Los autores agradecen el Dr. Lee Khai Pin, cabeza y Hospital Consultor Senior, emergencia de los niños, KK mujeres y niños, 100 Bukit Timah Road, Singapur para el apoyo administrativo con la producción de vídeo artículo. Artículo reproducido con permiso de «Mano Metal Detector para metálico exterior cuerpo ingestión en pediátrico emergencia»; Agosto 2018:85 (8): 618-624; Hazwani Binte Hamzah, James Vigil, Suraj Manickam, Sashikumar Ganapathy: copyright el diario indio de la pediatría.

El protocolo sigue las pautas del Comité de ética institucional de la investigación humana. La Junta de revisión institucional central proporciona la aprobación ética para el estudio.
1. participante de detección
<ol><li>Reclutar a los pacientes que presentan para el Departamento de emergencia pediátrico con historia de ingestión de cuerpo extraño.</li>
	<li>Excluir a los pacientes que están en señal de socorro respiratoria y ha implantado dispositivos como pace makers.</li>
</ol>2. prepare al paciente
<ol><li>Vestido del paciente en una bata de hospital.</li>
	<li>Quitar todos los elementos que pueden contener objetos metálicos tales como ropa, lentes, aretes, dijes y adornos antes del examen para evitar la interferencia con la proyección de HHMD.</li>
	<li>Colocar al paciente en el centro de la habitación de otros posibles interferencias metálicas.</li>
</ol>3. prepare el examinador y los cuidadores
<ol><li>Retire todos los accesorios metálicos como joyas y reloj.</li>
	<li>Solicitar el cuidador para quitar todos los accesorios metálicos como joyas y reloj.</li>
</ol>4. metal Detector proyección
<ol><li>Utilizar el HHMD (figura 1) para realizar la proyección.</li>
	<li>Para confirmar el estado de trabajo de lo HHMD provocando una positiva señal audiovisual moviendo sobre una pieza de metal.</li>
	<li>Realizar el HHMD exploración en bipedestación para niños más grandes.</li>
	<li>Realizar el HHMD exploración en posición erecta para niños por tener la bodega de cuidador del paciente adecuadamente.</li>
	<li>Sostener las manos del paciente hacia arriba y lejos del cuerpo.</li>
	<li>Posición de la cabeza del paciente en extensión para exponer el cuello.</li>
	<li>Análisis del nivel de la barbilla al nivel de la articulación de la cadera para cubrir las zonas anatómicas del cuello a la pelvis.</li>
	<li>Realizar el análisis en forma de zig-zag en todo el cuerpo para garantizar que cada área ha sido explorado.</li>
	<li>Escanear ambos anterior y posterior como el grosor del paciente debe tenerse en cuenta.</li>
	<li>Observe detenidamente la señal audiovisual positiva durante la exploración para la presencia de MFBs.</li>
</ol>5. realizar la pertinente radiografía
<ol><li>Solicitar la pertinente radiografía del cuello, pecho o abdomen, según el área detectado por la investigación HHMD para el MFBs.</li>
	<li>Proceder con todas las vistas de rayos x (cuello, tórax y abdomen) si la tecla MFB no fue detectado durante la proyección HHMD y el índice clínico de sospecha para ingestión de cuerpo extraño es alta.</li>
</ol>6. registrar los resultados de la prueba
<ol><li>Registrar los resultados en el cuaderno diario de pesca (figura 2) creado específicamente basan en el flujo de trabajo (figura 1).</li>
	<li>Correlacionar la señal positiva de HHMD audiovisual con la señal ósea subyacente en el cuerpo, palpando los siguientes hitos huesudos prominente - esternal, xiphisternum, sínfisis del pubis y cresta ilíaca bilateral.</li>
	<li>Grabar la señal en el esquema de referencia anatómica de la bitácora antes de proceder con el examen de rayos x.</li>
</ol>

<ol>
	<li>Lee, J., Ahmad, S., Gale, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+coins+ingested+by+children+using+a+handheld+metal+detector:+a+systematic+review.">Detection of coins ingested by children using a handheld metal detector: a systematic review.</a> Emergency Medicine Journal. 22 (12), 839-844 (2005).</li><li>Hesham A-Kader, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Foreign+body+ingestion:+children+like+to+put+objects+in+their+mouth.">Foreign body ingestion: children like to put objects in their mouth.</a> World Journal of Pediatrics. 6 (4), 301-310 (2010).</li><li>Muensterer, O. J., Joppich, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+and+topographic+localization+of+metallic+foreign+bodies+by+metal+detector.">Identification and topographic localization of metallic foreign bodies by metal detector.</a> Journal of Pediatric Surgery. 39 (8), 1245-1248 (2004).</li><li>Nation, J., Jiang, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+utility+of+a+handheld+metal+detector+in+detection+and+localization+of+pediatric+metallic+foreign+body+ingestion.">The utility of a handheld metal detector in detection and localization of pediatric metallic foreign body ingestion.</a> International Journal of Pediatric Otorhinolaryngology. 92, 1-6 (2017).</li><li>Arena, L., Baker, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+a+metal+detector+to+identify+ingested+metallic+foreign+bodies.">Use of a metal detector to identify ingested metallic foreign bodies.</a> American Journal of Roentgenology. 155 (4), 803-804 (1990).</li><li>Younger, R. M., Darrow, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Handheld+metal+detector+confirmation+of+radiopaque+foreign+bodies+in+the+esophagus.">Handheld metal detector confirmation of radiopaque foreign bodies in the esophagus.</a> Archives of Otolaryngology--Head &amp; Neck Surgery. 127 (11), 1371-1374 (2001).</li><li>Schalamon, J., Haxhija, E. Q., Ainoedhofer, H., G&#246;ssler, A., Schleef, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+a+hand-held+metal+detector+for+localisation+of+ingested+metallic+foreign+bodies+-+a+critical+investigation.">The use of a hand-held metal detector for localisation of ingested metallic foreign bodies - a critical investigation.</a> European Journal of Pediatrics. 163 (4-5), 257-259 (2004).</li><li>Kay, M., Wyllie, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pediatric+foreign+bodies+and+their+management.">Pediatric foreign bodies and their management.</a> Current Gastroenterology Reports. 7 (3), 212-218 (2005).</li><li>Rothmann, B. F., Boeckman, C. 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W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluoroscopic+foley+catheter+removal+of+esophageal+foreign+bodies+in+children:+experience+with+415+episodes.">Fluoroscopic foley catheter removal of esophageal foreign bodies in children: experience with 415 episodes.</a> Pediatrics. 94 (5), 709-714 (1994).</li><li>Kerschner, J. E., Beste, D. J., Conley, S. F., Kenna, M. A., Lee, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mediastinitis+associated+with+foreign+body+erosion+of+the+esophagus+in+children.">Mediastinitis associated with foreign body erosion of the esophagus in children.</a> International Journal of Pediatric Otorhinolaryngology. 59 (2), 89-97 (2001).</li><li>Ahn, D., Heo, S. J., Park, J. H., Sohn, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tracheoesophageal+fistula+with+tracheal+stenosis+resulting+from+retained+esophageal+foreign+body.">Tracheoesophageal fistula with tracheal stenosis resulting from retained esophageal foreign body.</a> Auris Nasus Larynx. 38 (6), 753-756 (2011).</li><li>Dahiya, M., Denton, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Esophagoaortic+perforation+by+foreign+body+(coin)+causing+sudden+death+in+a+3-year-old+child.">Esophagoaortic perforation by foreign body (coin) causing sudden death in a 3-year-old child.</a> The American Journal of Forensic Medicine and Pathology. 20 (2), 184-188 (1999).</li><li>O'Brien, G. C., Winter, D. C., Kirwan, W. O., Redmond, H. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ingested+foreign+bodies+in+the+paediatric+patient.">Ingested foreign bodies in the paediatric patient.</a> Irish Journal of Medical Science. 170 (2), 100-102 (2001).</li><li>Bassett, K. E., Schunk, J. E., Logan, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Localizing+ingested+coins+with+a+metal+detector.">Localizing ingested coins with a metal detector.</a> The American Journal of Emergency Medicine. 17 (4), 338-341 (1999).</li><li>Gooden, E. A., Forte, V., Papsin, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+a+commercially+available+metal+detector+for+the+localization+of+metallic+foreign+body+ingestion+in+children.">Use of a commercially available metal detector for the localization of metallic foreign body ingestion in children.</a> The Journal of Otolaryngology. 29 (4), 218-220 (2000).</li><li>Sacchetti, A., Carraccio, C., Lichenstein, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hand-held+metal+detector+identification+of+ingested+foreign+bodies.">Hand-held metal detector identification of ingested foreign bodies.</a> Pediatric Emergency Care. 10 (4), 204-207 (1994).</li><li>Hamzah, H. B., James, V., Manickam, S., Ganapathy, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Handheld+Metal+Detector+for+Metallic+Foreign+Body+Ingestion+in+Pediatric+Emergency.">Handheld Metal Detector for Metallic Foreign Body Ingestion in Pediatric Emergency.</a> Indian Journal of Pediatrics. 85 (8), 618-624 (2018).</li><li>Doraiswamy, N. V., Baig, H., Hallam, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metal+detector+and+swallowed+metal+foreign+bodies+in+children.">Metal detector and swallowed metal foreign bodies in children.</a> Journal of Accident &amp; Emergency Medicine. 16 (2), 123-125 (1999).</li><li>Seikel, K., Primm, P. A., Elizondo, B. J., Remley, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Handheld+metal+detector+localization+of+ingested+metallic+foreign+bodies:+accurate+in+any+hands.">Handheld metal detector localization of ingested metallic foreign bodies: accurate in any hands.</a> Archives of Pediatrics &amp; Adolescent Medicine. 153 (8), 853-857 (1999).</li><li>Lemberg, P. S., Darrow, D. H., Holinger, L. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aerodigestive+tract+foreign+bodies+in+the+older+child+and+adolescent.">Aerodigestive tract foreign bodies in the older child and adolescent.</a> The Annals of Otology, Rhinology, and Laryngology. 105 (4), 267-271 (1996).</li><li>Stack, L. B., Munter, 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=Foreign+bodies+in+the+gastrointestinal+tract.">Foreign bodies in the gastrointestinal tract.</a> Emergency Medicine Clinics of North America. 14 (3), 493-521 (1996).</li><li>Saz, E. U., Arikan, C., Ozgen&#231;, F., Duyu, M., Ozananar, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+utility+of+handheld+metal+detector+in+confirming+metallic+foreign+body+ingestion+in+the+pediatric+emergency+department.">The utility of handheld metal detector in confirming metallic foreign body ingestion in the pediatric emergency department.</a> The Turkish Journal of Gastroenterology: The Official Journal of Turkish Society of Gastroenterology. 21 (2), 135-139 (2010).</li></ol>

Los autores no tienen nada que revelar.

Metálico exterior ingestión del cuerpo en los niños es un problema común y en la mayoría de los pacientes, se trata de manejar conservador7,16. Las radiografías simples o repetidas pueden exponer a la población pediátrica de radio-sensible al riesgo de radiación no deseadas al intentar localizar el cuerpo extraño metálico ingerido. Varios estudios han descrito el uso de detectores de metal en ingestión de MFB en niños, con sensibilidad de más del 96...

Ingestión de cuerpo extraño es un problema muy extendido entre los niños que acuden al servicio de urgencias pediátrico debido a la naturaleza inquisitiva natural de los bebés y los niños para explorar objetos en cata y tragarlas1,2. Estudios previos han reportado que objetos metálicos constituyen 85% de toda ingestión de cuerpo extraño en su cohorte de pacientes3,4. El objetivo de este protocolo es evaluar la exactitud y la viabilidad de utilizar un detector de metales portátil para detectar cuerpos extraños metálicos ingeridos en niños. Proponemos que al introducir el detector de metales handheld detección protocolo temprano en el proceso de triage de los niños con alta sospecha de ingestión de cuerpo extraño metálico, el número de radiografías se ordenó localizar el cuerpo extraño metálico puede ser reducido en esta población sensible a la radio.
Detector de metales handheld (HHMD) es un complemento barato y de fácil acceso que puede ayudar a agilizar la gestión de los cuerpos extraños metálicos ingeridos. Detector de metales es un dispositivo de muy baja frecuencia, que tiene un receptor y un transmisor que detecta el cambio en su campo electromagnético causado por la presencia de un objeto metálico en sus alrededores3. Si un objeto metálico dentro del campo, se interrumpe el campo electromagnético y el sensor se partió de una alarma audiovisual en el dispositivo4. El examen del detector de metales se puede realizar completamente en menos de 2 minutos5. Por lo tanto, se puede introducir la proyección HHMD temprano en el proceso de triage de los niños con alta sospecha de ingestión de cuerpo extraño metálico para la localización topográfica.
Las monedas son las más comunes cuerpos extraños metálicos ingeridos1,4,6,7. Debido a la falta de bordes afilados y naturaleza no tóxica general, la mayoría de las ingestiones de moneda se manejan conservadoramente. Otros tipos de materiales comúnmente ingeridos son imanes, pilas de botón, pequeños juguetes, cacahuetes, joyas, botones, huesos, semillas y palomitas de maíz8,9. Impactación de cuerpos extraños ocurre más comúnmente en el esófago en tres sitios, es decir, esfínter esofágico superior, mediados de-esófago a nivel del arco aórtico y del esfínter esofágico inferior10. Cuerpos extraños metálicos (MFBs) situados en el esófago pueden ser asintomáticos en el 40% de pacientes11,12 y su retención prolongada puede llevar a varias complicaciones como perforación esofágica, mediastinitis, fístula traqueoesofágica, Estenosis traqueal y fístulas aorto-esofágica1,13,14,15. Otros lugares de la impactación de cuerpos extraños incluyen Unión ileo-cecal10, píloro y duodeno.
Realizar las radiografías de cuello, pecho y abdomen para diagnosticar sospecha de ingestión de cuerpo extraño es una práctica muy extendida en los departamentos de emergencias5. Entre todos los tipos de los cuerpos extraños ingeridos, sólo el 10% son radio-opacos10. Las pruebas radiológicas son lentas y tienen un riesgo de exposición a la radiación en los niños. Localización topográfica precisa de los objetos metálicos es muy importante guiar la terapia. La introducción de HHMD temprano en el proceso de triage puede ayudar a mitigar el extraño pedido de radiografías por localizar topográficamente el cuerpo extraño metálico ingerido.
Este estudio evaluó la exactitud de los HHMD localizar MFBs de manera sistemática y topográfica y determina la eficacia y viabilidad de HHMD en la población pediátrica. También examinamos cómo esta gratuita herramienta de detección de radiación fue aceptada por los pacientes y padres. Entre mayo y julio de 2016, se incluyeron todos los pacientes consecutivos que presentaron para el servicio de urgencias pediátrico con antecedentes de ingestión de cuerpo extraño en el estudio. Se obtuvo el consentimiento informado del padre o tutor legal. Pacientes con ingestión del cuerpo extranjero alegado sistemáticamente fueron escaneados usando HHMD. El estándar de oro de la atención para la identificación y localización de cuerpos extraños metálicos ingeridos realiza radiografías de la zona cervical, tórax y abdomen. Para facilitar la realización del estudio, un flujo de trabajo estandarizado fue creado (figura 1). Un libro de registro creado específicamente basado en el flujo de trabajo se utilizó para recoger los datos iniciales, que posteriormente fueron transcritas a una hoja de cálculo (figura 2). Los datos en la sección de resultados se describe como el número de casos seguido por porcentajes.

<table>
	<tbody>
		<tr>
			<td>Hand held metal detector</td>
			<td>Garrett Super Scanner V</td>
			<td>NA</td>
			<td>NA</td>
		</tr>
	</tbody>
</table>

handheld metal detector screening for metallic foreign body ingestion in children

Las monedas son los cuerpos extraños metálicos ingeridos más común entre los niños. El objetivo de este protocolo es evaluar la exactitud y la viabilidad de utilizar un detector de metales portátil para detectar cuerpos extraños metálicos ingeridos en niños. Proponemos que mediante la introducción de proyección de mano detector de metales en el proceso de triage de los niños con alta sospecha de ingestión de cuerpo extraño metálico, puede reducirse el número de radiografías se ordenó localizar el cuerpo extraño metálico en este Radio-sensible a la población. El protocolo de estudio requiere la selección de los participantes para la historia de la ingestión del cuerpo extranjero y la exclusión de pacientes con dificultad respiratoria o implantes metálicos. El paciente cambia a bata de hospital y artículos que pueden contener metales como gafas, pendientes, colgantes y adornos se quitan. El paciente se coloca en el centro de la habitación de otras interferencias metálicas. El estado de trabajo del computadora de mano detector de metales primero confirma que una positiva señal audiovisual. Entonces la proyección se realiza en una posición erecta con la cabeza en extensión para exponer el cuello, desde el nivel de la barbilla al nivel de la articulación de la cadera, para cubrir las zonas anatómicas del cuello a la pelvis en forma de zig-zag tanto anterior como posterior. Una señal positiva de audiovisual se observa cuidadosamente durante la exploración para la presencia de cuerpo extraño metálico. Las radiografías pertinentes se ordenan según la zona detectada en el examen del detector de metales. El detector de metales handheld fue capaz de identificar con precisión todas las monedas entre los cuerpos extraños metálicos ingeridos en nuestro estudio. El detector de metales handheld constantemente no podía detectar cuerpos extraños metálicos no moneda. Este protocolo demuestra la exactitud de la computadora de mano detector de metales en la identificación y localización de monedas y monedas como cuerpos extraños metálicos.

Los investigadores reclutaron a 36 pacientes para este estudio observacional. Estos pacientes habían presentado para el servicio de urgencias pediátrico con antecedentes de ingestión de cuerpo extraño. Entre estos, 28 pacientes tenían ingestas de cuerpos extraños metálicos (figura 3). Los restantes 8 pacientes habían ingerido cuerpos extraños no-metálico tales como huesos de pescado, palo satay y botón de plástico. Las monedas eran el tipo más co...

Watch this Scientific Journal Video about Handheld Metal Detector Screening for Metallic Foreign Body Ingestion in Children at JoVE.com

Handheld Metal Detector Screening for Metallic Foreign Body Ingestion in Children

Aquí, presentamos un protocolo para utilizar el detector de metales portátil, para detectar la presencia de cuerpos extraños metálicos ingeridos en niños que presentan al Departamento de medicina de emergencia pediátrica con antecedentes de ingestión de cuerpo extraño.

Detector de metales portátil detección de ingestión de cuerpo extraño metálico en niños

Coins are the most common ingested metallic foreign bodies among children. The goal of this protocol is to assess the accuracy and feasibility of using a ...

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10.6K vistas.  KK women's and Children's Hospital. Aquí, presentamos un protocolo para utilizar el detector de metales portátil, para detectar la presencia de cuerpos extraños metálicos ingeridos en niños que presentan al Departamento de medicina de emergencia pediátrica con antecedentes de ingestión de cuerpo extraño.

Video: Handheld Metal Detector Screening for Metallic Foreign Body Ingestion in Children

High Content Screening in Neurodegenerative Diseases

The functional annotation of genomes, construction of molecular networks and novel drug target identification, are important challenges that need to be addressed as a matter of great urgency1-4. Multiple complementary 'omics' approaches have provided clues as to the genetic risk factors and pathogenic mechanisms underlying numerous neurodegenerative diseases, but most findings still require functional validation5. For example, a recent genome wide association study for Parkinson's Disease (PD), identified many new loci as risk factors for the disease, but the underlying causative variant(s) or pathogenic mechanism is not known6, 7. As each associated region can contain several genes, the functional evaluation of each of the genes on phenotypes associated with the disease, using traditional cell biology techniques would take too long. 
	There is also a need to understand the molecular networks that link genetic mutations to the phenotypes they cause. It is expected that disease phenotypes are the result of multiple interactions that have been disrupted. Reconstruction of these networks using traditional molecular methods would be time consuming. Moreover, network predictions from independent studies of individual components, the reductionism approach, will probably underestimate the network complexity8. This underestimation could, in part, explain the low success rate of drug approval due to undesirable or toxic side effects. Gaining a network perspective of disease related pathways using HT/HC cellular screening approaches, and identifying key nodes within these pathways, could lead to the identification of targets that are more suited for therapeutic intervention.
	High-throughput screening (HTS) is an ideal methodology to address these issues9-12. but traditional methods were one dimensional whole-well cell assays, that used simplistic readouts for complex biological processes. They were unable to simultaneously quantify the many phenotypes observed in neurodegenerative diseases such as axonal transport deficits or alterations in morphology properties13, 14. This approach could not be used to investigate the dynamic nature of cellular processes or pathogenic events that occur in a subset of cells. To quantify such features one has to move to multi-dimensional phenotypes termed high-content screening (HCS)4, 15-17. HCS is the cell-based quantification of several processes simultaneously, which provides a more detailed representation of the cellular response to various perturbations compared to HTS. 
	HCS has many advantages over HTS18, 19, but conducting a high-throughput (HT)-high-content (HC) screen in neuronal models is problematic due to high cost, environmental variation and human error. In order to detect cellular responses on a 'phenomics' scale using HC imaging one has to reduce variation and error, while increasing sensitivity and reproducibility. 
	Herein we describe a method to accurately and reliably conduct shRNA screens using automated cell culturing20 and HC imaging in neuronal cellular models. We describe how we have used this methodology to identify modulators for one particular protein, DJ1, which when mutated causes autosomal recessive parkinsonism21. 
	Combining the versatility of HC imaging with HT methods, it is possible to accurately quantify a plethora of phenotypes. This could subsequently be utilized to advance our understanding of the genome, the pathways involved in disease pathogenesis as well as identify potential therapeutic targets.

We describe a methodology combining automated cell culturing with high-content imaging to visualize and quantify multiple cellular processes and structures, in a high-throughput manner. Such methods can aid in the further functional annotation of genomes as well as identify disease gene networks and potential drug targets.

Filtrado de contenido de alta en las enfermedades neurodegenerativas

The functional annotation of genomes, construction of molecular networks and novel drug target identification, are important challenges that need to be ...

Investigación

Medicina

Eye Tracking Young Children with Autism

The rise of accessible commercial eye-tracking systems has fueled a rapid increase in their use in psychological and psychiatric research. By providing a direct, detailed and objective measure of gaze behavior, eye-tracking has become a valuable tool for examining abnormal perceptual strategies in clinical populations and has been used to identify disorder-specific characteristics1, promote early identification2, and inform treatment3. In particular, investigators of autism spectrum disorders (ASD) have benefited from integrating eye-tracking into their research paradigms4-7. Eye-tracking has largely been used in these studies to reveal mechanisms underlying impaired task performance8 and abnormal brain functioning9, particularly during the processing of social information1,10-11. While older children and adults with ASD comprise the preponderance of research in this area, eye-tracking may be especially useful for studying young children with the disorder as it offers a non-invasive tool for assessing and quantifying early-emerging developmental abnormalities2,12-13. Implementing eye-tracking with young children with ASD, however, is associated with a number of unique challenges, including issues with compliant behavior resulting from specific task demands and disorder-related psychosocial considerations. In this protocol, we detail methodological considerations for optimizing research design, data acquisition and psychometric analysis while eye-tracking young children with ASD. The provided recommendations are also designed to be more broadly applicable for eye-tracking children with other developmental disabilities. By offering guidelines for best practices in these areas based upon lessons derived from our own work, we hope to help other investigators make sound research design and analysis choices while avoiding common pitfalls that can compromise data acquisition while eye-tracking young children with ASD or other developmental difficulties.

Eye tracking has long been used to study gaze patterns in typically-developing individuals, but recent technological advancements have made its use with clinical populations, including autism, more feasible. While eye-tracking young children with autism can offer insight into early symptom manifestations, it involves methodological challenges. Suggestions for best practices are provided.

Eye Tracking niños pequeños con autismo

The rise of accessible commercial eye-tracking systems has fueled a rapid increase in their use in psychological and psychiatric research. By providing a ...

Protocol for Long Duration Whole Body Hyperthermia in Mice

Hyperthermia is a general term used to define the increase in core body temperature above normal. It is often used to describe the increased core body temperature that is observed during fever. The use of hyperthermia as an adjuvant has emerged as a promising procedure for tumor regression in the field of cancer biology. For this purpose, the most important requirement is to have reliable and uniform heating protocols. We have developed a protocol for hyperthermia (whole body) in mice. In this protocol, animals are exposed to cycles of hyperthermia for 90 min followed by a rest period of 15 min. During this period mice have easy access to food and water. High body temperature spikes in the mice during first few hyperthermia exposure cycles are prevented by immobilizing the animal. Additionally, normal saline is administered in first few cycles to minimize the effects of dehydration. This protocol can simulate fever like conditions in mice up to 12-24 hr. We have used 8-12 weeks old BALB/Cj female mice to demonstrate the protocol.

This paper describes a protocol for whole body hyperthermia in mice that can stimulate fever like conditions up to 12-24 hr.

Protocolo para la hipertermia corporal Larga Duración Total en ratones

Hyperthermia is a general term used to define the increase in core body temperature above normal. It is often used to describe the increased core body ...

Measuring Cardiac Autonomic Nervous System (ANS) Activity in Children

The autonomic nervous system (ANS) controls mainly automatic bodily functions that are engaged in homeostasis, like heart rate, digestion, respiratory rate, salivation, perspiration and renal function. The ANS has two main branches: the sympathetic nervous system, preparing the human body for action in times of danger and stress, and the parasympathetic nervous system, which regulates the resting state of the body.
ANS activity can be measured invasively, for instance by radiotracer techniques or microelectrode recording from superficial nerves, or it can be measured non-invasively by using changes in an organ's response as a proxy for changes in ANS activity, for instance of the sweat glands or the heart. Invasive measurements have the highest validity but are very poorly feasible in large scale samples where non-invasive measures are the preferred approach. Autonomic effects on the heart can be reliably quantified by the recording of the electrocardiogram (ECG) in combination with the impedance cardiogram (ICG), which reflects the changes in thorax impedance in response to respiration and the ejection of blood from the ventricle into the aorta. From the respiration and ECG signals, respiratory sinus arrhythmia can be extracted as a measure of cardiac parasympathetic control. From the ECG and the left ventricular ejection signals, the preejection period can be extracted as a measure of cardiac sympathetic control. ECG and ICG recording is mostly done in laboratory settings. However, having the subjects report to a laboratory greatly reduces ecological validity, is not always doable in large scale epidemiological studies, and can be intimidating for young children. An ambulatory device for ECG and ICG simultaneously resolves these three problems.
Here, we present a study design for a minimally invasive and rapid assessment of cardiac autonomic control in children, using a validated ambulatory device 1-5, the VU University Ambulatory Monitoring System (VU-AMS, Amsterdam, the Netherlands,<a href="http://www.vu-ams.nl" target="_blank"> www.vu-ams.nl</a>).

Measuring Cardiac Autonomic Nervous System ANS Activity in Children

Measurement of autonomic nervous system activity usually confines the researcher and participant to the laboratory, which may provide an intimidating environment to children. The VU University Ambulatory Monitoring System (VU-AMS) device can record cardiac autonomic control in any setting. The VU-AMS proved very amenable to testing in children.

Sistema de medición de actividad nerviosa autonómica cardiaca (ANS) en los niños

The autonomic nervous system (ANS) controls mainly automatic bodily functions that are engaged in homeostasis, like heart rate, digestion, respiratory ...

Screening for Melanoma Modifiers using a Zebrafish Autochthonous Tumor Model

Genomic studies of human cancers have yielded a wealth of information about genes that are altered in tumors1,2,3. A challenge arising from these studies is that many genes are altered, and it can be difficult to distinguish genetic alterations that drove tumorigenesis from that those arose incidentally during transformation. To draw this distinction it is beneficial to have an assay that can quantitatively measure the effect of an altered gene on tumor initiation and other processes that enable tumors to persist and disseminate. Here we present a rapid means to screen large numbers of candidate melanoma modifiers in zebrafish using an autochthonous tumor model4 that encompasses steps required for melanoma initiation and maintenance. A key reagent in this assay is the miniCoopR vector, which couples a wild-type copy of the mitfa melanocyte specification factor to a Gateway recombination cassette into which candidate melanoma genes can be recombined5. The miniCoopR vector has a mitfa rescuing minigene which contains the promoter, open reading frame and 3'-untranslated region of the wild-type mitfa gene. It allows us to make constructs using full-length open reading frames of candidate melanoma modifiers. These individual clones can then be injected into single cell Tg(mitfa:BRAFV600E);p53(lf);mitfa(lf)zebrafish embryos. The miniCoopR vector gets integrated by Tol2-mediated transgenesis6 and rescues melanocytes. Because they are physically coupled to the mitfa rescuing minigene, candidate genes are expressed in rescued melanocytes, some of which will transform and develop into tumors. The effect of a candidate gene on melanoma initiation and melanoma cell properties can be measured using melanoma-free survival curves, invasion assays, antibody staining and transplantation assays.

A rapid way to screen for melanoma modifiers using a zebrafish autochthonous tumor model is presented. It takes advantage of the miniCoopR vector which allows for expression of candidate melanoma genes in melanocytes. A method to obtain melanoma-free survival curves, an invasion assay, a protocol for antibody staining of scale melanocytes and a melanoma transplantation assay are described.

La detección de Modificadores melanoma utilizando un modelo de pez cebra Tumor autóctona

Genomic studies of  human cancers have yielded a wealth of information about genes that are altered  in tumors1,2,3. A challenge arising from these ...

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

We have successfully integrated previously established Intracranial window (ICW) technology 1-4 with intravital 2-photon confocal microscopy to develop a novel platform that allows for direct long-term visualization of tissue structure changes intracranially. Imaging at a single cell resolution in a real-time fashion provides supplementary dynamic information beyond that provided by standard end-point histological analysis, which looks solely at 'snap-shot' cross sections of tissue.
Establishing this intravital imaging technique in fluorescent chimeric mice, we are able to image four fluorescent channels simultaneously. By incorporating fluorescently labeled cells, such as GFP+ bone marrow, it is possible to track the fate of these cells studying their long-term migration, integration and differentiation within tissue. Further integration of a secondary reporter cell, such as an mCherry glioma tumor line, allows for characterization of cell:cell interactions. Structural changes in the tissue microenvironment can be highlighted through the addition of intra-vital dyes and antibodies, for example CD31 tagged antibodies and Dextran molecules.
Moreover, we describe the combination of our ICW imaging model with a small animal micro-irradiator that provides stereotactic irradiation, creating a platform through which the dynamic tissue changes that occur following the administration of ionizing irradiation can be assessed.
Current limitations of our model include penetrance of the microscope, which is limited to a depth of up to 900 &mu;m from the sub cortical surface, limiting imaging to the dorsal axis of the brain. The presence of the skull bone makes the ICW a more challenging technical procedure, compared to the more established and utilized chamber models currently used to study mammary tissue and fat pads 5-7. In addition, the ICW provides many challenges when optimizing the imaging.

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

We describe a novel in vivo imaging technique that couples fluorescent chimeric mice with intracranial windows and high-resolution 2-photon microscopy. This imaging platform aids studies of dynamic changes in brain tissue and microvasculature, at a single-cell level, following pathological insults and is adaptable to assess intracranial drug delivery and distribution.

Una novela de alta resolución<em&gt; En vivo</em&gt; Técnica de escaneo para el Estudio de la respuesta dinámica de estructuras intracraneales al crecimiento del tumor y Terapéutica

We have successfully integrated previously established Intracranial window (ICW) technology 1-4 with intravital 2-photon confocal microscopy to develop a ...

A Novel in vivo Gene Transfer Technique and in vitro Cell Based Assays for the Study of Bone Loss in Musculoskeletal Disorders

Differentiation and activation of osteoclasts play a key role in the development of musculoskeletal diseases as these cells are primarily involved in bone resorption. Osteoclasts can be generated in vitro from monocyte/macrophage precursor cells in the presence of certain cytokines, which promote survival and differentiation. Here, both in vivo and in vitro techniques are demonstrated, which allow scientists to study different cytokine contributions towards osteoclast differentiation, signaling, and activation. The minicircle DNA delivery gene transfer system provides an alternative method to establish an osteoporosis-related model is particularly useful to study the efficacy of various pharmacological inhibitors in vivo. Similarly, in vitro culturing protocols for producing osteoclasts from human precursor cells in the presence of specific cytokines enables scientists to study osteoclastogenesis in human cells for translational applications. Combined, these techniques have the potential to accelerate drug discovery efforts for osteoclast-specific targeted therapeutics, which may benefit millions of osteoporosis and arthritis patients worldwide.

A Novel in vivo Gene Transfer Technique and in vitro Cell Based Assays for the Study of Bone Loss in Musculoskeletal Disorders

Differentiation of precursor cells into osteoclasts is regulated by cytokines and growth factors. Here, a novel gene transfer technique for differentiation of osteoclasts in vivo and cell culture protocols for differentiating precursor cells into osteoclasts in vitro as a method to study the effects of cytokines on osteoclastogenesis are described.

A Novel<em&gt; In vivo</em&gt; Técnica de transferencia de genes y<em&gt; In vitro</em&gt; ensayos basados ​​en células para el estudio de la pérdida ósea en los trastornos musculoesqueléticos

Differentiation and activation of osteoclasts play a key role in the development of musculoskeletal diseases as these cells are primarily involved in bone ...

Normothermic Ex Vivo Kidney Perfusion for the Preservation of Kidney Grafts prior to Transplantation

Kidney transplantation has become a well-established treatment option for patients with end-stage renal failure. The persisting organ shortage remains a serious problem. Therefore, the acceptance criteria for organ donors have been extended leading to the usage of marginal kidney grafts. These marginal organs tolerate cold storage poorly resulting in increased preservation injury and higher rates of delayed graft function. To overcome the limitations of cold storage, extensive research is focused on alternative normothermic preservation methods.
Ex vivo normothermic organ perfusion is an innovative preservation technique. The first experimental and clinical trials for ex vivo lung, liver, and kidney perfusions demonstrated favorable outcomes.
In addition to the reduction of cold ischemic injury, the method of normothermic kidney storage offers the opportunity for organ assessment and repair. This manuscript provides information about kidney retrieval, organ preservation techniques, and isolated ex vivo normothermic kidney perfusion (NEVKP) in a porcine model. Surgical techniques, set up for the perfusion solution and the circuit, potential assessment options, and representative results are demonstrated.

Normothermic Ex Vivo Kidney Perfusion for the Preservation of Kidney Grafts prior to Transplantation

The severe organ shortage has resulted in increased use of marginal kidney grafts for transplantation. This has triggered interest in alternative storage methods, since marginal grafts especially tolerate cold storage poorly. The technique of normothermic ex vivo kidney perfusion (NEVKP) represents a novel preservation method for kidney grafts prior to &#160;transplantation.

Normotérmica<em&gt; Ex Vivo</em&gt; Perfusión renal para la Preservación de riñón Injertos antes del trasplante

Kidney transplantation has become a well-established treatment option for patients with end-stage renal failure. The persisting organ shortage remains a ...

Heterotopic Renal Autotransplantation in a Porcine Model: A Step-by-Step Protocol

Kidney transplantation is the treatment of choice for patients suffering from end-stage renal disease. It offers better life expectancy and higher quality of life when compared to dialysis. Although the last few decades have seen major improvements in patient outcomes following kidney transplantation, the increasing shortage of available organs represents a severe problem worldwide. To expand the donor pool, marginal kidney grafts recovered from extended criteria donors (ECD) or donated after circulatory death (DCD) are now accepted for transplantation. To further improve the postoperative outcome of these marginal grafts, research must focus on new therapeutic approaches such as alternative preservation techniques, immunomodulation, gene transfer, and stem cell administration.
Experimental studies in animal models are the final step before newly developed techniques can be translated into clinical practice. Porcine kidney transplantation is an excellent model of human transplantation and allows investigation of novel approaches. The major advantage of the porcine model is its anatomical and physiological similarity to the human body, which facilitates the rapid translation of new findings to clinical trials. This article offers a surgical step-by-step protocol for an autotransplantation model and highlights key factors to ensure experimental success. Adequate pre- and postoperative housing, attentive anesthesia, and consistent surgical techniques result in favorable postoperative outcomes. Resection of the contralateral native kidney provides the opportunity to assess post-transplant graft function. The placement of venous and urinary catheters and the use of metabolic cages allow further detailed evaluation. For long-term follow-up studies and investigation of alternative graft preservation techniques, autotransplantation models are superior to allotransplantation models, as they avoid the confounding bias posed by rejection and immunosuppressive medication.

Porcine models of organ transplantation provide an important platform to study mechanisms of organ preservation. This article describes a heterotopic porcine renal autotransplantation model, which allows investigating new approaches to improve the outcome of transplantation using marginal kidney grafts.

Heterotópico renal autotrasplante en un modelo porcino: Un Protocolo de Paso a Paso

Kidney transplantation is the treatment of choice for patients suffering from end-stage renal disease. It offers better life expectancy and higher quality ...

Nerve-sparing Mid-urethral Obstruction (NeMO) in Female Small Rodents

Partial bladder outlet obstruction (pBOO) has a high prevalence, causes significant patient burden, and immense health care costs. The most common animal model to investigate bladder remodeling in pBOO are female rodents undergoing partial obstruction at the proximal urethra. Variability in the degree of obstruction and animal mortality are major concerns with proximal obstruction. Furthermore, dissecting around the proximal urethra and bladder neck jeopardizes bladder innervation.
We developed a nerve-sparing mid-urethral obstruction (NeMO) model for pBOO avoiding the disadvantages of the traditional model. We approached the urethra just inferior to the pubic symphysis, which obviated the need for laparotomy as well as for dissection in this area; also, the striated urethral sphincter remained untouched. We performed NeMO in female Sprague-Dawley rats (12 obstructions, 6 sham animals) as well as in female C57/bl6 mice (20 obstructions, 18 sham animals). After two weeks, we evaluated bladder function, bladder mass, and body mass.
We had no mortalities among obstructed- or sham-operated female rats; as described for the traditional proximal pBOO-method, we tied the suture around the proximal urethra and a temporarily placed 0.9 mm metal rod. NeMO induced an 85% increase in bladder mass after two weeks, average residual urine volume was 0.4 mL in partially obstructed rats while only 0.03 mL in sham animals. In mice, we tested 3 sizes of cannulas that we placed along the urethra when tying the suture. We found that using a 27-gauge cannula resulted in over 50% animal mortality; placing the 25-gauge cannula did not yield the desired response in increasing bladder mass; utilizing a 26-gauge cannula yielded favorable results with minimal animal mortality (1/8) yet a significant 2-fold increase in bladder mass.

Nerve-sparing Mid-urethral Obstruction NeMO in Female Small Rodents

Traditional modeling of partial bladder outlet obstruction in rodents is fraught with animal mortality. A denervation injury from dissection around the proximal urethra and bladder neck is also of major concern. We developed and evaluated a safe and reliable mid-urethral obstruction model, avoiding the shortcomings of the traditional model.

Preservación de los nervios de mitad de obstrucción uretral (NEMO) en femenino pequeños roedores

Partial bladder outlet obstruction (pBOO) has a high prevalence, causes significant patient burden, and immense health care costs. The most common animal ...