Die Autoren danken Dr. Lee Khai Pin, Kopf & Senior Consultant, Notfall, KK Frauen und Kinder Krankenhaus, 100 Bukit Timah Road, Singapur für die administrative Unterstützung bei der Produktion von video-Artikel. Artikel reproduziert mit freundlicher Genehmigung von "Handheld Metall Detektor für metallische ausländische Körper Einnahme in Pädiatrische Notfall"; August 2018:85 (8): 618-624; Hazwani Binte Hamzah, Vigil James, Suraj Manickam, Sashikumar Ganapathy: copyright The Indian Journal of Pediatrics.

Das Protokoll folgt den Richtlinien des institutionellen menschliche Forschung Ethikkommission. Das zentrale institutionelle Review Board hat die Ethik Zustimmung erteilt für das Studium.
1. Teilnehmer Screening
<ol><li>Rekrutieren Sie Patienten, die in der pädiatrischen Notaufnahme mit Geschichte der Fremdkörper Verschlucken zu präsentieren.</li>
	<li>Schließen Sie die Patienten, die in Atemnot und hat Geräte wie Herzschrittmacher implantiert.</li>
</ol>2. Vorbereitung des Patienten
<ol><li>Der Patient in einem krankenhauskleid Kleid.</li>
	<li>Entfernen Sie alle Elemente, die metallische Gegenstände wie Kleidung, Brillen, Ohrringe, Anhänger und Ornamente vor der Untersuchung zur Vermeidung von Interferenzen mit dem HHMD Screening enthalten können.</li>
	<li>Legen Sie den Patienten in der Mitte des Raumes von anderen potenziellen metallischen Interferenzen.</li>
</ol>3. bereiten Sie die Prüfer und Betreuer
<ol><li>Entfernen Sie alle metallischen Accessoires wie Schmuck und Uhren.</li>
	<li>Fordern Sie die Pflegeperson, alle metallischen Accessoires wie Schmuck und Uhren zu entfernen.</li>
</ol>(4) Metall-Detektor-Screening
<ol><li>Verwenden Sie die HHMD (Abbildung 1), das Screening durchzuführen.</li>
	<li>Bestätigen Sie den Arbeitsstatus der HHMD durch ein positives Audio-visuellen Signal durch winken über ein Stück Metall zu entlocken.</li>
	<li>Führen Sie die HHMD Scannen in eine stehende Position für größere Kinder.</li>
	<li>Führen Sie die HHMD Scannen in einer aufrechten Position für Säuglinge mit der Bezugsperson halten die Patienten angemessen.</li>
	<li>Halten Sie die Hände des Patienten, und vom Körper weg.</li>
	<li>Positionieren Sie den Kopf des Patienten auf den Hals zu entlarven.</li>
	<li>Scannen von der Ebene des Kinns auf die Ebene des Hüftgelenks anatomischen Bereich vom Hals bis Becken abdecken.</li>
	<li>Führen Sie den Scan Zick-Zack über den Körper, um sicherzustellen, dass jeder Bereich gescannt wurde.</li>
	<li>Scannen Sie beide vorn und nach hinten wie die Dicke des Patienten berücksichtigt werden muss.</li>
	<li>Beachten Sie sorgfältig das positive audiovisuelle Signal beim Scannen auf das Vorhandensein von MFBs.</li>
</ol>5. führen Sie die entsprechenden Röntgenaufnahme
<ol><li>Bestellen Sie der entsprechenden Röntgenaufnahme der Hals, Brust oder Bauch, nach der Gegend von HHMD Screening für die MFBs erkannt.</li>
	<li>Fahren Sie mit den x-ray-Ansichten (Hals, Brust und Bauch), wenn die MFB bei HHMD Screening und klinische Index des Misstrauens nicht erkannt wurde, für Fremdkörper Verschlucken hoch.</li>
</ol>6. zeichnen Sie die Testergebnisse
<ol><li>Rekord der Testergebnisse im Logbuch (Abbildung 2) speziell anhand der Workflow (Abbildung 1).</li>
	<li>Korrelieren Sie das positive Signal für audiovisuelle HHMD mit dem zugrunde liegenden knöchernen Wahrzeichen auf den Körper, durch Abtasten der folgenden prominenten knöchernen Wahrzeichen - sternalen Kerbe, so Schambeinfuge und bilateralen Beckenkamm.</li>
	<li>Notieren Sie das Wahrzeichen auf dem Logbuch anatomische Referenz Umriss bevor Sie mit der Röntgenuntersuchung.</li>
</ol>

<ol>
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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>

Die Autoren haben nichts preisgeben.

Metallische verwaltet ausländische Körper Einnahme bei Kindern ist ein häufiges Problem und in der Mehrzahl der Patienten ist dies konservativ7,16. Einzelne oder wiederholte Röntgenaufnahmen können die Radio-Sensitive pädiatrischen Population unerwünschte Strahlung Gefahr beim Versuch, den aufgenommenen metallische Fremdkörper lokalisieren aussetzen. Mehrere Studien haben den Einsatz von Metalldetektoren im MFB Einnahme bei Kindern, mehr als 96 % Sensitiv...

Fremdkörper Verschlucken ist ein weit verbreitetes Problem unter den Kindern präsentiert die pädiatrische Notaufnahme wegen der natürlichen neugierigen Natur von Säuglingen und Kindern Objekte erforschen durch Verkostung und schlucken sie1,2. Frühere Studien haben berichtet, dass Metallgegenstände 85 % der alle Fremdkörper-Ingestionen in ihre Kohorte von Patienten3,4konstituiert. Das Ziel dieses Protokolls ist zur Beurteilung der Genauigkeit und Machbarkeit mit einem handheld metallortungsgerät aufgenommenen metallische Fremdkörper bei Kindern zu erkennen. Wir schlagen vor, dass durch die Einführung des handheld Metalldetektors screening Protokoll früh in der Triage-Prozess von Kindern mit Verdacht auf metallische Fremdkörper Verschlucken, die Anzahl der Röntgenbilder bestellt, den metallische Fremdkörper zu lokalisieren sein kann in dieser Radio-Sensitive-Population reduziert.
Handheld-Metalldetektor (HHMD) ist kostengünstig, leicht zugängliche Ergänzung, die helfen können die Verwaltung von angesaugte metallischen Fremdkörpern zu beschleunigen. Metall-Detektor ist ein sehr niedrigen frequenzgerät, mit einem Empfänger und einem Sender, der die Änderung in seiner elektromagnetischen Feldes verursacht durch das Vorhandensein von einem Metallgegenstand in seiner Nähe3erkennt. Wenn ein metallisches Objekt innerhalb des Feldes ist, das elektromagnetische Feld ist gestört und der Sensor einen Audio-visuellen Alarm auf dem Gerät4eingestellt werden. Die Metalldetektor-Prüfung kann in weniger als 2 Minuten5gründlich durchgeführt werden. Daher kann der HHMD Screening früh in der Triage-Prozess von Kindern mit Verdacht auf metallische Fremdkörper Verschlucken für topographische Lokalisierung eingeführt werden.
Münzen sind die häufigsten geschluckt metallische Fremdkörper1,4,6,7. Aufgrund des Fehlens von scharfen Kanten und allgemeine ungiftige Natur sind die Mehrheit der Münze Ingestionen konservativ verwaltet. Andere Arten von Materialien, die häufig eingenommen gehören Magnete, Knopfbatterien, kleine Spielzeuge, Erdnüsse, Schmuck, Knöpfe, Knochen, Samen und Popcorn8,9. Impaktion von Fremdkörpern tritt am häufigsten in der Speiseröhre an drei Standorten, nämlich oberen Ösophagussphinkters, Mitte des Ösophagus auf der Ebene der Aortenbogen und unteren Ösophagussphinkters10. Metallische Fremdkörper (MFBs) befindet sich in der Speiseröhre können asymptomatisch in 40 % der Patienten11,12 und seiner längeren Impaktion führen zu verschiedenen Komplikationen wie esophageal Perforierung, Mediastinitis, tracheoesophageal Fistula, trachealen Stenosen und Aorto-und Speiseröhrenkrebs Fisteln1,13,14,15. Weitere Standorte der Fremdkörper Impaktion sind Pylorus, Duodenum und Ileo-Caecum Kreuzung10.
Durchführung von Röntgenaufnahmen von Hals, Brust und Bauch zu diagnostizieren, vermutete, dass Fremdkörper Verschlucken ist eine weit verbreitete Praxis in den Notaufnahmen5. Unter allen Arten von angesaugte Fremdkörper sind nur 10 % Radio-opak-10. Radiologischen Untersuchungen sind zeitaufwendig und haben ein Risiko der Strahlenbelastung bei Kindern. Präzise topographische Lokalisierung von angesaugte Gegenstände aus Metall ist sehr wichtig, um die Therapie zu führen. Die Einführung der HHMD screening früh in der Triage-Prozess kann helfen, überflüssige Bestellung von Röntgenbildern durch topographisch Lokalisierung des aufgenommenen metallische Fremdkörpers zu mildern.
Die Studie bewertet die Genauigkeit der HHMD MFBs topographische systematisch zu lokalisieren und bestimmt die Wirksamkeit und Durchführbarkeit der HHMD in der pädiatrischen Bevölkerung. Wir prüfen auch, wie gut diese Strahlung kostenlose screening-Tool von Patienten und Eltern angenommen wurde. Zwischen Mai und Juli 2016 wurden alle aufeinander folgenden Patienten, die pädiatrische Notaufnahme mit einer Geschichte von Fremdkörper Verschlucken präsentiert in die Studie einbezogen. Die Einwilligung wurde von den Eltern oder Erziehungsberechtigten. Patienten mit angeblichen Fremdkörper Verschlucken wurden systematisch mit HHMD gescannt. Der Goldstandard der Pflege zur Identifizierung und Lokalisierung von angesaugte metallischen Fremdkörpern führt einfache Röntgenaufnahmen von der zervikalen Bereich, Thorax und Abdomen. Um die Durchführung der Prüfung zu erleichtern, wurde ein standardisierter Workflow (Abbildung 1) erstellt. Ein Log-Buch speziell anhand der Workflow erstellt wurde verwendet, um die ursprünglichen Daten zu sammeln, die anschließend in eine Tabelle (Abbildung 2) transkribiert wurden. Die Daten im Abschnitt Ergebnis werden als die Zahl der Fälle, gefolgt von Prozentzahlen dargestellt.

<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

Münzen sind die am häufigsten aufgenommenen metallische Fremdkörper unter den Kindern. Das Ziel dieses Protokolls ist zur Beurteilung der Genauigkeit und Machbarkeit mit einem handheld metallortungsgerät aufgenommenen metallische Fremdkörper bei Kindern zu erkennen. Wir schlagen vor, dass durch die Einführung von Handheld-Metalldetektor Screening früh in der Triage-Prozess von Kindern mit Verdacht auf metallische Fremdkörper Verschlucken, die Anzahl der Röntgenaufnahmen den metallische Fremdkörper lokalisieren bestellt wird dabei reduziert werden kann Radio-Sensitive Bevölkerung. Das Studienprotokoll erfordert das Screening der Teilnehmer für die Geschichte des Fremdkörpers Aufnahme und Ausschluss von Patienten mit Atemnot oder metallische Implantate. Der Patient ändert sich krankenhemd und Elemente, die wie Brillen, Ohrringe, Anhänger Metall enthalten könnte, und Ornamente werden entfernt. Der Patient befindet sich in der Mitte des Raumes von anderen metallischen Interferenzen. Zuerst bestätigt den Arbeitsstatus des handheld Metalldetektors ein positives Audio-visuellen Signal zu entlocken. Dann erfolgt das Screening in einer aufrechten Position mit Kopf in Verlängerung um den Hals, von der Ebene des Kinns auf die Ebene des Hüftgelenks, die anatomische Bereiche von Hals bis Becken in einer Zick-Zack-Weise sowohl vorn als auch nach hinten decken verfügbar zu machen. Ein positives Audio-visuellen Signal ist aufmerksam zur Kenntnis genommen, während des Scanvorgangs auf das Vorhandensein von metallischen Fremdkörper. Relevanten Röntgenaufnahmen sind gemäß den Bereich auf der Metall-Detektor-Screening erkannt bestellt. Der handheld Metalldetektor konnte alle Münzen unter den aufgenommenen metallische Fremdkörper in unserer Studie genau zu bestimmen. Der handheld Metalldetektor konnte nicht konsequent-Münze metallische Fremdkörper erkennen. Dieses Protokoll zeigt die Genauigkeit der Handheld-Metalldetektor zur Identifizierung und Lokalisierung von Münzen und Münze wie metallische Fremdkörper.

Die Forscher rekrutiert 36 Patienten für diese Beobachtungsstudie. Diese Patienten hatten pädiatrische Notaufnahme mit einer Geschichte von Fremdkörper Verschlucken vorgestellt. Unter diesen waren 28 Patienten metallische Fremdkörper Ingestionen (Abbildung 3). Die übrigen 8 Patienten hatten nicht-metallische Fremdkörper wie Gräten, Satay Stick und Plastikknopf aufgenommen. Münzen waren die häufigste Art des Fremdkörpers eingenommen (Tabelle ...

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

Hier präsentieren wir ein Protokoll, um den handheld Metalldetektor zum Bildschirm für die Anwesenheit von angesaugte metallischen Fremdkörpern bei Kindern verwenden, die an die pädiatrische Notfallmedizin-Abteilung mit einer Geschichte von Fremdkörper Verschlucken zu präsentieren.

Handheld-Metalldetektor-Screening für metallische Fremdkörper Verschlucken bei Kindern

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 ...

handheld-metal-detector-screening-for-metallic-foreign-body-ingestion

Research

JoVE Journal

Medicine

10.6K Aufrufe.  KK women's and Children's Hospital. Hier präsentieren wir ein Protokoll, um den handheld Metalldetektor zum Bildschirm für die Anwesenheit von angesaugte metallischen Fremdkörpern bei Kindern verwenden, die an die pädiatrische Notfallmedizin-Abteilung mit einer Geschichte von Fremdkörper Verschlucken zu präsentieren.

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

Dissection of Human Vitreous Body Elements for Proteomic Analysis

The vitreous is an optically clear, collagenous extracellular matrix that fills the inside of the eye and overlies the retina. 1,2 Abnormal interactions between vitreous substructures and the retina underlie several vitreoretinal diseases, including retinal tear and detachment, macular pucker, macular hole, age-related macular degeneration, vitreomacular traction, proliferative vitreoretinopathy, proliferative diabetic retinopathy, and inherited vitreoretinopathies. 1,2 The molecular composition of the vitreous substructures is not known. Since the vitreous body is transparent with limited surgical access, it has been difficult to study its substructures at the molecular level. We developed a method to separate and preserve these tissues for proteomic and biochemical analysis. The dissection technique in this experimental video shows how to isolate vitreous base, anterior hyaloid, vitreous core, and vitreous cortex from postmortem human eyes. One-dimensional SDS-PAGE analyses of each vitreous component showed that our dissection technique resulted in four unique protein profiles corresponding to each substructure of the human vitreous body. Identification of differentially compartmentalized proteins will reveal candidate molecules underlying various vitreoretinal diseases.

This video shows an effective technique for differentiating and dissecting the various semi-transparent structures of the human vitreous body in post mortem eyes.

Dissection of Human Glaskörper Elemente für Proteomanalyse

The vitreous is an optically clear, collagenous extracellular matrix that fills the inside of the eye and overlies the retina. 1,2 Abnormal interactions ...

Forschung

Medizin

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.

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

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 kleine Kinder mit Autismus

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.

Protokoll für Long Duration Ganzkörper-Hyperthermie in Mäuse

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.

Mess-Cardiac Autonomic Nervous System (ANS) Aktivität bei Kindern

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.

Screening für Melanom Modifiers mit einem Zebrafisch Autochthone Tumormodell

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 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; Gentransfer-Technik und<em&gt; In-vitro-</em&gt; Zellbasierte Assays für das Studium der Knochenverlust in Skeletterkrankungen

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

Calibrated Forceps Model of Spinal Cord Compression Injury

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The calibrated forceps model of compression injury is a convenient, low cost, and very reproducible animal model for SCI. We used a pair of modified forceps in accordance with the method published by Plemel et al. (2008) to laterally compress the spinal cord to a distance of 0.35 mm. In this video, we will demonstrate a dorsal laminectomy to expose the spinal cord, followed by compression of the spinal cord with the modified forceps. In the video, we will also address issues related to the care of paraplegic laboratory animals. This injury model produces mice that exhibit impairment in sensation, as well as impaired hindlimb locomotor function. Furthermore, this method of injury produces consistent aberrations in the pathology of the SCI, as determined by immunohistochemical methods. After watching this video, viewers should be able to determine the necessary supplies and methods for producing SCI of various severities in the mouse for studies on SCI and/or treatments designed to mitigate impairment after injury.

Spinal cord injury models should be highly reproducible. We demonstrate that the calibrated forceps compression model of spinal cord injury is an easy to use surgical method for generating reproducible injuries to the murine spinal cord.

Calibrated Zange Modell Spinal Cord Injury Compression

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The ...

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.

Normothermen<em&gt; Ex Vivo</em&gt; Kidney Perfusion für die Erhaltung der Nierentransplantate vor der Transplantation

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.

Heterotope Renal Autotransplantation in einem Schweine-Modell: Eine Schritt-für-Schritt-Protokoll

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