Nous tenons à remercier Kun Lu et Ronald Kurz de l&#39;assistance technique. Ce travail a été soutenu par l&#39;ONR n ° Prix Muri: N00014-10-1-0072, NSF Grant # SBE-0542013 à la dynamique temporelle de Learning Center, une science de la NSF Learning Center, NIH et # 2 R01 NS036449-11.

1. En dehors de la salle du scanner <ol><li> Placez la table de roulement avec l&#39;extrémité libre soutenus et l&#39;extrémité extérieure de la poignée de longs détachés. </li><li> Vérifiez que le robot est éteint. </li><li> Placez le robot dans la prise de table et fixer la plaque de sécurité en aluminium sur le robot avec 2 vis. </li><li> Fixez le effecteur fin à la poignée de robot avec l&#39;adaptateur en aluminium et vérifier qu&#39;il se déplace librement. </li><li> Fixez la 10 &#39;câble parallèle avec de l&#39;aluminium de blindage pour le robot et vérifier que le blindage est intact. Ajouter une feuille supplémentaire si nécessaire. </li><li> Placez la boîte en aluminium de blindage sur le robot en prenant soin de placer les câbles d&#39;alimentation parallèle et dans la rainure sur le dos. </li><li> Soigneusement vis sur la boîte de blindage. </li><li> Pack de papier d&#39;aluminium dans la rainure du câble sur la boîte de blindage et être sûr que le film est en contact avec le blindage sur le câble parallèle. </li></ol> 2. S&#39;engager dans la salle du scanner avec deux personnes, A et B <ol><li> Préparez-vous à entrer dans un environnement champ magnétique élevé en supprimant toute et tous les objets métalliques non ferreux, y compris celles, par exemple, les téléphones cellulaires, des clés, pièces de monnaie, etc .. </li><li> Avec une personne A la fin tenant l&#39;extrémité libre de la table de robot et la personne B stabiliser la fin, en boîte il libre d&#39;abord que le robot passe juste la porte de la chambre. </li><li> Clips personne B de la corde de sécurité attaché à un trou d&#39;ancrage sur le dos de la sacoche de protection et vérifie que l&#39;autre extrémité est solidement attaché à une fixation murale. </li><li> En travaillant ensemble, rouler la table dans la salle et l&#39;attacher avec des sangles velcro au pied de la table IRMf. La fin du robot de la table doit rester aussi loin que possible du scanner. </li><li> Branchez le câble parallèle du robot au filtre personnalisé sur le passage à la salle de contrôle, et de brancher le robot. Le blindage sur le câble parallèle doivent prendre contact avec le filtre. </li><li> Fixez la partie extérieure de la longue poignée (effecteur final) et vérifiez que celle-ci entrera l&#39;alésage de l&#39;IRMf proprement. </li></ol> 3. Dans la salle de contrôle <ol><li> Démarrer l&#39;ordinateur de contrôle et de joindre un câble parallèle de la 6 &#39;au robot. Veillez à retirer tous les filtres supplémentaires qui peuvent être sur la partie intérieure de la passer à travers. </li><li> Vérifiez que les moteurs du robot sont éteints, le démarrage de la routine de calibrage Phantom et vérifier que les afficheurs de position des moteurs de la routine de calibrage du robot sont stables. </li><li> Vérifiez la connexion du câble parallèle et que seul le grand filtre personnalisé est attachée si la routine de calibrage ne peut pas voir le robot ou il ya de grandes variations dans la lecture du moteur. </li><li> Mettez le robot en ouvrant le port à l&#39;arrière de la boîte de blindage et appuyez sur les interrupteurs avec un bâton. </li><li> Redémarrez la routine de calibrage avec le effecteur final approximativement centré à l&#39;extrémité du guide d&#39;ondes conique. Étape par l&#39;étalonnage et vérifiez que la boîte de calibrage boîte a l&#39;interaction haptique approprié. </li><li> Fixez la sortie TTL du IRMf (connecteur BNC) à l&#39;ADC LabJack sur l&#39;ordinateur de contrôle. </li></ol> 4. Le sujet <ol><li> Préparer le sujet pour un environnement de haut champ magnétique avec les protocoles IRMf standard. </li><li> Mettre en place un équipement supplémentaire pour l&#39;expérience, par exemple, système d&#39;affichage visuel. Nous utilisons les NordicNeuro Lab, Vision System Inc qui fournit l&#39;observation stéréoscopique d&#39;affichages visuels, une caractéristique qui est particulièrement utile lors de la présentation des environnements virtuels. </li><li> Après le sujet s&#39;allonge sur la table et la tête de la bobine se trouve, régler la distance du robot en dévissant les vis à main et glisser le haut du tableau jusqu&#39;à ce que le sujet peut se déplacer confortablement. </li><li> Guide de la table manuellement en maintenant sur l&#39;extrémité du robot tandis que la table IRMf est déplaçant dans et hors de l&#39;alésage pour éviter les roulettes de vaciller. Assurez-vous que l&#39;effecteur final va dans le trou et ne pas attraper de l&#39;extérieur. </li><li> Exécutez l&#39;expérience. </li></ol> 5. Décomposer la configuration avec deux personnes A et B <ol><li> Après la sortie du patient, retirez la fin de l&#39;extérieur le long manche et se détacher de la table robot de la table IRMf en desserrant la sangle velcro. </li><li> Du robot, défaire le câble parallèle blindé et débranchez le câble d&#39;alimentation. </li><li> Avec une personne A tenant l&#39;extrémité libre de la table de robot et la personne B guider l&#39;extrémité du robot, de déplacer la table à la porte. À la porte, défaire l&#39;attache, et rouler sur la table du robot à la salle. </li><li> Annuler toutes les vis de la boîte de blindage et les deux vis de la plaque de la sécurité et retirer le robot. </li></ol> 6. Les résultats représentatifs: Idéalement, le robot haptique et l&#39;IRMf ne devrait pas affecter l&#39;autre. Nous pouvons dire en ligne si le robot est touché par l&#39;IRMf. Généralement, si un câble parallèle du robot n&#39;est pas correctement blindé et filtré, puis la lecture des moteurs sera oscillateurfin rapidement. Ceci peut être fixé par un double contrôle de l&#39;aluminium de blindage sur le câble, que le noyau ferreux est correctement placé sur le câble parallèle à proximité du robot, et que le filtre seulement pour le robot est le filtre personnalisé sur le côté salle du scanner de l&#39;passent à travers . Détection des erreurs dans les IRMf est vraiment possible qu&#39;après que les données ont été réduites et analysées, mais une analyse anatomique devraient être prises au début de l&#39;étude et vérifié les effets de fermeture éclair ou d&#39;autres artefacts indicatif de bruit corrélé (par exemple pic de bruit) 7. Souvent, un tel bruit vient du métal au contact de métal et peuvent être nettoyés en serrant les vis sur la table de robot, surtout les vis de réglage main sur le côté de la table. De nos tests le signal IRMf de référence pour bruit (SNR) est ~ 380 et avec le robot entièrement blindé dans la pièce qui tombe à un niveau encore raisonnable ~ 330. Si le bouclier n&#39;est pas en place sur le robot, puis la chute de SNR suite à ~ 250, et les effets sonores deviennent très importantes. Comme le montre le 4, le degré 3 de la liberté dans la commune centre de la poignée a peu d&#39;effet sur ​​la dynamique de l&#39;interaction robot / main, sauf pour le changement de distance du robot. La commune dans le centre de la poignée agit comme un point d&#39;appui et renverse le mouvement apparent dans deux directions (gauche-droite et haut-bas) mais pas le troisième (avant-arrière). Depuis le fantôme et la main sont aux extrémités opposées de la manette comme manche avec son point d&#39;appui dans le milieu, les gains sont appliquées dans le logiciel dans chacune des trois directions euclidienne: les gains négatifs dans les deux directions, commandé par la rotule et un gain positif dans le sens de l&#39;articulation du curseur. L&#39;effet net de la poignée et le pivot reproduit les trois degrés de liberté complète du robot Phantom, à seulement 9 pieds de distance. <img src="/files/ftp_upload/3364/3364fig1.jpg" alt="Figure 1"/> Figure 1 L&#39;appareil utilisé pour monter le robot haptique pour une utilisation dans l&#39;environnement IRMf. Haut montre le robot haptique montés dans le cas avant boîtier (en haut, à gauche) et le cardan / curseur conjointe à mi-chemin de la poignée (en haut, à droite). Bas, de gauche montre un sujet dans le scanner de manipuler la poignée. Bas, est juste une bande dessinée de l&#39;effecteur de blindage et de fin. <img src="/files/ftp_upload/3364/3364fig2.jpg" alt="Figure 2"/> Figure 2 Résultats de l&#39;épreuve BIRN pour le robot entièrement blindé avec le mouvement. Les trois images avec des croix indiquent les sections du modèle de tête sphérique, et le bas à droite montre une vue en trois dimensions. Les petits points sont des bulles dans le modèle de tête statique et sont toujours présents. Le manque de bandes ou fermetures à glissière grandes indique que le bruit du robot n&#39;est pas corrélé.

<ol>
	<li>Ogawa, S., Lee, T. M., Kay, A. R., Tank, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brain+magnetic+resonance+imaging+with+contrast+dependent+on+blood+oxygenation.">Brain magnetic resonance imaging with contrast dependent on blood oxygenation.</a> Proc. Natl. Acad. Sci. U.S.A. 87, 9868-9872 (1990).</li><li>Heeger, D. J., Ress, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=What+does+fMRI+tell+us+about+neuronal+activity.">What does fMRI tell us about neuronal activity.</a> Nat. Rev. Neurosci. 3, 142-151 (2002).</li><li>deCharms, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Applications+of+real-time+fMRI.">Applications of real-time fMRI.</a> Nat. Rev. Neurosci. 9, 720-729 (2008).</li><li>Hribar, A., Koritnik, B., Munih, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phantom+haptic+device+upgrade+for+use+in+fMRI.">Phantom haptic device upgrade for use in fMRI.</a> Medical and Biological Engineering and Computing. 47, 677-684 (2009).</li><li>Trommershauser, J., Gepshtein, S., Maloney, L. T., Landy, M. S., Banks, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimal+compensation+for+changes+in+task-relevant+movement+variability.">Optimal compensation for changes in task-relevant movement variability.</a> J. Neurosci. 25, 7169-7178 (2005).</li><li>Konczak, J., Li, K. Y., Tuite, P. J., Poizner, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Haptic+perception+of+object+curvature+in+Parkinson's+disease.">Haptic perception of object curvature in Parkinson's disease.</a> PLoS ONE. 3, e2625-e2625 (2008).</li><li>Lipton, M. L., Lipton, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Artifacts:+When+things+go+wrong,+it's+not+necessarily+all+bad.">Artifacts: When things go wrong, it's not necessarily all bad.</a> Totally Accessible MRI. , 139-153 (2008).</li><li>Rajh, M., Glodez, S., Flasker, J., Gotlih, K., Kostanjevec, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design+and+analysis+of+an+fMRI+compatible+haptic+robot.">Design and analysis of an fMRI compatible haptic robot.</a> Robotics and Computer-Integrated Manufacturing. 27, 267-275 (2011).</li><li>Burdet, E., Gassert, R., Gowrishankar, G., Chapuis, D., Bleuler, H., Ang, M., Khatib, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=fMRI+compatible+haptic+interfaces+to+investigate+human+motor+control.">fMRI compatible haptic interfaces to investigate human motor control.</a> Experimental Robotics IX, of Springer Tracts in Advanced Robotics. 21, 25-34 (2006).</li><li>Nakamura, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+networks+in+motor+sequence+learning:+abnormal+topographies+in+Parkinson's+disease.">Functional networks in motor sequence learning: abnormal topographies in Parkinson's disease.</a> Hum Brain Mapp. 12, 42-60 (2001).</li></ol>

Pas de conflits d&#39;intérêt déclarés.

Le robot IRMf compatibles ouvre de nouvelles possibilités pour des expériences en neuroscience de contrôle moteur. L&#39;étape la plus critique dans le setup est le blindage du robot pour éviter des artefacts dans l&#39;IRMf, que nous ne en deux étapes. D&#39;abord, le robot lui-même est d&#39;environ 9 &#39;loin de l&#39;ennuyer avec un long, léger, poignée soutenue en son milieu avec un degré 3 de la liberté commune. Deuxièmement, le robot est enfermé dans un 1 / 16 boîte en aluminium &quot;-1 / 4&quot; ...

<table border="1"><tbody><tr><td> Phantom prime 1.5/6dof, modèle de force de haute </td><td> SensAble </td><td> <a href="www.sensable.com">www.sensable.com</a> </td></tr></tbody></table>

adaptation of a haptic robot in a 3t fmri

Par résonance magnétique fonctionnelle (IRMf) permet une excellente imagerie cérébrale fonctionnelle via le signal BOLD 1 avec des avantages, y compris les rayonnements non ionisants, millimètre précision spatiale des données anatomiques et fonctionnelles 2, et presque en temps réel des analyses 3. Robots haptiques de fournir une mesure précise et un contrôle de la position et la force d&#39;un curseur dans un espace relativement confiné. Ici, nous combinons ces deux technologies pour permettre des expériences de précision impliquant le contrôle moteur avec haptique / interaction environnement tactiles telles que l&#39;atteinte ou la préhension. L&#39;idée de base est d&#39;attacher une extrémité 8 pieds effecteur pris en charge dans le centre pour le robot 4 permettant au sujet d&#39;utiliser le robot, mais c&#39;est le blindage et le garder hors de la partie la plus extrême du champ magnétique de la machine IRMf (figure 1 ). Le Fantôme Premium 3.0, 6DoF, grande force du robot (SensAble Technologies, Inc) est un excellent choix pour fournir à retour de force dans des expériences de réalité virtuelle 5, 6, mais il est intrinsèquement non-MR sûr, introduit du bruit significative à la sensibilité équipements IRMf, et ses moteurs électriques peuvent être touchés par le IRMf est fortement champ magnétique variable. Nous avons construit une table et un système de blindage qui permet au robot d&#39;être en sécurité introduits dans l&#39;environnement IRMf et les limites à la fois la dégradation du signal IRMf par les moteurs électriques bruyantes et la dégradation de la performance du moteur électrique par le champ magnétique variable fortement de l&#39; IRMf. Avec le bouclier, le rapport signal sur bruit (SNR: moyenne écart-type de signal / bruit) de l&#39;IRMf va partir d&#39;une base de ~ 380 à ~ 330 et ~ 250 sans blindage. Le bruit reste semble être corrélés et ne pas ajouter des artefacts à l&#39;IRMf d&#39;une sphère d&#39;essai (figure 2). La longue poignée rigide permet de placer le robot hors de portée des parties les plus fortes variations du champ magnétique donc il n&#39;ya pas d&#39;effet significatif de l&#39;IRMf sur le robot. L&#39;effet de la poignée sur la cinématique du robot est minime car il est léger (~ 2,6 lb), mais extrêmement rigide 3 / 4 &quot;de graphite et bien équilibrée sur la commune 3DOF au milieu. Le résultat final est une IRMf système compatible haptique, avec environ 1 pied cube d&#39;espace de travail, et, lorsqu&#39;il est combiné avec la réalité virtuelle, elle permet une nouvelle série d&#39;expériences à réaliser dans l&#39;environnement IRMf notamment naturalistes atteindre, le déplacement passif de la branche et la perception haptique, l&#39;apprentissage d&#39;adaptation dans les champs de force variable , l&#39;identification texture ou 5, 6.

Watch this Scientific Journal Video about Adaptation of a Haptic Robot in a 3T fMRI at JoVE.com

Adaptation of a Haptic Robot in a 3T fMRI

L&#39;adaptation et l&#39;utilisation d&#39;un robot haptique dans une IRMf 3T est décrite.

Adaptation d&#39;un robot haptique dans une IRMf 3T

Functional magnetic resonance imaging (fMRI) provides excellent functional brain imaging via the BOLD signal 1 with advantages including non-ionizing ...

adaptation-of-a-haptic-robot-in-a-3t-fmri

Research

JoVE Journal

Bioengineering

9.6K vues.  University of California. L'adaptation et l'utilisation d'un robot haptique dans une IRMf 3T est décrite.

Vidéo: Adaptation of a Haptic Robot in a 3T fMRI

Fabrication of Electrochemical-DNA Biosensors for the Reagentless Detection of Nucleic Acids, Proteins and Small Molecules

As medicine is currently practiced, doctors send specimens to a central laboratory for testing and thus must wait hours or days to 
receive the results. Many patients would be better served by rapid, bedside tests. To this end our laboratory and others have developed a versatile, reagentless 
biosensor platform that supports the quantitative, reagentless, electrochemical detection of nucleic acids (DNA, RNA), proteins (including antibodies) and small 
molecules analytes directly in unprocessed clinical and environmental samples. In this video, we demonstrate the preparation and use of several biosensors in this 
"E-DNA" class. In particular, we fabricate and demonstrate sensors for the detection of a target DNA sequence in a polymerase chain reaction mixture, an HIV-specific antibody and the drug cocaine. The preparation procedure requires only three hours of hands-on effort followed by an overnight incubation, and their use requires only minutes.

"E-DNA" sensors, reagentless, electrochemical biosensors that perform well even when challenged directly in blood and other complex matrices, have been adapted to the detection of a wide range of nucleic acid, protein and small molecule analytes. Here we present a general procedure for the fabrication and use of such sensors.

La fabrication des électrochimique de l&#39;ADN des Biocapteurs pour la détection sans réactif d&#39;acides nucléiques, protéines et petites molécules

As medicine is currently practiced, doctors send specimens to a central laboratory for testing and thus must wait hours or days to 
receive the results. ...

Recherche

Bio-ingénierie

Haptic/Graphic Rehabilitation: Integrating a Robot into a Virtual Environment Library and Applying it to Stroke Therapy

Recent research that tests interactive devices for prolonged therapy practice has revealed new prospects for robotics combined with graphical and other forms of biofeedback. Previous human-robot interactive systems have required different software commands to be implemented for each robot leading to unnecessary developmental overhead time each time a new system becomes available. For example, when a haptic/graphic virtual reality environment has been coded for one specific robot to provide haptic feedback, that specific robot would not be able to be traded for another robot without recoding the program. However, recent efforts in the open source community have proposed a wrapper class approach that can elicit nearly identical responses regardless of the robot used. The result can lead researchers across the globe to perform similar experiments using shared code. Therefore modular "switching out"of one robot for another would not affect development time. In this paper, we outline the successful creation and implementation of a wrapper class for one robot into the open-source H3DAPI, which integrates the software commands most commonly used by all robots.

Recently, a vast amount of prospects have come available for human-robot interactive systems. In this paper we outline the integration of a new robotic device with open source software that can rapidly make possible a library of interactive functionality. We then outline a clinical application for a neurorehabilitation application.

Haptique / Réhabilitation graphique: Intégration d&#39;un robot dans un environnement virtuel Bibliothèque et en l&#39;appliquant à la thérapie des maladies

Recent research that tests interactive devices for prolonged therapy practice has revealed new prospects for robotics combined with graphical and other ...

Creating Two-Dimensional Patterned Substrates for Protein and Cell Confinement

Microcontact printing provides a rapid, highly reproducible method for the creation of well-defined patterned substrates.1 While microcontact printing can be employed to directly print a large number of molecules, including proteins,2 DNA,3 and silanes,4 the formation of self-assembled monolayers (SAMs) from long chain alkane thiols on gold provides a simple way to confine proteins and cells to specific patterns containing adhesive and resistant regions. This confinement can be used to control cell morphology and is useful for examining a variety of questions in protein and cell biology. Here, we describe a general method for the creation of well-defined protein patterns for cellular studies.5 This process involves three steps: the production of a patterned master using photolithography, the creation of a PDMS stamp, and microcontact printing of a gold-coated substrate. Once patterned, these cell culture substrates are capable of confining proteins and/or cells (primary cells or cell lines) to the pattern.
The use of self-assembled monolayer chemistry allows for precise control over the patterned protein/cell adhesive regions and non-adhesive regions; this cannot be achieved using direct protein stamping. Hexadecanethiol, the long chain alkane thiol used in the microcontact printing step, produces a hydrophobic surface that readily adsorbs protein from solution. The glycol-terminated thiol, used for backfilling the non-printed regions of the substrate, creates a monolayer that is resistant to protein adsorption and therefore cell growth.6 These thiol monomers produce highly structured monolayers that precisely define regions of the substrate that can support protein adsorption and cell growth. As a result, these substrates are useful for a wide variety of applications from the study of intercellular behavior7 to the creation of microelectronics.8
While other types of monolayer chemistry have been used for cell culture studies, including work from our group using trichlorosilanes to create patterns directly on glass substrates,9 patterned monolayers formed from alkane thiols on gold are straight-forward to prepare. Moreover, the monomers used for monolayer preparation are commercially available, stable, and do not require storage or handling under inert atmosphere. Patterned substrates prepared from alkane thiols can also be recycled and reused several times, maintaining cell confinement.10

Self-assembled monolayers (SAMs) formed from long chain alkane thiols on gold provide well-defined substrates for the formation of protein patterns and cell confinement. Microcontact printing of hexadecanethiol using a polydimethylsiloxane (PDMS) stamp followed by backfilling with a glycol-terminated alkane thiol monomer produces a pattern where protein and cells adsorb only to the stamped hexadecanethiol region.

Créer deux dimensions Substrats motifs pour le confinement des protéines et de cellules

Microcontact printing provides a rapid, highly reproducible method for the creation of well-defined patterned substrates.1  While microcontact printing ...

Cellular Lipid Extraction for Targeted Stable Isotope Dilution Liquid Chromatography-Mass Spectrometry Analysis

The metabolism of fatty acids, such as arachidonic acid (AA) and linoleic acid (LA), results in the formation of oxidized bioactive lipids, including numerous stereoisomers1,2. These metabolites can be formed from free or esterified fatty acids. Many of these oxidized metabolites have biological activity and have been implicated in various diseases including cardiovascular and neurodegenerative diseases, asthma, and cancer3-7. Oxidized bioactive lipids can be formed enzymatically or by reactive oxygen species (ROS). Enzymes that metabolize fatty acids include cyclooxygenase (COX), lipoxygenase (LO), and cytochromes P450 (CYPs)1,8. Enzymatic metabolism results in enantioselective formation whereas ROS oxidation results in the racemic formation of products.
While this protocol focuses primarily on the analysis of AA- and some LA-derived bioactive metabolites; it could be easily applied to metabolites of other fatty acids. Bioactive lipids are extracted from cell lysate or media using liquid-liquid (l-l) extraction. At the beginning of the l-l extraction process, stable isotope internal standards are added to account for errors during sample preparation. Stable isotope dilution (SID) also accounts for any differences, such as ion suppression, that metabolites may experience during the mass spectrometry (MS) analysis9. After the extraction, derivatization with an electron capture (EC) reagent, pentafluorylbenzyl bromide (PFB) is employed to increase detection sensitivity10,11. Multiple reaction monitoring (MRM) is used to increase the selectivity of the MS analysis. Before MS analysis, lipids are separated using chiral normal phase high performance liquid chromatography (HPLC). The HPLC conditions are optimized to separate the enantiomers and various stereoisomers of the monitored lipids12. This specific LC-MS method monitors prostaglandins (PGs), isoprostanes (isoPs), hydroxyeicosatetraenoic acids (HETEs), hydroxyoctadecadienoic acids (HODEs), oxoeicosatetraenoic acids (oxoETEs) and oxooctadecadienoic acids (oxoODEs); however, the HPLC and MS parameters can be optimized to include any fatty acid metabolites13.
Most of the currently available bioanalytical methods do not take into account the separate quantification of enantiomers. This is extremely important when trying to deduce whether or not the metabolites were formed enzymatically or by ROS. Additionally, the ratios of the enantiomers may provide evidence for a specific enzymatic pathway of formation. The use of SID allows for accurate quantification of metabolites and accounts for any sample loss during preparation as well as the differences experienced during ionization. Using the PFB electron capture reagent increases the sensitivity of detection by two orders of magnitude over conventional APCI methods. Overall, this method, SID-LC-EC-atmospheric pressure chemical ionization APCI-MRM/MS, is one of the most sensitive, selective, and accurate methods of quantification for bioactive lipids.

This protocol will demonstrate the extraction and analysis of free and esterified bioactive fatty acids from cells. Fatty acids are accurately quantified using stable isotope dilution, chiral liquid chromatography, electron capture atmospheric chemical ionization multiple reaction monitoring mass spectrometry (SID-LC-ECAPCI-MRM/MS).

L&#39;extraction des lipides cellulaires pour ciblées dilution isotopique stable analyse de spectrométrie de masse chromatographie liquide-

The metabolism of fatty acids, such as arachidonic acid (AA) and linoleic acid (LA), results in the formation of oxidized bioactive lipids, including ...

Designing a Bio-responsive Robot from DNA Origami

Nucleic acids are astonishingly versatile. In addition to their natural role as storage medium for biological information1, they can be utilized in parallel computing2,3 , recognize and bind molecular or cellular targets4,5 , catalyze chemical reactions6,7 , and generate calculated responses in a biological system8,9. Importantly, nucleic acids can be programmed to self-assemble into 2D and 3D structures10-12, enabling the integration of all these remarkable features in a single robot linking the sensing of biological cues to a preset response in order to exert a desired effect.
Creating shapes from nucleic acids was first proposed by Seeman13, and several variations on this theme have since been realized using various techniques11,12,14,15 . However, the most significant is perhaps the one proposed by Rothemund, termed scaffolded DNA origami16. In this technique, the folding of a long (&gt;7,000 bases) single-stranded DNA 'scaffold' is directed to a desired shape by hundreds of short complementary strands termed 'staples'. Folding is carried out by temperature annealing ramp. This technique was successfully demonstrated in the creation of a diverse array of 2D shapes with remarkable precision and robustness. DNA origami was later extended to 3D as well17,18 .
 
The current paper will focus on the caDNAno 2.0 software19 developed by Douglas and colleagues. caDNAno is a robust, user-friendly CAD tool enabling the design of 2D and 3D DNA origami shapes with versatile features. The design process relies on a systematic and accurate abstraction scheme for DNA structures, making it relatively straightforward and efficient.
In this paper we demonstrate the design of a DNA origami nanorobot that has been recently described20. This robot is 'robotic' in the sense that it links sensing to actuation, in order to perform a task. We explain how various sensing schemes can be integrated into the structure, and how this can be relayed to a desired effect. Finally we use Cando21 to simulate the mechanical properties of the designed shape. The concept we discuss can be adapted to multiple tasks and settings.

DNA origami is a powerful method for fabricating precise nanoscale objects by programming the self-assembly of DNA molecules. Here, we describe how DNA origami can be utilized to design a robotic robot capable of sensing biological cues and responding by shape shifting, subsequently relayed to a desired effect.

Conception d&#39;un robot Bio-sensible de l&#39;ADN origami

Nucleic acids are astonishingly versatile. In addition to their natural role as storage medium for biological information1, they can be utilized in ...

Medical-grade Sterilizable Target for Fluid-immersed Fetoscope Optical Distortion Calibration

We have developed a calibration target for use with fluid-immersed endoscopes within the context of the GIFT-Surg (Guided Instrumentation for Fetal Therapy and Surgery) project. One of the aims of this project is to engineer novel, real-time image processing methods for intra-operative use in the treatment of congenital birth defects, such as spina bifida and the twin-to-twin transfusion syndrome. The developed target allows for the sterility-preserving optical distortion calibration of endoscopes within a few minutes. Good optical distortion calibration and compensation are important for mitigating undesirable effects like radial distortions, which not only hamper accurate imaging using existing endoscopic technology during fetal surgery, but also make acquired images less suitable for potentially very useful image computing applications, like real-time mosaicing. In this paper proposes a novel fabrication method to create an affordable, sterilizable calibration target suitable for use in a clinical setup. This method involves etching a calibration pattern by laser cutting a sandblasted stainless steel sheet. This target was validated using the camera calibration module provided by OpenCV, a state-of-the-art software library popular in the computer vision community.

This article describes the design and development of a sterilizable custom camera optical distortion calibration target for the peri-operative, fluid-immersed calibration of endoscopes during endoscopic interventions.

Qualité médicale cible stérilisable pour fœtoscope optique Distorsion Calibration Fluid immergé

We have developed a calibration target for use with fluid-immersed endoscopes within the context of the GIFT-Surg (Guided Instrumentation for Fetal ...

Optimizing the Use of a Liquid Handling Robot to Conduct a High Throughput Forward Chemical Genetics Screen of Arabidopsis thaliana

Chemical genetics is increasingly being employed to decode traits in plants that may be recalcitrant to traditional genetics due to gene redundancy or lethality. However, the probability of a synthetic small molecule being bioactive is low; therefore, thousands of molecules must be tested in order to find those of interest. Liquid handling robotics systems are designed to handle large numbers of samples, increasing the speed with which a chemical library can be screened in addition to minimizing/standardizing error. To achieve a high-throughput forward chemical genetics screen of a library of 50,000 small molecules on Arabidopsis thaliana (Arabidopsis), protocols using a bench-top multichannel liquid handling robot were developed that require minimal technician involvement. With these protocols, 3,271 small molecules were discovered that caused visible phenotypic alterations. 1,563 compounds induced short roots, 1,148 compounds altered coloration, 383 compounds caused root hair and other, non-categorized, alterations, and 177 compounds inhibited germination.

Optimizing the Use of a Liquid Handling Robot to Conduct a High Throughput Forward Chemical Genetics Screen of Arabidopsis thaliana

A high throughput screen of synthetic small molecules was conducted on the model plant species, Arabidopsis thaliana. This protocol, developed for a liquid handling robot, increases the speed of forward chemical genetics screens, accelerating the discovery of novel small molecules affecting plant physiology.

Optimisation de l’utilisation d’un Robot de manutention liquide pour mener un haut débit avant chimique génétique écran d’Arabidopsis thaliana

Chemical genetics is increasingly being employed to decode traits in plants that may be recalcitrant to traditional genetics due to gene redundancy or ...

Decellularization of Whole Human Heart Inside a Pressurized Pouch in an Inverted Orientation

The ultimate solution for patients with end-stage heart failure is organ transplant. But donor hearts are limited, immunosuppression is required, and ultimately rejection can occur. Creating a functional, autologous bio-artificial heart could solve these challenges. Biofabrication of a heart comprised of scaffold and cells is one option. A natural scaffold with tissue-specific composition as well as micro- and macro-architecture can be obtained by decellularizing hearts from humans or large animals such as pigs. Decellularization involves washing out cellular debris while preserving 3D extracellular matrix and vasculature and allowing &#34;cellularization&#34; at a later timepoint. Capitalizing on our novel finding that perfusion decellularization of complex organs is possible, we developed a more &#34;physiological&#34; method to decellularize non-transplantable human hearts by placing them inside a pressurized pouch, in an inverted orientation, under controlled pressure. The purpose of using a pressurized pouch is to create pressure gradients across the aortic valve to keep it closed and improve myocardial perfusion. Simultaneous assessment of flow dynamics and cellular debris removal during decellularization allowed us to monitor both fluid inflow and debris outflow, thereby generating a scaffold that can be used either for simple cardiac repair (e.g. as a patch or valve scaffold) or as a whole-organ scaffold.

This method enables decellularization of a complex solid organ using a simple protocol based on osmotic shock and perfusion of ionic detergent with minimal organ matrix disruption. It comprises a novel decellularization technique for human hearts inside a pressurized pouch with real-time monitoring of flow dynamics and cellular debris outflow.

DÉCELLULARISATION du cœur humain à l’intérieur d’une poche sous pression dans une Orientation inversée

The ultimate solution for patients with end-stage heart failure is organ transplant. But donor hearts are limited, immunosuppression is required, and ...

A Tripeptide-Stabilized Nanoemulsion of Oleic Acid

We describe a method to produce a nanoemulsion composed of an oleic acids-Pt(II) core and a lysine-tyrosine-phenylalanine (KYF) coating (KYF-Pt-NE). The KYF-Pt-NE encapsulates Pt(II) at 10 wt. %, has a diameter of 107 &#177; 27 nm and a negative surface charge. The KYF-Pt-NE is stable in water and in serum, and is biologically active. The conjugation of a fluorophore to KYF allows the synthesis of a fluorescent nanoemulsion that is suitable for biological imaging. The synthesis of the nanoemulsion is performed in an aqueous environment, and the KYF-Pt-NE forms via self-assembly of a short KYF peptide and an oleic acids-platinum(II) conjugate. The self-assembly process depends on the temperature of the solution, the molar ratio of the substrates, and the flow rate of the substrate addition. Crucial steps include maintaining the optimal stirring rate during the synthesis, permitting sufficient time for self-assembly, and pre-concentrating the nanoemulsion gradually in a centrifugal concentrator.

This protocol describes an efficient method to synthesize a nanoemulsion of an oleic acids-platinum(II) conjugate stabilized with a lysine-tyrosine-phenylalanine (KYF) tripeptide. The nanoemulsion forms under mild synthetic conditions via self-assembly of the KYF and the conjugate.

Une Nanoémulsion Tripeptide-stabilisé de l’acide oléique

We describe a method to produce a nanoemulsion composed of an oleic acids-Pt(II) core and a lysine-tyrosine-phenylalanine (KYF) coating (KYF-Pt-NE). The ...

Isokinetic Robotic Device to Improve Test-Retest and Inter-Rater Reliability for Stretch Reflex Measurements in Stroke Patients with Spasticity

Measuring spasticity is important in treatment planning and determining efficacy after treatment. However, the current tool used in clinical settings has been shown to be limited in inter-rater reliability. One factor in this poor inter-rater reliability is the variability of passive motion while measuring the angle of catch (AoC) measurements. Therefore, an isokinetic device has been proposed to standardize the manual joint motion; however, the benefits of isokinetic motion for AoC measurements has not been tested in a standardized manner. This protocol investigates whether isokinetic motion itself can improve inter-rater reliability for AoC measurements. For this purpose, a robotic isokinetic device was developed that is combined with surface electromyography (EMG). Two conditions, manual and isokinetic motions, are compared with the standardized method to measure the angle and subjective feeling of catch. It is shown that in 17 stroke patients with mild elbow flexor spasticity, isokinetic motion improved the intraclass correlation coefficient (ICC) for inter-rater reliability of AoC measurements to 0.890 [95% confidence interval (CI): 0.685&#8211;0.961] by the EMG criteria, and 0.931 (95% CI: 0.791&#8211;0.978) by the torque criteria, from 0.788 (95% CI: 0.493&#8211;0.920) by manual motion. In conclusion, isokinetic motion itself can improve inter-rater reliability of AoC measurements in stroke patients with mild spasticity. Given that this system may provide greater standardized angle measurements and catch of feeling, it may be a good option for the evaluation of spasticity in a clinical setting.

Using a robotic isokinetic device with electromyography (EMG) measurements, this protocol illustrates that isokinetic motion itself can improve inter-rater reliability for the angle of catch measurements in stroke patients with mild elbow flexor spasticity.

Dispositif robotique Isokinetic pour améliorer la fiabilité de test-retest et d'inter-rater pour des mesures de réflexe d'étirement dans les patients de course avec la spasticité

Measuring spasticity is important in treatment planning and determining efficacy after treatment. However, the current tool used in clinical settings has ...

Methods Article

Adaptation d'un robot haptique dans une IRMf 3T