Partie 1: Mise en place et préparer la peau <ol><li> Mettre en place les capteurs suivants respirer ceinture, EMG, ECG, conductance de la peau, température, sol; </li><li> Préparer la peau en frottant doucement avec un tampon imbibé d&#39;alcool dans les endroits suivants: le front au-dessus des yeux pour l&#39;EMG, à l&#39;intérieur des deux poignets pour l&#39;ECG, le dos de la main gauche pour le sol (emplacement de l&#39;électrode de masse peut varier selon le matériel utilisé); </li><li> Attachez des capteurs pour la peau dans les endroits suivants: la respiration de ceinture autour de la taille, l&#39;EMG sur le front au-dessus de chaque œil, ECG à l&#39;intérieur de chaque poignet conductance de la peau, sur le doigt du milieu de la main gauche, un thermomètre sur le pointeur de la main gauche, au sol sur le dos de la main gauche; </li><li> Sélectionnez la session pour la respiration VRC +; </li><li> Assurez-vous que Impédance est acceptable pour tous les capteurs et les capteurs sont sensibles au mouvement de client et produire des lectures raisonnables. </li></ol> Partie 2: Enregistrement <ol><li> Expliquer la procédure pour le client. </li><li> (Min 0-2) Demander au client de s&#39;asseoir tranquillement avec les yeux ouverts. Enregistrer une base de deux minutes. </li><li> (Min 2-4) Au bout de deux minutes, commencer un stresseur deux minutes cognitifs - des problèmes d&#39;arithmétique augmentant graduellement en difficulté d&#39;être résolu sans l&#39;utilisation d&#39;un stylo et du papier ou calculatrice (si l&#39;arithmétique est particulièrement difficile pour le client, demandez-leur à plusieurs reprises soustrait sept à partir de 100, c&#39;est à dire, «sept en série»). Donnez les instructions suivantes pour le client: «Dans les 2 prochaines minutes, je vais vous poser pour résoudre certains problèmes mathématiques. S&#39;il vous plaît travailler aussi vite que vous pouvez et dites-moi la réponse &quot;. Ne pas fournir de la rétroaction quant à savoir si réponse du client est correcte ou incorrecte et fournir des encouragements minimale (par exemple, &quot;juste faire du mieux que vous pouvez») afin de ne pas éliminer les propriétés douces induire un stress de cet exercice. </li><li> (Min 4-6) faire savoir au client que cette partie de l&#39;évaluation est terminée et demandez-lui / elle de s&#39;asseoir tranquillement avec les yeux ouverts pendant 2 minutes. </li><li> (Min 6-8) Commencez stress physique: tourner sur le bruit nocif bruyant (les bébés pleurer accidents de voiture, etc) et demander au client d&#39;écouter les sons sans se désintéressent-ils. </li><li> (Min 8-10) Eteignez le bruit, et de demander au client de s&#39;asseoir tranquillement avec les yeux ouverts pendant 2 minutes. </li><li> (Min 10-12) Commencez stress émotionnel: demander au client de parler d&#39;un incident stressant qu&#39;ils me souviens bien, inclure les détails de l&#39;événement, ce qu&#39;ils pensaient et de sentiment. Fournir un minimum de réponse empathique afin de ne pas limiter la réponse au stress. </li><li> (Min 12-14) Interrompre l&#39;histoire du client, même si il / elle n&#39;a pas fini, et de demander au client de s&#39;asseoir tranquillement avec les yeux ouverts. </li><li> Arrêter l&#39;enregistrement, enregistrer les données, et faire savoir au client que l&#39;évaluation est faite. </li></ol> Partie 3: Compte rendu et la planification du traitement <ol><li> Demander au client de leur expérience lors de l&#39;évaluation. Quelles sont les parties ont été particulièrement stressante et pourquoi? At-il / elle se sent toute changements physiologiques qui se passe? </li><li> Examen des données selon les critères décrits dans l&#39;annexe I. </li><li> Choisissez les paramètres physiologiques qui doivent être abordées basée sur l&#39;analyse ci-dessus. Si la lecture du capteur au départ est plus élevé ou plus bas que la normale, le but du traitement est d&#39;aider le client à mettre ce paramètre physiologique à la normale dans un environnement non stressant. Si la lecture du capteur pendant le stress est supérieur ou inférieur à la normale, le but du traitement est d&#39;aider le client à identifier les facteurs de stress et être capable de maintenir le niveau normal de fonctionnement physiologique sous stress. Si la lecture du capteur pendant le temps vers le bas indique l&#39;absence de récupération, le but du traitement est d&#39;aider le client à mettre ce paramètre physiologique à un niveau normal rapidement après un stress. Tout ou partie de ces objectifs peuvent être inclus dans le plan de traitement. </li></ol> Annexe I Lignes directrices soulignent l&#39;évaluation d&#39;interprétation (Tous les &quot;gras&quot; réponses indiquent les zones à traiter) <table border="0"><tbody><tr><td colspan="3"> EMG (lecture normale &lt;3 microvolts) </td></tr><tr><td> 1. Est-EMG élevé au départ? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 2. Est-EMG élevées après le départ à l&#39;un des trois facteurs de stress? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 3. Y at-il reprise de l&#39;EMG au niveau de base pendant les périodes de repos? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td colspan="3"> La fréquence cardiaque (plage normale de 60 à 80 battements par minute) </td></tr><tr><td> 1. Est-ce le rythme cardiaque élevé au départ? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 2. Est-ce une fréquence cardiaque élevée passé la ligne de base à l&#39;un des trois facteurs de stress? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 3. Est-il la récupération de la fréquence cardiaque au niveau de base pendant les périodes de repos? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td colspan="3"> Le rythme respiratoire (taux normal &lt;12 respirations par minute) </td></tr><tr><td> 1. Est-ce le rythme respiratoire élevé au départ? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 2. Taux est passé de respiration élevés de la ligne de base pour tout facteur de stress? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 3. Y at-il reprise de la respiration taux à base pendant les périodes de repos? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td colspan="3"> Conductance de la peau </td></tr><tr><td> 1. Est-conductance de la peau élevé au départ? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 2. Est-conductance de la peau surélevée passé la ligne de base pour tout facteur de stress? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 3. Y at-il reprise de conductance de la peau au niveau de base pendant les périodes de repos? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td colspan="3"> La température des doigts (lecture normale&gt; 90F) </td></tr><tr><td> 1. Est-ce la température des doigts faibles au départ? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 2. Est diminution de la température doigt passé la ligne de base pour tout facteur de stress? </td><td> Y es _ </td><td> Non _ </td></tr><tr><td> 3. Y at-il reprise de la température des doigts au niveau de base pendant les périodes de repos? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td colspan="3"> La variabilité du rythme cardiaque (normes varient selon l&#39;âge) </td></tr><tr><td> 1. Est faible variabilité du rythme cardiaque au départ? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 2. Est diminution de la variabilité du rythme cardiaque passé la ligne de base pour tout facteur de stress? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 3. Est-il la récupération de la variabilité du rythme cardiaque pendant les périodes de repos? </td><td> Oui _ </td><td> Non _ </td></tr></tbody></table> Annexe I L&#39;interprétation d&#39;évaluation du stress: <table border="0"><tbody><tr><td colspan="3"> EMG (lecture normale &lt;3 microvolts) </td></tr><tr><td> 1. Est-EMG élevé au départ? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 2. Est-EMG élevées après le départ à l&#39;un des trois facteurs de stress? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 3. Y at-il reprise de l&#39;EMG au niveau de base pendant les périodes de repos? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> La fréquence cardiaque (plage normale de 60 à 80 battements par minute) </td></tr><tr><td> 1. Est-ce le rythme cardiaque élevé au départ? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 2. Est-ce une fréquence cardiaque élevée passé la ligne de base à l&#39;un des trois facteurs de stress? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 3. Est-il la récupération de la fréquence cardiaque au niveau de base pendant les périodes de repos? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> Le rythme respiratoire (taux normal &lt;12 respirations par minute) </td></tr><tr><td> 1. Est-ce le rythme respiratoire élevé au départ? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 2. Taux est passé de respiration élevés de la ligne de base pour tout facteur de stress? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 3. Y at-il reprise de la respiration taux à base pendant les périodes de repos? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> Conductance de la peau (lecture normale &lt;5 micro ohms) </td></tr><tr><td> 1. Est-conductance de la peau élevé au départ? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 2. Est-conductance de la peau surélevée passé la ligne de base pour tout facteur de stress? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 3. Y at-il reprise de conductance de la peau au niveau de base pendant les périodes de repos? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> La température des doigts (lecture normale&gt; 90F) </td></tr><tr><td> 1. Est-ce la température des doigts faibles au départ? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 2. Est diminution de la température doigt passé la ligne de base pour tout facteur de stress? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 3. Y at-il reprise de la température des doigts au niveau de base pendant les périodes de repos? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> La variabilité du rythme cardiaque (normes varient selon l&#39;âge) </td></tr><tr><td> 1. Est faible variabilité du rythme cardiaque au départ? </td><td> Oui _ </td><td> Non _ </td></tr><tr><td> 2. Est diminution de la variabilité du rythme cardiaque passé la ligne de base pour tout facteur de stress? </td><td> Oui _ </td><td> Non _ </td></tr><tr> &lt;td&gt; 3. Est-il la récupération de la variabilité du rythme cardiaque pendant les périodes de repos? </td><td> Oui _ </td><td> Non _ </td></tr></tbody></table> La planification du traitement: Si une lecture est supérieure ou inférieure à la normale au départ, l&#39;objectif du traitement est d&#39;aider le client à mettre ce paramètre physiologique à la normale dans toutes les circonstances Si une lecture est supérieure ou inférieure à la normale pendant le facteur de stress, l&#39;objectif du traitement est d&#39;aider le client à identifier les facteurs de stress et être capable de maintenir le niveau normal de fonctionnement physiologique sous stress Si une lecture indique l&#39;absence de récupération pendant les temps morts, le but du traitement est d&#39;aider le client à mettre ce paramètre physiologique à un niveau normal rapidement après un stress.

<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.

Psychophysiologiques Terminé dans l&#39;évaluation du stress appendi fournit un praticien biofeedback avec les informations nécessaires à la formulation d&#39;un plan de traitement. Cette évaluation fournit des informations sur les stéréotypies réponse de l&#39;individu, ou l&#39;uniformité de certaines réactions physiologiques à tous les facteurs de stress, et la spécificité du stimulus-réponse, ou un individu est unique réponses physiologiques à différents types de stresseurs. Avec cette information, ...

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psychophysiological stress assessment using biofeedback 

Dans le dernier demi-siècle, la recherche dans le biofeedback a montré à quel point l&#39;esprit humain peut influencer le fonctionnement du système nerveux autonome, que l&#39;on croyait hors du contrôle conscient. En laissant les gens observent les signaux de leur propre corps, le biofeedback leur permet de développer une plus grande prise de conscience de leurs réactions physiologiques et psychologiques, comme le stress, et d&#39;apprendre à modifier ces réactions. Praticiens biofeedback peut faciliter ce processus en évaluant les réactions des gens s à des événements stressants et légèrement la formulation d&#39;un plan de traitement basé sur le biofeedback. Pendant l&#39;évaluation du stress du praticien premiers enregistrements de référence pour les lectures physiologiques, puis présente le client avec plusieurs facteurs de stress légers, comme une cognitif, facteur de stress physique et émotionnel. Variété des facteurs de stress sont présentés afin de déterminer stimulus-réponse d&#39;une personne spécificité, ou des différences dans la réaction de chaque personne aux stimuli qualitativement différents. Cette vidéo va démontrer le processus d&#39;évaluation du stress psychophysiologiques utilisant le biofeedback et de présenter des lignes directrices générales pour la planification du traitement.

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

Psychophysiological Stress Assessment Using Biofeedback

Dans cette vidéo, nous allons décrire une méthode d&#39;évaluation de la réaction psychophysiologiques personne au stress en utilisant le biofeedback. Nous présenterons également des directives générales pour la planification du traitement.

Évaluation du stress psychophysiologiques utilisant le biofeedback

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

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Biology

Psychophysiological Stress Assessment Using Biofeedback 

13.3K vues.  Cambridge Health Alliance. Dans cette vidéo, nous allons décrire une méthode d'évaluation de la réaction psychophysiologiques personne au stress en utilisant le biofeedback. Nous présenterons également des directives générales pour la planification du traitement.

Vidéo: 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.

Surveillance des hormones végétales Durant les réponses au stress

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

Recherche

Biologie

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.

Oblitération clip microchirurgie de l&#39;anévrisme cérébral Moyen Utilisation de l&#39;évaluation de débit peropératoire

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.

Une évaluation quantitative du lipidome levure en utilisant la spectrométrie de masse électrospray ionisation

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.

L&#39;utilisation du biofeedback dans la Réalité Virtuelle clinique: Le projet INTREPID

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

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.

Analyse du stress oxydatif dans embryons de poissons zèbres

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.

Méthode d&#39;évaluation des effets d&#39;une gamme de longueurs d&#39;onde et des intensités de Luminothérapie Rouge / proche infrarouge sur le stress oxydatif<em&gt; In Vitro</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.

Un protocole pour l’utilisation de gènes série d’analyses d’enrichissement pour identifier le modèle Animal approprié de recherche translationnelle

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.

Évaluation de sperme de poissons à l’aide de logiciels et de dispositifs de refroidissement

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.

Un modèle épithélial bronchique d’interface air-liquide pour l’exposition réaliste et répétée par inhalation aux particules aéroportées pour l’essai de toxicité

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

In Vitro Analysis of E3 Ubiquitin Ligase Function

The covalent attachment of ubiquitin (Ub) to internal lysine residue(s) of a substrate protein, a process termed ubiquitylation, represents one of the most important post-translational modifications in eukaryotic organisms. Ubiquitylation is mediated by a sequential cascade of three enzyme classes including ubiquitin-activating enzymes (E1 enzymes), ubiquitin-conjugating enzymes (E2 enzymes), and ubiquitin ligases (E3 enzymes), and sometimes, ubiquitin-chain elongation factors (E4 enzymes). Here, in vitro protocols for ubiquitylation assays are provided, which allow the assessment of E3 ubiquitin ligase activity, the cooperation between E2-E3 pairs, and substrate selection. Cooperating E2-E3 pairs can be screened by monitoring the generation of free poly-ubiquitin chains and/or auto-ubiquitylation of the E3 ligase. Substrate ubiquitylation is defined by selective binding of the E3 ligase and can be detected by western blotting of the in vitro reaction. Furthermore, an E2~Ub discharge assay is described, which is a useful tool for the direct assessment of functional E2-E3 cooperation. Here, the E3-dependent transfer of ubiquitin is followed from the corresponding E2 enzyme onto free lysine amino acids (mimicking substrate ubiquitylation) or internal lysines of the E3 ligase itself (auto-ubiquitylation). In conclusion, three different in vitro protocols are provided that are fast and easy to perform to address E3 ligase catalytic functionality.

In Vitro Analysis of E3 Ubiquitin Ligase Function

The present study provides detailed in vitro ubiquitylation assay protocols for the analysis of E3 ubiquitin ligase catalytic activity. Recombinant proteins were expressed using prokaryotic systems such as Escherichia coli culture.

In vitro Analyse de la fonction E3 Ubiquitin Ligase

The covalent attachment of ubiquitin (Ub) to internal lysine residue(s) of a substrate protein, a process termed ubiquitylation, represents one of the ...