1 부 : 설정 및 피부 준비 <ol><li> 벨트, EMG, ECG, 피부 전도성, 온도,지면 호흡 다음과 같은 센서를 설정; </li><li> 부드럽게 다음 위치에 닦아 알코올로 문지르면하여 피부 준비 : EMG를위한 눈 위의 이마, 내부 ECG에 대한 두 손목, 다시 왼쪽의 지상을 위해 (사용하는 장비에 따라 달라질 수 있습니다 접지 전극의 위치); </li><li> 다음 위치에서 피부에 센서를 부착 : EMG는 ECG 각 손목의 안쪽에 각 눈 위의 이마, 허리 벨트를 호흡, 피부 왼손의 가운데 손가락에 전도성, 왼손의 포인터에 온도계, 지상 왼쪽 손등에; </li><li> + HRV를 호흡에 대한 세션을 선택합니다; </li><li> 들이닥침이 센서는 고객의 움직임과 합리적인 판독을 생산하기 위해 반응하는 모든 센서에 대해 허용하고 있는지 확인하십시오. </li></ol> 2 부 : 녹음 <ol><li> 클라이언트에 프로 시저를 설명합니다. </li><li> (최소 0-2) 열린 눈으로 처박혀 있어야하는 클라이언트를 지시한다. 2 분 기준을 기록합니다. </li><li> (최소 2-4) 2 분 끝에, 2 분인지 스트레스를 시작 - 산술 문제가 점차 펜 &amp; 종이 또는 계산기 (중 사용하지 않고 해결될 수 난이도 증가 연산이 클라이언트에 특히 어려운 경우, 그들에게 물어 반복 즉), 100에서 &quot;일련 벌리&quot;를 시작 일곱 빼야합니다. 클라이언트에 다음과 같은 지침을 제공 : &quot;다음 2 분 후면, 난 당신이 몇 가지 수학 문제를 해결하기 위해 요청하는 것입니다. 최대한 빨리 작업하고 &quot;나에게 해답을 주시기를 바랍니다. 고객의 답변이 정확하지 않거나 올바르지 않습니다 여부를 피드백을 제공하고 있으므로 본 운동의 약한 스트레스 유도 속성을 제거하지 (예, &quot;단지 당신이 할 수있는 최선을 다하라&quot;) 최소한의 격려를 제공하지 않습니다. </li><li> (최소 4-6) 클라이언트가 평가의이 부분은 ​​끝났다는 걸 알아요 그리고 / 그 2 분 동안 열린 눈으로 조용히 앉아서 그에게 물어 보자. </li><li> (최소 6-8) 물리적인 스트레스를 시작 : 소리 유해 소음을 턴 (아기 울음, 교통 사고 등) 및 조정없이 소리로 들어 클라이언트를 부탁드립니다. </li><li> (최소 80-10) 노이즈를 끄고, 2 분 동안 열린 눈으로 처박혀 있어야하는 클라이언트를 부탁드립니다. </li><li> (최소 10-12) 정서적 스트레스를 시작 : 그들이 잘 기억 스트레스 사건에 대해 이야기하기 위해 클라이언트를 요청, 그들이 생각하고 느낌이 있었는지 이벤트의 세부 정보를 포함합니다. 스트레스 반응을 완화하지 않는만큼 최소한의 empathic 반응을 제공합니다. </li><li> (최소 12-14)는 그 / 그녀가 완료되지 않은 경우에도 고객의 이야기를 방해하고 열린 눈으로 처박혀 있어야하는 클라이언트를 지시합니다. </li><li> 녹음을 중지 데이터를 저장하고, 클라이언트가 심사가 완료 사실을 이미 알려드 렸습니다. </li></ol> 파트 3 : 보고서 및 치료 계획 <ol><li> 평가 기간 동안 그들의 경험에 대해 클라이언트를 부탁드립니다. 어떤 부분은 특히 스트레스 왜 있었어? 그 / 그녀가 모든 생리적 변화가 일어나고있는 게 느껴지나요? </li><li> 부록 I.에서 설명한 기준에 따라 검토 자료 </li><li> 위의 분석을 바탕으로 해결해야 생리 매개 변수를 선택할 수 있습니다. 기준에 센서를 읽는가 높거나 정상보다 낮은 경우, 치료 목표는 클라이언트가 아닌 스트레스 환경에 생리적 매개 변수 정상을 유치하는 데 도움이됩니다. 스트레스 동안 센서 읽기가 높거나 정상보다 낮은 경우, 치료의 목표는 클라이언트가 스트레스를 파악하고 스트레스 생​​리 기능의 정상적인 수준을 유지할 수 있도록 돕는 것입니다. 아래 시간 동안 센서 독서 복구 부족을 나타내는 경우, 치료의 목표는 클라이언트가 그 생리적 매개 변수를 정상 수준으로 빠르게 스트레스를 다음을 유치하는 데 도움이됩니다. 이러한 목표 중 어느 하나 또는 모두가 치료 계획에 포함될 수 있습니다. </li></ol> 부록 I 스트레스 평가 해석 지침 (모든 &quot;굵은&quot;답변 영역이 주소로 표시) <table border="0"><tbody><tr><td colspan="3"> EMG (일반 읽기 &lt;3 microvolts) </td></tr><tr><td> 1. EMG는 기준에서 높은가요? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 2. EMG는 3 스트레스의에서 기준 과거 고가인가​​요? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 3. 휴식 기간 동안 기본 수준으로 EMG의 복구가 있습니까? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td colspan="3"> 심장 박동 (정상 범위 분당 60-80 비트) </td></tr><tr><td> 1. 기준에서 높은 심장 박동인가? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 2. 심장 박동수가 3 스트레스의에서 기준 과거 고가인가​​요? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 3. 휴식 기간 동안 기본 수준으로 심장 박동의 회복이 있습니까? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td colspan="3"> 호흡 속도 (일반 속도 &lt;분당 12 번 호흡할) </td></tr><tr><td> 1. 베이스 라인에서 고가 호흡 속도인가요? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 2. 호흡입니다 요금은 스트레스에 대한 기준 과거 고가? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 3. 휴식 기간 동안 기준에 속도를 호흡 회수가 있습니까? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td colspan="3"> 피부 전도성 </td></tr><tr><td> 1. 베이스 라인에서 고가 피부 전도인가? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 2. 피부 전도성은 스트레스에 대한 기준 과거 고가인가​​요? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 3. 기본으로 피부 전도성의 복구 휴식 기간 동안 있습니까? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td colspan="3"> 핑거 온도 (정상 읽기&gt; 90F) </td></tr><tr><td> 1. 기본 낮은 손가락 온도인가요? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 2. 손가락 온도 감소는 스트레스에 대한 기준 과거합니까? </td><td> 응 _ </td><td> 노 _ </td></tr><tr><td> 3. 기준에 손가락 온도의 회복은 휴식 기간 동안 있습니까? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td colspan="3"> 심장 박동의 변화 (규범은 연령에 따라 다릅니다) </td></tr><tr><td> 1. 기준에 심장 박동의 변화의 낮은가요? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 2. 어떤 스트레스에 대한 기준 과거 심장 박동의 변화 감소합니까? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 3. 휴식 기간 동안 심장 박동의 변화의 복구가 있습니까? </td><td> 예 _ </td><td> 노 _ </td></tr></tbody></table> 부록 I 스트레스 평가 해석 : <table border="0"><tbody><tr><td colspan="3"> EMG (일반 읽기 &lt;3 microvolts) </td></tr><tr><td> 1. EMG는 기준에서 높은가요? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 2. EMG는 3 스트레스의에서 기준 과거 고가인가​​요? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 3. 휴식 기간 동안 기본 수준으로 EMG의 복구가 있습니까? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 심장 박동 (정상 범위 분당 60-80 비트) </td></tr><tr><td> 1. 기준에서 높은 심장 박동인가? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 2. 심장 박동수가 3 스트레스의에서 기준 과거 고가인가​​요? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 3. 휴식 기간 동안 기본 수준으로 심장 박동의 회복이 있습니까? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 호흡 속도 (일반 속도 &lt;분당 12 번 호흡할) </td></tr><tr><td> 1. 베이스 라인에서 고가 호흡 속도인가요? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 2. 호흡입니다 요금은 스트레스에 대한 기준 과거 고가? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 3. 휴식 기간 동안 기준에 속도를 호흡 회수가 있습니까? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 피부 전도성 (일반 읽기 &lt;5 마이크로 ohms) </td></tr><tr><td> 1. 베이스 라인에서 고가 피부 전도인가? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 2. 피부 전도성은 스트레스에 대한 기준 과거 고가인가​​요? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 3. 기본으로 피부 전도성의 복구 휴식 기간 동안 있습니까? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 핑거 온도 (정상 읽기&gt; 90F) </td></tr><tr><td> 1. 기본 낮은 손가락 온도인가요? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 2. 손가락 온도 감소는 스트레스에 대한 기준 과거합니까? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 3. 기준에 손가락 온도의 회복은 휴식 기간 동안 있습니까? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 심장 박동의 변화 (규범은 연령에 따라 다릅니다) </td></tr><tr><td> 1. 기준에 심장 박동의 변화의 낮은가요? </td><td> 예 _ </td><td> 노 _ </td></tr><tr><td> 2. 어떤 스트레스에 대한 기준 과거 심장 박동의 변화 감소합니까? </td><td> 예 _ </td><td> 노 _ </td></tr><tr> &lt;TD&gt; 3. 휴식 기간 동안 심장 박동의 변화의 복구가 있습니까? </td><td> 예 _ </td><td> 노 _ </td></tr></tbody></table> 치료 계획 : 독서 기준에 정상보다 높거나 낮은 경우, 치료의 목표는 클라이언트가 모든 상황에서 생리적 매개 변수 정상을 유치하는 데 도움이됩니다 독서가 스트레스 동안 정상보다 높거나 낮은 경우, 치료의 목표는 클라이언트가 스트레스를 파악하고 스트레스 생​​리 기능의 정상적인 수준을 유지할 수 있도록 돕는 것입니다 독서 아래 시간 동안 복구 부족을 나타냅니다 경우, 치료의 목표는 클라이언트가 그 생리적 매개 변수를 정상 수준으로 빠르게 스트레스를 다음과 같이 유치하는 데 도움이됩니다.

<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 스트레스 평가에 완료 psychophysiological는 치료 계획을 공식화하는 데 필요한 정보와 함께 바이오 피드백 개업을 제공합니다. 이 평가는 개인의 반응 stereotypy 또는 모든 스트레스 특정 생리적 반응의 균일하고, 자극 - 반응 특이성, 또는 스트레스의 종류에 개인의 고유한 생리 반응 모두에 대한 정보를 제공합니다. 이 정보를 통해 클라이언트는 다음 생리 반응의 종류는 어떤 상황에서 기대하?...

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psychophysiological stress assessment using biofeedback 

지난 반세기에 바이오 피드백의 연구가 어떤 인간의 정신은 자율 신경계의 기능에 영향을 미칠 수있는 범위를 표시했다 이전에 의식 제어 밖에 생각. 사람들이 자신의 신체의 신호를 관찰시키는함으로써, 바이오 피드백은 그들이 같은 스트레스로 자신의 생리와 심리적 반응의 큰 의식을 개발할 수있게하고, 이러한 반응을 수정할 수 배울 수 있습니다. 바이오 피드백 실무자는 약간 스트레스 사건 사람의 반응을 평가하고 바이오 피드백 기반 치료 계획을 공식화하여이 프로세스를 용이하게하실 수 있습니다. 스트레스 평가하는 동안 먼저 의사는 생리 판독을위한 기준을 기록하고, 다음과 같은 인지적, 신체적, 정서적 스트레스와 같은 몇 가지 가벼운 스트레스와 클라이언트를 제공합니다. 스트레스의 다양한 사람의 자극 - 반응 특이성, 또는 질적으로 다른 자극에 각 사람의 반응의 차이를 결정하기 위해 제공됩니다. 이 동영상은 치료 계획에 대한 바이오 피드백과 현재의 일반적인 지침을 사용하여 psychophysiological 스트레스 평가의 과정을 보여주는 것입니다.

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

Psychophysiological Stress Assessment Using Biofeedback

이 비디오에서는 바이오 피드백을 사용하여 스트레스 사람의 psychophysiological 반응을 평가 한 방법을 설명합니다. 우리는 또한 치료 계획을위한 일반적인 지침을 제시합니다.

생체 자기 제어를 사용 Psychophysiological 스트레스 평가

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 ...

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Research

JoVE Journal

Biology

Psychophysiological Stress Assessment Using Biofeedback 

13.2K 조회수.  Cambridge Health Alliance. 이 비디오에서는 바이오 피드백을 사용하여 스트레스 사람의 psychophysiological 반응을 평가 한 방법을 설명합니다. 우리는 또한 치료 계획을위한 일반적인 지침을 제시합니다.

비디오: 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.

스트레스 응답하는 동안 식물 호르몬을 모니터링

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

연구

생물학

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.

Intraoperative 흐름 평가를 사용하여 중간 뇌동맥류야의 Microsurgical 클립 말살

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.

전기 분무 이온화 질량 분광법을 사용하여 효모 Lipidome의 양적 평가

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.

임상 가상 현실의 생체 자기 제어의 사용 인트 리피드 프로젝트

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.

Tali 이미지 기반 Cytometer를 사용하여 GFP의 표현과 생존 능력의 평가

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.

제브라 피쉬 배아에서 산화 스트레스의 분석

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.

파장과 산화 스트레스에 레드 / 근적외선 치료의 강렬의 범위의 효과의 평가를위한 방법<em&gt; 체외</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.

유전자를 사용 하 여 프로토콜 변환 연구에 대 한 적절 한 동물 모델을 식별 하기 위해 농축 분석 설정

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.

생선 정자 평가 소프트웨어를 사용 하 여 및 냉각 장치

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.

독성 테스트를 위해 공기 중 입자에 대한 사실적이고 반복적인 흡입 노출을 위한 공기 액체 인터페이스 기관지 상피 모델

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