著者らは、全米科学財団（NSF）からの財政支援は、「健康と環境のための中間赤外線テクノロジー（MIRTHE）」と題されたEEC-0540832を付与認める

楽器の1。準備<ol><li>アンモニア光学センサープラットフォーム、レーザーダイオード·コントローラ、カスタム構築された制御電子ユニット（CEU）、ブレスサンプラー、空気ポンプ、およびラップトップに外部電源をオンにします。 </li><li>アンモニアセンサーの両方の排気と冷却ファンが動作していることを確認してください。注：一つは、センサの背面に位置しており、第二は、簡単にアクセスでき、センサ内に発見された。 </li><li>音響検出モジュールとニードル弁の温度は、アンモニアセンサボックスの側に位置するデジタル表示を確認することによって38.0℃であることを確認します。センサーは、温度が安定するまでの電力を供給されてから約35分待ちます。 </li><li>息サンプラーのデスクトップに見られるアイコン名「ChangeT」をクリックして、55℃に入口チューブとマウスピースの温度を設定します。その後、温度を「UPDをクリックして、続いて、上または下矢印をクリックすることで変更することができます&quot;温度を食べた。デスクトップに戻るには、「終了」をクリックしてください。温度が安定するためのシステムは、少なくとも5分を許可します。 55℃でのサンプラとチューブの温度を維持することは、アンモニアの表面損失を最小限にする。 </li><li>アンモニアセンサーを制御ノートパソコンにソフトウェアを開きます。プログラムは、デスクトップ上の「NH 3ブレスセンサープログラム」という名前のフォルダ内でアクセスすることができます。このフォルダの中には、ユーザーはアイコン「主LabVIEWソフトウェア」を選択する必要があります。このフォルダには、いくつかのアプリケーションが含まれていますが、ユーザは、所望のインタフェースにアクセスするには、 &quot;Mainsequence.vi」を選択する必要があります。画面の左上隅に「ファイル名を指定して実行」を選択します。注：これは、キャリブレーション·シーケンスをロックする行を開始します。 QEPASモニターのレーザーは、自動化ラインロック手順の間に選択された最適な電流、で動作します。このプロセスには、約25分かかります。 </li><li>サンプラー上で新しいセッションを作成するように、EACH対象セッションが接続されたフラッシュドライブに保存されている独自のファイルを持っています。注：これは、サンプラーのデスクトップ上に見られるプログラム「ブレスサンプラー」を開くことによって実行されます。適切にセッションを識別するためにスペースがあります。セッション中に生成されたすべてのデータは、この識別子の下にフラッシュドライブに保存されます。実験の日は、典型的には、ファイル名の一部として使用される。入口とマウスピースの温度が息サンプリングプログラムに入る前に調整する必要があります。 </li><li>インレットパイプに新たな使い捨てのマウスピースを挿入します。指からのアンモニアでマウスピースの汚染を避けるため使い捨て手袋を着用してください。 </li></ol> 2。呼気サンプル集注：関連する治験審査委員会（倫理委員会）は、ヒト被験者を含む任意の研究を承認する必要があります。大幅に息アンモニアに影響を与えることができる多くの要因がある。これらの要因は、直接で息アンモニア測定を変更することができます全身アンモニアレベルをffectingまたは計測への肺からの呼気の代謝産物の旅に影響を与えることによって。 <ol><li>被験者が到着時に約12時間、彼らが試験前に練習から朝を控えていることは、食品を消費していないた、絶食状態で研究室に到着したことを確認してください。 </li><li>全く物質がデータ収集の前に、少なくとも1時間、口の中に導入されていないことを確認してください。被験者は試験前より大きく、1時間歯を磨くことを確認してください。 </li><li>アンモニアセンサの前方に被写体を着座。インレットパイプを保持し、彼らはアンモニアの混入を避けるために、マウスピースには触れていないことを確認するために対象を指示する。 </li><li>サンプラーインターフェイス上で「スタート」をクリックします。対象は限りができますようにのためにマウスピースに息を吐く、またはオペレータは、試料が十分であることがあると判断されるまできた。注：これは、少なくとも10秒間持続する単一のフル呼気である。口の圧力が代理としてリアルタイムで測定される流量用。色分けされた圧力計は、対象の食材を助けるし、50ml /秒の呼気流量を表し水10cmの所望の呼気圧力を維持する。それは、Fe NOを決定するためのプロトコルのためにATS / ERSによって採用されているので、この流量を選択した。この呼気流量は、子供と大人で達成可能である。同様に再現可能な呼気3は、10％未満だけ異なることを得られるべきである。 </li><li>呼気サンプルが完了すると、サンプラーインターフェイス上で「停止」をクリックしてください。 </li></ol> 3。呼気サンプル測定<ol><li>呼気サンプルが分析された後、モニタを動作させる検査技師は、その呼吸プロファイルの任意のセグメントを分析する能力を有する。関心のある息の一部が第III相セグメントです。これは、二酸化炭素の濃度の「プラトー」を特徴とし、ティンバーの後期に中央に見出されるATH。 </li><li>二酸化炭素台地を開始し、息の圧力低下が前に右停止するサンプラーインターフェイス上で縦線をドラッグすることで、試料の第III相の部分を選択します。明確化のため、図1を参照してください。 </li><li>サンプラータッチスクリーンインターフェース上で「ストア」をクリックして、フラッシュドライブにデータを保存します。 </li><li>呼気データが格納されると、ユーザは、新たな呼気サンプルを開始するために「開始」を選択することができる。 </li></ol> 4。ブレスアンモニアに口リンスし、pHの影響を示す<ol><li>ベースラインのアンモニアレベルを確立する3呼吸をサンプリングします。呼吸が離れてから少なくとも5分を取っていることを確認します。 </li><li> 60秒間、30mlの水の分量で十分に口をすすぐ。 </li><li>リンス（第2）の60秒以内に呼気サンプルを収集します。時間をかけてアンモニアの変化を観察するために次の1時間にわたって呼気サンプルを収集します。注：サムplesは毎分ほど頻繁に撮影することができますが、長い間隔は、一般的に使用されている。 </li><li> 60秒を繰り返し4.2.1のための基本的な溶液（水中の炭酸水素ナトリウム）の30ミリリットル分量で十分に口をすすぐ。 </li><li> 60秒を繰り返し4.2.1酸性溶液30ml分量で十分に口をすすぐ。 </li></ol> 5。ブレスアンモニアに入口と輸送管温度の影響を示す<ol><li>入口温度を15分かけてサンプルを3呼吸、体温、約30℃未満に設定</li><li>息サンプラーのタッチスクリーンインターフェース上でデスクトップアイコンを使用して、それぞれ55℃に入口と輸送管の温度を上げる。少なくとも5分間、システムが定常状態に達することを可能にする。 </li><li>サンプル3呼吸、加熱された注入口に離れて5分。 </li></ol>

<ol>
	<li>Adeva, M. M., Souto, G., Blanco, N., Donapetry, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ammonium+metabolism+in+humans.">Ammonium metabolism in humans.</a> Metabolism: clinical and experimental. 61 (11), 1495-1511 (2012).</li><li>Auron, A., Brophy, P. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hyperammonemia+in+review:+pathophysiology,+diagnosis,+and+treatment.">Hyperammonemia in review: pathophysiology, diagnosis, and treatment.</a> Pediatric nephrology. 27 (2), 207-222 (2012).</li><li>Blanco Vela, C. I., Bosques Padilla, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+ammonia+concentrations+in+cirrhosis+patients-still+confusing+after+all+these+years.">Determination of ammonia concentrations in cirrhosis patients-still confusing after all these years.</a> Annals of hepatology. 10 Suppl 2, (2011).</li><li>Mpabanzi, L., Ol de Damink, S. W. M., van de Poll, M. C. G., Soeters, P. B., Jalan, R., Dejong, C. H. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=To+pee+or+not+to+pee:+ammonia+hypothesis+of+hepatic+encephalopathy+revisited.">To pee or not to pee: ammonia hypothesis of hepatic encephalopathy revisited.</a> European journal of gastroenterology &amp; hepatology. 23 (6), 449-454 (2011).</li><li>Goggs, R., Serrano, S., Szladovits, B., Keir, I., Ong, R., Hughes, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+investigation+of+a+point-of-care+blood+ammonia+analyzer.">Clinical investigation of a point-of-care blood ammonia analyzer.</a> Veterinary clinical pathology / American Society for Veterinary Clinical Pathology. 37 (2), 198-206 (2008).</li><li>Huizenga, J. R., Tangerman, A., Gips, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+ammonia+in+biological+fluids.">Determination of ammonia in biological fluids.</a> Annals of clinical biochemistry. 31 (Pt 6), 529-543 (1994).</li><li>Lewicki, R., 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=Real+time+ammonia+detection+in+exhaled+human+breath+with+a+quantum+cascade+laser+based+sensor.">Real time ammonia detection in exhaled human breath with a quantum cascade laser based sensor.</a> 2009 Conference on Lasers and ElectroOptics and 2009 Conference on Quantum electronics and Laser Science Conference. 1, (2009).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=American+Thoracic+Society.+European+Respiratory+Society.+Recommendations+for+Standardized+Procedures+for+the+Online+and+Offline+Measurement+of+Exhaled+Lower+Respiratory+Nitric+Oxide+and+Nasal+Nitric+Oxide.">American Thoracic Society. European Respiratory Society. Recommendations for Standardized Procedures for the Online and Offline Measurement of Exhaled Lower Respiratory Nitric Oxide and Nasal Nitric Oxide.</a> American journal of respiratory and critical care medicine. 171 (8), 912-930 (2005).</li><li>Solga, S. F., 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=Factors+influencing+breath+ammonia+determination.">Factors influencing breath ammonia determination.</a> Journal of breath research. 7 (3), (2013).</li><li>Schmidt, F. M., 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=Ammonia+in+breath+and+emitted+from+skin.">Ammonia in breath and emitted from skin.</a> Journal of breath research. 7 (1), (2013).</li><li>Span&#283;l, P., Dryahina, K., Smith, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+quantitative+study+of+the+influence+of+inhaled+compounds+on+their+concentrations+in+exhaled+breath.">A quantitative study of the influence of inhaled compounds on their concentrations in exhaled breath.</a> Journal of breath research. 7 (1), (2013).</li><li>Boots, A. W., van Berkel, J. J. B. N., Dallinga, J. W., Smolinska, A., Wouters, E. F., van Schooten, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+versatile+use+of+exhaled+volatile+organic+compounds+in+human+health+and+disease.">The versatile use of exhaled volatile organic compounds in human health and disease.</a> Journal of breath research. 6 (2), (2012).</li><li>Hibbard, T., Killard, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Breath+ammonia+levels+in+a+normal+human+population+study+as+determined+by+photoacoustic+laser+spectroscopy.">Breath ammonia levels in a normal human population study as determined by photoacoustic laser spectroscopy.</a> Journal of breath research. 5 (3), (2011).</li><li>Wang, T., Pysanenko, A., Dryahina, K., Span&#283;l, P., Smith, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+breath,+exhaled+via+the+mouth+and+nose,+and+the+air+in+the+oral+cavity.">Analysis of breath, exhaled via the mouth and nose, and the air in the oral cavity.</a> Journal of breath research. 2 (3), (2008).</li><li>Amann, A., Smith, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Volatile Biomarkers. 1st Edition. , (2013).</li></ol>

Dr. Risby serves as a consultant for Loccioni Humancare the manufacturer of the breath sampler.

リアルタイムで微量代謝物を検出することが可能な非侵襲的手順の利点は明白である。しかし、息研究のフィールドは、この潜在能力を発揮するために苦労している。息の測定は、多くの交絡因子に対して脆弱動的なプロセスである。我々のアプローチは重要な強みを持っている：具体的には、息をサンプラーに接続されたライスQEPAS基づくアンモニアモニターの感度および速度を正確に測?...

アンモニアは、タンパク質代謝の1ユビキタス副産物である。アンモニア測定は、したがって、臨床医は、様々な疾患と健康状態2の評価に役立つことができます。しかし、アンモニアは、非常に反応性であるので、血液または呼気を介して、正確に測定することは困難である。一般的に使用されるものの、血アッセイ精度3についての基本的な問題を含む多数の欠点を有する。しかし、血アッセイの主要な問題は、彼らが唯一のこれまで気まぐれに収集現実である。アンモニア生理学は、多くの血中グルコースおよび他の多くの代謝プロセスと同様に、流体、これまで4を変化させるので、これは重要である。対照的に、息アッセイは容易反復測定を可能にする、完全に非侵襲的かつ迅速である。それはユニークな方法で深刻なアンメットニーズに対処することができるので、このようにして、息のアンモニア測定は魅力的です。 呼気収集しかしながら、ユニークな問題を提示する。瀉血は本質jeoを運ぶ一方、呼吸の変動、口腔粘膜による汚染：いくつかの予測できない方法（などなど 、止血時間、汗汚れ、血球溶血、実験室での測定が遅れ、5）、呼気測定の研究者が新たな挑戦の別のグループと競合しなければならないの誤差のpardyまたは細菌、アンモニア、周囲の空気と装置湿度や温度の影響など6。確かに、それは未知の生物を発見する実験手順を使用して人間に実験装置を接続する作業を過小評価することは賢明ではありません。これらの障害に起因する部分では、息のアンモニアは、まだその可能性を満たしていません。 ここで、我々は、高速かつ正確な結果を得るために私たちの息のアンモニア測定プロトコルを提示する。モニター、インターフェイスサンプラー、人間の影響に注意：我々のプロトコルは、3つの分野での強さを持っています。モニターは、前述の7のようにライス大学の同僚によって建てられました。 MEAの基礎surementは、石英強化光音響分光法（QEPAS）音響トランスデューサ圧電水晶音叉を採用している手法である。音響波が目標微量ガス種によって変調されたレーザー放射の吸収により生成されるときに光音響効果が生じる。微量気体を音響的に変調された周波数に共振する音響セルを用いて検出される。アンモニアの吸収波長は息種を妨害するからスペクトル干渉がないことを選択しました。人間の呼気測定の目的のためには、モニターの主な特徴は、広い測定範囲（十億分〜50部から、少なくとも5000 PPBにPPB）と速度（1秒の測定）が含まれています。モニターの速度は呼吸サイクル全体で時間分解能を可能にします。 モニタは、特別に設計された呼気サンプラに結合される。サンプラーは、圧力センサとカプノグラフから構成されています。これは、リアルタイムで表示され、アーカイブ口の圧力と二酸化炭素だけでなく、センサーによって決定アンモニア濃度を測定。このサンプラは、したがって、呼気が収集されるように呼吸努力の品質を評価する技術者を可能にする。これは米国胸部学会/欧州呼吸器学会（ATS / ERS）8のタスクフォースによって提案された呼気の一酸化窒素（NO鉄） を分析するための勧告を超えてすることを可能にします。すべての息サンプリングのため、使い捨てのワンウェイのインラインバルブが息サンプラーの口ポートで使用されました。 なぜならサンプラーが提供するモニターと品質管理の速度、我々は慎重に9人の影響を評価することができました。呼吸するように指示されると、ほとんどの被験者は、例えば、当初は過換気。このような経口のpHや口リンス、サンプラー、モニターおよび関連するすべての配管の温度、および呼吸のモードなど、他の重要な影響は、後に研究し、FO基礎である下記R実例実験。 最後に、そしておそらく最も顕著には、複数の経験豊富な基は完全に異なるセンサ及び測定手順を用いて呼気アンモニアを測定していることを強調しなければならない。これらは重要な利点と有効性を持つことができる。完全な比較は本研究10,11,12の範囲を超えている。

<table border="0" cellpadding="0" cellspacing="0" width="666">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:211px;">Rice Ammonia Monitor System</td>
			<td style="width:171px;">N/A</td>
			<td style="width:119px;">N/A</td>
			<td style="width:167px;">Not available for commercial purchase</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:211px;">Loccioni Breath Sampler</td>
			<td style="width:171px;">Loccioni Humancare</td>
			<td style="width:119px;">N/A</td>
			<td>Single breath version</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:211px;">Disposable Mouth Piece</td>
			<td style="width:171px;">WestPrime Healthcare</td>
			<td style="width:119px;">G011-200</td>
			<td>Manufacturer is AlcoQuant</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:211px;">Laptop</td>
			<td style="width:171px;">Lenovo</td>
			<td style="width:119px;">N/A</td>
			<td>Old model no longer sold by manufacturer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:211px;">Acid Rinse</td>
			<td style="width:171px;">N/A</td>
			<td style="width:119px;">N/A</td>
			<td>Household acidic drink (coffee, soft drink, citrus juices, etc)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:211px;">Base Rinse</td>
			<td style="width:171px;">N/A</td>
			<td style="width:119px;">N/A</td>
			<td>Water mixed with a nonexact amount of sodium bicarbonate (Arm &#38; Hammer Baking Soda)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:211px;">Neutral Rinse</td>
			<td style="width:171px;">N/A</td>
			<td style="width:119px;">N/A</td>
			<td>Water</td>
		</tr>
	</tbody>
</table>

fast and accurate exhaled breath ammonia measurement

This exhaled breath ammonia method uses a fast and highly sensitive spectroscopic method known as quartz enhanced photoacoustic spectroscopy (QEPAS) that uses a quantum cascade based laser. The monitor is coupled to a sampler that measures mouth pressure and carbon dioxide. The system is temperature controlled and specifically designed to address the reactivity of this compound. The sampler provides immediate feedback to the subject and the technician on the quality of the breath effort. Together with the quick response time of the monitor, this system is capable of accurately measuring exhaled breath ammonia representative of deep lung systemic levels. 
Because the system is easy to use and produces real time results, it has enabled experiments to identify factors that influence measurements. For example, mouth rinse and oral pH reproducibly and significantly affect results and therefore must be controlled. Temperature and mode of breathing are other examples. As our understanding of these factors evolves, error is reduced, and clinical studies become more meaningful. This system is very reliable and individual measurements are inexpensive. 
The sampler is relatively inexpensive and quite portable, but the monitor is neither. This limits options for some clinical studies and provides rational for future innovations.

被験者は、ベースライン呼気アンモニアレベルの広い範囲を生成することが期待できる。健康な人は、100〜1,000 PPBの呼気アンモニア測定で一日を始めることができる。任意の流体で口をすすぐことは、すぐに検出可能な呼気アンモニアの量を変更します。中性および酸性の液は、通常、半分以上で観察されるアンモニアの量を削減する。すすぎの効果が切れるようにこれらのレベルは、その?...

Watch this Scientific Journal Video about Fast and Accurate Exhaled Breath Ammonia Measurement at JoVE.com

Fast and Accurate Exhaled Breath Ammonia Measurement

Ammonia is an important physiologic metabolite relevant to various disease and wellness states. It is also a difficult molecule to measure in breath, which demands particular precautions be taken to obtain accurate results. Not all factors influencing ammonia are known, but progress can be difficult without accounting for these factors.

高速かつ正確な呼気アンモニア測定

This exhaled breath ammonia method uses a fast and highly sensitive spectroscopic method known as quartz enhanced photoacoustic spectroscopy (QEPAS) that ...

fast-and-accurate-exhaled-breath-ammonia-measurement

Research

JoVE Journal

Medicine

13.1K ビュー.  St. Luke's University Hospital. Ammonia is an important physiologic metabolite relevant to various disease and wellness states. It is also a difficult molecule to measure in breath, which demands particular precautions be taken to obtain accurate results. Not all factors influencing ammonia are known, but progress can be difficult without accounting for these factors.

ビデオ: Fast and Accurate Exhaled Breath Ammonia Measurement

Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1&beta; using a Whole Blood Stimulation Assay

Inflammatory processes resulting from the secretion of soluble mediators by immune cells, lead to various manifestations in skin, joints and other tissues as well as altered cytokine homeostasis. The innate immune system plays a crucial role in recognizing pathogens and other endogenous danger stimuli. One of the major cytokines released by innate immune cells is Interleukin (IL)-1. Therefore, we utilize a whole blood stimulation assay in order to measure the secretion of inflammatory cytokines and specifically of the pro-inflammatory cytokine IL-1&beta; 1, 2, 3.
Patients with genetic dysfunctions of the innate immune system causing autoinflammatory syndromes show an exaggerated release of mature IL-1&beta; upon stimulation with LPS alone. In order to evaluate the innate immune component of patients who present with inflammatory-associated pathologies, we use a specific immunoassay to detect cellular immune responses to pathogen-associated molecular patterns (PAMPs), such as the gram-negative bacterial endotoxin, lipopolysaccharide (LPS). These PAMPs are recognized by pathogen recognition receptors (PRRs), which are found on the cells of the innate immune system 4, 5, 6, 7. A primary signal, LPS, in conjunction with a secondary signal, ATP, is necessary for the activation of the inflammasome, a multiprotein complex that processes pro-IL-1&beta; to its mature, bioactive form 4, 5, 6, 8, 9, 10.
The whole blood assay requires minimal sample manipulation to assess cytokine production when compared to other methods that require labor intensive isolation and culturing of specific cell populations. This method differs from other whole blood stimulation assays; rather than diluting samples with a ratio of RPMI media, we perform a white blood cell count directly from diluted whole blood and therefore, stimulate a known number of white blood cells in culture 2. The results of this particular whole blood assay demonstrate a novel technique useful in elucidating patient cohorts presenting with autoinflammatory pathophysiologies.

Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1&#946; using a Whole Blood Stimulation Assay

We describe a simple immunoassay to measure the production of pro-inflammatory cytokines, such as IL-1 beta production, in patients presenting with autoinflammatory phenotypes. By activating cells in whole blood cultures with pathogen-associated molecular patterns, specifically with lipopolysaccharide, cytokine secretion can be conveniently evaluated in whole blood supernatants.

全血刺激のためのアッセイを用いて炎症性サイトカインIL -1βの正確かつ簡単に測定

Inflammatory processes resulting from the secretion of soluble mediators by immune cells, lead to various manifestations in skin, joints and other tissues ...

医学

Accurate and Simple Evaluation of Vascular Anastomoses in Monochorionic Placenta using Colored Dye

The presence of placental vascular anastomoses is a conditio sine qua non for the development of twin-to-twin transfusion syndrome (TTTS) and twin anemia polycythemia sequence (TAPS)1,2. Injection studies of twin placentas have shown that such anastomoses are almost invariably present in monochorionic twins and extremely rare in dichorionic twins1. Three types of anastomoses have been documented: from artery to artery, from vein to vein and from artery to vein. Arterio-venous (AV) anastomoses are unidirectional and are referred to as "deep" anastomoses since they proceed through a shared placental cotyledon, whereas arterio-arterial (AA) and veno-venous (VV) anastomoses are bi-directional and are referred to as "superficial" since they lie on the chorionic plate. Both TTTS and TAPS are caused by net imbalance of blood flow between the twins due to AV anastomoses. Blood from one twin (the donor) is pumped through an artery into the shared placental cotyledon and then drained through a vein into the circulation of the other twin (the recipient). Unless blood is pumped back from the recipient to the donor through oppositely directed deep AV anastomoses or through superficial anastomoses, an imbalance of blood volumes occurs, gradually leading to the development of TTTS or TAPS. The presence of an AA anastomosis has been shown to protect against the development of TTTS and TAPS by compensating for the circulatory imbalance caused by the uni-directional AV anastomoses1,2. 
Injection of monochorionic placentas soon after birth is a useful mean to understand the etiology of various (hematological) complications in monochorionic twins and is a required test to reach the diagnosis of TAPS2. In addition, injection of TTTS placentas treated with fetoscopic laser surgery allows identification of possible residual anastomoses3-5. This additional information is of paramount importance for all perinatologists involved in the management and care of monochorionic twins with TTTS or TAPS. Several placental injection techniques are currently being used. We provide a simple protocol to accurately evaluate the presence of (residual) vascular anastomoses using colored dye injection.

Twin-to-twin transfusion syndrome and twin anemia polycythemia sequence are two potentially devastating problems in perinatal medicine. Both disorders occur only in monochorionic twins and result from unbalanced blood flow through placental vascular anastomoses. We provide a simple protocol to accurately evaluate the presence of vascular anastomoses using colored dye injection of placental vessels after birth.

カラー染料を使用して単一絨毛膜性の胎盤における血管吻合の正確かつ簡単に評価

The presence of placental vascular anastomoses is a conditio sine qua non for the development of twin-to-twin transfusion syndrome (TTTS) and twin anemia ...

Remote Magnetic Navigation for Accurate, Real-time Catheter Positioning and Ablation in Cardiac Electrophysiology Procedures

New remote navigation systems have been developed to improve current limitations of conventional manually guided catheter ablation in complex cardiac substrates such as left atrial flutter. This protocol describes all the clinical and invasive interventional steps performed during a human electrophysiological study and ablation to assess the accuracy, safety and real-time navigation of the Catheter Guidance, Control and Imaging (CGCI) system. Patients who underwent ablation of a right or left atrium flutter substrate were included. Specifically, data from three left atrial flutter and two counterclockwise right atrial flutter procedures are shown in this report. One representative left atrial flutter procedure is shown in the movie. This system is based on eight coil-core electromagnets, which generate a dynamic magnetic field focused on the heart. Remote navigation by rapid changes (msec) in the magnetic field magnitude and a very flexible magnetized catheter allow real-time closed-loop integration and accurate, stable positioning and ablation of the arrhythmogenic substrate.

This report provides a detailed description of a new remote navigation system based on magnetic driven forces, which has been recently introduced as a new robotic tool for human cardiac electrophysiology procedures.

心臓電気生理学プロシージャでの正確なリアルタイムカテーテルポジショニングとアブレーション用のリモート磁気ナビゲーション

New remote navigation systems have been developed to improve current limitations of conventional manually guided catheter ablation in complex cardiac ...

The Measurement and Treatment of Suppression in Amblyopia

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current estimates put the prevalence of amblyopia at approximately 1-3%1-3, the majority of cases being monocular2. Amblyopia is most frequently caused by ocular misalignment (strabismus), blur induced by unequal refractive error (anisometropia), and in some cases by form deprivation.
Although amblyopia is initially caused by abnormal visual input in infancy, once established, the visual deficit often remains when normal visual input has been restored using surgery and/or refractive correction. This is because amblyopia is the result of abnormal visual cortex development rather than a problem with the amblyopic eye itself4,5 . Amblyopia is characterized by both monocular and binocular deficits6,7 which include impaired visual acuity and poor or absent stereopsis respectively. The visual dysfunction in amblyopia is often associated with a strong suppression of the inputs from the amblyopic eye under binocular viewing conditions8. Recent work has indicated that suppression may play a central role in both the monocular and binocular deficits associated with amblyopia9,10 .
Current clinical tests for suppression tend to verify the presence or absence of suppression rather than giving a quantitative measurement of the degree of suppression. Here we describe a technique for measuring amblyopic suppression with a compact, portable device11,12 . The device consists of a laptop computer connected to a pair of virtual reality goggles. The novelty of the technique lies in the way we present visual stimuli to measure suppression. Stimuli are shown to the amblyopic eye at high contrast while the contrast of the stimuli shown to the non-amblyopic eye are varied. Patients perform a simple signal/noise task that allows for a precise measurement of the strength of excitatory binocular interactions. The contrast offset at which neither eye has a performance advantage is a measure of the "balance point" and is a direct measure of suppression. This technique has been validated psychophysically both in control13,14 and patient6,9,11 populations.
In addition to measuring suppression this technique also forms the basis of a novel form of treatment to decrease suppression over time and improve binocular and often monocular function in adult patients with amblyopia12,15,16 . This new treatment approach can be deployed either on the goggle system described above or on a specially modified iPod touch device15.

Amblyopia is a developmental disorder of the visual cortex that is often accompanied by strong suppression of one eye. We present a new technique for measuring and treating interocular suppression in patients with amblyopia that can be deployed using virtual reality goggles or a portable iPod Touch device.

弱視における抑制の測定と治療

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current ...

Assessment of Gastric Emptying in Non-obese Diabetic Mice Using a [13C]-octanoic Acid Breath Test

Gastric emptying studies in mice have been limited by the inability to follow gastric emptying changes in the same animal since the most commonly used techniques require killing of the animals and postmortem recovery of the meal1,2. This approach prevents longitudinal studies to determine changes in gastric emptying with age and progression of disease. The commonly used [13C]-octanoic acid breath test for humans3 has been modified for use in mice4-6 and rats7 and we previously showed that this test is reliable and responsive to changes in gastric emptying in response to drugs and during diabetic disease progression8. In this video presentation the principle and practical implementation of this modified test is explained. As in the previous study, NOD LtJ mice are used, a model of type 1 diabetes9. A proportion of these mice develop the symptoms of gastroparesis, a complication of diabetes characterized by delayed gastric emptying without mechanical obstruction of the stomach10.
This paper demonstrates how to train the mice for testing, how to prepare the test meal and obtain 4 hr gastric emptying data and how to analyze the obtained data. The carbon isotope analyzer used in the present study is suitable for the automatic sampling of the air samples from up to 12 mice at the same time. This technique allows the longitudinal follow-up of gastric emptying from larger groups of mice with diabetes or other long-standing diseases.

Assessment of Gastric Emptying in Non-obese Diabetic Mice Using a [13C]-octanoic Acid Breath Test

Determination of gastric emptying with a non-invasive [13C]-octanoic acid breath test for tracking gastroparesis in female NOD LtJ mice.

[を使用した非肥満糖尿病マウスにおける胃排出能の評価<sup&gt; 13</sup&gt; C] - オクタン酸呼気テスト

Gastric emptying studies in mice have been limited by the inability to follow gastric emptying changes in the same animal since the most commonly used ...

Lesion Explorer: A Video-guided, Standardized Protocol for Accurate and Reliable MRI-derived Volumetrics in Alzheimer's Disease and Normal Elderly

Obtaining in vivo human brain tissue volumetrics from MRI is often complicated by various technical and biological issues. These challenges are exacerbated when significant brain atrophy and age-related white matter changes (e.g. Leukoaraiosis) are present. Lesion Explorer (LE) is an accurate and reliable neuroimaging pipeline specifically developed to address such issues commonly observed on MRI of Alzheimer's disease and normal elderly. The pipeline is a complex set of semi-automatic procedures which has been previously validated in a series of internal and external reliability tests1,2. However, LE's accuracy and reliability is highly dependent on properly trained manual operators to execute commands, identify distinct anatomical landmarks, and manually edit/verify various computer-generated segmentation outputs.
LE can be divided into 3 main components, each requiring a set of commands and manual operations: 1) Brain-Sizer, 2) SABRE, and 3) Lesion-Seg. Brain-Sizer's manual operations involve editing of the automatic skull-stripped total intracranial vault (TIV) extraction mask, designation of ventricular cerebrospinal fluid (vCSF), and removal of subtentorial structures. The SABRE component requires checking of image alignment along the anterior and posterior commissure (ACPC) plane, and identification of several anatomical landmarks required for regional parcellation. Finally, the Lesion-Seg component involves manual checking of the automatic lesion segmentation of subcortical hyperintensities (SH) for false positive errors.
While on-site training of the LE pipeline is preferable, readily available visual teaching tools with interactive training images are a viable alternative. Developed to ensure a high degree of accuracy and reliability, the following is a step-by-step, video-guided, standardized protocol for LE's manual procedures.

Lesion Explorer (LE) is a semi-automatic, image-processing pipeline developed to obtain regional brain tissue and subcortical hyperintensity lesion volumetrics from structural MRI of Alzheimer's disease and normal elderly. To ensure a high level of accuracy and reliability, the following is a video-guided, standardized protocol for LE's manual procedures.

病変エクスプローラ：アルツハイマー病における正確で信頼性のMRI誘導Volumetrics用ビデオガイド下、標準化されたプロトコルおよび標準高齢者

Obtaining in vivo human brain tissue volumetrics from MRI is often complicated by various technical and biological issues. These challenges are ...

Voluntary Breath-hold Technique for Reducing Heart Dose in Left Breast Radiotherapy

Breath-holding techniques reduce the amount of radiation received by cardiac structures during tangential-field left breast radiotherapy. With these techniques, patients hold their breath while radiotherapy is delivered, pushing the heart down and away from the radiotherapy field. Despite clear dosimetric benefits, these techniques are not yet in widespread use. One reason for this is that commercially available solutions require specialist equipment, necessitating not only significant capital investment, but often also incurring ongoing costs such as a need for daily disposable mouthpieces. The voluntary breath-hold technique described here does not require any additional specialist equipment. All breath-holding techniques require a surrogate to monitor breath-hold consistency and whether breath-hold is maintained. Voluntary breath-hold uses the distance moved by the anterior and lateral reference marks (tattoos) away from the treatment room lasers in breath-hold to monitor consistency at CT-planning and treatment setup. Light fields are then used to monitor breath-hold consistency prior to and during radiotherapy delivery.

The current priority in breast cancer radiotherapy is to reduce cardiac doses without compromising target tissue coverage. The voluntary breath-hold technique described here is a simple, inexpensive solution to this problem and capable of being instituted widely without the need for specialized equipment.

左胸放射線治療における心臓線量を削減するための自​​主的な息止め法

Breath-holding techniques reduce the amount of radiation received by cardiac structures during tangential-field left breast radiotherapy. With these ...

Creating Anatomically Accurate and Reproducible Intracranial Xenografts of Human Brain Tumors

Orthotopic tumor models are currently the best way to study the characteristics of a tumor type, with and without intervention, in the context of a live animal &#8211;&#160;particularly in sites with unique physiological and architectural qualities such as the brain. In vitro and ectopic models cannot account for features such as vasculature, blood brain barrier, metabolism, drug delivery and toxicity, and a host of other relevant factors. Orthotopic models have their limitations too, but with proper technique tumor cells of interest can be accurately engrafted into tissue that most closely mimics conditions in the human brain. By employing methods that deliver precisely measured volumes to accurately defined locations at a consistent rate and pressure, mouse models of human brain tumors with predictable growth rates can be reproducibly created and are suitable for reliable analysis of various interventions. The protocol described here focuses on the technical details of designing and preparing for an intracranial injection, performing the surgery, and ensuring successful and reproducible tumor growth and provides starting points for a variety of conditions that can be customized for a range of different brain tumor models.

The brain is a unique site with qualities that are not well represented by in vitro or ectopic analyses. Orthotopic mouse models with reproducible location and growth characteristics can be reliably created with intracranial injections using a stereotaxic fixation instrument and a low pressure syringe pump.

解剖学的に正確な作成と人間の脳の腫瘍の再現性頭蓋内異種移植片

Orthotopic tumor models are currently the best way to study the characteristics of a tumor type, with and without intervention, in the context of a live ...

3D Ultrasound Imaging: Fast and Cost-effective Morphometry of Musculoskeletal Tissue

The developmental goal of 3D ultrasound imaging (3DUS) is to engineer a modality to perform 3D morphological ultrasound analysis of human muscles. 3DUS images are constructed from calibrated freehand 2D B-mode ultrasound images, which are positioned into a voxel array. Ultrasound (US) imaging allows quantification of muscle size, fascicle length, and angle of pennation. These morphological variables are important determinants of muscle force and length range of force exertion. The presented protocol describes an approach to determine volume and fascicle length of m.&#160;vastus lateralis and m.&#160;gastrocnemius medialis. 3DUS facilitates standardization using 3D anatomical references. This approach provides a fast and cost-effective approach for quantifying 3D morphology in skeletal muscles. In healthcare and sports, information on the morphometry of muscles is very valuable in diagnostics and/or follow-up evaluations after treatment or training.

3D ultrasound imaging (3DUS) allows fast and cost-effective morphometry of musculoskeletal tissues. We present a protocol to measure muscle volume and fascicle length using 3DUS.

3 D の超音波画像診断: 筋骨格系組織の迅速かつコスト効果の高い計測

The developmental goal of 3D ultrasound imaging (3DUS) is to engineer a modality to perform 3D morphological ultrasound analysis of human muscles. 3DUS ...

Fast Inspection of Quality of Indigo Naturalis by Multiple Light Scattering

Quality control of Chinese herbal medicine is a crucial component of Chinese herbal medicine research and development. Faced with the challenges of modernization and internationalization of Chinese herbal medicine, it is urgent to establish thorough and effective procedures for quality identification of Chinese herbal medicine, and there is an urgent need for new analytical and testing techniques that are efficient, accurate, and environmentally friendly. 
Multiple light scattering is a cutting-edge and analytical method that can accurately and rapidly assess the quality of Chinese herbal medicine without altering the nature or state of the sample or using organic reagents. Indigo Naturalis is considered a good remedy for pediatric hyperthermia, psoriasis, leukemia, and ulcerative colitis. In this study, the process of addition of Indigo Naturalis powder in water was recorded precisely using a multiple light scattering instrument. 
The qualitative and quantitative measurements of the instrument can be used to accurately capture the overall trajectory and sinking behavior of Indigo Naturalis powder into water and to establish a rapid evaluation method for the quality of Indigo Naturalis with the transmission and backscattering spectrograms of the sample as qualitative indicators and stability index as a quantitative indicator. The analytical technique based on multiple light scattering provides a fast, accurate, green, and environmentally friendly method for the quality evaluation of Indigo Naturalis and supports the development and transformation of high-quality Indigo Naturalis.

Here, we present a quality evaluation method based on a multiple light scattering technique to evaluate the quality of Indigo Naturalis.

多重光散乱によるIndigo Naturalisの品質の高速検査

Quality control of Chinese herbal medicine is a crucial component of Chinese herbal medicine research and development. Faced with the challenges of ...