Este trabalho foi apoiado por uma bolsa da Fundação Simons (SFARI # 89638 para RB).

O protocolo seguinte adere às diretrizes do Conselho de Revisão Institucional da Universidade de Washington. 1. Avaliação eletrofisiológica <ol><li> Preparação de Sessão <ol><li> Preparação da sala: coloque o manipulandum (ver Figura 1), um bloco de madeira com um sensor ligado, que envia um marcador com carimbo de tempo para aquisição de software quando ele é apreendido, em cima da mesa dentro de agarrar alcance do participante. Ative software de aquisição de EEG e começar a &quot;nova sessão&quot; (Figura S1). </li><li> Preparação útil: solução morna de água destilada (1 L), cloreto de potássio (1 colher de sopa) e xampu (1 colher de chá) a 104 ° F. Mergulhe 128-eletrodo densa-array sistema de EEG na solução salina aquecida. </li><li> Preparação Participante: garantir que o participante está sentado confortavelmente cerca de 75 cm do monitor de apresentação do estímulo e em plena vista da câmera de vídeoera. Encontre e marque o vértice sobre a cabeça do participante com um marcador de pele. Meça o vértice por encontrar a interseção do ponto médio entre o násio eo ínion e ponto médio entre preauriculars. </li><li> Net aplicação: Coloque a tampa de EEG na cabeça do participante de tal forma que o eletrodo vértice é colocado diretamente sobre a marca de vértice. Verificar impedâncias e assegurar que as impedâncias são abaixo do limiar apropriado para o sistema de EEG em uso (Figura S2). </li><li> Comece a sessão de vídeo gravação. </li></ol></li><li> Configuração de gravação: Sinal de referência para o eletrodo de vértice. Filtro analógico entre 0,1 e 100 Hz, amplificar o sinal e digitalizar a 500 amostras / seg. </li><li> Apresentação do estímulo: presente participante com três condições: observar, executar e descanso, adaptados do paradigma desenvolvido por Muthukumaraswamy e colegas 5. <ol><li> Observe condição: Instrua participante para sentar-se calmamente e assistirum vídeo de uma pessoa segurando o manipulandum. Cada ensaio deve durar 6 segundos. Tempo de vídeo pré-gravado para os ensaios observar com precisão para assegurar que o aperto observada ocorre exactamente 3 seg. Monitorar atenção visual do participante durante a tarefa, e marcar ensaios durante a qual eles não frequentam a tela para ser descartado no pós-processamento. </li><li> Executar condição: Instrua participante para sentar-se calmamente com descanso mão direita, logo abaixo do manipulandum e, ao ouvir um sinal sonoro pré-gravado, a imitar a garra manipulandum do observar videoclipe condição. Cada ensaio deve durar 6 segundos. Verifique se o sinal sonoro é apresentado em exatamente 3 segundos por prerecording uma faixa auditivo que mantém um consistente cronometrado executar sugestão e intervalo inter-julgamento. Utilize um sensor na manipulandum para gravar com precisão o tempo que compreensão do participante ocorre (Figura S3). </li><li> Condição de repouso: Instrua participante para sentar-se quietly com os olhos abertos e passivamente observar uma pequena cruz no monitor estímulo. Registro EEG contínuo durante a condição de descanso para 3 min. </li><li> Para tanto observar e executar as condições, apresentam blocos casualizados de dez ensaios, para um total de quarenta ensaios por condição. Certifique-se de que a imagem do manipulandum permanece na tela ao longo dos blocos de observar e executar, inclusive entre os ensaios. Administrar a condição de repouso após a conclusão das condições de observar e executar. </li></ol></li><li> Informática <ol><li> Após a coleta de dados, verifique novamente impedâncias. Observe quaisquer alterações aos níveis de impedância. Terminar gravação software de aquisição. </li><li> Pós-processamento: sinal Rereference EEG à média. Segmento de dados de EEG contínuo em quarenta ensaios de 6 seg para cada condição (Figura S4). </li><li> Realizar detecção artefato automatizado. Use algoritmos automatizados para inspecionar segmentos para artefatos de movimento, identificando rápidas amplitudes média exceeding 200 mV, diferencial amplitudes médias superiores a 100 mV e zero variância através de um dado ensaio (Figura S5). </li><li> Realizar a detecção manual do artefato inspecionando visualmente dados e confirmando com análise de vídeo da sessão para remover todos os ensaios na condição de observação contaminado com qualquer artefato do movimento e todos os ensaios na condição de execução contaminado com qualquer artefato movimento relacionado ao gesto alcance. Excluir ensaios com artefato importante da análise. Descarte qualquer ensaios que foram sinalizados durante a aquisição como não compareceram. Examinar e taxa de rejeição julgamento para cada grupo de diagnóstico em análise, note. </li></ol></li><li> A análise dos dados <ol><li> Por Muthukumaraswamy et al. 5, segmento ensaios limpos em 2 épocas seg consistindo de 1 seg de dados antes que o segure e 1 segundo depois, tanto para as condições de observar (como marcado pela fotocélula) e executar (conforme marcado pelo sensor manipulandum). Segmento cinclinou-se duas épocas de seg a condição de repouso. </li><li> Fast Fourier Transform (FFT) cada segmento. Selecione um grupo de oito eletrodos em cada hemisfério em torno das posições padrão C3 e C4 para análises estatísticas (seguindo Muthukumaraswamy et al. 5 e Bernier et al. 3) (Figura 2). Para cada condição, em média, o poder através dos estudos incluídos para calcular espectros de potência. </li><li> Calcule atenuação mu examinando a potência média durante quer a execução ou a observação de uma ação motora, em relação à potência média durante a condição de repouso, em toda a gama Hz 8-13. Use o log dessa razão para determinar o grau de atenuação. Nota: um valor negativo representa atenuação durante a execução ou observação, enquanto que um valor positivo representa aumento. Esta metodologia leva em conta a variabilidade entre os indivíduos, ea não normalidade dos valores expressos em forma de razão. &lt;br /&gt; Nota: Este protocolo foi desenvolvido usando um 128-eletrodo densa-array sistema de EEG com software Net Station versão 4.1. Enquanto os passos básicos são semelhantes em todos os sistemas de EEG, protocolos de aquisição e análise pode variar. </li></ol></li></ol> 2. Caracterização da amostra <ol><li> Identificar população potencial de pacientes para participação no paradigma através de registros de pesquisas, listas de participantes anteriores, ou referências de clínicas e médicos da região. </li><li> Potenciais participantes de tela para a probabilidade de atender os critérios de diagnóstico para a construção clínica (por exemplo, Transtorno do Espectro do Autismo) e para identificar quaisquer critérios de exclusão, como a presença de traumatismo craniano, tumor, história apreensão, ou uso de medicação anti-convulsivante ou barbitúricos que pode distorcer a sinal eletrofisiológico. </li><li> Confirme o estatuto de diagnóstico da população de pacientes através do uso de instrumentos de diagnóstico padrão ouro (por exemplo, Autism Diagnostic Interview-ReVised (ADI-R 11) e do Autism Diagnostic Observation Schedule-Generic (ADOS-G, 12), administrada pelo médico especialista seguinte Manual Diagnóstico e Estatístico - 5 ª Edição (DSM-5) Critérios de 13. </li><li> Identificar amostra controle pareado em variáveis ​​relevantes de interesse, tais como idade, sexo, capacidade cognitiva, etc </li></ol>

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Os autores declaram não haver interesses financeiros concorrentes.

A bem-sucedida aquisição, processamento e análise de dados eletrofisiológicos relacionados ao ritmo mu ea aplicação de populações clínicas requer 1) a aplicação de ferramentas metodológicas de EEG, 2) detecção de artefato cuidadoso e redução de dados, 3) a identificação precisa do ritmo mu, e 4) a caracterização precisa da população clínica e identificação de grupos de controle apropriados. Metodologia apropriada EEG requer bom funcionamen...

Eletroencefalografia (EEG) é um método eficaz, eficiente e não invasivo de avaliar e registrar a atividade cerebral. Como os neurónios fogo no cérebro, a tensão resultante pode ser amplificado, gravada, e representada graficamente. A resolução temporal do EEG permite a análise do mesmo breves alterações nos padrões de oscilação do cérebro, assim como a análise da resposta do cérebro a estímulos específicos. Apesar de ser a mais antiga técnica de imagens do cérebro, que remonta ao final do século 19, EEG ainda tem ampla aplicabilidade. Embora a ressonância magnética funcional (fMRI) tem excelente resolução espacial, tem relativamente baixa resolução temporal. Isto representa uma grande limitação da avaliação fMRI dada a incrível velocidade com que os processos ocorrem no cérebro. EEG tem a capacidade de avaliar a atividade elétrica do cérebro ao nível de milissegundos, proporcionando potencial iNsight nas fases de processamento do cérebro. Tecnologias em desenvolvimento também têm expandido a aplicabilidade do EEG. Um aumento na densidade de sistemas de gravação tem permitido o desenvolvimento de técnicas de localização de fonte, mitigar alguns dos limites de EEG em relação resolução espacial. Além disso, os sistemas modernos têm reduzido o participante set-up tempo individual significativamente, permitindo a avaliação das populações anteriormente indisponíveis, tais como amostras clínicas 1-3,28-30 infantil e. Dada a excelente resolução temporal, o EEG pode ser usado para examinar a resposta neuronal relacionada com comportamentos específicos, estados ou estímulos externos. Um exemplo deste utilitário é a avaliação do sistema de neurônios-espelho (MNS) em seres humanos. Os neurônios-espelho foram inicialmente identificados em macacos usando única gravação neurônio 4, evidenciando um grupo deneurônios que responderam simultaneamente a execução ea observação de ações motoras. Este método de gravação direta de colocar eletrodos no cérebro raramente é utilizado em seres humanos, e apenas em casos clínicos terríveis. EEG tem proporcionado um método para avaliar os MNS pela monitorização do ritmo EEG mu. Este padrão de oscilação no intervalo de 8-12 Hz demonstrou atenuar potência EEG em resposta à execução de acções e observação do motor, semelhantes ao padrão de activação observada em macacos de 5-7. Da mesma forma, a estimulação das regiões cerebrais putativos MNS através de Estimulação Magnética Transcraniana (por exemplo, o giro frontal inferior) abole EEG ritmo mu 8 e supressão ritmo EEG mu se correlaciona com sinais BOLD de fMRI em regiões de neurônios espelho putativos em assuntos 9, fornecendo apoio adicional de que este ritmo índices, pelo menos em parte, a atividade MNS. Avaliação do ritmo EEG mu permitiu uma avaliação não invasiva de ato de neurônios-espelhoivity em humanos. EEG fornece uma metodologia única para examinar a atividade cerebral e quando combinado com ensaios baseados comportamentalmente, ele pode ser uma ferramenta poderosa para elucidar aspectos da cognição social, como a imitação, em populações clínicas. Além disso, a aplicabilidade do EEG para uso com populações com deficiências cognitivas ou de linguagem permite a introspecção em habilidades dos indivíduos para os quais outras técnicas de imagem ou paradigmas comportamentais podem ser utilizar menos sucesso. O protocolo descrito fornece uma avenida para examinar funções sociais cognitivos teoricamente ligados ao sistema de neurônios-espelho em indivíduos com desenvolvimento típico e atípico, como o Transtorno do Espectro do Autismo.

<table border="0" cellpadding="0" cellspacing="0" width="994">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:267px;">Geodesic EEG System</td>
			<td style="width:71px;">EGI</td>
			<td style="width:119px;">N/A</td>
			<td style="width:539px;">Any EEG system, not only EGI based systems, is applicable for the described study</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:267px;">MATLAB software</td>
			<td style="width:71px;">MATLAB</td>
			<td style="width:119px;">N/A</td>
			<td>Any mathematical, statistical software that can work with matrices is applicable</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:267px;">Netstation software</td>
			<td style="width:71px;">EGI</td>
			<td style="width:119px;">N/A</td>
			<td>Any EEG acquisition software is applicable for the described study</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:267px;">Manipulandum</td>
			<td style="width:71px;">custom</td>
			<td style="width:119px;">N/A</td>
			<td>Any object that is co-registered with data acquisition software to signal a successful grasp</td>
		</tr>
	</tbody>
</table>

eeg mu rhythm in typical and atypical development

Eletroencefalografia (EEG) é um método eficaz, eficiente e não invasivo de avaliar e registrar a atividade cerebral. Dada a excelente resolução temporal, o EEG pode ser usado para examinar a resposta neuronal relacionada com comportamentos específicos, estados ou estímulos externos. Um exemplo deste utilitário é a avaliação do sistema de neurônios-espelho (MNS) em seres humanos através do exame do ritmo EEG mu. O ritmo EEG mu, atividade oscilatório na faixa de freqüência Hz 8-12 gravado a partir de eletrodos localizados centralmente, é suprimida quando um executa individuais, ou simplesmente observa, ações voltadas gol. Como tal, tem sido proposto que reflectem a actividade das MNS. Foi teorizado que a disfunção do sistema de neurônios-espelho (MNS) desempenha um papel que contribui para os déficits sociais do transtorno do espectro do autismo (ASD). Os MNS pode então ser examinado de forma não invasiva em populações clínicos usando EEG mu ritmo atenuação como um índice para a sua actividade. O prot descritoocol fornece uma avenida para examinar funções sociais cognitivos teoricamente ligados aos MNS em indivíduos com desenvolvimento típico e atípico, como ASD.

Adultos típicos, crianças e bebés têm consistentemente demonstrado ritmo mu tanto durante a execução e observação de ações através de uma variedade de paradigmas e estímulos 5, 14-30. Atenuação nesta faixa de frequências é consistentemente localizada através de eletrodos centrais (Figura 3), indicando que esta não é a redução do poder de alfa que é registrado em outras regiões do couro cabeludo. Da mesma forma, a atenuação nesta frequência durante a observação de m...

Watch this Scientific Journal Video about EEG Mu Rhythm in Typical and Atypical Development at JoVE.com

EEG Mu Rhythm in Typical and Atypical Development

Avaliação do ritmo EEG mu fornece uma metodologia única para examinar a atividade cerebral e quando combinado com ensaios baseados comportamental, pode ser uma ferramenta poderosa para elucidar aspectos da cognição social, como a imitação, em populações clínicas.

EEG Mu Ritmo no desenvolvimento típico e atípico

Electroencephalography (EEG) is an effective, efficient, and noninvasive method of assessing and recording brain activity. Given the excellent temporal ...

eeg-mu-rhythm-in-typical-and-atypical-development

Research

JoVE Journal

Medicine

25.4K visualizações.  University of Washington.  Avaliação do ritmo EEG mu fornece uma metodologia única para examinar a atividade cerebral e quando combinado com ensaios baseados comportamental, pode ser uma ferramenta poderosa para elucidar aspectos da cognição social, como a imitação, em populações clínicas. 

Vídeo: EEG Mu Rhythm in Typical and Atypical Development

Direct Pressure Monitoring Accurately Predicts Pulmonary Vein Occlusion During Cryoballoon Ablation

Cryoballoon ablation (CBA) is an established therapy for atrial fibrillation (AF). Pulmonary vein (PV) occlusion is essential for achieving antral contact and PV isolation and is typically assessed by contrast injection. We present a novel method of direct pressure monitoring for assessment of PV occlusion.
Transcatheter pressure is monitored during balloon advancement to the PV antrum. Pressure is recorded via a single pressure transducer connected to the inner lumen of the cryoballoon. Pressure curve characteristics are used to assess occlusion in conjunction with fluoroscopic or intracardiac echocardiography (ICE) guidance. PV occlusion is confirmed when loss of typical left atrial (LA) pressure waveform is observed with recordings of PA pressure characteristics (no A wave and rapid V wave upstroke). Complete pulmonary vein occlusion as assessed with this technique has been confirmed with concurrent contrast utilization during the initial testing of the technique and has been shown to be highly accurate and readily reproducible.
We evaluated the efficacy of this novel technique in 35 patients. A total of 128 veins were assessed for occlusion with the cryoballoon utilizing the pressure monitoring technique; occlusive pressure was demonstrated in 113 veins with resultant successful pulmonary vein isolation in 111 veins (98.2%). Occlusion was confirmed with subsequent contrast injection during the initial ten procedures, after which contrast utilization was rapidly reduced or eliminated given the highly accurate identification of occlusive pressure waveform with limited initial training.
Verification of PV occlusive pressure during CBA is a novel approach to assessing effective PV occlusion and it accurately predicts electrical isolation. Utilization of this method results in significant decrease in fluoroscopy time and volume of contrast.

Effective pulmonary vein isolation utilizing a cryoballoon depends on complete pulmonary vein occlusion. The point of occlusion can be effectively predicted by direct analysis of pulmonary vein pressure waveform analysis during balloon inflation using a simple and reproducible technique.

Monitorização da Pressão direta é capaz de predizer oclusão da veia pulmonar durante ablação Cryoballoon

Cryoballoon  ablation (CBA) is an established therapy for atrial fibrillation (AF).  Pulmonary vein (PV) occlusion is essential for achieving antral ...

Medicina

Development of an Audio-based Virtual Gaming Environment to Assist with Navigation Skills in the Blind

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind. Using only audio based cues and set within the context of a video game metaphor, users gather relevant spatial information regarding a building's layout. This allows the user to develop an accurate spatial cognitive map of a large-scale three-dimensional space that can be manipulated for the purposes of a real indoor navigation task. After game play, participants are then assessed on their ability to navigate within the target physical building represented in the game. Preliminary results suggest that early blind users were able to acquire relevant information regarding the spatial layout of a previously unfamiliar building as indexed by their performance on a series of navigation tasks. These tasks included path finding through the virtual and physical building, as well as a series of drop off tasks. We find that the immersive and highly interactive nature of the AbES software appears to greatly engage the blind user to actively explore the virtual environment. Applications of this approach may extend to larger populations of visually impaired individuals.

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind.

Desenvolvimento de um ambiente de jogo de áudio baseada Virtual para ajudar com habilidades de navegação em Cegos

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind. Using only audio ...

Clinical Examination Protocol to Detect Atypical and Classical Scrapie in Sheep

The diagnosis of scrapie, a transmissible spongiform encephalopathy (TSEs) of sheep and goats, is currently based on the detection of disease-associated prion protein by post mortem tests. Unless a random sample of the sheep or goat population is actively monitored for scrapie, identification of scrapie cases relies on the reporting of clinical suspects, which is dependent on the individual&#39;s familiarization with the disease and ability to recognize clinical signs associated with scrapie. Scrapie may not be considered in the differential diagnosis of neurological diseases in small ruminants, particularly in countries with low scrapie prevalence, or not recognized&#160;if it presents as nonpruritic form like atypical scrapie. To aid in the identification of clinical suspects, a short examination protocol is presented to assess the display of specific clinical signs associated with pruritic and nonpruritic forms of TSEs in sheep, which could also be applied to goats. This includes assessment of behavior, vision (by testing of the menace response), pruritus (by testing the response to scratching), and movement (with and without blindfolding). This may lead to a more detailed neurologic examination&#160;of reporting animals as scrapie suspects. It&#160;could also be used in experimental TSE studies of sheep or goats to evaluate disease progression or to identify clinical end-point.

A short clinical examination protocol is presented to aid in the detection of classical and atypical scrapie in sheep and goats.

Protocolo de Exame Clínico para Detectar Scrapie Atípico e Clássico em Ovelhas

The diagnosis of scrapie, a transmissible spongiform encephalopathy (TSEs) of sheep and goats, is currently based on the detection of disease-associated ...

Real-time Cytotoxicity Assays in Human Whole Blood

A live cell-based whole blood cytotoxicity assay (WCA) that allows access to temporal information of the overall cell cytotoxicity is developed with high-throughput cell positioning technology. The targeted tumor cell populations are first preprogrammed to immobilization into an array format, and labeled with green fluorescent cytosolic dyes. Following the cell array formation, antibody drugs are added in combination with human whole blood. Propidium iodide (PI) is then added to assess cell death. The cell array is analyzed with an automatic imaging system. While cytosolic dye labels the targeted tumor cell populations, PI labels the dead tumor cell populations. Thus, the percentage of target cancer cell killing can be quantified by calculating the number of surviving targeted cells to the number of dead targeted cells. With this method, researchers are able to access time-dependent and dose-dependent cell cytotoxicity information. Remarkably, no hazardous radiochemicals are used. The WCA presented here has been tested with lymphoma, leukemia, and solid tumor cell lines. Therefore, WCA allows researchers to assess drug efficacy in a highly relevant ex&#160;vivo condition.

The whole blood cytotoxicity assay (WCA) is a cytotoxicity assay developed by incorporating high-throughput cell positioning technology with fluorescence microscopy and automated image processing. Here, we describe how lymphoma cells treated with an anti-CD20 antibody can be analyzed real-time in human whole blood to provide quantitative cellular cytotoxicity analysis.

Em tempo real de Citotoxicidade em Sangue Total Humano

A live cell-based whole blood cytotoxicity assay (WCA) that allows access to temporal information of the overall cell cytotoxicity is developed with ...

Brain Source Imaging in Preclinical Rat Models of Focal Epilepsy using High-Resolution EEG Recordings

Electroencephalogram (EEG) has been traditionally used to determine which brain regions are the most likely candidates for resection in patients with focal epilepsy. This methodology relies on the assumption that seizures originate from the same regions of the brain from which interictal epileptiform discharges (IEDs) emerge. Preclinical models are very useful to find correlates between IED locations and the actual regions underlying seizure initiation in focal epilepsy. Rats have been commonly used in preclinical studies of epilepsy1; hence, there exist a large variety of models for focal epilepsy in this particular species. However, it is challenging to record multichannel EEG and to perform brain source imaging in such a small animal. To overcome this issue, we combine a patented-technology to obtain 32-channel EEG recordings from rodents2 and an MRI probabilistic atlas for brain anatomical structures in Wistar rats to perform brain source imaging. In this video, we introduce the procedures to acquire multichannel EEG from Wistar rats with focal cortical dysplasia, and describe the steps both to define the volume conductor model from the MRI atlas and to uniquely determine the IEDs. Finally, we validate the whole methodology by obtaining brain source images of IEDs and compare them with those obtained at different time frames during the seizure onset.

This video introduces the preparation, recording, and source analysis procedures of high-resolution EEG on sedated rats with a particular preclinical model of focal epilepsy under noninvasive conditions.

Cérebro Fonte Imaging no rato pré-clínica Todos os modelos de Focal Epilepsia usando de alta resolução EEG Recordings

Electroencephalogram (EEG) has been traditionally used to determine which brain regions are the most likely candidates for resection in patients with ...

Determination of the Relative Potency of an Anti-TNF Monoclonal Antibody (mAb) by Neutralizing TNF Using an In Vitro Bioanalytical Method

This protocol shows the measurement of the apoptotic activity neutralization of TNF&#945; in a mouse fibroblast cell model (WEHI 164) using an anti-TNF&#945; mAb. In addition, this protocol can be used to evaluate other anti-TNF&#945; molecules, such as fusion proteins. The cellular model employed here is sensitive to TNF&#945;-mediated apoptosis when an additional stress factor is induced in cell culture conditions (e.g., serum deprivation). This procedure exemplifies how to execute this analytical assay, highlighting the key operations relating to the sample preparation, cell dilution, apoptosis induction, and spectrophotometric measurements that are critical to ensure successful results. This protocol reveals the best-performance conditions relating to apoptosis induction and efficient signal recording, leading to low uncertainty values.

Determination of the Relative Potency of an Anti-TNF Monoclonal Antibody mAb by Neutralizing TNF Using an In Vitro Bioanalytical Method

A protocol for the determination of the relative anti-apoptotic activity of an anti-TNF&#945; mAb using a neutralization mechanism with WEHI 164 cells is presented here. This protocol is useful for comparing the neutralization strength of different molecules with the same biological functionality.

Determinação da potência relativa de um anti-TNF anticorpo Monoclonal (mAb) por neutralizando TNF usando um método de Bioanalytical In Vitro

This protocol shows the measurement of the apoptotic activity neutralization of TNF&#945; in a mouse fibroblast cell model (WEHI 164) using an ...

Assessment of Spontaneous Alternation, Novel Object Recognition and Limb Clasping in Transgenic Mouse Models of Amyloid-&#946; and Tau Neuropathology

Here we describe a staged, behavioral testing approach that can be used to screen for compounds that exhibit in vivo efficacy on cognitive and functional motor behaviors in transgenic mouse models of &#946;-amyloidosis and tauopathy. The paradigm includes tests for spontaneous alternation in a Y-maze, novel object recognition, and limb clasping. These tests were selected because they: 1) interrogate function of cognitive or motor domains and the correlate neural circuitry relevant to the human disease state, 2) have clearly defined endpoints, 3) have easily implementable quality control checks, 4) can be run in a moderate throughput format, and 5) require little intervention by the investigator. These methods are designed for investigators looking to screen compounds for activity in short-term and working memory tasks, or functional motor behaviors associated with Alzheimer's disease mouse models. The methods described here use behavioral tests that engage a number of different brain regions including hippocampus and various cortical areas. Investigators that desire cognitive tests that specifically assess cognition mediated by a single brain region could use these techniques to supplement other behavioral tests.

Described here is a staged, behavioral screening approach that can be used to screen for compounds that exhibit in vivo efficacy on cognitive and functional motor behaviors in transgenic mouse models of &#946;-amyloidosis and tauopathy. These methods are optimized to screen compounds for activity in short-term and working memory tasks.

Avaliação de Alternância Espontânea, Reconhecimento de Objetos Novos e Clasping de Membros em Modelos de Rato Transgênico de Neuropatologia Amilóide-β e Tau

Here we describe a staged, behavioral testing approach that can be used to screen for compounds that exhibit in vivo efficacy on cognitive and functional ...

The Pilocarpine Model of Temporal Lobe Epilepsy and EEG Monitoring Using Radiotelemetry System in Mice

Temporal lobe epilepsy (TLE) is a common neurological disorder in adulthood. For translational studies of chronic epilepsy, pilocarpine-induced status epilepticus (SE) is frequently selected to recapitulate spontaneous recurrent seizures (SRS). Here we present a protocol of SE induction by intraperitoneal (i.p.) injection of pilocarpine and monitoring of chronic recurring seizures in live animals using a wireless telemetry video and electroencephalogram (EEG) system. We demonstrated notable behavioral changes that need attention after pilocarpine injection and their correlation with hippocampal neuronal loss at 7 days and 6 weeks post-pilocarpine. We also describe the experimental procedures of electrode implantation for video and EEG recording, and analysis of the frequency and duration of chronic recurrent seizures. Finally, we discuss the possible reasons why the expected results are not achieved in each case. This provides a basic overview of modeling chronic epilepsy in mice and guidelines for troubleshooting. We believe this protocol can serve as a baseline for suitable models of chronic epilepsy and epileptogenesis.

This manuscript describes a method of inducing status epilepticus by systemic pilocarpine injection and of monitoring spontaneous recurrent seizures in live animals using a wireless telemetry video and electroencephalogram system. This protocol can be utilized for studying the pathophysiologic mechanisms of chronic epilepsy, epileptogenesis, and acute seizures.

O modelo de pilocarpina de epilepsia do lobo Temporal e EEG monitoramento usando sistema de radiotelemetria em ratos

Temporal lobe epilepsy (TLE) is a common neurological disorder in adulthood. For translational studies of chronic epilepsy, pilocarpine-induced status ...

Contractions of Human-iPSC-derived Cardiomyocyte Syncytia Measured with a Ca-sensitive Fluorescent Dye in Temperature-controlled 384-well Plates

Spontaneously contracting syncytia of cardiomyocytes derived from human-induced pluripotent stem cells (hiPSC-CM) are a useful model of human cardiac physiology and pharmacology. Various methods have been proposed to record this spontaneous activity and to evaluate drug effects, but many of these methods suffer from limited throughput and/or physiological relevance. We developed a high-throughput screening system to quantify the effects of exogenous compounds on hiPSC-CM's beating frequency, using a Ca-sensitive fluorescent dye and a temperature-controlled imaging multi-well plate reader. We describe how to prepare the cell plates and the compound plates and how to run the automated assay to achieve high sensitivity and reproducibility. We also describe how to transform and analyze the fluorescence data to provide reliable measures of drug effects on spontaneous rhythm. This assay can be used in drug discovery programs to guide chemical optimization away from, or toward, compounds affecting human cardiac function.

Spontaneously contracting syncytia of cardiomyocytes derived from human-induced pluripotent stem cells are useful models of human cardiac physiology and pharmacology. Here we present a high-throughput screening system to quantify the effects of exogenous compounds on beating frequency, using a Ca-sensitive fluorescent dye and a temperature-controlled imaging multi-well plate reader.

Contrações do citoplasma de casos humanos-iPSC-derivado medida com um corante fluorescente de Ca-sensíveis em placas de 384 poços de temperatura controlada

Spontaneously contracting syncytia of cardiomyocytes derived from human-induced pluripotent stem cells (hiPSC-CM) are a useful model of human cardiac ...

A Training and Testing System for Performing Vascular Reconstruction In Vitro

Manual vascular reconstruction training is essential for a beginner surgeon. However, an optimal training system for vascular reconstruction in vitro has yet to be developed. In this study, we introduce an in vitro training and testing system using a magnetic anchoring technique with which a trainee can practice manual vascular reconstruction individually. Additionally, this system can also be used to test the quality of the reconstruction. The described system includes a vascular reconstruction training machine, magnetic tractors, and a magnetic suture puller. In this manuscript, we detail an end-to-end vein anastomosis using porcine right and left iliac veins. To identify the potential damage caused by a magnetic suture puller on the suture, we created three groups with six segments of 4-0 polypropylene sutures each: a control group with no intervention on the polypropylene suture, a group in which the polypropylene suture is manually pulled with sterile gloves 20x, and a magnetic puller group in which the magnetic puller pulled the polypropylene suture 20x. These groups were tested by light microscopy and breaking strength tests, and the effect of reconstruction was assessed. In the light microscopy test, the control group was less likely to be damaged (p &#60; 0.05) and the number of damaged points of the manual group and magnetic puller group were similar (p &#62; 0.05). The results of the breaking strength test were compared across groups and no significant difference was observed (p &#62; 0.05). The end-to-end anastomosis of the porcine iliac veins was successfully performed using this training system, and the reconstructed veins could undergo 2.0 kPa perfusion pressure. Using this training and testing system the trainee can practice manual vascular reconstruction in vitro individually with the aid of magnetic tractors and a magnetic suture puller, and the quality of the reconstruction can be tested.

Here we present a training and testing system where a trainee can complete manual vascular reconstruction in vitro individually using a magnetic anchoring technique. The system can also be used to test the quality of reconstruction.

Um sistema de treinamento e teste para a realização de reconstrução vascular in vitro

Manual vascular reconstruction training is essential for a beginner surgeon. However, an optimal training system for vascular reconstruction in vitro has ...