この作品は、ドイツ学術振興（SFB 656、ミュンスター、ドイツ、プロジェクトC7＆C6）とIZKFミュンスター（コアユニットSMAP）によってサポートされていました。著者らは、放射性トレーサーを製造するための優れた技術支援とダニエルビュルケルトとスベンFatumのためのチュックヴァンル、アンKanzog、ウテノイゲバウアー、Wiebke Gottschlich、ローマPriebeに感謝しています。

1。ドナー臓器の回復<ol><li>定位顕微鏡、8-10週齢ラットの記録量（ドナーとレシピエントの体重が一致している必要があります）を設定します。 </li><li>酸素/イソフルラン吸入を（イソフルラン4％/ 2 L /分の酸素）を使用して、ドナーラット（ルイスブラウンノルウェーF1、LBN F1）を麻酔。 2から2.5パーセントにイソフルラン下げることで麻酔を維持します。 </li><li>手術パッド上麻酔ラットを置きます。パッド上に粘着テープで、ラットの四肢を固定し、ラットの目に眼軟膏（Bepanthen、バイエル）を適用。 </li><li>剃るとラットの腹部を消毒し、恥骨から肝臓の尾国境に腹側正中切開を行います。慎重に右側に腸を動かすことにより左腎とその血管を公開します。左腎上のガーゼの薄いシートを置き、乾燥を防ぐために温めておいた等張食塩水でそれを湿ら。 </li><li>慎重に（デュモンアングルティップ鉗子で脂肪組織を解剖デュモンSS-45鉗子イノックス医療、ファイン科学ツール、カタログ番号11203から25）、左尿管を包む。尿管との直接の接触を避け、代わりに、十分な周囲の脂肪組織でそれを動員する。 </li><li>デュモン斜めヒントピンセットを用いて腎動脈から左腎静脈を分離。両方の血管からすべての脂肪と結合組織を削除して、副腎や精巣血管を焼灼する。 </li><li>それぞれ、左腎動脈と静脈との接合部の上下大動脈と下大静脈（IVC）を解剖すると、すべての発信動脈を焼灼。 </li><li>上腸間膜動脈にできるだけ近くに、左腎動脈上顕微クランプ（マイクロSerrefines、ファイン科学ツール、カタログ番号18055から04まで）と副腎大動脈をクランプ。外科絹（5-0、Vömel、アートなし。14739）左腎動脈下約5mmを使って大動脈大動脈を連結。 </li><li>外科絹（5-0、Vömel）で大動脈IVCを連結し、副腎IVCをクランプ顕微クランプ付き（マイクロSerrefines、ファイン科学ツール、カタログ番号18055から03）。 </li><li>細かいハサミを（ - ToughCutストレート11.5センチメートル、ファイン科学ツール、カタログ番号14058から11アイリスはさみ）を使用してできるだけ近くIVCに腎静脈を切った。 </li><li>ゆっくり大動脈大動脈カニューレ（Microlance 3 27G¾、BD、カタログ番号302200）を挿入して2ミリリットルの氷冷HTK灌流液（CUSTODIOL HTK、博士はフランツ·ケーラーChemie社）とその場で腎臓灌流。腎臓の色の変化（現在はオリーブ彩色）ことを確認してください。 </li><li>まず、最終的に大動脈大動脈と膀胱にできるだけ近い尿管その後、副腎大動脈を横断する。 </li><li>氷の上でHTK灌流液中尿管や店舗とともに腎臓とその血管供給を削除します。 </li><li>麻酔下で心臓の血管のクランプと連続した切除を除去することにより、ドナーラットを安楽死させる。 </li></ol> 2。受信者の準備とTransplantaる<ol><li>定位顕微鏡、記録、ラットの体重を設定します。 </li><li>イソフルラン4％を使用したレシピエントラット（ルイス）を麻酔。 2から2.5パーセントにイソフルラン下げることで麻酔を維持します。 </li><li>手術パッド上麻酔ラットを置きます。パッド上に粘着テープで、ラットの四肢を固定し、ラットの `sの目に眼軟膏を適用します。直腸温度センサ加温パッドを使用して監視および制御中核体温。手術中に繰り返し温度、呼吸や動物の心拍数を制御します。 </li><li>剃ると腹部を消毒し、恥骨から肝臓の尾国境に腹側正中切開を行います。皮膚の外傷を最小限にするためにメスforcuttingのスキンを使用。右側に腸を動かすことにより左腎とその血管を公開します。 </li><li>注意深くデュモン斜めヒント鉗子や綿棒を使って腎カプセルを削除します。 </li><li>腎近左腎動脈、静脈と尿管を閉塞手術糸を（4-0 Mersilene、エチコン、カタログ番号EH6732H）を使用して2つのライゲーションと門。物品だけ腎門を残して腎臓の大部分は、。綿棒で切断面をきれいにし、出血を確認してください。必要に応じて別の字を追加します。 </li><li>慎重に綿棒と尾サイトに大動脈大動脈と大静脈をカバーする結合組織を通してぶっきらぼうに解剖する。 </li><li>デュモン斜めヒントピンセットを用いて大動脈大動脈と大静脈からの目に見える神経を分離。 </li><li>最初精巣血管の分岐下、および総腸骨血管の分岐上記の2番目の1：慎重に2〜4ミリメートルの長さの2つの開口部を形成し、大動脈大動脈と大静脈の間に結合組織シートを解剖する。間に腸腰動脈と静脈を焼灼。 </li><li>二つの開口部のそれぞれを通して、シングル手術糸（エチコン、猫no.EH6665E。Mersilene 0）を描画します。これらのスレッドを使用すると、大動脈を引っ張るそっと背サイトに分岐血管へのアクセスを得るための動物の右側に大動脈と大静脈。二つの開口部の間にこれらの血管のいずれかを焼灼し、その後手術のスレッドを削除します。 </li><li>次に、大動脈と上部開口部から始まり、上部開口（マイクロSerrefinesで下大静脈で終わる4顕微クランプ（マイクロSerrefines、ファイン科学ツール、カタログ番号18055から04）を時計回りに、アプリケーションによって大動脈とIVCを閉塞、ファイン科学ツール、カタログ番号18055から03）。 </li><li>その後慎重に穴を開ける針で大動脈の孤立部分の上部エンディング（Microlance 3 27G¾、BD、カタログ番号302200）は、細かい春のはさみ（Vannasスプリングはさみの下刃を挿入する - 3ミリメートルブレード、ファイン穿刺開口を通して科学ツール、cat.no 15000から00）と短い縦切開を行います。残りの血液を除去するために氷冷HTK灌流液で孤立大動脈をフラッシュします。 </li><li>同様に、慎重に穴を開ける針で静脈の隔離部の下部結末は、穿刺口から細かい春のはさみの下刃を挿入し、まもなく大動脈切開下に終わる縦切開を行います。繰り返しになりますが、残りの血液を除去するために容器をフラッシュします。 </li><li>ストレージソリューションから腎臓移植を削除します。左腎の元の位置の下に配置し、それが正しい方向であることを確認します。 </li><li>移植腎動脈の向きを確認し、ねじれがないことを確認。その後、移植上のガーゼの薄いシートを配置し、それを冷却するために氷のように冷たいHTK灌流液でそれを湿らせて、乾燥を防ぐために。 </li><li>アッパーで最初の、その後の下端 - 移植の腎動脈が、手術糸（。Ethilon 9-0、エチコン、猫なし2809G）とに開口する大動脈部分の副腎部の開口部を固定大動脈切開。手術糸を使用して連続縫合で時計回りに切開を閉じ（Ethilon 9-0）、デュモン斜めヒント鉗子と湾曲Castroviejo針ホルダ（ファイン科学ツール、カタログ番号12061から01）。その後、反時計回りに第2の連続縫合糸を通して結合組織を持つ最初の縫合糸をカバーしています。さて、グラフトの大動脈部分の唯一の下部には、閉鎖されていない。定期的に氷のように冷たいHTK灌流液で移植片を冷却する。 </li><li>縫合糸の気密性と整合性の検証、ならびに移植片の残りの血液を除去グラフトの大動脈片の開放端に挿入された鈍針を通して氷冷灌流HTK溶液1mlで移植片を灌流する。その後、外科絹（5-0、Vömel）と大動脈の部分の開放端を連結。 </li><li>移植腎静脈の向きを確認し、ねじれがないことを確認。 </li><li>大静脈に切開の下端にその後、アッパーで最初の - 手術糸（Ethilon 9-0）とグラフトの静脈の開口部を固定。切開を閉じ手術糸（Ethilon 9-0）、デュモン斜めヒント鉗子と湾曲Castroviejo針ホルダーを用いて連続縫合で時計回りに。定期的に氷のように冷たいHTK灌流液で移植片を冷却する。 </li><li>最大静脈クランプで始まる、顕微クランプを取り外し反時計回り。最後の大動脈クランプを除去した後、慎重に綿棒で、最終的に軽度の出血を止める。数秒後に移植はバラ色の彩色になり、そしてuretric収縮が表示されるはずです。 </li><li>受信者の `sの膀胱に移植尿管の挿入は慎重Vannasスプリングシザー（学生Vannasスプリングシザーストレート、ファイン科学ツール、カタログ番号91500から09を使用して膀胱の上部にある筋肉の小さなパッチを削除するための）。その後、デュモン角度の先端鉗子を使用して移植尿管から周囲の組織（主に脂肪）を取り外します。したがって、慎重に尿管の先端をつかむと、優しく周囲の脂肪、結合組織を分離し、埋め込まれた血管あちこち反対方向にそれらを引っ張ることによってそれをメートル。手術糸（PROLENE 6-0、エチコン、カタログ番号8697H）の末尾uretric先端を固定し、付属の針で膀胱の予め準備上部を穿孔。膀胱を通して尿管を引いて、第二の穿孔を介してベースでそれを終了します。 </li><li>手術糸（Ethilon 9-0）で膀胱の上部にある組織を分離した尿管から以前にこだわる。添付された手術糸と尿管の先端を切って静かに下から押して、膀胱に戻し尿管を引っ張る。その後、手術糸（Ethilon 9-0）と第二膀胱穿刺を閉じます。 </li><li>手術用縫合糸（Mersilene 0）を使用して、連続する二つの独立したが、縫合によって腹壁と皮膚を閉じます。 Pividonヨウ素を使用して傷を消毒し、傷の痛みを制御するために、手術後三日間ブプレノルフィン量（kg / BW /入札当たり0.1 mg）を皮下投与する。 </li></ol> 3。イメージング同種移植の拒絶<ol><li>イソフルラン2から2.5パーセントを使用して非空腹ラットを麻酔。 </li><li> 30 MBqのFDG（0.​​1ミリリットル0.9％NaCl中30 MBqのFDG）が24 Gカテーテル（ブラウン、Introcan、4252500から01）を使用して、横方向の尾静脈をcatheterizing介して静脈内投与する。その後、0.9ミリリットル0.9％NaCl溶液でカテーテルをパージ。 </li><li>毎時0.9％NaCl溶液1mlを静脈注射により動物のスキャンの開始とハイドまで麻酔なしで拘束ラットのままにしておきます。 </li><li>スキャンが開始される直前にイソフルラン2％を使用してラットの再麻酔。 </li><li>高解像度のマルチワイヤ室ベースの動物PETカメラquadHIDAC（オックスフォード陽電子システムズ株式会社、オックスフォード、英国）で開催麻酔ラット。 PETスキャナの空間分解能が1.0mmで、全FOV（直径165ミリメートル、軸方向長さ280 mm）の上で一定である。パルスオキシメータと直腸温度センサーと制御麻酔を使用してスキャン中にモニタと制御体温。 </li><li> 60メートルの動的取得を開始FDGの腎排泄に起因する腎臓でトレーサーの蓄積を減らすためにFDG-注入後180分を開始した。 </li><li> 18 F-フッ化物IVの5 MBqのを適用し、腎実質の識別のために、18 F-クリアランス5の計算のためのスキャナでラットの位置を移動することなくすぐに60分間、18 F-フッ化物注射後に別のPETスキャンを実行します。両方のスキャンから画像を再構成する。手動で2分腎皮質の周りに18 F-フッ化物注射液（灌流相）後に再構成された画像上で関心体積（VOI）をトレースし、FDG画像にVOIを転送します。慎重にVOIから腎盂を除外します。両方のスキャンから画像を再構成して、手動で腎皮質の周り関心体積（VOI）をトレース。リストモードデータは0.4×0.4×0.4ミリメートル3のボクセルサイズで画像に再構成された。慎重にVOIから腎盂を除外します。 </li><li>比OによるFDG集積を計算fの合計数と量と注入された投与量（％ID）の割合を計算します。私たちは、画像解析のためのMATLAB（バージョンR2011b、MathWorks社、ナティック、MA、USA）と断層画像（TIM）バージョン2.8を使用していました。 </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>

利害の衝突は宣言されていない。

FDG-PETイメージングは​​、急性拒絶反応の診断のための新しいオプションです。なぜなら、その非侵襲的かつ特異的な性質のため、FDG-PETは、コア針生検による古典的な診断と比較して利点がある。生検、FDG-PET解析移植片全体の限られたサンプルサイズとは対照的である。また、人は抗凝固療法を受けている患者に適用することができ、もう一方は繰り返し処理効率4を監視するため...

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 <td></td>
 <td>Equipment</td>
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 <td>Mathieu Needle Holder - 14 cm</td>
 <td>Fine Science Tools</td>
 <td>12010-14</td>
 <td></td>
 </tr>
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 <td>Castroviejo Micro Needle Holder - 9 cm</td>
 <td>Fine Science Tools</td>
 <td>12061-01</td>
 <td></td>
 </tr>
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 <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>
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 <td>Student Vannas Spring Scissors </td>
 <td>Fine Science Tools</td>
 <td>91500-09</td>
 <td></td>
 </tr>
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 <td>Vannas Spring Scissors - 3 mm Blades</td>
 <td>Fine Science Tools</td>
 <td>15000-00</td>
 <td></td>
 </tr>
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 <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>
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 <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

末期腎疾患の患者の数、腎同種移植レシピエントの数が増加し続ける。急性細胞の同種移植片拒絶反応（AR）のエピソードは、長期的な同種移植片生存のための負の予後因子であり、そのタイムリーな診断は、同種移植片機能1のために極めて重要である。現時点では、ARのみ間違い侵襲的な方法として、移植傷害あるいは損失の重大なリスクをむく、コア針生検によって診断することができる。また、生検は、抗凝固薬を服用した患者で実現不可能であり、ARが焦点または斑状である場合は、この手法の限られたサンプリングサイトは偽陰性の結果になることがあります。結果として、これはしばしば、様々な移植モデルの使用を含む動物に行わなければならない新しいAR検出方法に対する継続的な検索をもたらした。 60代前半ラット腎移植はexaminための十分に確立された実験的な方法であるため、AR 2 ATIONと分析。さらに小動物陽電子放射断層撮影（PET）が同種片側腎摘出ラット腎移植モデルでARを評価し、非侵襲的に、特定のための新たな選択肢として移植FDG-PETイメージングを提案する18 F-フルオロデオキシグルコース（FDG）を使用して、我々はここに存在し人間の状況3にもARのと早期診断。さらに、この方法は、移植片拒絶4の監視を改善するためのフォローアップのために適用することができる。

組織学 AR白血球の間に、 すなわち 、主にTリンパ球拒絶反応の重症度は、炎症の程度によって反射されるのに対し、移植に補充される。ここに示されている定期的な酸シッフ（PAS）染色（ 図1）では、腎同種移植片はAR、すなわち糸球体炎、尿細管炎、endothelialitisとグラフト浸潤（ 図1、ATX POD4）（POD =病日）の重要な組織学的兆候を示してい?...

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

我々はここに非侵襲的に18F-フルオロデオキシと陽電子放射断層撮影法を用いた急性同種移植片拒絶反応を評価するために、ラット腎移植モデルを提示する。

使用したラット腎移植後の急性拒絶反応の非侵襲的イメージング<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 ビュー.  University of Münster. 我々はここに非侵襲的に18F-フルオロデオキシと陽電子放射断層撮影法を用いた急性同種移植片拒絶反応を評価するために、ラット腎移植モデルを提示する。

ビデオ: 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.

乳幼児の脳代謝と血行動態の非侵襲的光計測

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

医学

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.

縦用の眼の前房に移植、非侵襲<emインビボ&gt;</emリアルタイムでのシングルセル分解能&gt;イメージング

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.

と陽電子放射断層撮影法を用いた非侵襲的イメージングとリビングラットにおける脳虚血の分析<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.

卵巣がん再発を検出し、モニターするための非侵襲的イメージングのためのマウスモデル

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.

マウス腎臓移植：同種移植片拒絶のモデル

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.

使用するグルコース代謝動態を決定します<sup&gt; 18</sup&gt; F-FDGマイクロ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.

ラットモデルにおける部分的な術後後の漏れを減らすために脂肪組織由来幹細胞シートの移植

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.

人間の活動の地域の定量化と全身褐色脂肪組織18F-FDG PET/CT を使用

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.

重症脳損傷患者 [18F] FDG トレーサーを用いた定量的評価

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.

免疫標識腎組織の光学クリアリングとイメージング

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