Gostaríamos de agradecer a Lu Kun e Ronald Kurz para assistência técnica. Este trabalho foi financiado pelo ONR n º Prêmio MURI: N00014-10-1-0072, a concessão do NSF # SBE-0542013 a dinâmica temporal do Centro de Aprendizagem, uma Ciência NSF do Centro de Aprendizagem, e conceder NIH # 2 NS036449 R01-11.

1. Fora da sala de scanner <ol><li> Colocar a tabela de rolamento com a extremidade livre suportada eo fim exterior do punho longo destacados. </li><li> Verifique se o robô está desligado. </li><li> Coloque o robô no soquete de mesa e segura a placa de segurança de alumínio sobre o robô com dois parafusos. </li><li> Anexar o effecter fim ao lidar com robô com o adaptador de alumínio e verifique se ela se move livremente. </li><li> Anexar o 10 &#39;cabo paralelo com blindagem em alumínio para o robô e verifique se a proteção está intacta. Adicionar folha extra se necessário. </li><li> Coloque a caixa de blindagem em alumínio sobre o robô tomando cuidado para colocar os paralelos e os cabos de alimentação na ranhura na parte de trás. </li><li> Apertar novamente com cuidado na caixa de blindagem. </li><li> Pacote de papel alumínio na ranhura de cabo na caixa de blindagem e não se esqueça da folha entra em contato com a blindagem do cabo paralelo. </li></ol> 2. Movendo para a sala de scanner com duas pessoas, A e B <ol><li> Prepare-se para entrar em um ambiente de alto campo magnético através da remoção de todos e quaisquer objetos metálicos não ferrosos, incluindo aqueles, por exemplo, telefones celulares, chaves, moedas, etc .. </li><li> Uma pessoa com final segurando a extremidade livre da tabela robô e B pessoa estabilizar a caixa final, roll-lo livre até que o primeiro robô só entra pela porta da sala. </li><li> Clips B pessoa a corda de segurança presa a uma âncora buraco na parte de trás do caso blindagem e verifica que a outra extremidade está firmemente presa a uma âncora de parede. </li><li> Trabalhando juntos, rola a tabela para o quarto e anexá-lo com tiras de velcro para o pé da mesa fMRI. O fim robô da tabela deve ficar tão longe do scanner possível. </li><li> Conecte o cabo paralelo do robô para o filtro personalizado no passar para a sala de controle, e ligar o robô. A folha de blindagem no cabo paralelo deve fazer contato com o filtro. </li><li> Anexar a parte exterior da alça longa (effecter final) e verificar se ele vai entrar no furo do fMRI limpa. </li></ol> 3. Na sala de controle <ol><li> Inicie o computador de controle e anexar o cabo paralelo 6 &#39;para o robô. Certifique-se de remover qualquer filtro extra que pode ser na parte interior da passagem. </li><li> Verificar se os motores do robô são desligados, comece a rotina de calibração Fantasma e verificar se as leituras posição dos motores da rotina do robô de calibração são estáveis. </li><li> Verifique a conexão do cabo paralelo e que só filtrar os grandes personalizado é anexado se a rotina de calibração não pode ver o robô ou há grandes variações na leitura motor. </li><li> Ligar o robô, abrindo a porta na parte traseira da caixa de blindagem e pressione as chaves com um pau. </li><li> Reiniciar a rotina de calibração com o fim effecter aproximadamente centrado no final do guia de ondas cônico. Passo através da calibração e verificação de que a caixa de calibração caixa tem a interação háptica apropriado. </li><li> Anexar a saída TTL da fMRI (conector BNC) para o ADC LABJACK no computador controle. </li></ol> 4. O assunto <ol><li> Preparar o tema para um ambiente de alto campo magnético com os protocolos fMRI padrão. </li><li> Configurar qualquer equipamento extra para o experimento, por exemplo, sistema de exibição visual. Usamos o NordicNeuro Lab, Vision System Inc., que proporciona uma visualização estéreo de displays visuais, uma característica que é particularmente útil quando apresenta ambientes virtuais. </li><li> Depois que o sujeito se deita sobre a mesa e da bobina a sua sede, ajustar a distância do robô soltando os parafusos de mão e deslizando o topo da tabela até que o sujeito pode mover-se confortavelmente. </li><li> Guia da tabela manualmente, segurando a extremidade do robô enquanto a tabela fMRI está se movendo para dentro e para fora do furo para evitar a oscilação de rodas. Verifique se o effecter final vai para o furo e não pegar do lado de fora. </li><li> Executar o experimento. </li></ol> 5. Quebrar a configuração com duas pessoas A e B <ol><li> Após a saída do paciente, remova o fim exterior da alça de comprimento e retire a tabela de robô da tabela fMRI, desfazendo a cinta de velcro. </li><li> Do robô, desfazer o cabo blindado em paralelo e desconecte o cabo de alimentação. </li><li> Uma pessoa com segurando a extremidade livre da tabela robô e B pessoa guiando o fim robô, mover a tabela para a porta. Na porta, desfazer a corda, and roll tabela robô até o hall. </li><li> Desfazer todos os parafusos da caixa de blindagem e os dois parafusos da placa de segurança e remover o robô. </li></ol> 6. Resultados representativos: Idealmente, o robô haptic e fMRI não deve afetar o outro. Podemos dizer on-line se o robô está sendo afetado pela fMRI. Geralmente, se o cabo paralelo do robô não é devidamente blindado e filtrada, em seguida, a leitura dos motores vai oscilaçãofinal rapidamente. Isso pode ser corrigido com um duplo verificar a blindagem em alumínio no cabo, que o núcleo ferroso é devidamente colocado no cabo paralelo perto do robô, e que o filtro só para o robô é o filtro personalizado no lado sala de scanner da passagem . Detecção de erros no fMRI é realmente só é possível depois que os dados foram reduzidos e analisados, mas um exame de anatomia deve ser tomada no início do estudo e verificado para efeitos de zíper ou outros artefatos indicativos de ruído correlacionados (por exemplo, o ruído pico) 7. Freqüentemente, esse ruído vem de metal em contato com metal e pode ser limpo, apertando todos os parafusos em cima da mesa robô, especialmente os parafusos de mão de ajuste do lado da mesa. De nossos testes o sinal fMRI base-ruído (SNR) é de ~ 380 e com o robô totalmente blindados na sala que cai para um ainda razoável ~ 330. Se o escudo não está em vigor no robô, então a queda SNR ainda mais a ~ 250, e os efeitos de ruído se tornar muito significativo. Como mostrado em 4, o grau 3 de articular a liberdade no centro da alça tem pouco efeito sobre a dinâmica da interação do robô / mão, exceto para transferi-lo longe do robô. A junta no centro da alça funciona como um ponto de apoio e inverte o movimento aparente em duas das direções (esquerda-direita e de cima para baixo), mas não o terceiro (frente-verso). Desde o Fantasma ea mão estão em extremos opostos da alavanca como lidar com o seu fulcro no meio, os ganhos são aplicados em software em cada uma das três direções euclidiana: ganhos negativos nos dois sentidos controlada pela junta giratória e um ganho positivo na direção da junta deslizante. O efeito líquido da alça e giratória reproduz o total de 3 graus de liberdade do robô Phantom, apenas &quot;nove fora. <img src="/files/ftp_upload/3364/3364fig1.jpg" alt="Figura 1"/> Figura 1 O aparelho utilizado para montar o robô haptic para uso no ambiente de fMRI. Superior mostra o robô haptic montado no caso antes de gabinete (em cima, à esquerda) e o controle deslizante / cardan conjunta no ponto médio do punho (topo, direita). Baixo, da esquerda mostra um objeto no scanner manipular a alça. Inferior, à direita é um cartoon do effecter blindagem e fim. <img src="/files/ftp_upload/3364/3364fig2.jpg" alt="Figura 2"/> Figura 2 Resultados do teste BIRN para o robô totalmente blindado com o movimento. As três imagens com cruzes mostram seções do modelo de cabeça esférica, e na parte inferior direita mostra uma visão tridimensional. Os pequenos pontos são bolhas no modelo de cabeça estático e sempre estão presentes. A falta de listras grandes ou zíperes indica que o ruído do robô é não correlacionadas.

<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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O robô fMRI compatível abre novas possibilidades para experimentos na neurociência de controle motor. O passo mais crítico na instalação é a blindagem do robô para evitar artefatos no fMRI, o que fazemos em duas etapas. Primeiro, o robô em si é de cerca de 9 &quot;fora do furo com um longo e leve, alça apoiada no seu meio com um grau 3 da articulação liberdade. Segundo, o robô é envolto em um 1 / 16 caixa de alumínio &quot;-1 / 4&quot; com um cone de plástico (13 &quot;de diâmetro de base, 6&quot; de d...

<table border="1"><tbody><tr><td> Fantasma prêmio 1.5/6dof, modelo elevada força </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

Ressonância magnética funcional (fMRI) fornece imagiologia cerebral funcional excelente através do sinal BOLD 1 com vantagens, incluindo radiação não-ionizante, precisão espacial milímetro de dados anatômicos e funcionais 2, e quase em tempo real, análises 3. Robôs Haptic proporcionar a medição precisa e controle de posição e força de um cursor em um espaço razoavelmente confinados. Aqui nós combinamos essas duas tecnologias para permitir que experimentos de precisão envolvendo o controle motor com haptic / interação tátil ambiente, tais como alcançar ou agarrar. A idéia básica é para anexar um effecter final 8 pés apoiados no centro para o robô quatro permitindo ao indivíduo usar o robô, mas protegê-lo e mantê-lo fora da parte mais extrema do campo magnético da máquina de fMRI (Figura 1 ). O Fantasma premium 3.0, 6DOF, alta força de robô (SensAble Technologies, Inc.) é uma excelente escolha para a prestação de force-feedback em experiências de realidade virtual 5, 6, mas é inerentemente não seguro MR, introduz ruído significativo para o sensível fMRI, equipamentos e seus motores elétricos podem ser afetadas pela fMRI fortemente variando o campo magnético. Construímos uma tabela e sistema de blindagem que permite que o robô para ser introduzida com segurança o ambiente fMRI e limita tanto a degradação do sinal de fMRI pelos motores eletricamente ruidosos ea degradação do desempenho motor elétrico pelo campo magnético variável fortemente do fMRI. Com o escudo, a relação sinal-ruído (SNR: média de sinal / ruído desvio padrão) da fMRI passa de uma linha de base de ~ 380 a ~ 330 e ~ 250 sem a blindagem. O ruído restante parece ser não correlacionadas e não adicionar artefatos à ressonância magnética funcional de uma esfera de teste (Figura 2). A alça longa, dura permite a colocação do robô fora do alcance das partes mais fortemente variando do campo magnético que não há efeito significativo da fMRI no robô. O efeito da alça sobre cinemática do robô é mínima, pois é leve (~ 2,6 lbs), mas extremamente dura 3 / 4 &quot;de grafite e bem equilibrado na articulação 3DoF no meio. O resultado final é um sistema háptico fMRI compatível, com cerca de 1 metro cúbico de espaço de trabalho, e, quando combinado com a realidade virtual, que permite um novo conjunto de experimentos a serem realizados no ambiente de fMRI incluindo naturalista deslocamento, atingindo passiva do membro e da percepção tátil, o aprendizado de adaptação em diferentes campos de força , ou identificação textura 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

A adaptação e utilização de um robô haptic em um fMRI 3T é descrito.

Adaptação de um robô Haptic em um 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 visualizações.  University of California. A adaptação e utilização de um robô haptic em um fMRI 3T é descrito.

Vídeo: 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.

Fabricação de Eletroquímica-DNA Biossensores para Detecção Reagentless de ácidos nucléicos, proteínas e pequenas moléculas

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. ...

Bioengenharia

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 / Reabilitação Graphic: Integrando um robô em um ambiente virtual Biblioteca e aplicando-a Terapia curso

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.

Criando Two-Dimensional Substratos modelado para Confinamento de proteína e celular

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).

Extração de lipídeos celulares para Targeted diluição de isótopos estáveis ​​Líquido Análise Espectrometria de Massa-Cromatografia

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.

Projetando um robô Bio-responsivo de DNA 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.

Alvo esterilizável de grau médico para fetoscópio Distortion Calibração óptica imerso-Fluid

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.

Otimizando o uso de um robô de manipulação líquidos para conduzir a uma alta taxa de transferência para a frente química genética tela 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.

Decellularization do coração humana dentro de uma bolsa pressurizada em uma orientação 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.

Uma Nanoemulsão estabilizada tripeptídeo de 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 Isokinetic para melhorar a confiabilidade do teste-reteste e do Inter-rater para medidas do reflexo do estiramento em pacientes do curso com spasticity

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