著者らは、スキャンソフトウェアの改善との継続的なサポートのためのマスター参照モデルとAlicona会社を製造するための歯科技工士ニコラLanfranconiに感謝します。

1。試料の準備/ベース<ol><li>標本の周りフラットベースを適用します。スキャンテーブルの上に試料を置きます。水平面（ 図1）に咬合面を配向。 </li></ol> 2。ソフトウェア解析<ol><li>プログラムを起動した後、実験室測定モジュール（ 図2） </li><li>スキャンテーブルの中心に試料を置きます。 </li><li>正しい倍率レンズを選択してください。大きなオブジェクトの場合は、完全なアーチスキャンのように、5倍の対物レンズを使用しています。 </li><li>試験片の表面はライブビューウィンドウ（ 図3）上に表示されるまで、3Dマウスを使って、スキャン光学系を移動します。 </li><li>最適な走査パラメータを達成するための露出とコントラストを設定するためにセンサ制御を使用してください。金属表面の場合は、400および800秒と0.3と0.8の間のコントラストの間に露出します（ 図4）を使用します。 </li><li>ボタンをチェックすることにより、画像品質を確認「ライブプレビューで表示画質」。 </li><li>の「測定制御」で正しいパラメータを設定します。測定タイプが「3Dデータセット」である。に、ImageFieldタイプ」は、一般に、ImageField」（ 図5）である。 </li><li> 「新しいのImageField」をクリックします。 </li><li>スキャンボリュームを定義します。ソフトウェアは、メジャーボリュームの境界座標を必要とします。 <ol><li>最高と最低のスキャンレベルに試料を移動し、両方の点で、「追加位置」をクリックしてください。 「Z範囲」の値は、スキャンボリューム（ 図6）の実際の高さを示している。 </li><li>走査空間のXY boundingsに試料を移動します。 X軸およびY軸の長さを定義するために、「追加位置」をクリックしてください。スキャンボリュームの寸法は、「情報」タブで示され、X軸及びY軸が1cmで試験片の寸法を超えるべきである。 </li></ol></li><li> 「ポイント」の番号を確認してください。ソフトウェアは、1つ &quot;のImageField「スキャンにおける走査億表面の点が可能である。実数この点の制限が超えています。ポイントサイズの「横ダウンサンプリング」によって計測点の数を「デシメーション」。 <ol><li> 「詳細設定」（ 図7）をクリックします。 </li><li> 「ポイント」数は100、M（ 図7）以下に減少するまで、右に「ラテラルダウンサンプリング」のスライダを移動します。ダウンサンプリングは、走査対象のより大きな画素サイズで、その結果、表面の点の横方向の解像度を低下させる。最適なスキャン結果を確実にするために、垂直方向の解像度が増加する。 </li></ol></li><li> 「測定開始」をクリックします。これは「プレビューモード」を起動します。ソフトウェアは、選択されたXおよびY軸寸法のプレスキャンを実行する。 </li><li>プリスキャンが完了した後、関心領域を選択する。これは、ファイルサイズと走査時間（ 図8）を低減するのに役立つ。 </li><li>フラットベース（ 図9）を含む、試料の周りの試料と2小節のフィールドを除いて、プレスキャンのすべての部分を選択します。 </li><li>スキャンを開始するには「スタート」をクリックします。 </li><li>マウスを使って「ImageViewerの」でスキャンを制御し、マウスの左ボタン（ 図10）を押します。 </li><li>ウィンドウを閉じ、「オンリービュー疑似色」をクリックしてください。 </li><li> 「設定」をクリックして、「擬似カラーリング」と「再現性」を選択します。 </li><li> 「最大」に設定0.2に付加価値と「適用範囲」をクリックしてください。 </li><li>再現性を制御するには、同じ傾斜と材料と地域のために等しくなければなりません。具体的には、試料の周りのベースは、均質な再現性（ 図11）を表示する必要があります。 </li><li> 「データベース」をクリックし、適切なフォルダにスキャンを保存します。 </li><li>必要に応じて、さまざまなファイル形式へのスキャンをエクスポートします。 「File/Export/3Dデータとして/ ...」をクリックしますプロトコルは、その時点で一時停止し、後に継続することができる。 </li></ol> 3。差分解析<ol><li>ベースを離れてカットする「3D-Editor &quot;を使用してください。このエリアには、違いのために使用されることはありません分析。 </li><li> 2つのスキャンを比較し、分析するために、「差測定」ソフトウェア（ 図12）を起動します。 </li><li>比較する2番目のモデルを選択します。 </li><li>モデルの最初の一致を実行するために「自動ラフアライメント」をクリックします。 </li><li> 「手動調整」をクリックして、それらが同じ向き（ 図13）になるまで回転させて移動してモデルを揃える。 </li><li> 「自動アライメント」をクリックして、最適なモデルマッチングに最適なアルゴリズムを開始するには「適用」。 </li><li> 2マッチしたモデルの差分マップを表示するには、 &quot;違い&quot;をクリックしてください。 </li><li> （ 図14）の偏差を表示するには、適切な色の範囲を選択します。分析用のスクリーンショットのような視覚偏差パターンを保存します。 </li></ol>差の統計値を表示する「統計」をクリックします。 1ミクロンクラスのサイズを選択して、統計的な比較のためにテキストファイルにヒストグラムデータを保存する（ 図15）。

<ol>
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Mater. 19 (1), 19-24 (2003).</li><li>Mehl, A., Ender, A., M&#246;rmann, W., Attin, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accuracy+testing+of+a+new+intraoral+3D+camera.">Accuracy testing of a new intraoral 3D camera.</a> International Journal of Computerized Dentistry. 12 (1), 11-28 (2009).</li><li>Fickl, S., 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=Dimensional+changes+of+the+ridge+contour+after+socket+preservation+and+buccal+overbuilding:+an+animal+study.">Dimensional changes of the ridge contour after socket preservation and buccal overbuilding: an animal study.</a> J. Clin. Periodontol. 36 (5), 442-448 (2009).</li><li>Vlaar, S. T., vander Zel, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accuracy+of+dental+digitizers.">Accuracy of dental digitizers.</a> Int. Dent. J. 56 (5), 301-309 (2006).</li><li>Ender, A., Mehl, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accuracy+of+complete-arch+dental+impressions:+a+new+method+of+measuring+trueness+and+precision.">Accuracy of complete-arch dental impressions: a new method of measuring trueness and precision.</a> 109 (2), 121-128 (2013).</li><li>Ender, A., Mehl, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+scanning+strategies+on+the+accuracy+of+digital+intraoral+scanning+systems.">Influence of scanning strategies on the accuracy of digital intraoral scanning systems.</a> Int. J. Comput. Dent. 16 (1), 11-21 (2013).</li><li>Ender, A., Mehl, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Full+arch+scans:+conventional+versus+digital+impressions--an+in-vitro+study.">Full arch scans: conventional versus digital impressions--an in-vitro study.</a> Int. J. Comput. Dent. 14 (1), 11-21 (2011).</li><li>Meer, W. J., Andriessen, F. S., Wismeijer, D., Ren, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+intra-oral+dental+scanners+in+the+digital+workflow+of+implantology.">Application of intra-oral dental scanners in the digital workflow of implantology.</a> PLoS One. 7 (8), (2012).</li></ol>

著者は、彼らが競合する経済的利益やその他の利害関係がないことを宣言します。

精度は、歯科医学の基本的な需要がある。基準スキャナは、高真度と精度で小規模および大規模なオブジェクトをスキャンすることができます。最適な走査方法であっても個々の歯の表面の詳細な形態学は、解像度が高く再現性でスキャンすることができる。スキャナの異なった倍率レベルで、マクロとミクロの形態学的構造を取得することができる。これは、モデルの様々な材料を走査す?...

精度は、歯科医学の多くの分野での主要な関心である。歯の硬組織を交換すると、適切な機能を確保し、さらに残りの歯の構造1,2を破壊防ぐために、正確なフィッティングプロテーゼを必要とします。固定部分義歯と総補綴物は、準備された歯やインプラント3のような構造を支援する正確なフィッティングのために特に重要である。非常に正確な再現が必要な理由は、特に、歯科印象と歯科技工所のワークフローの分野である。しかし、歯科治療の他の分野は、治療の成功を確認し、新たな治療戦略などソフトとハードの組織増強、腐食や磨耗監視、歯周治療、および矯正治療4,5を評価するために、真実かつ正確な韻律成果から利益を得る。これらの分野の多くでは、現在の検証手順は、キャリパーや顕微鏡6,7との直線距離の測定値である。これらのメタODSは、少数の測定点と検査部位の三次元（3D）の変化の限定された情報に限定される。新しい測定方法は、試験対象物8,9の表面全体の光または放射線撮影が挙げられる。ここで、全表面又は体積を測定し、コンピュータ画面上の3Dオブジェクトとして表示される。リニア測定が可能であるだけでなく、様々なスキャン時間からモデルの重ね合わせ。この重ね合わせにより、各スキ​​ャン点における表面変化の評価が可能である。これは、特定の領域を監視したり、すべての3座標軸の変形を表示する可能性を可能にします。また、体積変化が10を測定することができる。これらの新しい方法に限定する点は、試験対象物を捕捉するために使用されるスキャナの精度である。基準スキャナの精度内の変更のいずれも、試験対象または走査誤差の変化に分割することができない。スキャン精度は、多くの場合、MANUによって与えられた値であり、小さ ​​な、校正されたオブジェクト11をスキャンすることに由来しで経済。歯列弓のような大きなオブジェクトをスキャンする場合、この最小限のスキャンエラーが異なります。精度は、真度と精度で構成されています。真実は、その本当のジオメトリからスキャンされたオブジェクトの偏差です。精度を繰り返しスキャン（ISO 5725から1）との偏差である。本研究では、焦点変動走査技術に基づく新しい光学的基準スキャナは、最高の精度で完全なアーチモデルに単一の歯から検体をスキャンするために導入した。この基準スキャナは、従来とデジタル技術12月14日からと歯科咬合と歯科材料の摩耗に関する実際のプロジェクトのための歯科用印象の精度を比較した、いくつかの研究のためのベースとして使用された。本研究の目的は、基準スキャナおよび歯科研究の分野においてこのデバイスを使用するいくつかの可能性の精度の基本的な情報を提供することであった。

<table border="0" cellpadding="0" cellspacing="0" width="669">
	<tbody>
		<tr>
			<td>Reference model</td>
			<td></td>
			<td></td>
			<td>individual non-precious metal model, derived from a patient impression</td>
		</tr>
		<tr>
			<td>Araldit repair</td>
			<td>Huntsmen Advanced Material, Basel, Switzerland</td>
			<td></td>
			<td>used for making the base of the reference model</td>
		</tr>
		<tr>
			<td>CamBase</td>
			<td>Dentona, Dortmund, Germany</td>
			<td></td>
			<td>Type IV dental ston for pouring conventional impressions</td>
		</tr>
		<tr>
			<td>Identium</td>
			<td>Kettenbach, Eschenburg, Germany</td>
			<td></td>
			<td>Vinylsiloxanether impression material for conventional impression</td>
		</tr>
		<tr>
			<td>inEOS model holder</td>
			<td>Sirona Dental Systems, Bensheim, Germany</td>
			<td></td>
			<td>used for fixing stone models at the reference scanner</td>
		</tr>
		<tr>
			<td>Accutrans</td>
			<td>Coltene Whaledent, Altst&#228;tten, Switzerland</td>
			<td></td>
			<td>used for making the base of thestone models</td>
		</tr>
		<tr>
			<td>President putty</td>
			<td>Coltene Whaledent, Altst&#228;tten, Switzerland</td>
			<td></td>
			<td>mix with accutrans for betterstability of the base</td>
		</tr>
		<tr>
			<td>Alicona Infinite Focus</td>
			<td>Alicona Imaging, Graz, Austria</td>
			<td></td>
			<td>Reference scanner&#160;</td>
		</tr>
	</tbody>
</table>

accuracy in dental medicine, a new way to measure trueness and precision

基準スキャナは、手順の多くを確認するために、歯科医療において使用されている。主な関心は、彼らが歯科修復のための拠点として機能したような印象の方法を確認することです。多くの参考スキャナの電流制限は、完全な歯列弓のような大きな物体を走査し、精度の不足、または詳細歯の表面を評価するための限定された可能性である。焦点変動走査技術に基づいて、新たな基準スキャナは、最も一般的なローカルおよび正確性に関して評価した。特定の走査プロトコルは、歯科印象から元の歯の表面を走査するために試験した。また、別のモデル材料を確認した。結果は、完全なアーチスキャンの場合の精度のための真度および1.6±0.6μmの5.3±1.1ミクロンの平均偏差と基準スキャナの高いスキャニング精度を示した。 T現在の歯科用印象方法ははるかに高い偏差（12.5±2.5ミクロン：20.4±2.2μmで、精度の正確さ）を示したハン基準スキャナの内部スキャンの精度。単一の歯の表面のような小さなオブジェクトは、びらん性および研磨歯の表面損失を評価するためのシステムを可能にする、より高精度で走査することができる。基準スキャナは、歯科研究分野の多くのための違いを測定するために使用することができる。高い局所および全身精度と組み合わさ種の異なった倍率レベルは、完全なアーチに変化するまで、単一の歯または修復の変化を評価するために使用することができる。

図16aは、石膏試料のスキャンを示しています。最適なスキャンパラメータを確認するには、材料のための再現性を制御します。標本のセクションでは、さまざまなコントラストと明るさの設定でスキャンされ、再現性は、最適なスキャン設定を見つけるために、各スキャン後にチェックされている。金属表面は、高いコントラストでコントラストも低い1.0石膏や樹脂材料で最高?...

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Accuracy in Dental Medicine, A New Way to Measure Trueness and Precision

精度は、歯科医療の主要な需要である。正確性を検証するために、リファレンス·スキャナが必要とされている。この記事では、高い真度と精度を有する歯科用の形態の多種多様なを取得するように調整スキャン方法を使用して新しい基準スキャナを提示します。

歯科医学、真度と精度を測定するための新しい方法の精度

Reference scanners are used in dental medicine to verify a lot of procedures. The main interest is to verify impression methods as they serve as a base ...

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13.3K ビュー.  University of Zürich. 精度は、歯科医療の主要な需要である。正確性を検証するために、リファレンス·スキャナが必要とされている。この記事では、高い真度と精度を有する歯科用の形態の多種多様なを取得するように調整スキャン方法を使用して新しい基準スキャナを提示します。 

ビデオ: Accuracy in Dental Medicine, A New Way to Measure Trueness and Precision

Technique to Collect Fungiform (Taste) Papillae from Human Tongue

The sense of taste is critical for human life. It informs the body about the quality of food that will be potentially ingested and stimulates metabolic processes that prepare the alimentary canal for digestion. Steady progress is being made towards understanding the early biochemical and molecular events underlying taste transduction (for a review, Breslin and Spector, 20081). However, progress to date has largely resulted from animal models. Yet, since marked differences in receptor specificity and receptor density vary among species, human taste transduction will only be understood by using human taste tissue. Here we describe a biopsy technique to collect human fungiform papillae, visible as rounded pink anterior structures, about 0.5 mm in diameter that contain taste buds. These biopsied papillae are used for several purposes including the isolation of viable taste bud cells, in situ hybridization, immunohistochemistry and, through techniques of molecular biology, the identification of taste-specific novel proteins.

Technique to Collect Fungiform Taste Papillae from Human Tongue

Knowledge of molecular mechanisms underlying gustatory transduction has recently enjoyed significant advances, largely due to using animal models. However, the wide diversity in taste sensitivity and specificity among mammals warrants studies in human tissue. We describe a biopsy technique to collect living taste cells from the papillae on human tongue.

人間の舌から茸（味）乳頭を収集する手法

The sense of taste is critical for human life. It informs the body about the quality of food that will be potentially ingested and stimulates metabolic ...

医学

Construction of Vapor Chambers Used to Expose Mice to Alcohol During the Equivalent of all Three Trimesters of Human Development

Exposure to alcohol during development can result in a constellation of morphological and behavioral abnormalities that are collectively known as Fetal Alcohol Spectrum Disorders (FASDs). At the most severe end of the spectrum is Fetal Alcohol Syndrome (FAS), characterized by growth retardation, craniofacial dysmorphology, and neurobehavioral deficits. Studies with animal models, including rodents, have elucidated many molecular and cellular mechanisms involved in the pathophysiology of FASDs. Ethanol administration to pregnant rodents has been used to model human exposure during the first and second trimesters of pregnancy. Third trimester ethanol consumption in humans has been modeled using neonatal rodents. However, few rodent studies have characterized the effect of ethanol exposure during the equivalent to all three trimesters of human pregnancy, a pattern of exposure that is common in pregnant women. Here, we show how to build vapor chambers from readily obtainable materials that can each accommodate up to six standard mouse cages. We describe a vapor chamber paradigm that can be used to model exposure to ethanol, with minimal handling, during all three trimesters. Our studies demonstrate that pregnant dams developed significant metabolic tolerance to ethanol. However, neonatal mice did not develop metabolic tolerance and the number of fetuses, fetus weight, placenta weight, number of pups/litter, number of dead pups/litter, and pup weight were not significantly affected by ethanol exposure. An important advantage of this paradigm is its applicability to studies with genetically-modified mice. Additionally, this paradigm minimizes handling of animals, a major confound in fetal alcohol research.

We demonstrate the construction of alcohol vapor chambers using readily available materials that simultaneously house 6 mouse cages. We further describe their use in a mouse model of fetal alcohol exposure equivalent to all 3&#160;trimesters of human pregnancy. This paradigm exposes animals during gestation and postnatal days 1-12.

人間開発の3つのすべての学期の等価時にはアルコールにマウスを公開するために使用蒸気チェンバースの建設

Exposure to alcohol during development can result in a constellation of morphological and behavioral abnormalities that are collectively known as Fetal ...

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. Despite the discovery of key regulators of uterine contractility, this process is still not fully understood. Transgenic mice provide an ideal model in which to study parturition. Previously, the only method to study uterine contractility in the mouse was ex vivo isometric tension recordings, which are suboptimal for several reasons. The uterus must be removed from its physiological environment, a limited time course of investigation is possible, and the mice must be sacrificed. The recent development of radiometric telemetry has allowed for longitudinal, real-time measurements of in vivo intrauterine pressure in mice. Here, the implantation of an intrauterine telemeter to measure pressure changes in the mouse uterus from mid-pregnancy until delivery is described. By comparing differences in pressures between wild type and transgenic mice, the physiological impact of a gene of interest can be elucidated. This technique should expedite the development of therapeutics used to treat myometrial disorders during pregnancy, including preterm labor.

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

This manuscript provides a protocol for implanting telemeters in the mouse for the purpose of measuring intrauterine pressures during pregnancy.

マウス収縮圧力を測定するために子宮内テレメトリ<em&gt;インビボ</em

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. ...

Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor initiation, metastasis and resistance to radio- and chemotherapy. Here we describe a specific methodology for culturing primary human pancreatic CSCs as tumor spheres in anchorage-independent conditions. Cells are grown in serum-free, non-adherent conditions in order to enrich for CSCs while their more differentiated progenies do not survive and proliferate during the initial phase following seeding of single cells. This assay can be used to estimate the percentage of CSCs present in a population of tumor cells. Both size (which can range from 35 to 250 micrometers) and number of tumor spheres formed represents CSC activity harbored in either bulk populations of cultured cancer cells or freshly harvested and digested tumors 1,2. Using this assay, we recently found that metformin selectively ablates pancreatic CSCs; a finding that was subsequently further corroborated by demonstrating diminished expression of pluripotency-associated genes/surface markers and reduced in vivo tumorigenicity of metformin-treated cells. As the final step for preclinical development we treated mice bearing established tumors with metformin and found significantly prolonged survival. Clinical studies testing the use of metformin in patients with PDAC are currently underway (e.g., NCT01210911, NCT01167738, and NCT01488552). Mechanistically, we found that metformin induces a fatal energy crisis in CSCs by enhancing reactive oxygen species (ROS) production and reducing mitochondrial transmembrane potential. In contrast, non-CSCs were not eliminated by metformin treatment, but rather underwent reversible cell cycle arrest. Therefore, our study serves as a successful example for the potential of in vitro sphere formation as a screening tool to identify compounds that potentially target CSCs, but this technique will require further in vitro and in vivo validation to eliminate false discoveries.

Pancreatic cancer stem cells (CSCs) can be expanded in vitro using the anchorage-independent sphere culture technique, which represents a powerful tool to study CSC biology and can serve as the first step to develop novel CSC-targeting therapies. Here the methodology for expanding, analyzing and targeting of pancreatic CSCs is provided.

新しい治療戦略を開発するための膵臓癌幹細胞特性を研究

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor ...

Boldness, Aggression, and Shoaling Assays for Zebrafish Behavioral Syndromes

A behavioral syndrome exists when specific behaviors interact under different contexts. Zebrafish have been test subjects in recent studies and it is important to standardize protocols to ensure proper analyses and interpretations. In our previous studies, we have measured boldness by monitoring a series of behaviors (time near surface, latency in transitions, number of transitions, and darts) in a 1.5 L trapezoidal tank. Likewise, we quantified aggression by observing bites, lateral displays, darts, and time near an inclined mirror in a rectangular 19 L tank. By dividing a 76 L tank into thirds, we also examined shoaling preferences. The shoaling assay is a highly customizable assay and can be tailored for specific hypotheses. However, protocols for this assay also must be standardized, yet flexible enough for customization. In previous studies, end chambers were either empty, contained 5 or 10 zebrafish, or 5 pearl danios (D. albolineatus). In the following manuscript, we present a detailed protocol and representative data that accompany successful applications of the protocol, which will allow for replication of behavioral syndrome experiments.

This manuscript describes the setup, implementation, and analysis of boldness, aggression, and shoaling in zebrafish and testing for the presence of a behavioral syndrome. A standardized approach for behavioral quantification will allow for easier comparison across studies. Modifications to this protocol are possible as each assay can be run individually.

ゼブラフィッシュ行動症候群のための大胆、攻撃的、および浅水アッセイ

A behavioral syndrome exists when specific behaviors interact under different contexts. Zebrafish have been test subjects in recent studies and it is ...

Open Tracheostomy Gastric Acid Aspiration Murine Model of Acute Lung Injury Results in Maximal Acute Nonlethal Lung Injury

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute respiratory distress syndrome (ARDS), characterized by neutrophilic alveolitis, and injury to both alveolar epithelium and vascular endothelium. Clinically, ARDS is defined by acute onset of hypoxemia, bilateral patchy pulmonary infiltrates and non-cardiogenic pulmonary edema. Human studies have provided us with valuable information about the physiological and inflammatory changes in the lung caused by ARDS, which has led to various hypotheses about the underling mechanisms. Unfortunately, difficulties determining the etiology of ARDS, as well as a wide range of pathophysiology have resulted in a lack of critical information that could be useful in developing therapeutic strategies.
Translational animal models are valuable when their pathogenesis and pathophysiology accurately reproduce a concept proven in both in vitro and clinical settings. Although large animal models (e.g., sheep) share characteristics of the anatomy of human trachea-bronchial tree, murine models provide a host of other advantages including: low cost; short reproductive cycle lending itself to greater data acquisition; a well understood immunologic system; and a well characterized genome leading to the availability of a variety of gene deletion and transgenic strains. A robust model of low pH induced ARDS requires a murine ALI that targets mainly the alveolar epithelium, secondarily the vascular endothelium, as well as the small airways leading to the alveoli. Furthermore, a reproducible injury with wide differences between different injurious and non-injurious insults is important.
The murine gastric acid aspiration model presented here using hydrochloric acid employs an open tracheostomy and recreates a pathogenic scenario that reproduces the low pH pneumonitis injury in humans. Additionally, this model can be used to examine interaction of a low pH insult with other pulmonary injurious entities (e.g., food particles, pathogenic bacteria).

This protocol induces acute lung injury in a mouse that has close fidelity to the pathogenesis of acid pneumonitis observed in humans. We generate a maximal acute nonlethal low pH lung injury and account for differences in rodent-human anatomic respiratory structure using an open tracheostomy coupled with circumferential pressure release.

最大急性非致死肺損傷における急性肺損傷の結果のオープン気管切開胃酸吸引マウスモデル

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute ...

Conducting Maximal and Submaximal Endurance Exercise Testing to Measure Physiological and Biological Responses to Acute Exercise in Humans

Regular physical activity has a positive effect on human health, but the mechanisms controlling these effects remain unclear. The physiologic and biologic responses to acute exercise are predominantly influenced by the duration and intensity of the exercise regimen. As exercise is increasingly thought of as a therapeutic treatment and/or diagnostic tool, it is important that standardizable methodologies be utilized to understand the variability and to increase the reproducibility of exercise outputs and measurements of responses to such regimens. To that end, we describe two different cycling exercise regimens that yield different physiologic outputs. In a maximal exercise test, exercise intensity is continually increased with a greater workload resulting in an increasing cardiopulmonary and metabolic response (heart rate, stroke volume, ventilation, oxygen consumption and carbon dioxide production). In contrast, during endurance exercise tests, the demand is increased from that at rest, but is raised to a fixed submaximal exercise intensity resulting in a cardiopulmonary and metabolic response that typically plateaus. Along with the protocols, we provide suggestions on measuring physiologic outputs that include, but are not limited to, heart rate, slow and forced vital capacity, gas exchange metrics, and blood pressure to enable the comparison of exercise outputs between studies. Biospecimens can then be sampled to assess cellular, protein, and/or gene expression responses. Overall, this approach can be easily adapted into both short- and long-term effects of two distinct exercise regimens.

To assess the influence of exercise intensity on physiologic and biologic responses, two different exercise testing protocols were utilized. Methods outlining exercise testing on a cycle ergometer as an incremental maximal oxygen consumption test and endurance, steady state submaximal endurance test are described.

最大及び最大下運動生理・生物学的ヒトにおける急性運動レスポンスを測定するテストを実施

Regular physical activity has a positive effect on human health, but the mechanisms controlling these effects remain unclear. The physiologic and biologic ...

Precision Implementation of Minimal Erythema Dose (MED) Testing to Assess Individual Variation in Human Inflammatory Response

Minimal erythema dose (MED) testing is frequently used in clinical settings for determining the smallest amount of ultraviolet (UV) irradiation necessary to produce erythema (inflammatory reddening) on the surface of the skin. In this context, the MED is regarded as a key factor in determining starting doses for UV phototherapy for common skin conditions such as psoriasis and eczema. In research settings, MED testing also has potential to be a powerful tool for assessing within- and between-persons variation in inflammatory responses. However, MED testing has not been widely adopted for use in research settings, likely owing to a lack of published guidelines, which is a barrier to obtaining reproducible results from this assay. Also, protocols and equipment for establishing MED vary widely, making it difficult to compare results across laboratories. Here, we describe a precise and reproducible method to induce and measure superficial erythema using newly designed protocols and methods that can easily be adapted to other equipment and laboratory environments. The method described here includes detail on procedures that will allow extrapolation of a standardized dosage schedule to other equipment so that this protocol can be adapted to any UV radiation source.

Precision Implementation of Minimal Erythema Dose MED Testing to Assess Individual Variation in Human Inflammatory Response

Minimal erythema dose (MED) testing is used to establish dosage schedules for ultraviolet radiation phototherapy. It can assess individual variation in inflammatory response but lacks methodology for achieving reproducible results. Here, we present a precision implementation of MED and demonstrate its ability to capture individual variation in inflammatory response.

ヒト炎症反応の個々の変動を評価するための最小紅斑用量(MED)試験の精密実施

Minimal erythema dose (MED) testing is frequently used in clinical settings for determining the smallest amount of ultraviolet (UV) irradiation necessary ...

Murine Precision-Cut Liver Slices as an Ex Vivo Model of Liver Biology

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic fatty liver disease, metabolic liver disease, and liver cancer) requires experimental manipulation of animal models and in vitro cell cultures. Both techniques have limitations, such as the requirement of large numbers of animals for in vivo manipulation. However, in vitro cell cultures do not reproduce the structure and function of the multicellular hepatic environment. The use of precision-cut liver slices is a technique in which uniform slices of viable mouse liver are maintained in laboratory tissue culture for experimental manipulation. This technique occupies an experimental niche that exists between animal studies and in vitro cell culture methods. The presented protocol describes a straightforward and reliable method to isolate and culture precision-cut liver slices from mice. As an application of this technique, ex vivo liver slices are treated with bile acids to simulate cholestatic liver injury and ultimately assess the mechanisms of hepatic fibrogenesis.

This protocol provides a simple and reliable method for the production of viable precision-cut liver slices from mice. The ex vivo tissue samples can be maintained under laboratory tissue culture conditions for multiple days, providing a flexible model to examine liver pathobiology.

肝臓生物学のEx Vivoモデルとしてのマウス精密カット肝臓スライス

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic ...

Intraoperative Assessment of Resection Margins in Oral Cavity Cancer: This is the Way

The goal of head and neck oncological surgery is complete tumor resection with adequate resection margins while preserving acceptable function and appearance. For oral cavity squamous cell carcinoma (OCSCC), different studies showed that only 15%-26% of all resections are adequate. A major reason for the low number of adequate resections is the lack of information during surgery; the margin status is only available after the final histopathologic assessment, days after surgery.
The surgeons and pathologists at the Erasmus MC University Medical Center in Rotterdam started the implementation of specimen-driven intraoperative assessment of resection margins (IOARM) in 2013, which became the standard of care in 2015. This method enables the surgeon to turn an inadequate resection into an adequate resection by performing an additional resection during the initial surgery. Intraoperative assessment is supported by a relocation method procedure that allows accurate identification of inadequate margins (found on the specimen) in the wound bed.
The implementation of this protocol resulted in an improvement of adequate resections from 15%-40%. However, the specimen-driven IOARM is not widely adopted because grossing fresh tissue is counter-intuitive for pathologists. The fear exists that grossing fresh tissue will deteriorate the anatomical orientation, shape, and size of the specimen and therefore will affect the final histopathologic assessment. These possible negative effects are countered by the described protocol. Here, the protocol for specimen-driven IOARM is presented in detail, as performed at the institute.

The goal of this protocol is to provide a clear overview of specimen-driven intraoperative assessment of resection margins. It is encouraged to implement this protocol to improve patient care at other institutes.