パート1：セットアップと皮膚の準備<ol><li>ベルト、EMG、ECG、皮膚コンダクタンス、温度、地面を呼吸以下のセンサーを設定します。 </li><li>軽くアルコール次の場所にワイプでこすって肌を準備します：バックグラウンドのための左手の内側に心電図のための手首の両方の筋電図のための目の上の額、、（接地電極の位置が使用される機器によって異なります）。 </li><li>腰周りの呼吸ベルト、それぞれの目の上の額の筋電図、各手首の内側に心電図、左手の中指の皮膚コンダクタンス、左側のポインターで温度計、接地：次の場所の皮膚にセンサーを取り付けます左側の背面にある。 </li><li>呼吸+ HRVのためのセッションを選択します。 </li><li> impendenceはすべてのセンサーに対して許容可能であることとセンサは、クライアントの動きと生産合理的な測定値に応答することを確認してください。 </li></ol>パート2：録音<ol><li>クライアントに手順を説明します。 </li><li> （分0-2）開いた目で静かに座ってするようにクライアントに指示します。 2分間のベースラインを記録します。 </li><li> 2分の終わりに（分2-4）、2分間の認知のストレッサーを開始-徐々に難易度が増加計算の問題は、ペン＆紙または電卓（計算はクライアントのために特に困難である場合、それらを求めるを使用せずに解決するために繰り返し100から始まる7を減算する、すなわち、&quot;シリアルセブン&quot;）。クライアントに次の指示を与える：&quot;次の2分で、私はいくつかの数学の問題を解決するかどうかを確認するされます。早くすることができますように動作すると、&quot;私に答えを教えてください。ようにこの運動の軽度のストレス誘導特性を除去するためではない（例えば、&quot;ちょうどあなたができるベストを尽くす&quot;）クライアントの答えが正しいか間違っていると最小限の励ましを提供するかどうかのフィードバックを提供しないでください。 </li><li> （分4-6）クライアントは評価のこの部分が上であることを知っていると/彼女が2分間開いた目で静かに座って彼に聞いてみよう。 </li><li> （分6-8）物理的なストレッサーを開始します。大声で有害なノイズをオンにする（赤ちゃん泣いて、車がクラッシュする、など）、チューニングなしで音に耳を傾けるようにクライアントに依頼。 </li><li> （分8-10）ノイズの電源をオフにし、2分間開いた目で静かに座って、クライアントに依頼してください。 </li><li> （分10-12）感情的なストレッサーを開始：彼らは思考と感じていたか、イベントの詳細を含む、クライアントはそれらがよく覚えているストレスの多い事件について話を頼む。ストレス応答を軽減しないように最小限の共感的応答を提供します。 </li><li> （分12-14）彼/彼女が完了していない場合でも、クライアントの話を中断し、開いた目で静かに座ってするようにクライアントを指示する。 </li><li>録画を停止、データを保存し、クライアントは評価が行われていることを知らせる。 </li></ol>パート3：報告会および治療計画<ol><li>アセスメントの間に彼らの経験についてクライアントに尋ねる。 どの部分が特にストレスだったとなぜですか？ 彼は/彼女はどんな生理的変化が起こっ感じましたか？ </li><li>付録Iに記載の基準に従って審査データ</li><li>上記の分析に基づいて対処する必要が生理学的パラメータを選んだ。 ベースライン時にセンサーの読み取り値が正常よりも高いまたは低い場合は、治療の目標は、クライアントが非ストレスの多い環境で正常にその生理​​的パラメータを持って支援することです。 ストレッサーの間にセンサーの読み取り値が正常値よりも高い値や低い場合は、治療目標は、クライアントがストレッサーを特定し、ストレス下生理機能の正常レベルを維持することができるよう支援することです。 ダウンタイム時のセンサーの読み取り値が回復の欠如を示している場合、治療目標は、クライアントがすぐにストレッサーを、以下の通常のレベルにその生理​​的パラメータを持って支援することです。 これらの目標のいずれかまたはすべてが治療計画に含まれていてもよい。 </li></ol>付録I ストレスのアセスメントの解釈のガイドライン （すべての&quot;太字&quot;答えは対処する領域を示す） <table border="0"><tbody><tr><td colspan="3"> EMG（通常の読書&lt;3マイクロボルト） </td></tr><tr><td> 1。 EMGはベースラインで上昇している？ </td><td> はい _ </td><td>ない_ </td></tr><tr><td> 2。 EMGは3ストレッサーのいずれかでベースラインを越えて上昇している？ </td><td> はい _ </td><td>ない_ </td></tr><tr><td> 3。休憩時間にベースラインレベルにEMGの回復はありますか？ </td><td>はい_ </td><td> ない _ </td></tr><tr><td colspan="3"> 心拍数（正常範囲毎分60〜80拍） </td></tr><tr><td> 1。ベースライン時に高心拍数です？ </td><td> はい _ </td><td>ない_ </td></tr><tr><td> 2。心拍数は3ストレッサーのいずれかでベースラインを越えて上昇していますか？ </td><td> はい _ </td><td>ない_ </td></tr><tr><td> 3。休憩時間にベースラインレベルに心拍数の回復はありますか？ </td><td>はい_ </td><td> ない _ </td></tr><tr><td colspan="3"> 呼吸速度（通常の速度&lt;1分あたり12呼吸） </td></tr><tr><td> 1。ベースラインで上昇呼吸率です？ </td><td> はい _ </td><td>ない_ </td></tr><tr><td> 2。どんなストレッサーのためのベースラインを越えて上昇呼吸率です？ </td><td> はい _ </td><td>ない_ </td></tr><tr><td> 3。残りの期間中にベースラインに速度を呼吸の回復はありますか？ </td><td>はい_ </td><td> ない _ </td></tr><tr><td colspan="3"> 皮膚コンダクタンス </td></tr><tr><td> 1。ベースラインで上昇皮膚コンダクタンスです？ </td><td> はい _ </td><td>ない_ </td></tr><tr><td> 2。皮膚コンダクタンスは、どんなストレッサーのためにベースラインを越えて上昇していますか？ </td><td> はい _ </td><td>ない_ </td></tr><tr><td> 3。休憩時間にベースラインに皮膚コンダクタンスの回復はありますか？ </td><td>はい_ </td><td> ない _ </td></tr><tr><td colspan="3"> 指の温度（通常の読書&gt; 90F） </td></tr><tr><td> 1。指の温度は、ベースラインで低でしょうか？ </td><td> はい _ </td><td>ない_ </td></tr><tr><td> 2。指の温度低下は、どのストレッサーのベースラインを通過していますか？ </td><td> Y ES _ </td><td>ない_ </td></tr><tr><td> 3。休憩時間にベースラインに指温度の回復はありますか？ </td><td>はい_ </td><td> ない _ </td></tr><tr><td colspan="3"> 心拍数の変動（規範は年齢によって異なります） </td></tr><tr><td> 1。ベースラインで低心拍変動ですか。 </td><td> はい _ </td><td>ない_ </td></tr><tr><td> 2。心拍変動の減少は、どんなストレッサーのベースラインを通過していますか？ </td><td> はい _ </td><td>ない_ </td></tr><tr><td> 3。休憩時間に心拍変動の回復はありますか？ </td><td>はい_ </td><td> ない _ </td></tr></tbody></table>付録I ストレスのアセスメントの解釈： <table border="0"><tbody><tr><td colspan="3"> EMG（通常の読書&lt;3マイクロボルト） </td></tr><tr><td> 1。 EMGはベースラインで上昇している？ </td><td> はい_ </td><td>ない_ </td></tr><tr><td> 2。 EMGは3ストレッサーのいずれかでベースラインを越えて上昇している？ </td><td> はい_ </td><td>ない_ </td></tr><tr><td> 3。休憩時間にベースラインレベルにEMGの回復はありますか？ </td><td> はい_ </td><td>ない_ </td></tr><tr><td> 心拍数（正常範囲毎分60〜80拍） </td></tr><tr><td> 1。ベースライン時に高心拍数です？ </td><td> はい_ </td><td>ない_ </td></tr><tr><td> 2。心拍数は3ストレッサーのいずれかでベースラインを越えて上昇していますか？ </td><td> はい_ </td><td>ない_ </td></tr><tr><td> 3。休憩時間にベースラインレベルに心拍数の回復はありますか？ </td><td> はい_ </td><td>ない_ </td></tr><tr><td> 呼吸速度（通常の速度&lt;1分あたり12呼吸） </td></tr><tr><td> 1。ベースラインで上昇呼吸率です？ </td><td> はい_ </td><td>ない_ </td></tr><tr><td> 2。どんなストレッサーのためのベースラインを越えて上昇呼吸率です？ </td><td> はい_ </td><td>ない_ </td></tr><tr><td> 3。残りの期間中にベースラインに速度を呼吸の回復はありますか？ </td><td> はい_ </td><td>ない_ </td></tr><tr><td> 皮膚コンダクタンス（通常の読書&lt;5マイクロオーム） </td></tr><tr><td> 1。ベースラインで上昇皮膚コンダクタンスです？ </td><td> はい_ </td><td>ない_ </td></tr><tr><td> 2。皮膚コンダクタンスは、どんなストレッサーのためにベースラインを越えて上昇していますか？ </td><td> はい_ </td><td>ない_ </td></tr><tr><td> 3。休憩時間にベースラインに皮膚コンダクタンスの回復はありますか？ </td><td> はい_ </td><td>ない_ </td></tr><tr><td> 指の温度（通常の読書&gt; 90F） </td></tr><tr><td> 1。指の温度は、ベースラインで低でしょうか？ </td><td> はい_ </td><td>ない_ </td></tr><tr><td> 2。指の温度低下は、どのストレッサーのベースラインを通過していますか？ </td><td> はい_ </td><td>ない_ </td></tr><tr><td> 3。休憩時間にベースラインに指温度の回復はありますか？ </td><td> はい_ </td><td>ない_ </td></tr><tr><td> 心拍数の変動（規範は年齢によって異なります） </td></tr><tr><td> 1。ベースラインで低心拍変動ですか。 </td><td> はい_ </td><td>ない_ </td></tr><tr><td> 2。心拍変動の減少は、どんなストレッサーのベースラインを通過していますか？ </td><td> はい_ </td><td>ない_ </td></tr><tr> &lt;TD&gt; 3。休憩時間に心拍変動の回復はありますか？ </td><td> はい_ </td><td>ない_ </td></tr></tbody></table>治療計画： 読書は、ベースライン時、通常よりも高いまたは低い場合、治療の目標は、クライアントがすべての状況で正常にその生理​​的パラメータをもたらすことを支援することです。 読書がストレス時に通常よりも高いまたは低い場合、治療の目標は、クライアントがストレッサーを特定し、ストレス下生理機能の正常レベルを維持できるよう支援することです 読書は、ダウンタイムの間に回復の欠如を示している場合、治療の目標は、クライアントがすぐにストレッサーを、以下の通常のレベルにその生理​​的パラメータを持って支援することです。

<ol>
	<li>Arena, J., Schwartz, M. S., Schwartz, M. S., Andrasik, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Psychophysiological+assessment+and+biofeedback+baselines.">Psychophysiological assessment and biofeedback baselines.</a> A Practitioners Guide. , 128-158 (2005).</li><li>Schwartz, N. M., Schwartz, M. S., Schwartz, M. S., Andrasik, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Definitons+of+biofeedback+and+applied+psychophysiology.">Definitons of biofeedback and applied psychophysiology.</a> A Practitioners Guide. , 27-42 (2005).</li></ol>

The authors have nothing to disclose.

appendiストレスの評価に完成した精神生理学的には、治療計画を策定するために必要な情報とバイオフィードバック開業医を提供します。この評価は、個々の応答アポモルフィン、またはすべてのストレッサー、および刺激応答の特異性、またはストレス要因の異なる種類の個々のユニークな生理的応答に特定の生理学的反応の均一性の両方に関する情報を提供します。この情報を使用して?...

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psychophysiological stress assessment using biofeedback 

過去半世紀では、バイオフィードバックの研究は、どの人間の心は、自律神経系の機能に影響を与えることができる程度を示している、以前に意識的なコントロールの外側であると考えられて。人々は自分の体からの信号を観測できるようにすることで、バイオフィードバックは、それらがそのようなストレスのようなそれらの生理学的および心理的反応を、より意識を開発するために、これらの反応を変更することを学ぶことができます。バイオフィードバック専門家は、軽度のストレスの多い出来事に人々の反応を評価し、バイオフィードバックベースの治療計画を策定することにより、このプロセスを容易にすることができます。ストレスのアセスメント時に最初の医師は、生理学的測定値のベースラインを記録し、そのような認知、物理的および感情的なストレスなどのいくつかの軽度のストレス因子、をクライアントに提示。ストレス要因の様々な人の刺激応答の特異性、または質的に異なる刺激に対する各人の反応の違いを決定するために提示されます。このビデオでは、治療計画のためのバイオフィードバックと現在の一般的なガイドラインを使用して、精神生理学的ストレスの評価のプロセスのデモンストレーションを行います。

Watch this Scientific Journal Video about Psychophysiological Stress Assessment Using Biofeedback at JoVE.com

Psychophysiological Stress Assessment Using Biofeedback

このビデオでは、バイオフィードバックを使用して強調して人の精神生理学的反応を評価する一つの方法を説明します。我々はまた、治療計画のための一般的なガイドラインを紹介します。

バイオフィードバックを用いた精神生理学的ストレス評価

In the last half century, research in biofeedback has shown the extent to which the human mind can influence the functioning of the autonomic nervous ...

psychophysiological-stress-assessment-using-biofeedback

Research

JoVE Journal

Biology

Psychophysiological Stress Assessment Using Biofeedback 

13.2K ビュー.  Cambridge Health Alliance. このビデオでは、バイオフィードバックを使用して強調して人の精神生理学的反応を評価する一つの方法を説明します。我々はまた、治療計画のための一般的なガイドラインを紹介します。

ビデオ: Psychophysiological Stress Assessment Using Biofeedback

Monitoring Plant Hormones During Stress Responses

Plant hormones and related signaling compounds play an important role in the regulation of plant responses to various environmental stimuli and stresses. Among the most severe stresses are insect herbivory, pathogen infection, and drought stress. For each of these stresses a specific set of hormones and/or combinations thereof are known to fine-tune the responses, thereby ensuring the plant's survival. The major hormones involved in the regulation of these responses are jasmonic acid (JA), salicylic acid (SA), and abscisic acid (ABA). To better understand the role of individual hormones as well as their potential interaction during these responses it is necessary to monitor changes in their abundance in a temporal as well as in a spatial fashion. For the easy, sensitive, and reproducible quantification of these and other signaling compounds we developed a method based on vapor phase extraction and gas chromatography/mass spectrometry (GC/MS) analysis (1, 2, 3, 4). After extracting these compounds from the plant tissue by acidic aqueous 1-propanol mixed with dichloromethane the carboxylic acid-containing compounds are methylated, volatilized under heat, and collected on a polymeric absorbent. After elution into a sample vial the analytes are separated by gas chromatography and detected by chemical ionization mass spectrometry. The use of appropriate internal standards then allows for the simple quantification by relating the peak areas of analyte and internal standard.

A simple method is provided that allows for the rapid extraction and analysis of multiple plant hormones from small tissue samples. The procedure uses vapor phase extraction as the solemn purification step. Samples are analyzed by GC/MS with chemical ionization that produces mainly (M+1)+ ions.

ストレス応答時には植物ホルモンのモニタリング

Plant hormones and related signaling compounds play an important role in the regulation of plant responses to various environmental stimuli and stresses. ...

生物学

Microsurgical Clip Obliteration of Middle Cerebral Aneurysm Using Intraoperative Flow Assessment

Cerebral aneurysms are abnormal widening or ballooning of a localized segment of an intracranial blood vessel.  Surgical clipping is an important treatment for aneurysms which attempts to exclude blood from flowing into the aneurysmal segment of the vessel while preserving blood flow in a normal fashion.  Improper clip placement may result in residual aneurysm with the potential for subsequent aneurysm rupture or partial or full occlusion of distal arteries resulting in cerebral infarction.  Here we describe the use of an ultrasonic flow probe to provide quantitative evaluation of arterial flow before and after microsurgical clip placement at the base of a middle cerebral artery aneurysm.  This information helps ensure adequate aneurysm reconstruction with preservation of normal distal blood flow.

Description of the surgical obliteration of a cerebral aneurysm utilizing an ultrasonic flow probe to assess arterial flow prior to and after aneurysm clip placement.

術中フローアセスメントを用いた中大脳動脈瘤の顕微クリップ閉塞

Cerebral aneurysms are abnormal widening or ballooning of a localized segment of an intracranial blood vessel.  Surgical clipping is an important ...

A Quantitative Assessment of The Yeast Lipidome using Electrospray Ionization Mass Spectrometry

Lipids are one of the major classes of biomolecules and play important roles membrane dynamics, energy storage, and signalling1-4. The budding yeast Saccharomyces cerevisiae, a genetically and biochemically manipulable unicellular eukaryote with annotated genome and very simple lipidome, is a valuable model for studying biological functions of various lipid species in multicellular eukaryotes2,3,5. S. cerevisiae has 10 major classes of lipids with chain lengths mainly of 16 or 18 carbon atoms and either zero or one degree of unsaturation6,7. Existing methods for lipid identification and quantification - such as high performance liquid chromatography, thin-layer chromatography, fluorescence microscopy, and gas chromatography followed by MS - are well established but have low sensitivity, insufficiently separate various molecular forms of lipids, require lipid derivitization prior to analysis, or can be quite time consuming. Here we present a detailed description of our experimental approach to solve these inherent limitations by using survey-scan ESI/MS for the identification and quantification of the entire complement of lipids in yeast cells. The described method does not require chromatographic separation of complex lipid mixtures recovered from yeast cells, thereby greatly accelerating the process of data acquisition. This method enables lipid identification and quantification at the concentrations as low as g/ml and has been successfully applied to assessing lipidomes of whole yeast cells and their purified organelles. Lipids extraction from whole yeast cells for using this method of lipid analysis takes two to three hours. It takes only five to ten minutes to run each sample of extracted and dried lipids on a Q-TOF mass spectrometer equipped with a nano-electrospray source.

We describe a new quantitative lipidomics method for identifying numerous lipid species in yeast using survey-scan electrospray ionization mass spectrometry (ESI/MS). This method exceeds currently available methods for lipid identification and quantification in the ability to resolve various molecular forms of lipids, sensitivity, and speed.

エレクトロスプレーイオン化質量分析法を用いて酵母Lipidomeの定量評価

Lipids are one of the major classes of biomolecules and play important roles membrane dynamics, energy storage, and signalling1-4. The budding yeast ...

The use of Biofeedback in Clinical Virtual Reality: The INTREPID Project

Generalized anxiety disorder (GAD) is a psychiatric disorder characterized by a constant and unspecific anxiety that interferes with daily-life activities. Its high prevalence in general population and the severe limitations it causes, point out the necessity to find new efficient strategies to treat it. Together with the cognitive-behavioral treatments, relaxation represents a useful approach for the treatment of GAD, but it has the limitation that it is hard to be learned. The INTREPID project is aimed to implement a new instrument to treat anxiety-related disorders and to test its clinical efficacy in reducing anxiety-related symptoms. The innovation of this approach is the combination of virtual reality and biofeedback, so that the first one is directly modified by the output of the second one. In this way, the patient is made aware of his or her reactions through the modification of some features of the VR environment in real time. Using mental exercises the patient learns to control these physiological parameters and using the feedback provided by the virtual environment is able to gauge his or her success. The supplemental use of portable devices, such as PDA or smart-phones, allows the patient to perform at home, individually and autonomously, the same exercises experienced in therapist's office. The goal is to anchor the learned protocol in a real life context, so enhancing the patients' ability to deal with their symptoms. The expected result is a better and faster learning of relaxation techniques, and thus an increased effectiveness of the treatment if compared with traditional clinical protocols.

The Project on Virtual Reality Intelligent Multi-sensor System for the Treatment of Anxiety-related Disorders (INTREPID) is aimed at developing a multi-sensor context-aware virtual reality system for the treatment of anxiety-related disorders.

臨床バーチャルリアリティにおけるバイオフィードバックの利用：INTREPIDプロジェクト

Generalized anxiety disorder (GAD) is a psychiatric disorder characterized by a constant and unspecific anxiety that interferes with daily-life ...

Assessment of GFP Expression and Viability Using the Tali Image-Based Cytometer

Single-cell and population information are commonly obtained either by flow cytometry or fluorescence microscopy. However, these two methods provide different information. Flow cytometry gives quantitative multi-parametric information about physical characteristics and staining or expression, but doesn't allow for visualization. Stand-alone fluorescence microscopy provides visual data, but doesn't allow for straightforward quantitative measurements1.
Image-based cytometry bridges the gap between these two methods, enabling the quick visualization and simultaneous quantitative analysis of thousands of cells in heterogeneous populations2. Here, we present a method for performing cell viability and green fluorescent protein (GFP) expression assays using the Tali Image-Based Cytometer3. The Tali instrument is a 3-channel (bright field, green fluorescence, red fluorescence) benchtop assay platform that offers several advantages over flow cytometry and fluorescence microscopy. The Tali cytometer is less expensive, takes up less bench space, requires less maintenance, and the work flow has been simplified so that the operation and analysis is much simpler and quicker. The Tali cytometer is capable of performing a range of suspension cell-based assays, including GFP and red fluorescent protein (RFP) expression, apoptosis4-6 and cell viability analysis with propidium iodide (PI)7-11.
Here, we demonstrate the use of the Tali instrument in performing a cell viability assay in cells expressing GFP. GFP-transduced cells are stained using the Tali Viability Kit - Dead Cell Red. The cells are then pipetted into a Tali Cellular Analysis Slide and loaded into the cytometer. Bright field, red fluorescence and green fluorescence images are captured and analyzed using assay specific algorithms. Histograms are then generated to display cell size, PI fluorescence intensity, and GFP fluorescence intensity. These parameters can then be thresholded to home in on a specific cell population.
A side-by side comparison of the Tali Image-Based Cytometer and traditional flow cytometry demonstrates that the two methods provide comparable data regarding cell viability and protein expression. However, the Tali instrument provides additional visual information about the cell population that cannot be obtained using a flow cytometer.

This protocol describes how to perform cell viability and fluorescence expression assays using the Tali Image-Based Cytometer.

タリイメージベースのメ​​ーターを用いたGFPの発現と生存率の評価

Single-cell and population information are commonly obtained either by flow cytometry or fluorescence microscopy. However, these two methods provide ...

Analysis of Oxidative Stress in Zebrafish Embryos

High levels of reactive oxygen species (ROS) may cause a change of cellular redox state towards oxidative stress condition. This situation causes oxidation of molecules (lipid, DNA, protein) and leads to cell death. Oxidative stress also impacts the progression of several pathological conditions such as diabetes, retinopathies, neurodegeneration, and cancer. Thus, it is important to define tools to investigate oxidative stress conditions not only at the level of single cells but also in the context of whole organisms. Here, we consider the zebrafish embryo as a useful in vivo system to perform such studies and present a protocol to measure in vivo oxidative stress. Taking advantage of fluorescent ROS probes and zebrafish transgenic fluorescent lines, we develop two different methods to measure oxidative stress in vivo: i) a &#8220;whole embryo ROS-detection method&#8221; for qualitative measurement of oxidative stress and ii) a &#8220;single-cell&#160;ROS detection method&#8221; for quantitative measurements of oxidative stress. Herein, we demonstrate the efficacy of these procedures by increasing oxidative stress in tissues by oxidant agents and physiological or genetic methods. This protocol is amenable for forward genetic screens and it will help address cause-effect relationships of ROS in animal models of oxidative stress-related pathologies such as neurological disorders and cancer.

Here we report a protocol to measure oxidative stress in living zebrafish embryos. This procedure allows reactive oxygen species (ROS) detection in both whole embryo tissues and single-cell populations. This protocol will accomplish both qualitative and quantitative analyses.

ゼブラフィッシュ胚における酸化ストレスの分析

High levels of reactive oxygen species (ROS) may cause a change of cellular redox state towards oxidative stress condition. This situation causes ...

Method for the Assessment of Effects of a Range of Wavelengths and Intensities of Red/near-infrared Light Therapy on Oxidative Stress In Vitro

Red/near-infrared light therapy (R/NIR-LT), delivered by laser or light emitting diode (LED), improves functional and morphological outcomes in a range of central nervous system injuries in vivo, possibly by reducing oxidative stress. However, effects of R/NIR-LT on oxidative stress have been shown to vary depending on wavelength or intensity of irradiation. Studies comparing treatment parameters are lacking, due to absence of commercially available devices that deliver multiple wavelengths or intensities, suitable for high through-put in vitro optimization studies. This protocol describes a technique for delivery of light at a range of wavelengths and intensities to optimize therapeutic doses required for a given injury model. We hypothesized that a method of delivering light, in which wavelength and intensity parameters could easily be altered, could facilitate determination of an optimal dose of R/NIR-LT for reducing reactive oxygen species (ROS) in vitro.
Non-coherent Xenon light was filtered through narrow-band interference filters to deliver varying wavelengths (center wavelengths of 440, 550, 670 and 810nm) and fluences (8.5 x 10-3 to 3.8 x 10-1 J/cm2) of light to cultured cells. Light output from the apparatus was calibrated to emit therapeutically relevant, equal quantal doses of light at each wavelength. Reactive species were detected in glutamate stressed cells treated with the light, using DCFH-DA and H2O2 sensitive fluorescent dyes.&#160;
We successfully delivered light at a range of physiologically and therapeutically relevant wavelengths and intensities, to cultured cells exposed to glutamate as a model of CNS injury. While the fluences of R/NIR-LT used in the current study did not exert an effect on ROS generated by the cultured cells, the method of light delivery is applicable to other systems including isolated mitochondria or more physiologically relevant organotypic slice culture models, and could be used to assess effects on a range of outcome measures of oxidative metabolism.

Method for the Assessment of Effects of a Range of Wavelengths and Intensities of Red/near-infrared Light Therapy on Oxidative Stress In Vitro

Non-coherent Xenon light was passed through narrow-band interference and neutral density filters to deliver light of varying wavelength and intensity to cultured cells. This protocol was used to assess the effects of red/near-infrared light therapy on production of reactive species in vitro: no effects were observed using the tested parameters.

酸化ストレスに赤/近赤外光療法の波長および強度の範囲の効果の評価のための手法<em&gt;インビトロ</em

Red/near-infrared light therapy (R/NIR-LT), delivered by laser or light emitting diode (LED), improves functional and morphological outcomes in a range of ...

A Protocol for Using Gene Set Enrichment Analysis to Identify the Appropriate Animal Model for Translational Research

Recent studies that compared transcriptomic datasets of human diseases with datasets from mouse models using traditional gene-to-gene comparison techniques resulted in contradictory conclusions regarding the relevance of animal models for translational research. A major reason for the discrepancies between different gene expression analyses is the arbitrary filtering of differentially expressed genes. Furthermore, the comparison of single genes between different species and platforms often is limited by technical variance, leading to misinterpretation of the con/discordance between data from human and animal models. Thus, standardized approaches for systematic data analysis are needed. To overcome subjective gene filtering and ineffective gene-to-gene comparisons, we recently demonstrated that gene set enrichment analysis (GSEA) has the potential to avoid these problems. Therefore, we developed a standardized protocol for the use of GSEA to distinguish between appropriate and inappropriate animal models for translational research. This protocol is not suitable to predict how to design new model systems a-priori, as it requires existing experimental omics data. However, the protocol describes how to interpret existing data in a standardized manner in order to select the most suitable animal model, thus avoiding unnecessary animal experiments and misleading translational studies.

We provide a standardized protocol for the use of gene set enrichment analysis of transcriptomic data to identify an ideal mouse model for translational research. 
This protocol can be used with DNA microarray and RNA sequencing data and can further be extended to other omics data if data are available.

遺伝子を使用するプロトコル設定トランスレーショナルリサーチの適切な動物モデルを識別するために濃縮分析

Recent studies that compared transcriptomic datasets of human diseases with datasets from mouse models using traditional gene-to-gene comparison ...

Fish Sperm Assessment Using Software and Cooling Devices

For gamete quality evaluation, there are innovative, rapid, and quantitative techniques that can provide useful data for aquaculture. Computerized systems for sperm analysis were developed to measure several parameters and one of the most commonly measured is the sperm motility.
Initially, this computer technology was designed for mammalian species, although it can also be used for fish sperm analysis. Fish have specific features that can affect sperm assessment such as a short motility time after activation and, in some cases, adaptation to lower temperatures. Thus, it is necessary to modify both software and hardware components to make motility analysis more efficient for fish sperm analysis. For mammalian sperm, the heating plate is used to maintain optimal temperatures of spermatozoa. However, for some fish species, it is advantageous to use a lower temperature to prolong the duration of motility, since the sperm remain active for less than 2 min. Therefore, cooling devices are necessary to refrigerate samples at constant temperature over the time of analysis, including on the optical microscope. This protocol describes the analysis of fish sperm motility using software for sperm analysis and new cooling devices to optimize the results.

The present protocol describes a procedure of fish sperm assessment using computer-assisted sperm analysis and cooling devices. The software gives a rapid, accurate and quantitative analysis of fish sperm quality based on spermatozoa motility, which can be a useful tool in aquaculture to improve reproduction success.

魚の精子の評価ソフトウェアを使用して、冷却装置

For gamete quality evaluation, there are innovative, rapid, and quantitative techniques that can provide useful data for aquaculture. Computerized systems ...

An Air-liquid Interface Bronchial Epithelial Model for Realistic, Repeated Inhalation Exposure to Airborne Particles for Toxicity Testing

For toxicity testing of airborne particles, air-liquid interface (ALI) exposure systems have been developed for in vitro tests in order to mimic realistic exposure conditions. This puts specific demands on the cell culture models. Many cell types are negatively affected by exposure to air (e.g., drying out) and only remain viable for a few days. This limits the exposure conditions that can be used in these models: usually relatively high concentrations are applied as a cloud (i.e., droplets containing particles, which settle down rapidly) within a short period of time. Such experimental conditions do not reflect realistic long-term exposure to low concentrations of particles. To overcome these limitations the use of a human bronchial epithelial cell line, Calu-3 was investigated. These cells can be cultured at ALI conditions for several weeks while retaining a healthy morphology and a stable monolayer with tight junctions. In addition, this bronchial model is suitable for testing the effects of repeated exposures to low, realistic concentrations of airborne particles using an ALI exposure system. This system uses a continuous airflow in contrast to other ALI exposure systems that use a single nebulization producing a cloud. Therefore, the continuous flow system is suitable for repeated and prolonged exposure to airborne particles while continuously monitoring the particle characteristics, exposure concentration, and delivered dose. Taken together, this bronchial model, in combination with the continuous flow exposure system, is able to mimic realistic, repeated inhalation exposure conditions that can be used for toxicity testing.

Described is the cell culture and exposure method of an in vitro bronchial model for realistic, repeated inhalation exposure to particles for toxicity testing.

毒性試験のための空気中の粒子への現実的な反復吸入暴露のための気管支上皮モデル

For toxicity testing of airborne particles, air-liquid interface (ALI) exposure systems have been developed for in vitro tests in order to mimic realistic ...