Gli autori ringraziano il Dr. Lee Khai Pin, testa & ospedale Senior Consultant, emergenza bambini, KK donne e bambini, 100 Bukit Timah Road, Singapore per il supporto amministrativo con la produzione di video articolo. Articolo riprodotto con il permesso di "Palmare Metal Detector per metallico stranieri corpo ingestione in emergenza pediatrica"; Agosto 2018:85 (8): 618-624; Augusto Binte Hamzah, Giacomo di Veglia, Suraj Manickam, Sashikumar Ganapathy: copyright The Indian Journal of Pediatrics.

Il protocollo segue le linee guida del Comitato di etica della ricerca umana istituzionale. Il Comitato di revisione istituzionale centrale fornito l'approvazione etica per lo studio.
1. il partecipante di Screening
<ol><li>Reclutare pazienti che presentano al reparto pediatrico di emergenza con storia di ingestione dell'ente straniero.</li>
	<li>Escludere i pazienti che sono nell'afflizione respiratoria e ha impiantato dispositivi come pace-maker.</li>
</ol>2. preparare il paziente
<ol><li>Vestire il paziente in un abito dell'ospedale.</li>
	<li>Rimuovere tutti gli elementi che possono contenere oggetti metallici quali vestiti, occhiali, orecchini, ciondoli e ornamenti prima dell'esame per evitare interferenze con la proiezione di HHMD.</li>
	<li>Posizionare il paziente al centro della stanza lontano da altri potenziali interferenze metallici.</li>
</ol>3. preparare l'esaminatore e Caregivers
<ol><li>Rimuovere tutti gli accessori metallici come gioielli e orologi.</li>
	<li>Richiedere il caregiver per rimuovere tutti gli accessori metallici come gioielli e orologi.</li>
</ol>4. il metal Detector Screening
<ol><li>Utilizzare il HHMD (Figura 1) per condurre lo screening.</li>
	<li>Confermare la condizione di funzionamento dell'HHMD suscitando un positivo segnale audiovisivo fluttuando sopra un pezzo di metallo.</li>
	<li>Eseguire il HHMD scansione in posizione eretta per i bambini più grandi.</li>
	<li>Eseguire il HHMD scansione in posizione eretta per neonati avendo la stiva di caregiver del paziente in modo appropriato.</li>
	<li>Tenere le mani del paziente su e lontano dal corpo.</li>
	<li>Posizionare la testa del paziente in estensione per esporre il collo.</li>
	<li>Scansione dal livello del mento al livello dell'articolazione dell'anca per coprire le zone anatomiche dal collo al bacino.</li>
	<li>Eseguire la scansione in modo a zig-zag attraverso il corpo per garantire che ogni area è stato scansionato.</li>
	<li>Scansione sia anteriormente e posteriormente, come lo spessore del paziente dovrà essere presi in considerazione.</li>
	<li>Osservare attentamente il segnale audio-video positivo durante la scansione per la presenza di MFBs.</li>
</ol>5. eseguire la radiografia pertinente
<ol><li>Ordinare la radiografia pertinente del collo, petto, addome o secondo l'area rilevata dallo screening HHMD per il MFBs.</li>
	<li>Procedere con tutte le viste di raggi x (collo, torace e addome) se il MFB non venne rilevato durante lo screening HHMD e l'indice clinico di sospetto per ingestione dell'ente straniero è alta.</li>
</ol>6. registrare i risultati dei Test
<ol><li>Registrare i risultati del test nel giornale di bordo (Figura 2) creato appositamente sulla basano del flusso di lavoro (Figura 1).</li>
	<li>Correlare il segnale positivo di HHMD audiovisivo con il punto di riferimento ossea sottostante sul corpo, di palpare i seguenti monumenti ossuti prominenti - incavo sternale, xiphisternum, sinfisi pubica e la cresta iliaca bilaterale.</li>
	<li>Registrare il punto di riferimento sul contorno di riferimento anatomico di giornale di bordo prima di procedere con l'esame di raggi 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. 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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>

Gli autori non hanno nulla a rivelare.

Metallico stranieri ingestione del corpo nei bambini è un problema comune e nella maggior parte dei pazienti, questo è diretto conservativamente7,16. Singole o ripetute le radiografie possono esporre la popolazione pediatrica di radio-sensibili al rischio di radiazioni indesiderate durante il tentativo di localizzare il corpo estraneo metallico ingerito. Diversi studi hanno descritto l'uso di rivelatori di metalli a ingestione di MFB nei bambini, con sensibilit...

Ingestione dell'ente straniero è un problema diffuso tra i bambini che presentano al reparto pediatrico di emergenza a causa della natura inquisitive naturale dei lattanti e dei bambini di esplorare gli oggetti da degustazione e inghiottirle1,2. Gli studi precedenti hanno segnalato che oggetti metallici costituivano l'85% di tutte le ingestioni di corpo estraneo in loro coorte di pazienti3,4. L'obiettivo del presente protocollo è quello di valutare l'accuratezza e la possibilità di utilizzare un palmare metal detector per rilevare corpi estranei metallici ingeriti in bambini. Proponiamo che introducendo il metal detector palmare protocollo presto nel processo di valutazione dei bambini con alto sospetto di ingestione di corpo estraneo metallico di screening, il numero delle radiografie essendo ordinato per localizzare il corpo estraneo metallico può essere ridotto in questa popolazione di radio-sensibile.
Metal detector palmare (HHMD) è un'aggiunta di poco costoso, facilmente accessibile che possa aiutare ad accelerare la gestione dei corpi estranei metallici ingeriti. Metal detector è un dispositivo molto a bassa frequenza, che ha un ricevitore e un trasmettitore che rileva il cambiamento nel suo campo elettromagnetico causato dalla presenza di un oggetto metallico nelle sue vicinanze3. Se un oggetto metallico è all'interno del campo, il campo elettromagnetico è interrotta e il sensore farà scattare un allarme audiovisivo sul dispositivo4. L'esame rivelatore di metallo può essere accuratamente eseguita in meno di 2 minuti5. Quindi, lo screening HHMD può essere introdotto all'inizio del processo di valutazione dei bambini con alto sospetto di ingestione di corpo estraneo metallico per la localizzazione topografica.
Le monete sono il più comune ingestione di corpi estranei metallici1,4,6,7. A causa della mancanza di spigoli vivi e natura non tossico generale, la maggior parte delle ingestioni di moneta è conservativamente. Altri tipi di materiali comunemente ingeriti includono magneti, batterie a bottone, piccoli giocattoli, arachidi, gioielli, bottoni, ossa, semi e popcorn8,9. Compressione di corpi estranei si verifica più comunemente nell'esofago in tre siti, cioè sfintere esofageo superiore, metà di-esofago a livello dell'arco aortico e di sfintere esofageo inferiore10. Corpi estranei metallici (MFBs) situati nell'esofago possono essere asintomatici nel 40% dei pazienti11,12 e sua compressione prolungata può condurre alle varie complicazioni come perforazione esofagea, Mediastinite, fistola esofago-tracheale, stenosi tracheale e fistole aorto-esofageo1,13,14,15. Piloro, duodeno e giunzione ileo-caecal10includono altre posizioni di compressione del corpo estraneo.
Esecuzione di radiografie del collo, torace e addome per diagnosticare sospetta ingestione dell'ente straniero è una pratica molto diffusa nei reparti di emergenza5. Tra tutti i tipi di corpi stranieri ingeriti, solo il 10% sono radio-opaco10. I test radiologici sono che richiede tempo e hanno un rischio di esposizione a radiazioni nei bambini. Precisa localizzazione topografica degli oggetti metallici ingeriti è molto fondamentale per guidare la terapia. L'introduzione di screening nelle prime fasi del processo di triage HHMD può contribuire a mitigare estranei l'ordinazione delle radiografie localizzando topograficamente il corpo estraneo metallico ingerito.
Questo studio ha valutato l'accuratezza del HHMD per localizzare MFBs in modo sistematico e topografico e determinato l'efficacia e la fattibilità del HHMD nella popolazione pediatrica locale. Inoltre abbiamo esaminato quanto bene questa radiazione libera strumento di screening è stata accettata dai pazienti e genitori. Tra maggio e luglio 2016, tutti i pazienti consecutivi che hanno presentato al reparto pediatrico di emergenza con una storia di ingestione dell'ente straniero sono stati inclusi nello studio. Consenso informato è stato ottenuto dal genitore o tutore legale. Pazienti con ingestione dell'ente straniero presunta sistematicamente analizzati utilizzando HHMD. Il gold standard di cura per l'identificazione e la localizzazione di corpi estranei metallici ingeriti sta eseguendo le radiografie normali della zona cervicale, torace e addome. Per facilitare lo svolgimento dello studio, un flusso di lavoro standardizzato è stato creato (Figura 1). Un libro di registro appositamente creato in base al flusso di lavoro utilizzato per raccogliere i dati iniziali, che successivamente sono state trascritte in un foglio di calcolo (Figura 2). I dati nella sezione risultato sono raffigurati come il numero di casi seguiti da percentuali.

<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

Le monete sono i corpi estranei metallici ingeriti più comuni tra i bambini. L'obiettivo del presente protocollo è quello di valutare l'accuratezza e la possibilità di utilizzare un palmare metal detector per rilevare corpi estranei metallici ingeriti in bambini. Proponiamo che introducendo lo screening di metal detector palmare presto nel processo di valutazione dei bambini con alto sospetto di ingestione di corpo estraneo metallico, il numero delle radiografie essendo ordinato per localizzare il corpo estraneo metallico può essere ridotto in questo Radio-sensibili popolazione. Il protocollo di studio richiede la selezione dei partecipanti per la storia di ingestione dell'ente straniero e l'esclusione dei pazienti con afflizione respiratoria o gli impianti metallici. Il paziente cambia in abito dell'ospedale e gli elementi che potrebbero contenere metallo come occhiali, orecchini, ciondoli e ornamenti vengono rimossi. Il paziente è posizionato al centro della stanza lontano da altre interferenze metallici. La condizione di funzionamento del palmare metal detector è in primo luogo confermata suscitando un positivo segnale audio-video. Quindi lo screening avviene in posizione eretta con la testa in estensione per esporre il collo, dal livello del mento al livello dell'articolazione dell'anca, per coprire le zone anatomiche dal collo al bacino in un modo a zig-zag sia anteriormente che posteriormente. Un segnale positivo audiovisivo è osservato attentamente durante la scansione per la presenza di corpo estraneo metallico. Le radiografie pertinenti sono ordinate secondo l'area rilevata sullo screening del metal detector. Il metal detector palmare è stato in grado di identificare con precisione tutte le monete tra i corpi estranei metallici ingeriti nel nostro studio. Il metal detector palmare non potrebbe rilevare costantemente non-moneta metallici corpi estranei. Questo protocollo dimostra l'accuratezza dei metal detector palmare nell'identificazione e localizzazione di monete e moneta come corpi estranei metallici.

I ricercatori hanno reclutato 36 pazienti per questo studio d'osservazione. Questi pazienti avévano presentato al reparto pediatrico di emergenza con una storia di ingestione dell'ente straniero. Fra questi, 28 pazienti hanno avuti ingestioni di corpo estraneo metallico (Figura 3). I restanti 8 pazienti avevano ingerito corpi estranei non metallici come lische di pesce, bastone di carne satay e bottone di plastica. Le monete erano il tipo più comune di corp...

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

Qui, presentiamo un protocollo per utilizzare il palmare metal detector, alla schermata per la presenza di corpi estranei metallici ingeriti in bambini che presentano al reparto di medicina d'urgenza pediatrica con una storia di ingestione dell'ente straniero.

Metal Detector palmare Screening per ingestione di corpo estraneo metallico nei bambini

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

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JoVE Journal

Medicine

10.6K Views.  KK women's and Children's Hospital. Qui, presentiamo un protocollo per utilizzare il palmare metal detector, alla schermata per la presenza di corpi estranei metallici ingeriti in bambini che presentano al reparto di medicina d'urgenza pediatrica con una storia di ingestione dell'ente straniero.

Video: 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.

Dissezione di Human Elements corpo vitreo per l&#39;analisi proteomica

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

Ricerca

Medicina

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.

Contenuto di screening ad alto contenuto di Malattie Neurodegenerative

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 bambini piccoli 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.

Protocollo per lungo Ipertermia Corpo intera durata in Mouse

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.

Misurazione cardiaca Sistema Nervoso Autonomo (SNA) Attività nei bambini

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 per Modificatori melanoma utilizzando un modello Zebrafish Tumore autoctone

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; Tecnica Gene Transfer e<em&gt; In vitro</em&gt; test cellulari per lo studio della perdita ossea nei disturbi muscoloscheletrici

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.

Calibrato forcipe Modello di 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.

Normotermica<em&gt; Ex Vivo</em&gt; Rene perfusione per la Preservazione della Kidney Innesti prima del trapianto

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.

Eterotopico renale Autotrapianto in un modello di suino: un protocollo Step-by-Step

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