我々は、この作品でのサポートにファイザー製薬から契約（WS1763368）を認識したい。我々はまた彼らのサポートのために高等教育のサウジアラビア省（モーホー）に感謝したいと思います。

1。エクササイズ<ol><li>この実験のための関心の筋肉は、肘の屈筋（上腕二頭筋）であった。 </li><li>筋肉の強さは個々に適切な抵抗を与えることができるように各参加者のために測定した。これは、彼らの抵抗の最大値（RM）のための各参加者をテストすることによって決定した。 </li><li> RMをテストするため、我々は、筋肉の強さを測定するためにBIOPAC（DA - 100C）生体電気増幅器モジュール（モンテシステム、ゴレタ、カリフォルニア州）を介してコンピュータとインターフェースひずみゲージのデバイスを使用する。モジュールは、MP - 100 1秒あたり1,000ヘルツの周波数でデジタル変換器のサンプリングのアナログ、および24ビット（図10）の解像度で接続されました。 </li><li>ひずみゲージのデバイスは、45 °の角度でベンチに固定した。被験者は運動の力が自分の手首を介しているように、デバイスの背後に座ると、パディング領域に自分の肘を休ませるように指示された。これは、対象がないことを保証するために最善の方法だったtは上腕二頭筋（図11）以外の筋肉を募集。 </li><li>強度は、それぞれの収縮は、収縮を分離する約45秒で持続時間が3秒であることで3度も決定した。 3つの測定の平均値はRMだ。 </li><li>各参加者の二頭筋のためにRMを決定した後、運動の対象とするセッションは、そのRMの35％で行った。 </li><li>すべての被験者は、筋肉痛（DOMS）を誘導するために適切な重み付けダンベルを使用して同じ運動を施行した。これは、椅子に座っている間二頭筋の濃度のカールの25回の繰り返しの4セットを行うことにより実施し、肘で自分の太もも（図12）でサポートされました。 </li><li>各被験者は、各セットの間に90秒の休息期間を与えられた。被験者は、25台の繰り返しのフルセットをやったか、彼らは着実に運動中に体重を制御するために失敗した場合に停止するように指示されました。 </li></ol> 2。赤外のCaメラの準備とセットアップ<ol><li>赤外線イメージングが行われる部屋は、偽熱測定値になりかねない、室温の違い、から任意の外部バイアスを最小限に抑えるために一定の温度に設定した。この実験の目的のために我々は約23℃に維持した温度管理された部屋を持っていた° C（+ / - 0.5 ° C）。 </li><li>カメラが離れて1メートルの距離に設定し、垂直の角度で肌に（9A図）測定される*. </li><li>必要な距離が設定された後、被験者は画像が撮影されるまでじっと立ってするように勧められた。これが数秒以上かかることはありませんが、これは撮影画像の精度を保証するために動きを最小限に抑えるために非常に重要です。 </li><li>これは、任意の赤外線干渉を最小限に抑えるため、部屋が暗い色の塗料ではなく、明るい色を有することが好ましい。 </li><li>赤外線画像、becaを扱うときの照明にも重要です。蛍光灯やタングステン灯など​​の赤外線波が偽の高い測定値を与えることができる発する光源を使用してください。 LEDライトは、ほとんど何の赤外線干渉（図9B）を生成しないように最善の照明のオプションは、均一なLEDライトを装備したお部屋となります* </li></ol>テストの*シリーズは、我々は別の角度（0（垂直）、15、30、45、および60度）で、皮膚の画像を比較してFLIR 660 IRカメラ（図8）を使用して、私たちのラボで行われた、と皮膚からの距離が異なる（1、2、および5メートル）で、正確に皮膚の温度を検出する。すべての画像は校正済みの熱電対と比較し、画像と熱電対の測定値の間に最高の相関は、垂直の角度と、肌から1メートル（R = 0.93）の距離であった。別の角度と距離は、ピクセレーションの損失が発生し、画像と熱電対の測定値間の全体​​的な相関を減少させた。 3。画像アックイテルメrement <ol><li>この実験の目的のために、行使筋肉のイメージは運動前、および24で撮影、そして48時間後に運動した。 </li><li>ターゲット以外のソースからの体温は、熱画像を乱し、誤った測定値を与えることができる。このような理由から、誰が意図したターゲットの横や背後に立ってはならない。 </li><li>この調査では、行使し、非行使のアームの両方の画像は、比較目的のために採取した。我々は、腕のいずれかを行使前述したように、そして他のアームは、（図4と5）を対照として使用した。 </li><li>それが誰に属している画像を識別することは困難になる可能性があるため、赤外線カメラからの画像の番号は、別のスプレッドシート上で即座に記録した。 </li></ol> 4。画像の処理と分析<ol><li> 1.10.2：取得したIR画像は&quot;ThermoVision ExaminIR&quot;ソフトウェアのバージョンを用いて処理した。 </li><li>必要な画像を選択した後R分析、関心の4つの領域は、ソフトウェアのインタフェース（図6）の統計ボックスを使用して、アームの取得画像上で同定された。 </li><li>アーム間の必要な地域の位置が特定されているときに、ソフトウェアは選択された領域のそれぞれのための温度の平均と標準偏差を示しています。我々は、どちらのクロス個別に各地域を比較したり、腕全体の平均気温（図7）を得ることができる。 </li></ol> 5。ビジュアルアナログスケール＆血液分析<ol><li>視覚的アナログスケール（VAS）は、腕の主観的な痛みを評価するために使用されていました。スケールは10cm（100ミリメートル）一方の端に&quot;痛み&quot;、そして反対側の端に&quot;非常に痛い&quot;とマークしない長い行を持っていた。各参加者は、痛みへの反応を示すために、10cmの線に沿って印を作るために指示した。 </li><li> VASのは、24時間運動した後、および48時間で、練習前に被験者に投与した。 </li><li>末梢血は、COLていた血液中のミオグロビンの濃度レベルを測定するために被験者からlected。 </li><li>血液が運動の前に被験者肘静脈から描かれた、運動後30分は終わった、とで48時間。 </li><li>血液は、細胞から血清を分離するために10分間4000rpmで遠心分離した。ミオグロビンの分析が行われたまで、次いで、試料を-80℃で保存した。 </li><li>ミオグロビンは、TOSOH&quot;AIA - 360&quot;自動酵素免疫分析装置（東ソー株式会社、東京、日本）を用いて測定した。ミオグロビンのアッセイキット（ミョー025297、ST AIA - PACKミオグロビンは）命令を製造するために従って使用した。 </li></ol> 6。代表的な結果この調査中に撮影した赤外線熱画像の結果は明らかに図1に表されます。 41被験者の行使腕を3期間（運動前、24時間後の運動、および48時間後の運動）で撮影した画像は、明らか運動前の温度、および48時間で撮影された温度と比較して2日目の温度の顕著な増加（24時間後の運動）。図1に示すように、平均皮膚温度は32.80でした+ / - 1.03 °日1 C（運動前）、および33.96 + / - 1.46 °日2用C（24時間後の運動）、および32.82 + / - 3日目のための1.29（48時間後の運動）。 1日目から2日目に皮膚の温度の違いは、（ANOVA、P &lt;0.01）有意であった。 しかし、未行使の腕のために、3時間の期間の間で変化は明らかではなかった。平均皮膚温が33.08であることを図1に示して+ / - 0.83 °日1 C（運動前）、および32.79 + / - 1.42 °日2（24時間後の運動）、および33.17のためのC + / - 0.95のための3日目（48時間後の運動）。 3日間の皮膚の温度でこの差は有意（ANOVA、P = 0.38）はなかった。 VASによる痛みの測定値の結果を図2に示す。図に見られるがFigure 2は、報告された痛みは、2日目および3の劇的な増加が認められた。行使の筋肉の痛みのレベルは3.6から+ /増加 - 36.3へ、6.1日目1日+ / - 2日目で22.8、、37.5 + / - 25.3 3日目。 1日目から、この増加は有意であった（ANOVA、P &lt;0.01）。 ミオグロビンの濃度レベルの結果を図3に示されています。この図に見られるように、1日目の2ミオグロビン濃度（事前に、＆30分後の運動）の間の任意の変化はほとんどなかった。しかし3日目に、ミオグロビンの増加は非常に大きかった。 1日目の最初の2濃度と比較した場合、3日目のこの増加は、血液のミリリットル当たり約147ナノグラム（ng / mLの）であった。 3日目で249.51 ng / mLの - ベースラインで7.66 ng / mLに、31.66 + / - - 11.89 ng / mLの30分後の運動、および178.96 + /ミオグロビン濃度は、+ / 30.12であった。 3日目のこの増加は、（ANOVA、P &lt;0.01）、高度に有意であった。 相関分析は、皮膚の温度との間で行われたIR画像、およびVASの痛みのレベルから得られる。それは2日目、そして2日目に皮膚の温度測定上のVAS測定値の間にかなりの相関があることがわかった。この相関は有意であった（R = 0.312、P &lt;0.05）。しかし、VASの測定値と3日目の皮膚温との間には明らかな相関はなかった。この相関は有意であった（R = 0.047、P = 0.77）。 <img src="/files/ftp_upload/3551/3551fig1.jpg" alt="図1"/> 図1。行使腕（ダイヤモンド）で、皮膚の温度の違い、そして3日間の期間にわたって41科目の未行使の武器（四角）の代表的なグラフ。 <img src="/files/ftp_upload/3551/3551fig2.jpg" alt="図2"/> 図2。全41科目の3日間の期間にわたるVASで測定した知覚筋肉痛の違いの代表的なグラフ。 図 3の時間間隔毎に、全41科目のミオグロビン濃度の違い3。代表的なグラフ。 <img src="/files/ftp_upload/3551/3551fig4.jpg" alt="図4"/> 図4。運動の前に対象者の行使腕の）典型的なIRのイメージ。 24時間練習した後、同じ被験者の腕のB）IR画像。 <img src="/files/ftp_upload/3551/3551fig5.jpg" alt="図5"/> 図5。運動前に被写体の未行使の腕の）典型的なIRのイメージ。 24時間練習した後、同じ被験者の腕のB）IR画像。 <img src="/files/ftp_upload/3551/3551fig6.jpg" alt="図6"/> 図6。THERMを分析するための関心の4地域の実例腕のアルイメージ。 <img src="/files/ftp_upload/3551/3551fig7.jpg" alt="図7"/> 図7。行使腕のIR画像に興味の4箱を示す&quot;ThermoVision ExaminIR&quot;のためのソフトウェアインタフェース。また、各ボックスの統計的解釈も表示されます。 <img src="/files/ftp_upload/3551/3551fig8.jpg" alt="図8"/> 図8。この調査（FLIR 660）のために使用される赤外線熱カメラ。 <img src="/files/ftp_upload/3551/3551fig9.jpg" alt="図9"/> 図9。）離れて被験者の腕から赤外線カメラ1メーターのセットアップ。 B）LEDが点灯し、画像が撮影された実験室で使用。 <img src="/files/ftp_upload/3551/3551fig10.jpg" alt="図10"/> 図10。）筋力を測定するために使用されるBIOPACモジュール。 B）ひずみ​​ゲージのデバイスは4に固定5 °傾斜ベンチとBIOPACシステムへフック。 <img src="/files/ftp_upload/3551/3551fig11.jpg" alt="図11"/> 図11。ひずみゲージのデバイス上で力を発揮する典型的な主題。 <img src="/files/ftp_upload/3551/3551fig12.jpg" alt="図12"/> 図12。筋肉痛を誘発するための運動のプロトコルを受けている対象。

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K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Noninvasive+imaging+techniques+in+the+assessment+of+scleroderma+spectrum+disorders.">Noninvasive imaging techniques in the assessment of scleroderma spectrum disorders.</a> Arthritis &#38; Rheumatism. 61, 1103-1111 (2009).</li><li>Zaproudina, N., Ming, Z., Hanninen, O. O. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plantar+Infrared+Thermography+Measurements+and+Low+Back+Pain+Intensity.">Plantar Infrared Thermography Measurements and Low Back Pain Intensity.</a> Journal of Manipulative and Physiological Therapeutics. 29, 219-223 (2006).</li><li>Kim, Y. -. C., Bahk, J. -. H., Lee, S. -. C., Lee, Y. -. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Infrared+Thermographic+Imaging+in+the+Assessment+of+Successful+Block+on+Lumbar+Sympathetic+Ganglion.">Infrared Thermographic Imaging in the Assessment of Successful Block on Lumbar Sympathetic Ganglion.</a> Yonsei Medical Journal. 44, 119-124 (2003).</li><li>Brancaccio, P., Lippi, G., Maffulli, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biochemical+markers+of+muscular+damage.">Biochemical markers of muscular damage.</a> Clinical Chemistry and Laboratory Medicine. 48, 757-767 (2010).</li><li>Neubauer, O., K&#246;nig, D., Wagner, K. -. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recovery+after+an+Ironman+triathlon:+sustained+inflammatory+responses+and+muscular+stress.">Recovery after an Ironman triathlon: sustained inflammatory responses and muscular stress.</a> European Journal of Applied Physiology. 104, 417-426 (2008).</li></ol>

利害の競合が宣言されない。

この調査の主な目的は、激しい運動後の筋肉痛を検出および測定に熱赤外イメージングの有用性を評価することであった、そして我々の結果は、IRイメージングでは、特に運動の最初の24時間以内に、検出DOMSのための有効な手法であることが示唆された。 Pennes 18は手足の筋肉から皮膚への熱の流れの非常に詳細なモデルを提供するので、これは、驚くべきことではない。このモデル?...

<table border="1"><tbody><tr><td> デバイスの名前 </td><td> 会社 </td><td> カタログ番号 </td><td> コメント </td></tr><tr><td>赤外線サーマルカメラ</td><td> FLIR </td><td> FLIR SC660 </td><td></td></tr><tr><td>熱赤外線解析ソフトウェア</td><td> ThermoVision ExaminIR </td><td>ソフトウェアバージョン1.10.2 </td><td></td></tr><tr><td>生体アンプモジュール</td><td> BIOPAC </td><td> DA100C </td><td> DA100Cは、可変ゲインの設定、および調整可能な電圧リファレンスを提供します。 </td></tr><tr><td>デジタルコンバータモジュールへのアナログ</td><td> BIOPAC </td><td> MP100 </td><td></td></tr><tr><td>自動化された酵素イムノアッセイアナライザー</td><td>東ソー</td><td> AIA -360 </td><td>この装置は、血液サンプルを分析し、ミオグロビンの測定値を得るために使用された。 </td></tr>
 </tbody>
</table>

the use of thermal infra-red imaging to detect delayed onset muscle soreness

また、運動誘発性筋損傷（EIMD）として知られる遅発性筋痛（DOMS）は、、一般に時間の長時間物理的に非アクティブになっている人に経験し、練習1-4の予想外の試合で始まっているが、こともできますトレーニング5、通常の限界を超えて行使する選手で起こる。この痛みを伴う現象に伴う症状は、重度の衰弱痛み1,3,5に、わずかな筋肉の圧痛の範囲であり得る。これらの症状と24〜72時間後に運動1,3の間に関連する不快感の練習の終了に続く最初の24時間以内に増加し、ピークの強度。このような理由から、DOMSは、個々のパフォーマンスに影響を与える、と多くの1,4のための威圧的になることができるスポーツの傷害の最も一般的な再発形態の一つです。 最後の30年間の場合は、DOMSの現象は、相当な量を得て運動生理学、スポーツ、リハビリテーションの分野6の研究者、専門家の間で関心の。その基礎となるメカニズム、治療介入、および予防戦略1-5,7-12に関しては、この痛みの発生を調査して発表された研究の様々があった。しかし、それはこの条件6を定量化するために使用される測定ツールとメソッドの間でばらつきの広い額があるので、DOMSは、定量化する簡単な病理ではないことを文献から明らかである。それは合意は、それが困難な特定の介入が実際にこのタイプの痛みのかどうかに関連する症状を減少させることに役立つかどうかを確認するために行うDOMS用の最高の評価尺度、に行われていないことは明らかである。多くの研究が主観的ではなく、客観的な尺度である視覚的アナログスケール（VAS）10,13-15を 、使用して痛みを測定することに依存するためこのように、DOMSは、ややあいまいであると見ることができます。にもかかわらず針筋肉の生検、およびmyofibreタンパク質の血中濃度が約6のゴールドスタンダードと考えられるかもしれない、これらの血液タンパク質のいくつかの大きな変動がしばしば侵襲技術に関連する高いリスクに加えて、6,16の文書化されている。 したがって、現在の調査では、我々は、関連する筋肉の痛みを検出するために行使筋肉上記の皮膚の熱赤外（IR）イメージング技術をテストした。赤外線サーモグラフィを使用し、1950年の17以来、病気や感染症の様々なタイプの検出に成功したことが判明している。しかし驚くべきことに、何も近くには、DOMSと皮膚温度の変化に行われています。この調査の主な目的は、この安全かつ非侵襲的方法を用いてDOMSの変化を調べることであった。

Watch this Scientific Journal Video about The Use of Thermal Infra-Red Imaging to Detect Delayed Onset Muscle Soreness at JoVE.com

The Use of Thermal Infra-Red Imaging to Detect Delayed Onset Muscle Soreness

この調査の目的は、赤外線熱カメラを使用すると、運動後の筋肉痛を検出および定量するための有効なツールであるかどうか評価することであった。

遅発性筋痛を検出する熱赤外線イメージングの使用

Delayed onset muscle soreness (DOMS), also known as exercise induced muscle damage (EIMD), is commonly experienced in individuals who have been physically ...

the-use-thermal-infra-red-imaging-to-detect-delayed-onset-muscle

Research

JoVE Journal

Medicine

19.1K ビュー.  Loma Linda University. この調査の目的は、赤外線熱カメラを使用すると、運動後の筋肉痛を検出および定量するための有効なツールであるかどうか評価することであった。

ビデオ: The Use of Thermal Infra-Red Imaging to Detect Delayed Onset Muscle Soreness

Thermal Ablation for the Treatment of Abdominal Tumors

Percutaneous thermal ablation is an emerging treatment option for many tumors of the abdomen not amenable to conventional treatments. During a thermal ablation procedure, a thin applicator is guided into the target tumor under imaging guidance. Energy is then applied to the tissue until temperatures rise to cytotoxic levels (50-60 &deg;C). Various energy sources are available to heat biological tissues, including radiofrequency (RF) electrical current, microwaves, laser light and ultrasonic waves. Of these, RF and microwave ablation are most commonly used worldwide.
During RF ablation, alternating electrical current (~500 kHz) produces resistive heating around the interstitial electrode. Skin surface electrodes (ground pads) are used to complete the electrical circuit. RF ablation has been in use for nearly 20 years, with good results for local tumor control, extended survival and low complication rates1,2. Recent studies suggest RF ablation may be a first-line treatment option for small hepatocellular carcinoma and renal-cell carcinoma3-5. However, RF heating is hampered by local blood flow and high electrical impedance tissues (eg, lung, bone, desiccated or charred tissue)6,7. Microwaves may alleviate some of these problems by producing faster, volumetric heating8-10. To create larger or conformal ablations, multiple microwave antennas can be used simultaneously while RF electrodes require sequential operation, which limits their efficiency. Early experiences with microwave systems suggest efficacy and safety similar to, or better than RF devices11-13.
Alternatively, cryoablation freezes the target tissues to lethal levels (-20 to -40 &deg;C). Percutaneous cryoablation has been shown to be effective against RCC and many metastatic tumors, particularly colorectal cancer, in the liver14-16. Cryoablation may also be associated with less post-procedure pain and faster recovery for some indications17. Cryoablation is often contraindicated for primary liver cancer due to underlying coagulopathy and associated bleeding risks frequently seen in cirrhotic patients. In addition, sudden release of tumor cellular contents when the frozen tissue thaws can lead to a potentially serious condition known as cryoshock 16.
Thermal tumor ablation can be performed at open surgery, laparoscopy or using a percutaneous approach. When performed percutaneously, the ablation procedure relies on imaging for diagnosis, planning, applicator guidance, treatment monitoring and follow-up. Ultrasound is the most popular modality for guidance and treatment monitoring worldwide, but computed tomography (CT) and magnetic resonance imaging (MRI) are commonly used as well. Contrast-enhanced CT or MRI are typically employed for diagnosis and follow-up imaging.

A thermal tumor ablation procedure is described. The entire procedure is detailed, including pretreatment planning and imaging studies, anesthesia, adjuvant techniques to facilitate a percutaneous approach, imaging guidance of the ablation device to the tumor, thermal treatment, post-treatment care and follow-up.

腹部腫瘍の治療のための熱的アブレーション

Percutaneous thermal ablation is an emerging treatment option for many tumors of the abdomen not amenable to conventional treatments. During a thermal ...

医学

Quantitative Visualization and Detection of Skin Cancer Using Dynamic Thermal Imaging

In 2010 approximately 68,720 melanomas will be diagnosed in the US alone, with around 8,650 resulting in death 1. To date, the only effective treatment for melanoma remains surgical excision, therefore, the key to extended survival is early detection 2,3. Considering the large numbers of patients diagnosed every year and the limitations in accessing specialized care quickly, the development of objective in vivo diagnostic instruments to aid the diagnosis is essential. New techniques to detect skin cancer, especially non-invasive diagnostic tools, are being explored in numerous laboratories. Along with the surgical methods, techniques such as digital photography, dermoscopy, multispectral imaging systems (MelaFind), laser-based systems (confocal scanning laser microscopy, laser doppler perfusion imaging, optical coherence tomography), ultrasound, magnetic resonance imaging, are being tested. Each technique offers unique advantages and disadvantages, many of which pose a compromise between effectiveness and accuracy versus ease of use and cost considerations. Details about these techniques and comparisons are available in the literature 4.
Infrared (IR) imaging was shown to be a useful method to diagnose the signs of certain diseases by measuring the local skin temperature. There is a large body of evidence showing that disease or deviation from normal functioning are accompanied by changes of the temperature of the body, which again affect the temperature of the skin 5,6. Accurate data about the temperature of the human body and skin can provide a wealth of information on the processes responsible for heat generation and thermoregulation, in particular the deviation from normal conditions, often caused by disease. However, IR imaging has not been widely recognized in medicine due to the premature use of the technology 7,8 several decades ago, when temperature measurement accuracy and the spatial resolution were inadequate and sophisticated image processing tools were unavailable. This situation changed dramatically in the late 1990s-2000s. Advances in IR instrumentation, implementation of digital image processing algorithms and dynamic IR imaging, which enables scientists to analyze not only the spatial, but also the temporal thermal behavior of the skin 9, allowed breakthroughs in the field.
In our research, we explore the feasibility of IR imaging, combined with theoretical and experimental studies, as a cost effective, non-invasive, in vivo optical measurement technique for tumor detection, with emphasis on the screening and early detection of melanoma 10-13. In this study, we show data obtained in a patient study in which patients that possess a pigmented lesion with a clinical indication for biopsy are selected for imaging. We compared the difference in thermal responses between healthy and malignant tissue and compared our data with biopsy results. We concluded that the increased metabolic activity of the melanoma lesion can be detected by dynamic infrared imaging.

We demonstrated that malignant pigmented lesions with increased metabolic activity generate quantifiable amounts of heat and the measurement of the transient thermal response of the skin to a cooling excitation allows quantitative identification of melanoma and other skin cancers (vs. non-proliferative nevi) at an early stage of the disease.

定量的な可視化と動的な熱画像を用いた皮膚癌の発見

In 2010 approximately 68,720 melanomas will be diagnosed in the US alone, with around 8,650 resulting in death 1. To date, the only effective treatment ...

Use of Animal Model of Sepsis to Evaluate Novel Herbal Therapies

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several techniques, including infusion of exogenous bacterial toxin (endotoxemia) or bacteria (bacteremia), as well as surgical perforation of the cecum by cecal ligation and puncture (CLP)1-3. CLP allows bacteria spillage and fecal contamination of the peritoneal cavity, mimicking the human clinical disease of perforated appendicitis or diverticulitis. The severity of sepsis, as reflected by the eventual mortality rates, can be controlled surgically by varying the size of the needle used for cecal puncture2. In animals, CLP induces similar, biphasic hemodynamic cardiovascular, metabolic, and immunological responses as observed during the clinical course of human sepsis3. Thus, the CLP model is considered as one of the most clinically relevant models for experimental sepsis1-3.
Various animal models have been used to elucidate the intricate mechanisms underlying the pathogenesis of experimental sepsis. The lethal consequence of sepsis is attributable partly to an excessive accumulation of early cytokines (such as TNF, IL-1 and IFN-&gamma;)4-6 and late proinflammatory mediators (e.g., HMGB1)7. Compared with early proinflammatory cytokines, late-acting mediators have a wider therapeutic window for clinical applications. For instance, delayed administration of HMGB1-neutralizing antibodies beginning 24 hours after CLP, still rescued mice from lethality8,9, establishing HMGB1 as a late mediator of lethal sepsis. The discovery of HMGB1 as a late-acting mediator has initiated a new field of investigation for the development of sepsis therapies using Traditional Chinese Herbal Medicine. In this paper, we describe a procedure of CLP-induced sepsis, and its usage in screening herbal medicine for HMGB1-targeting therapies.

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection, and can be simulated by a surgical technique termed cecal ligation and puncture (CLP). Here we describe a method to use CLP-induced animal model to screen medicinal herbs for therapeutic agents.

新しいハーブ治療法を評価するために敗血症の動物モデルの使用

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several ...

The Use of Pharmacological-challenge fMRI in Pre-clinical Research: Application to the 5-HT System

Pharmacological MRI (phMRI) is a new and promising method to study the effects of substances on brain function that can ultimately be used to unravel underlying neurobiological mechanisms behind drug action and neurotransmitter-related disorders, such as depression and ADHD. Like most of the imaging methods (PET, SPECT, CT) it represents a progress in the investigation of brain disorders and the related function of neurotransmitter pathways in a non-invasive way with respect of the overall neuronal connectivity. Moreover it also provides the ideal tool for translation to clinical investigations. MRI, while still behind in molecular imaging strategies compared to PET and SPECT, has the great advantage to have a high spatial resolution and no need for the injection of a contrast-agent or radio-labeled molecules, thereby avoiding the repetitive exposure to ionizing radiations. Functional MRI (fMRI) is extensively used in research and clinical setting, where it is generally combined with a psycho-motor task. phMRI is an adaptation of fMRI enabling the investigation of a specific neurotransmitter system, such as serotonin (5-HT), under physiological or pathological conditions following activation via administration of a specific challenging drug.
The aim of the method described here is to assess brain 5-HT function in free-breathing animals. By challenging the 5-HT system while simultaneously acquiring functional MR images over time, the response of the brain to this challenge can be visualized. Several studies in animals have already demonstrated that drug-induced increases in extracellular levels of e.g. 5-HT (releasing agents, selective re-uptake blockers, etc) evoke region-specific changes in blood oxygenation level dependent (BOLD) MRI signals (signal due to a change of the oxygenated/deoxygenated hemoglobin levels occurring during brain activation through an increase of the blood supply to supply the oxygen and glucose to the demanding neurons) providing an index of neurotransmitter function. It has also been shown that these effects can be reversed by treatments that decrease 5-HT availability16,13,18,7. In adult rats, BOLD signal changes following acute SSRI administration have been described in several 5-HT related brain regions, i.e. cortical areas, hippocampus, hypothalamus and thalamus9,16,15. Stimulation of the 5-HT system and its response to this challenge can be thus used as a measure of its function in both animals and humans2,11.

The goal of this technique is to assess serotonin (5-HT) neurotransmitter function in the live and free-breathing animal with pharmacological magnetic resonance imaging (phMRI) and an intravenous challenge with a selective serotonin reuptake inhibitor (SSRI), fluoxetine.

前臨床研究における薬理学的チャレンジのfMRIの利用：5-HTシステムへの応用

Pharmacological MRI (phMRI) is a new and promising method to study the effects of substances on brain function that can ultimately be used to unravel ...

Molecular Imaging to Target Transplanted Muscle Progenitor Cells

Duchenne muscular dystrophy (DMD) is a severe genetic neuromuscular disorder that affects 1 in 3,500 boys, and is characterized by progressive muscle degeneration1, 2. In patients, the ability of resident muscle satellite cells (SCs) to regenerate damaged myofibers becomes increasingly inefficient4. Therefore, transplantation of muscle progenitor cells (MPCs)/myoblasts from healthy subjects is a promising therapeutic approach to DMD. A major limitation to the use of stem cell therapy, however, is a lack of reliable imaging technologies for long-term monitoring of implanted cells, and for evaluating its effectiveness. Here, we describe a non-invasive, real-time approach to evaluate the success of myoblast transplantation. This method takes advantage of a unified fusion reporter gene composed of genes (firefly luciferase [fluc], monomeric red fluorescent protein [mrfp] and sr39 thymidine kinase [sr39tk]) whose expression can be imaged with different imaging modalities9, 10. A variety of imaging modalities, including positron emission tomography (PET), single-photon emission computed tomography (SPECT), magnetic resonance imaging (MRI), optical imaging, and high frequency 3D-ultrasound are now available, each with unique advantages and limitations11. Bioluminescence imaging (BLI) studies, for example, have the advantage of being relatively low cost and high-throughput. It is for this reason that, in this study, we make use of the firefly luciferase (fluc) reporter gene sequence contained within the fusion gene and bioluminescence imaging (BLI) for the short-term localization of viable C2C12 myoblasts following implantation into a mouse model of DMD (muscular dystrophy on the X chromosome [mdx] mouse)12-14. Importantly, BLI provides us with a means to examine the kinetics of labeled MPCs post-implantation, and will be useful to track cells repeatedly over time and following migration. Our reporter gene approach further allows us to merge multiple imaging modalities in a single living subject; given the tomographic nature, fine spatial resolution and ability to scale up to larger animals and humans10,11, PET will form the basis of future work that we suggest may facilitate rapid translation of methods developed in cells to preclinical models and to clinical applications.

A non-invasive means to evaluate the success of myoblast transplantation is described. The method takes advantage of a unified fusion reporter gene composed of genes whose expression can be imaged with different imaging modalities. Here, we make use of a fluc reporter gene sequence to target cells via bioluminescence imaging.

移植筋前駆細胞を標的とする分子イメージング

Duchenne muscular dystrophy (DMD) is a severe genetic neuromuscular disorder that affects 1 in 3,500 boys, and is characterized by progressive muscle ...

High-throughput Flow Cytometry Cell-based Assay to Detect Antibodies to N-Methyl-D-aspartate Receptor or Dopamine-2 Receptor in Human Serum

Over the recent years, antibodies against surface and conformational proteins involved in neurotransmission have been detected in autoimmune CNS diseases in children and adults. These antibodies have been used to guide diagnosis and treatment. Cell-based assays have improved the detection of antibodies in patient serum. They are based on the surface expression of brain antigens on eukaryotic cells, which are then incubated with diluted patient sera followed by fluorochrome-conjugated secondary antibodies. After washing, secondary antibody binding is then analyzed by flow cytometry. Our group has developed a high-throughput flow cytometry live cell-based assay to reliably detect antibodies against specific neurotransmitter receptors. This flow cytometry method is straight forward, quantitative, efficient, and the use of a high-throughput sampler system allows for large patient cohorts to be easily assayed in a short space of time. Additionally, this cell-based assay can be easily adapted to detect antibodies to many different antigenic targets, both from the central nervous system and periphery. Discovering additional novel antibody biomarkers will enable prompt and accurate diagnosis and improve treatment of immune-mediated disorders.

Over the recent years, live cell-based assays have been used successfully to detect antibodies against surface and conformational antigens. Here, we describe a method using high-throughput flow cytometry enabling the analysis of large cohorts of patients. Detection of novel antibodies will improve diagnosis and treatment of immune-mediated disorders.

ハイスループットフローサイトメトリー細胞ベースのアッセイN-メチル-D-アスパラギン酸受容体またはヒト血清中のドーパ​​ミン-2受容体に対する抗体を検出する

Over the recent years, antibodies against surface and conformational proteins involved in neurotransmission have been detected in autoimmune CNS diseases ...

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 ...

Quantitative Magnetic Resonance Imaging of Skeletal Muscle Disease

Quantitative magnetic resonance imaging (qMRI) describes the development and use of MRI to quantify physical, chemical, and/or biological properties of living systems. Neuromuscular diseases often exhibit a temporally varying, spatially heterogeneous, and multi-faceted pathology. The goal of this protocol is to characterize this pathology using qMRI methods. The MRI acquisition protocol begins with localizer images (used to locate the position of the body and tissue of interest within the MRI system), quality control measurements of relevant magnetic field distributions, and structural imaging for general anatomical characterization. The qMRI portion of the protocol includes measurements of the longitudinal and transverse relaxation time constants (T1 and T2, respectively). Also acquired are diffusion-tensor MRI data, in which water diffusivity is measured and used to infer pathological processes such as edema. Quantitative magnetization transfer imaging is used to characterize the relative tissue content of macromolecular and free water protons. Lastly, fat-water MRI methods are used to characterize fibro-adipose tissue replacement of muscle. In addition to describing the data acquisition and analysis procedures, this paper also discusses the potential problems associated with these methods, the analysis and interpretation of the data, MRI safety, and strategies for artifact reduction and protocol optimization.

Neuromuscular diseases often exhibit a temporally varying, spatially heterogeneous, and multi-faceted pathology. The goal of this protocol is to characterize this pathology using non-invasive magnetic resonance imaging methods.

骨格筋疾患の定量的磁気共鳴イメージング

Quantitative magnetic resonance imaging (qMRI) describes the development and use of MRI to quantify physical, chemical, and/or biological properties of ...

Calcification of Vascular Smooth Muscle Cells and Imaging of Aortic Calcification and Inflammation

Cardiovascular disease is the leading cause of morbidity and mortality in the world. Atherosclerotic plaques, consisting of lipid-laden macrophages and calcification, develop in the coronary arteries, aortic valve, aorta, and peripheral conduit arteries and are the hallmark of cardiovascular disease. In humans, imaging with computed tomography allows for the quantification of vascular calcification; the presence of vascular calcification is a strong predictor of future cardiovascular events. Development of novel therapies in cardiovascular disease relies critically on improving our understanding of the underlying molecular mechanisms of atherosclerosis. Advancing our knowledge of atherosclerotic mechanisms relies on murine and cell-based models. Here, a method for imaging aortic calcification and macrophage infiltration using two spectrally distinct near-infrared fluorescent imaging probes is detailed. Near-infrared fluorescent imaging allows for the ex vivo quantification of calcification and macrophage accumulation in the entire aorta and can be used to further our understanding of the mechanistic relationship between inflammation and calcification in atherosclerosis. Additionally, a method for isolating and culturing animal aortic vascular smooth muscle cells and a protocol for inducing calcification in cultured smooth muscle cells from either murine aortas or from human coronary arteries is described. This in vitro method of modeling vascular calcification can be used to identify and characterize the signaling pathways likely important for the development of vascular disease, in the hopes of discovering novel targets for therapy.

Vascular calcification is an important predictor of and contributor to human cardiovascular disease. This protocol describes methods for inducing calcification of cultured primary vascular smooth muscle cells and for quantifying calcification and macrophage burden in animal aortas using near-infrared fluorescence imaging.

血管平滑筋細胞と大動脈石灰化と炎症のイメージングの石灰化

Cardiovascular disease is the leading cause of morbidity and mortality in the world.  Atherosclerotic plaques, consisting of lipid-laden macrophages and ...

In Vivo Evaluation of the Mechanical and Viscoelastic Properties of the Rat Tongue

The tongue is a highly innervated and vascularized muscle hydrostat on the floor of the mouth of most vertebrates. Its primary functions include supporting mastication and deglutition, as well as taste-sensing and phonetics. Accordingly, the strength and volume of the tongue can impact the ability of vertebrates to accomplish basic activities such as feeding, communicating, and breathing. Human patients with sleep apnea have enlarged tongues, characterized by reduced muscle tone and increased intramuscular fat that can be visualized and quantified by magnetic resonance imaging (MRI). The abilities to measure force generation and viscoelastic properties of the tongue constitute important tools for obtaining functional information to correlate with imaging data. Here, we present techniques for measuring tongue force production in anesthetized Zucker rats via electrical stimulation of the hypoglossal nerves and for determining the viscoelastic properties of the tongue by applying passive Lissajous force/deformation curves.

In Vivo Evaluation of the Mechanical and Viscoelastic Properties of the Rat Tongue

We describe a surgical procedure in an anesthetized rat model for determining the muscle tone and viscoelastic properties of the tongue. The procedure involves specific stimulation of the hypoglossal nerves and application of passive Lissajous force/deformation curves to the muscle.