belas Tools ciência, Cat.No. 91500-09 ). Em seguida, remova o tecido circundante (principalmente gordura) do ureter enxerto Dumont usando uma pinça ponta angular. Portanto, cuidadosamente agarrar a ponta do ureter e suavemente separar a gordura, o tecido conjuntivo envolvente e os vasos incorporados from que, puxando-os no sentido oposto. Fixar a ponta uretric na extremidade de um fio cirúrgico (Prolene 6-0, Ethicon, o gato. Não. 8697H) e perfurar o topo previamente preparada da bexiga com a agulha montada. Puxar o ureter através da bexiga e sai dele na base através de uma segunda perfuração. </li><li> Fixar a anteriormente a partir do ureter tecido separado no topo da bexiga com fio cirúrgico (9-0 Ethilon). Cortar a ponta do ureter com o fio cirúrgico em anexo e puxar o ureter volta para a bexiga empurrando cuidadosamente abaixo. Em seguida, feche a segunda punção da bexiga com fio cirúrgico (Ethilon 9-0). </li><li> Feche a parede abdominal e pele por duas suturas contínuas, mas em separado, utilizando fio de sutura cirúrgica (Mersilene 0). Desinfectar a ferida com iodo Pividon e administrar a buprenorfina (0,1 mg por kg / PV / dia) sc por três dias após a cirurgia para controlar a dor da ferida. </li></ol> 3. Imagem Allograft Rejeição<ol><li> Anestesiar ratos não submetidos a jejum utilizando isoflurano 2-2,5%. </li><li> 30 MBq de FDG (30 MBq de FDG em 0,1 ml de NaCl a 0,9%) são administradas iv através de cateterização de uma veia lateral da cauda utilizando um cateter de 24 L (Braun, Introcan, 4252500-01). Depois, remover o cateter com 0,9 ml de solução de NaCl a 0,9%. </li><li> Deixe o rato em um limitador sem anestesia até o início da digitalização e hidratar o animal por iv injeção de 1 ml de solução de NaCl 0,9% por hora. </li><li> Rat Re-anestesiar utilizando isoflurano 2% imediatamente antes do exame começar. </li><li> Coloque o rato anestesiado em uma câmara de alta resolução baseada em multi-wire animais PET câmera quadHIDAC (Oxford Positron Systems Ltd, Oxford, Reino Unido). A resolução espacial do aparelho de PET é de 1,0 mm e é constante ao longo de todo o FOV (diâmetro, 165 mm, comprimento axial, a 280 mm). Monitorar e controlar a temperatura do corpo durante a varredura usando um sensor de temperatura retal e anestesia controle com um oxímetro de pulso. </li><li> Comece aquisição dinâmico para 60 mno arranque 180 min após a injecção de FDG para reduzir a acumulação de marcador nos rins provocados pela excreção renal de FDG. </li><li> Aplicar 5 MBq de 18F-fluoreto iv e executar outra PET scan 18, imediatamente após a injecção de F-flúor durante 60 min, sem mover a posição do rato no scanner para identificação de parênquima renal e para o cálculo de 18 F-5 apuramento. Reconstruir imagens de ambos os exames. Traçar manualmente um volume de interesse (VOI) em imagens reconstruídas 2 min após injeção de 18 F-flúor (em fase de perfusão) em torno do córtex renal e transferir o VOI às imagens FDG. Excluir cuidadosamente a pélvis renal do VOI. Reconstruir imagens de ambos os scans e traçar manualmente um volume de interesse (VOI) em torno do córtex renal. Dados em modo lista foram reconstruídos em imagens com um tamanho de voxel de 0,4 × 0,4 × 0,4 milímetros 3. Excluir cuidadosamente a pélvis renal do VOI. </li><li> Calcule captação de FDG pela razão of contagem total e volume e calcular a porcentagem de dose injetada (% ID). Usamos MATLAB (Versão R2011b, MathWorks, Natick, MA, EUA) e tomográficas (TIM) Versão 2.8 para análise de imagem. </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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FDG-PET imagem é uma nova opção para o diagnóstico de rejeição aguda. Devido à sua natureza não-invasivo e específico, FDG-PET é vantagens em comparação com os diagnósticos clássicos por biópsia de agulha. Em contraste com o tamanho de uma biópsia, a análise de FDG-PET, todo o enxerto amostra limitada. Além disso, pode-se aplicar a pacientes em terapia anticoagulante, e pode-se realizar medidas de PET repetitivamente por exemplo, para monitorizar a eficácia do tratamento 4. Além d...

<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

O número de pacientes com doença renal em estágio final, eo número de pacientes transplantados renais aumenta continuamente. Episódios de rejeição do enxerto celular aguda (AR) são um fator prognóstico negativo para a sobrevivência do enxerto a longo prazo, e seu diagnóstico atempado é fundamental para a função do enxerto 1. No momento, AR só pode ser definitivamente diagnosticada por biópsia por agulha, que, como um método invasivo, descobre risco significativo de lesões ou até mesmo a perda do enxerto. Além disso, as biópsias não são viáveis ​​em pacientes que tomam medicamentos anticoagulantes e no site amostragem limitada desta técnica pode resultar em resultados falso-negativos se a AR é focal ou irregular. Como consequência, isto deu origem a uma busca permanente de novos métodos de detecção de AR, o que muitas vezes tem de ser feito em animais, incluindo a utilização de vários modelos de transplante. Desde o início dos anos 60 rato transplante renal é um método experimental bem estabelecido para o exação e análise de AR 2. Nós aqui presente, além de pequenos animais tomografia por emissão de pósitrons (PET), utilizando 18 F-(FDG) para avaliar a AR em um modelo alogênico uninefrectomizados rato transplante renal e propor enxerto imagem FDG-PET como uma nova opção para a não-invasivo, específico e diagnóstico precoce de AR também para a situação humana 3. Além disso, este método pode ser aplicado para o follow-up para melhorar o monitoramento da rejeição do transplante 4.

Histologia Durante AR leucócitos, isto é, principalmente os linfócitos T são recrutados para o transplante, ao passo que a severidade da rejeição é reflectido pelo grau de inflamação. Nas Periódica-Acid-Schiff (PAS) de coloração descrito aqui (Figura 1), o enxerto renal mostra sinais histológicos significativos de AR, a saber glomerulite, tubulite, endothelialitis e infiltração do enxerto (Figura 1, ATX POD4) (POD = dia pós-operatório)...

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

Apresentamos um modelo de transplante renal de ratos para não-invasiva avaliar a rejeição do enxerto aguda usando tomografia por emissão de pósitrons com 18F-fluorodeoxyglucose.

Imagem não invasivo de rejeição do enxerto aguda após Rat Transplante Renal Usando<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 visualizações.  University of Münster. Apresentamos um modelo de transplante renal de ratos para não-invasiva avaliar a rejeição do enxerto aguda usando tomografia por emissão de pósitrons com 18F-fluorodeoxyglucose.

Vídeo: 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.

Não-invasivo de medição óptica do metabolismo cerebral e Hemodinâmica em Lactentes

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

Medicina

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.

Transplante para a câmara anterior do olho para longitudinal, não-invasivo<em&gt; Em vivo</em&gt; Imaging com uma única célula de resolução em tempo real

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.

Imagem não invasivo e Análise de Isquemia Cerebral em Ratos de Vida Usando Positron Emission Tomography com<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.

Modelo murino de imagem não invasivo para detectar e monitorar o cancro do ovário Recorrência

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.

Rato Transplante Renal: Modelos de rejeição de enxertos

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.

Determinando Glucose Metabolismo Kinetics Usando<sup&gt; 18</sup&gt; F-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). ...

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.

Transplante de células-tronco derivadas de tecido adiposo folha para reduzir o vazamento após colectomia parcial em um modelo do rato

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.

Todo o corpo e quantificação Regional de ativo humano marrom tecido adiposo usando 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.

Avaliação semi-quantitativa usando [18F] FDG Tracer em pacientes com lesão cerebral grave

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.

Esclarecimento ótico e imagem latente do tecido Immunolabeled do rim

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