Este trabajo fue apoyado por la Deutsche Forschungsgemeinschaft (SFB 656, Münster, Alemania, los proyectos de C7 y C6) y el IZKF Münster (Core unidad SMAP). Los autores agradecen a Truc Van Le, Anne Kanzog, Ute Neugebauer, Wiebke Gottschlich y Priebe romana por su excelente asistencia técnica y Daniel Burkert y Sven Fatum para la producción de radiotrazadores.

1. La recuperación del donante de órganos <ol><li> Configure el microscopio estereotáctica, peso récord de la vieja rata de 8-10 semanas (donante y el peso corporal del receptor debe coincidir). </li><li> Anestesiar la rata donante (Lewis Brown Noruega F1, F1 LBN) el uso de oxígeno / inhalación de isoflurano (isoflurano 4% / 2 L / min de oxígeno). Mantener la anestesia mediante la reducción de isoflurano al 2-2,5%. </li><li> Coloque la rata anestesiada en la plataforma de la cirugía. Fijar extremidades de la rata con cinta adhesiva sobre la almohadilla y aplicar pomada oftálmica (Bepanthen, Bayer) a los ojos de la rata. </li><li> Shave y desinfectar el abdomen de la rata y realizar una incisión de línea media ventral desde el pubis hasta el borde caudal del hígado. Exponer el riñón izquierdo y sus vasos moviendo cuidadosamente el intestino hacia el lado derecho. Colocar una hoja delgada de gasa sobre el riñón izquierdo y humedecerlo con solución salina isotónica calentada para evitar la desecación. </li><li> Diseccionar cuidadosamente el tejido graso con Dumont Pinzas punta en ángulo (SS-Dumont45 Pinzas Inox Médico, Herramientas de Bellas Ciencia, n º de cat 11203-25) que envuelve el uréter izquierdo. Evite el contacto directo con el uréter, en cambio, movilizar con suficiente tejido graso circundante. </li><li> Separe la vena renal izquierda de la arteria renal con pinzas Dumont punta en ángulo. Quite toda grasa y tejido conectivo de los dos vasos y cauterizar los vasos adrenales y testiculares. </li><li> Diseccionar la aorta y la vena cava inferior (VCI) por encima y por debajo de su unión con la arteria renal izquierda y la vena, respectivamente, y todas las arterias cauterizar salientes. </li><li> Sujete la aorta suprarrenal con una pinza de microcirugía (Micro Serrefines, Herramientas de Bellas Ciencia, n º de cat 18055-04) por encima de la arteria renal izquierda, lo más cerca posible a la arteria mesentérica superior. Ligar la aorta infrarrenal con seda quirúrgica (5-0, Vömel, ref. 14739) aproximadamente 5 mm por debajo de la arteria renal izquierda. </li><li> Ligar la VCI infrarrenal con seda quirúrgica (5-0, Vömel) y sujetar el IVC suprarrenalcon una pinza de microcirugía (Serrefines Micro, Herramientas de Bellas Ciencia, art 18055-03). </li><li> Cortar la vena renal lo más cerca posible de la IVC con tijeras finas (Iris Scissors - ToughCut rectas 11,5 cm, Herramientas de Bellas Ciencia, art 14058-11). </li><li> Poco a poco la perfusión del riñón in situ con 2 solución de perfusión HTK ml de helado (CUSTODIOL HTK, Dr. Franz Köhler Chemie) mediante la inserción de una cánula (Microlance 3 27G ¾, BD, cat.no. 302200) en la aorta infrarrenal. Confirme que los cambios de color renales (ahora oliva teñido). </li><li> En primer lugar, seccionar la aorta suprarrenal, a continuación, la aorta infrarrenal y, finalmente, el uréter lo más cerca posible a la vejiga urinaria. </li><li> Retire el riñón y en la irrigación vascular junto con el uréter y la tienda en solución de perfusión HTK en hielo. </li><li> La eutanasia a la rata donante mediante la eliminación de las pinzas vasculares y la escisión consecutiva del corazón bajo anestesia. </li></ol> 2. Receptor Preparación y Transplantación <ol><li> Configure el microscopio estereotáctica, peso record de la rata. </li><li> Anestesiar la rata receptora (Lewis) con isoflurano 4%. Mantener la anestesia mediante la reducción de isoflurano al 2-2,5%. </li><li> Coloque la rata anestesiada en la plataforma de la cirugía. Fijar las extremidades de la rata con cinta adhesiva sobre la almohadilla y aplicar el ungüento oftálmico de los ojos de la rata `s. Monitor y control de la temperatura corporal central usando un sensor de temperatura rectal y una almohadilla de calentamiento. Durante la cirugía repetitivamente controlar la temperatura, la respiración y la frecuencia cardiaca del animal. </li><li> Shave y desinfectar el abdomen y realizar una incisión de línea media ventral desde el pubis hasta el borde caudal del hígado. Utilice un forcutting escalpelo de la piel con el fin de minimizar el trauma de la piel. Exponer el riñón izquierdo y sus vasos moviendo el intestino hacia el lado derecho. </li><li> Retire con cuidado la cápsula renal con pinzas Dumont punta en ángulo y los hisopos de algodón. </li><li> Ocluir la arteria renal izquierda, vena y uréter cerca de la renalhilo con dos ligaduras con hilo quirúrgico (Mersilene 4-0, Ethicon, cat. no. EH6732H). Impuestos Especiales de la mayor parte del riñón, dejando sólo el hilio renal. Limpie la superficie de corte con hisopos de algodón y compruebe si el sangrado. Añadir otra ligadura si es necesario. </li><li> Diseccionar cuidadosamente sin rodeos a través del tejido conectivo que cubre la aorta infrarrenal y la vena cava caudal en el sitio con hisopos de algodón. </li><li> Separe cualquier nervio visible desde la aorta infrarrenal y la vena cava utilizando pinzas Dumont punta en ángulo. </li><li> Diseccionar cuidadosamente a través de la hoja de tejido conectivo entre la aorta infrarrenal y la vena cava, formando dos aberturas de 2-4 mm de longitud: El primero por debajo de la ramificación de los vasos testiculares, y el segundo por encima de la bifurcación de los vasos ilíacos comunes. Cauterize iliolumbar las arterias y las venas en el medio. </li><li> Dibuje un solo hilo quirúrgico (Mersilene 0;. Ethicon, gato no.EH6665E) a través de cada una de las dos aberturas. El uso de estos hilos tiran de la infrarrenalaorta y la vena cava suavemente hacia el lado derecho del animal para obtener acceso a los vasos de ramificación en el sitio dorsal. Cauterize cualquiera de estos vasos entre las dos aberturas y quitar los hilos quirúrgicos a partir de entonces. </li><li> A continuación, ocluir la aorta y la vena cava inferior mediante la aplicación de las agujas del reloj cuatro pinzas de microcirugía (Micro Serrefines, Fine Science Tools, n º de cat 18055-04) que comienzan en la apertura superior con la aorta y terminando con la vena cava en la abertura superior (micro Serrefines , Science Tools Bellas, art 18055-03). </li><li> Después cuidadosamente perforar el final superior de la parte aislada de la aorta con una aguja (27G Microlance 3 ¾, BD, cat.no. 302200), inserte la hoja inferior de un resorte de tijera fina (Vannas Tijeras primavera - 3 mm Hojas, Fine Ciencia Herramientas, cat.no 15000-00) a través de la abertura de punción y realizar una incisión longitudinal corto. Enjuague la aorta aislada con solución de perfusión HTK-helada para eliminar la sangre restante. </li><li> Del mismo modo, cuidadosamente perforarel final inferior de la parte aislada de la vena cava con una aguja, insertar la cuchilla inferior de un resorte de tijera fina a través de la abertura de punción y realizar una incisión longitudinal que termina poco por debajo de la incisión de la aorta. Otra vez, enjuague el recipiente para eliminar cualquier resto de sangre. </li><li> Retire el injerto renal de la solución de almacenamiento. Colóquelo debajo de la posición anterior del riñón izquierdo, y asegúrese de que está correctamente orientado. </li><li> Compruebe la orientación de la arteria renal de los injertos y asegurarse de que no haya torceduras. A continuación, colocar una hoja delgada de gasa sobre el injerto y humedecerlo con solución de perfusión HTK enfriado con hielo para que se enfríe, y para evitar la desecación. </li><li> Fijar la abertura de la parte de la pieza suprarrenal de la aorta en la que la arteria renal del injerto se abre en con hilo quirúrgico (9-0 Ethilon, Ethicon, n º de cat 2809G..) - Primero en la parte superior, a continuación, en el extremo inferior de la incisión de la aorta. Cierre la incisión hacia la derecha con una sutura continua con hilo quirúrgicoEthilon (9-0), Dumont forceps punta en ángulo y un porta-agujas Castroviejo curva (Herramientas de Bellas Ciencia, cat.. No 12061-01). Luego, cubra la primera sutura de tejido conectivo a través de una segunda sutura continua hacia la izquierda. Ahora, sólo la parte inferior de la pieza aórtica del injerto sigue siendo a ser cerrado. Regularmente enfriar la solución de perfusión del injerto con HTK enfriado con hielo. </li><li> Perfundir el injerto con 1 ml de solución de perfusión HTK enfriado en hielo a través de una aguja de punta roma se inserta en el extremo abierto de la pieza aórtica injertos de validación de la estanqueidad y la integridad de la sutura, así como la eliminación de la sangre restante en el injerto. Después, ligar el extremo abierto de la pieza aórtica con seda quirúrgica (5-0, Vömel). </li><li> Compruebe la orientación de la vena renal injertos y asegurarse de que no haya torceduras. </li><li> Fijar la abertura de la vena del injerto con hilo quirúrgico (Ethilon 9-0) - primero en la parte superior, a continuación, en el extremo inferior de la incisión en la vena cava. Cierre de la incisiónhacia la derecha con una sutura continua con hilo quirúrgico (Ethilon 9-0), Dumont pinzas punta en ángulo y un porta-agujas Castroviejo curvo. Regularmente enfriar la solución de perfusión del injerto con HTK enfriado con hielo. </li><li> Antihorario eliminar las abrazaderas de microcirugía, comenzando con la vena cava superior abrazadera. Después de quitar el último pinzamiento aórtico, deje cuidadosamente sangrado leve finalmente con hisopos de algodón. Después de unos segundos, el injerto se volverá rosa teñida, y debe aparecer contracciones uretric. </li><li> Para la inserción del uréter en la vejiga injertos `s destinatario retire con cuidado una pequeña porción de la musculatura en la parte superior de la vejiga usando una tijera primavera Vannas (Estudiante Vannas tijera primavera rectas, Herramientas de Bellas Ciencia, n º de cat 91500-09 ). A continuación, retire el tejido circundante (principalmente grasa) del uréter de injerto utilizando pinzas Dumont punta en ángulo. Por lo tanto, tomar cuidadosamente la punta del uréter y suavemente separar el tejido circundante de grasa, conectivo y de los vasos incrustados lado a otrom que tirando de ellas en la dirección opuesta. Fijar la punta uretric en el extremo de un hilo quirúrgico (6-0 Prolene, Ethicon, gato.. Ninguna 8697H) y perforar la parte superior previamente preparada de la vejiga con la aguja fija. Tire el uréter a través de la vejiga y salir de ella en la base a través de una segunda perforación. </li><li> Fijar el previamente del uréter separados tejido en la parte superior de la vejiga con hilo quirúrgico (Ethilon 9-0). Corte la punta del uréter con el hilo quirúrgico adjunto y tire el uréter hacia la vejiga, presionando suavemente desde abajo. A continuación, cierre la segunda punción vesical con hilo quirúrgico (Ethilon 9-0). </li><li> Cierre de la pared abdominal y la piel por dos suturas continuas pero separado utilizando hilo de sutura quirúrgica (Mersilene 0). Desinfectar la herida con yodo Pividon y administrar buprenorfina (0,1 mg por kg / peso corporal / día) sc durante tres días después de la cirugía para controlar el dolor de la herida. </li></ol> 3. Imágenes aloinjerto Rechazo<ol><li> Anestesiar la rata postprandiales utilizando isoflurano 2-2,5%. </li><li> 30 MBq de FDG (30 MBq de FDG en 0,1 ml de NaCl al 0,9%) se administró iv a través de la cateterización de una vena lateral de la cola con un G catéter 24 (Braun, Introcan, 4252500-01). Después, purgar el catéter con 0,9 ml de solución de NaCl al 0,9%. </li><li> Agregar la rata en una inmovilización sin anestesia hasta el inicio de la exploración y el hidrato de animales por inyección iv de 1 ml de solución de NaCl al 0,9% por hora. </li><li> Rata reanestesiar utilizando isoflurano 2% inmediatamente antes de que comience la exploración. </li><li> Coloque rata anestesiada en una alta resolución de la cámara basada en múltiples hilos animales cámara quadHIDAC PET (Positron Systems Ltd Oxford, Oxford, Reino Unido). La resolución espacial del escáner de PET es de 1,0 mm y es constante en todo el campo de visión (diámetro, 165 mm de longitud axial, 280 mm). Supervisión y control de la temperatura corporal durante el análisis utilizando un sensor de temperatura rectal y la anestesia de control con un oxímetro de pulso. </li><li> Inicio adquisición dinámico para 60 men el inicio de 180 minutos después de la FDG-inyección para reducir la acumulación de trazador en los riñones causado por la excreción renal de FDG. </li><li> Aplicar 5 MBq de 18 iv F-fluoruro y realizar otra exploración PET inmediatamente después de la inyección de 18 F-fluoruro durante 60 min sin mover la posición de la rata en el escáner para la identificación de parénquima renal y para el cálculo de 18 F-5 aclaramiento. Reconstruir imágenes de ambas exploraciones. Trazar manualmente un volumen de interés (VOI) en las imágenes reconstruidas 2 minutos después de la inyección de 18 F-fluoruro (perfusión de fase) en torno a la corteza renal y transferir el VOI a las imágenes FDG. Excluir cuidadosamente la pelvis renal del VOI. Reconstruir imágenes de ambos análisis y trazar manualmente un volumen de interés (VOI) alrededor de la corteza renal. De datos en modo de lista fueron reconstruidas en imágenes con un tamaño de voxel de 0,4 × 0,4 × 0,4 mm 3. Excluir cuidadosamente la pelvis renal del VOI. </li><li> Calcular la captación de FDG por la relación Of recuentos totales y volumen y calcular el porcentaje de dosis inyectada (% ID). Se utilizó MATLAB (versión R2011b, MathWorks, Natick, MA, EE.UU.) y la obtención de imágenes tomográficas (TIM) Versión 2.8 para el análisis de imágenes. </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>

No hay conflictos de interés declarado.

FDG-PET es una nueva opción para el diagnóstico de rechazo agudo. Debido a su naturaleza no invasiva y específicos, la FDG-PET es ventajas en comparación con los diagnósticos clásicos de la biopsia con aguja gruesa. En contraste con el limitado tamaño de muestra de una biopsia, el análisis PET-FDG todo el injerto. Además, se puede aplicar a los pacientes en tratamiento con anticoagulantes, y se puede llevar a cabo medidas de PET repetidamente por ejemplo, para controlar la eficacia del tratamiento <sup...

<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

El número de pacientes con enfermedad renal en etapa terminal, y el número de receptores de aloinjertos renales aumenta continuamente. Los episodios de rechazo del injerto celular agudo (AR) son un factor pronóstico negativo para la supervivencia del injerto a largo plazo, y su diagnóstico oportuno es crucial para la función del injerto 1. En la actualidad, AR puede sólo puede ser diagnosticado por biopsia central con aguja, la cual, como un método invasivo, descubre riesgo significativo de lesión del injerto o incluso la pérdida. Por otra parte, las biopsias no son factibles en los pacientes que toman medicamentos anticoagulantes y el sitio de muestreo limitado de esta técnica puede dar lugar a resultados falsos negativos si el AR es focal o en parches. Como consecuencia, esto dio lugar a una búsqueda constante de nuevos métodos de detección de AR, que a menudo tiene que hacerse en animales, incluyendo el uso de varios modelos de trasplante. Desde principios de los 60 la rata trasplante renal es un método experimental bien establecido para el exación y el análisis de AR 2. Estamos aquí presentes además pequeña tomografía por emisión de positrones animales (PET) con 18 F-FDG (FDG) para evaluar la AR en una rata renal modelo uninefrectomizadas trasplantes alogénicos, y proponer injerto imágenes FDG-PET como una nueva opción para un no-invasivo, específico y el diagnóstico precoz de la AR también para la situación humana 3. Además, este método puede ser aplicado para el seguimiento para mejorar el seguimiento de rechazo del trasplante 4.

Histología Durante los leucocitos AR, es decir, principalmente los linfocitos T son reclutados en el trasplante, mientras que la severidad del rechazo se refleja por el grado de inflamación. En la tinción de ácido periódico de Schiff (PAS) se muestra aquí (Figura 1), el aloinjerto renal muestra signos histológicos significativos de AR, es decir glomerulitis, tubulitis, endotelialitis e infiltración del injerto (Figura 1, ATX POD4) (POD = día d...

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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

Nos en este documento presentamos un modelo de trasplante renal de rata para evaluar de forma no invasiva el rechazo agudo del aloinjerto mediante tomografía por emisión de positrones con 18F-fluorodesoxiglucosa.

Imagen no invasiva de rechazo agudo del aloinjerto después Rat Trasplante Renal Uso<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

Human Biology

Osmoregulation and Excretion

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16.9K vistas.  University of Münster. Nos en este documento presentamos un modelo de trasplante renal de rata para evaluar de forma no invasiva el rechazo agudo del aloinjerto mediante tomografía por emisión de positrones con 18F-fluorodesoxiglucosa.

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.

No invasiva de medición óptica del metabolismo cerebral en lactantes y Hemodinámica

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

Investigación

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.

Trasplante en la cámara anterior del ojo para Longitudinal, no invasiva<em&gt; En vivo</em&gt; Imagen con una sola célula de resolución en tiempo 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.

De imagen no invasivas y Análisis de la isquemia cerebral en ratas vivas utilizando Tomografía por emisión de positrones 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.

Modelo murino de imagen no invasivas para detectar y vigilar cáncer ovárico de Recurrencia

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.

Ratón Trasplante de Riñón: Modelos de rechazo del aloinjerto

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.

La determinación de metabolismo de la glucosa Kinetics Uso<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.

Trasplante de células madre derivadas de tejido adiposo hoja para reducir fugas después de colectomía parcial en un modelo de rata

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.

Cuerpo entero y cuantificación Regional de activo humano marrón tejido adiposo utilizando 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.

Evaluación semicuantitativa usando [18F] FDG trazador en pacientes con lesión 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.

Limpieza óptica e imágenes del tejido renal inmunoetiquetado

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