Nos gustaría dar las gracias al Kun Lu y Kurz Ronald de asistencia técnica. Este trabajo fue apoyado por la ONR N º Premio MURI: N00014-10-1-0072, NSF subvención # SBE-0542013 a la dinámica temporal de Centro de Aprendizaje, una ciencia de la NSF Centro de Aprendizaje, y subvención del NIH # 2 R01 NS036449-11.

1. Fuera de la sala del escáner <ol><li> Coloque la mesa rodante con el extremo libre el apoyo y el extremo exterior del mango largo separado. </li><li> Compruebe que el robot está apagado. </li><li> Coloque el robot en el zócalo de la tabla y asegurar la placa de seguridad de aluminio sobre el robot con dos tornillos. </li><li> Conecte el efector final del robot manejar con el adaptador de aluminio y comprobar que se mueve libremente. </li><li> Conecte el 10 &#39;cable paralelo con blindaje de aluminio para el robot y comprobar que el blindaje está intacto. Añadir hojas adicionales si es necesario. </li><li> Coloque la caja de blindaje de aluminio sobre el robot teniendo cuidado de colocar los cables en paralelo y el poder dentro de la ranura en la parte posterior. </li><li> Cuidadosamente los tornillos de la caja de protección. </li><li> Paquete de papel de aluminio en la ranura del cable en la caja de protección y asegurarse de que el papel entra en contacto con el blindaje del cable paralelo. </li></ol> 2. Entrando en la sala del escáner con dos personas, A y B <ol><li> Prepárate para entrar en un entorno de alto campo magnético mediante la eliminación de cualquier y todos los objetos metálicos como los no ferrosos, por ejemplo, teléfonos celulares, llaves, monedas, etc .. </li><li> Con la persona A finales sujeta el extremo libre de la mesa del robot y la persona B estabilizar el extremo de la caja, rodar sin primero hasta que el robot sólo entra por la puerta de la habitación. </li><li> Clips persona B de la cuerda de seguridad atada a un agujero de anclaje en la parte posterior de la caja de blindaje y comprueba que el otro extremo está firmemente unido a un anclaje de pared. </li><li> Trabajando juntos, rollo de la mesa en la habitación y adjuntarlo con tiras de velcro al pie de la tabla de resonancia magnética funcional. El extremo del robot de la tabla debe estar tan lejos de el escáner como sea posible. </li><li> Conecte el cable paralelo del robot para el filtro personalizado en el paso a la sala de control, y conectar el robot. El blindaje del cable paralelo debe estar en contacto con el filtro. </li><li> Coloque la parte exterior del mango largo (efector final) y verificar que se entrará en el agujero de la fMRI limpiamente. </li></ol> 3. En la sala de control <ol><li> Inicie el equipo de control y conectar el cable paralelo de 6 &#39;para el robot. Asegúrese de quitar todos los filtros extra que puede ser en la parte interior del paso a través. </li><li> Verificar que los motores del robot se apagan, la puesta en marcha de la rutina de calibración Phantom y verificar que las lecturas de la posición de los motores de la rutina de calibración del robot son estables. </li><li> Compruebe la conexión del cable paralelo y que sólo el filtro personalizado grueso se une, si la rutina de calibración no se puede ver el robot o hay grandes variaciones en la lectura de motor. </li><li> Encienda el robot por la apertura del puerto en la parte posterior de la caja de protección y los interruptores de prensa con un palo. </li><li> Reiniciar la rutina de calibración con el efector final aproximadamente centrado en la final de la guía de onda cónica. Paso a través de la calibración y comprobar que la caja de calibración caja tiene la interacción háptica apropiado. </li><li> Conecte la salida TTL de la resonancia magnética funcional (conector BNC) a la ADC Labjack en el ordenador de control. </li></ol> 4. El tema <ol><li> Preparar al sujeto para un entorno de alto campo magnético con los protocolos estándar de resonancia magnética funcional. </li><li> La creación de cualquier equipo adicional para el experimento, por ejemplo, el sistema de representación visual. Usamos el NordicNeuro Lab, Sistema de Visión Inc. que ofrece una visualización estéreo de muestra visual, una característica que es especialmente útil cuando se presentan los entornos virtuales. </li><li> Después de que el sujeto se acuesta sobre la mesa y la bobina de la cabeza se encuentra, ajustar la distancia del robot aflojando los tornillos de mano y deslizando la parte superior de la tabla hasta que el sujeto puede moverse con soltura. </li><li> Guía de la tabla manualmente mediante la celebración en el extremo del robot, mientras que la tabla de resonancia magnética funcional se está moviendo dentro y fuera del agujero para evitar que las ruedas se tambalee. Asegúrese de que el efector final entra en el agujero y no recoge en el exterior. </li><li> Realizar el experimento. </li></ol> 5. Romper la configuración con dos personas A y B <ol><li> Después de la salida del paciente, retire el extremo exterior del mango largo y separar la mesa de robot de la mesa de resonancia magnética funcional por deshacer la correa de velcro. </li><li> Del robot, deshacer el cable paralelo apantallado y desconecte el cable de alimentación. </li><li> Una persona con la celebración de la extremidad libre de la tabla de robot y la persona B guiar el robot final, mover la tabla a la puerta. En la puerta, deshacer la atadura, y gire la mesa de robot a la sala. </li><li> Deshacer todos los tornillos de la caja de protección y los dos tornillos de la placa de seguridad y quitar el robot. </li></ol> 6. Los resultados representativos: Idealmente, el robot táctil y fMRI no debería afectar a los demás. Podemos decir en línea si el robot está siendo afectada por la fMRI. Por lo general, si el cable paralelo del robot no está correctamente aislado y se filtra, entonces la lectura de los motores se oscilaciónfinales con rapidez. Esto se puede solucionar con un doble control de la protección de aluminio en el cable, que el núcleo de hierro está correctamente colocado en el cable paralelo, cerca del robot, y que el filtro sólo a los que el robot es el filtro personalizado en el lado de la sala del escáner pasar a través de . Detección de errores en la fMRI es realmente sólo es posible después de los datos se han reducido y analizado, pero una exploración anatómica se deben tomar al principio del estudio y se analiza los efectos de cremallera u otros artefactos indicativo de ruido correlacionado (por ejemplo, el ruido pico) 7. Con frecuencia, el ruido que proceda de metal en contacto con el metal y se puede limpiar apretando todos los tornillos en la mesa robot, especialmente los tornillos de ajuste la mano del lado de la mesa. De nuestras pruebas la señal de resonancia magnética funcional de referencia a ruido (SNR) es de ~ 380 y con el robot totalmente blindado en el cuarto que cae a un todavía razonable ~ 330. Si el protector no esté en su lugar en el robot, entonces la caída de SNR más de ~ 250, y los efectos del ruido se vuelven muy importantes. Como se muestra en 4, el grado 3 de la articulación la libertad en el centro del mango tiene poco efecto sobre la dinámica de la interacción robot / la mano si no es para alejarse de la robot. La articulación en el centro de la palanca actúa como un punto de apoyo e invierte el movimiento aparente de dos de las direcciones (izquierda-derecha y arriba-abajo), pero no el tercero (de adelante hacia atrás). Dado que el fantasma y la mano se encuentran en extremos opuestos de la palanca como manejar con su punto de apoyo en el medio, las ganancias se aplican en el software en cada una de las tres direcciones euclidiana: ganancias negativas en las dos direcciones controladas por la articulación giratoria y una ganancia positiva en la dirección de la junta deslizante. El efecto neto de la manija giratoria y se reproduce la totalidad de 3 grados de libertad del robot Phantom, a sólo 9 &#39;de distancia. <img src="/files/ftp_upload/3364/3364fig1.jpg" alt="Figura 1"/> Figura 1 El aparato utilizado para montar el robot táctil para su uso en el entorno de resonancia magnética funcional. Superior muestra el robot táctil montada en el caso antes de la caja (arriba, izquierda) y el cardán / regulador común en el punto medio de la manija (arriba, derecha). Abajo y de izquierda se muestra un objeto en el escáner de la manipulación de la palanca. Abajo, derecha es una caricatura de la protección y efector final. <img src="/files/ftp_upload/3364/3364fig2.jpg" alt="Figura 2"/> Figura 2 Resultados de la prueba BIRN para el robot totalmente blindados con el movimiento. Las tres imágenes con cruces muestran las secciones de la modelo de cabeza esférica, y la parte inferior derecha muestra una vista tridimensional. Los puntos pequeños son burbujas en el modelo de carga estática y están siempre presentes. La falta de grandes bandas o cremalleras indica que el ruido de los robots no está correlacionado.

<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>

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El robot compatible fMRI abre nuevas posibilidades para los experimentos en la neurociencia de control de motores. El paso más importante en la configuración es la protección del robot para evitar artefactos en la fMRI, lo que hacemos en dos pasos. En primer lugar, el propio robot es de unos 9 &#39;de distancia de la perforación con una larga y ligera, manejar el apoyo en su centro con un grado 3 de la libertad común. En segundo lugar, el robot está encerrado en un 16.1 casilla &quot;-1 / 4&quot; de aluminio con u...

<table border="1"><tbody><tr><td> Phantom premium 1.5/6dof, modelo de gran fuerza </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

Resonancia magnética funcional (fMRI) proporciona imágenes de excelente funcional del cerebro a través de la señal BOLD 1 con ventajas, incluyendo la radiación no ionizante, precisión milimétrica espacial de los datos anatómicos y funcionales 2, y casi en tiempo real de análisis 3. Robots háptica proporciona una medición precisa y control de la posición y la fuerza de un cursor en un espacio bastante reducido. Aquí combinamos estas dos tecnologías para permitir que los experimentos de precisión implica el control del motor con háptico / interacción táctil medio ambiente tales como alcanzar o agarrar. La idea básica consiste en conectar un efector final de 8 pies apoyados en el centro para el robot de 4 lo que permite al sujeto a utilizar el robot, sino protegerlo y mantenerlo alejado de la parte más extrema del campo magnético de la máquina de resonancia magnética funcional (Figura 1 ). El fantasma de Premium 3.0, 6DOF, alta fuerza de robot (SensAble Technologies, Inc.) es una excelente opción para proporcionar retroalimentación de fuerza en los experimentos de realidad virtual 5, 6, sino que es esencialmente no-MR seguro, introduce ruido significativo a la sensibilidad equipo de resonancia magnética funcional, y sus motores eléctricos pueden ser afectados por la fMRI es muy variable el campo magnético. Hemos construido una mesa y un sistema de protección que permite que el robot de forma segura introducción en el medio fMRI y los límites tanto de la degradación de la señal de resonancia magnética funcional por los motores de ruido eléctrico y la degradación del rendimiento del motor eléctrico por el campo magnético que varía fuertemente de la fMRI. Con el escudo, la relación señal a ruido (SNR: media de la señal / ruido de desviación estándar) de la fMRI va desde un valor basal de ~ 380 a ~ 330 y ~ 250 sin blindaje. El ruido restante parece ser correlacionados y no agregar artefactos a la resonancia magnética funcional de una esfera de prueba (Figura 2). El mango largo y rígido permite la colocación del robot fuera del alcance de las partes más fuertemente variables del campo magnético lo que no hay efecto significativo de la resonancia magnética funcional en el robot. El efecto del mango en la cinemática del robot es mínima ya que es ligero (~ 2,6 libras), pero muy dura 3 / 4 &quot;de grafito y equilibrado en el conjunto 3DOF en el centro. El resultado final es una resonancia magnética compatible con el sistema táctil con alrededor de 1 pie cúbico de espacio de trabajo, y, cuando se combina con la realidad virtual, que permite un nuevo conjunto de experimentos que se realizan en el medio ambiente incluyendo fMRI naturalista desplazamiento alcance, pasiva de la extremidad y la percepción háptica, aprendizaje para la adaptación en los campos de fuerza variable , o la textura de identificación 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

La adaptación y el uso de un robot háptico en una fMRI 3T se describe.

Adaptación de un Robot Haptic en un fMRI 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 vistas.  University of California. La adaptación y el uso de un robot háptico en una fMRI 3T se describe.

Video: 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.

Fabricación de ADN electroquímico biosensores para la detección Reagentless de ácidos nucleicos, proteínas y moléculas pequeñas

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. ...

Investigación

Bioingeniería

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.

Haptic / Graphic Rehabilitación: La integración de un robot en un entorno virtual Biblioteca y su aplicación a la terapia de Carrera

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.

La creación de dos dimensiones con dibujos sustratos para el confinamiento de proteínas y células

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).

Extracción de los lípidos celulares orientadas por dilución de isótopos estables análisis de espectrometría de cromatografía de líquidos-Mass

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.

Diseño de un Robot Bio-respuesta a partir del 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.

Objetivo esterilizable de calidad médica para Fluid-sumergido Fetoscopio óptico Distorsión de calibración

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.

Optimizar el uso de un Robot de manipulación de líquidos para llevar a cabo un alto rendimiento adelante química genética pantalla de 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.

Descelularización de todo humano corazón dentro de una bolsa presurizada en una orientación invertida

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.

Una nanoemulsión tripéptido-estabilizado del ácido oleico

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.

Dispositivo robótico isocinético para mejorar la fiabilidad de la prueba y el intertasador para las mediciones de reflejo de estiramiento en pacientes con accidente cerebrovascular con espasticidad

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