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Tutte le procedure sono state approvate dal comitato etico della associazione medica dello Stato Renania-Palatinato e eseguite in conformità con la Dichiarazione di Helsinki. 1. atomizzatore 4 <ol><li> Rimuovere il set dalla confezione. Controllare il prodotto per garantire non vi siano danni. NOTA: Il dispositivo è destinato ad essere utilizzato una sola volta. Si prega di fare riferimento alle istruzioni per l&#39;uso per ulteriori informazioni su immagazzinamento e distribuzione (vedi Materiali e attrezzature Tabella). </li></ol> 2. Preparazione generale del paziente <ol><li> Amministrare premedicazione, ad esempio, Lorazepam 1 - 2,5 mg la sera prima e / o 2-4 mg 1 - 2 ore prima dell&#39;intervento (con attenzione ai controindicazioni). </li><li> Stabilire il monitoraggio di parametri quali pulsossimetria (SpO2), elettrocardiogramma (ECG) e la pressione arteriosa non invasiva (PNI). NOTA: Monitor sedazione del paziente con standard quali Ramsay sedazione Scala e Bispectmonitoraggio Index ral. Un attento monitoraggio del livello di sedazione può aiutare a prevenire potenziali complicazioni come la depressione respiratoria. </li><li> Fissare una periferica di accesso per via endovenosa. </li></ol> 3. topica anestesia delle vie aeree superiori <ol><li> Applicare 1 miscela mL di lidocaina 2% con fenilefrina 0,25% per ogni narice. </li><li> Applicare lidocaina 10% spruzzo due volte direttamente sulla mucosa dell&#39;orofaringe, spruzzo dalla punta alla parte posteriore della lingua. Chiedi al paziente di gargarismi lidocaina in bocca a lungo possibile. NOTA: anestesia delle vie respiratorie sopraglottiche può essere effettuata in alternativa con tamponi di cotone con punta o delle vie aeree nasofaringeo, gocce, gel, Swish e gargarismi, aspirando e spruzzare anestetici locali. </li></ol> 4. analgo-sedazione <ol><li> Inizia analgo-sedazione con farmaci IV, ad esempio in bolo sufentanile iniziale (&lt;60 kg: 5 mcg;&gt; 60 kg: 10 mg;&gt; 100 kg: 15 mcg). NOTA: Dose farmaco-indotta la sedazione al livello più basso possibile al fine di evitare l&#39;apnea o costrizione delle vie aeree. </li><li> Dare un ulteriore bolo di sufentanil IV, ad esempio, 5 mg, come trattamento di &#39;salvataggio&#39; fino a quando un Ramsay sedazione punteggio di 2 (cooperativa, orientata, tranquillo) è raggiunto. </li></ol> 5. Apparecchiatura Preparazione <ol><li> Assemblare e verificare l&#39;endoscopio flessibile intubazione secondo gli standard istituzionali. </li><li> Collegare il tubo dell&#39;ossigeno dell&#39;atomizzatore all&#39;ossigeno flussometro direttamente. Attenzione! misuratori di portata di ossigeno con bottiglie umidificatore non devono essere utilizzati. </li><li> Attaccare il tubo di collegamento al Luer sul canale di lavoro dell&#39;endoscopio. Impostare la velocità di flusso di ossigeno al 10 L / min, come raccomandato nel manuale di istruzioni. </li><li> Collegare una siringa da 1 ml contenente la soluzione di lidocaina al 2% a tre vie Raccordo arma da fianco. NOTA: lidocaina 2% con il suo profilo di efficacia favorevole è un un idoneoestetico. </li></ol> Tecnica 6. atomizzatore <ol><li> Chiudere l&#39;apertura di controllo di flusso con un dito. Rapidamente iniettare il farmaco. Rilasciare immediatamente l&#39;apertura di controllo di flusso. NOTA: Una siringa deve essere collegato a tre vie Raccordo sidearm per ottenere un flusso di ossigeno basso continuo attraverso il canale di lavoro. </li><li> Mentre avanza l&#39;endoscopio flessibile nelle vie aeree, iniettare l&#39;anestetico locale utilizzando l&#39;atomizzatore nei seguenti siti: all&#39;interno narice, sulla narici posteriori; epiglottide; glottide; e corde vocali. </li><li> Ripetere questi passaggi numero di volte necessario, mentre avanza l&#39;endoscopio ( &quot;spray-as-you-go&quot;). Prima di passare attraverso le corde vocali, attendere 2 minuti per consentire effetto di droga sufficiente al sito. NOTA: Monitorare livello di sedazione di pazienti e rabboccare la sedazione con farmaci IV (ad es sufentanil 5 mg), a seconda delle necessità. </li></ol> 7. flessibile endoscopica intubazione <ol><li> Far avanzare il endoscope attentamente la glottide nella trachea e la posizione della punta dell&#39;endoscopio appena sopra la carena. Ferrovia un dimensioni, tubo endotracheale adatto ammanettato e lubrificato flessibile sopra l&#39;endoscopio nella trachea in rotazione. NOTA: Evitare l&#39;avanzamento accidentale della punta dell&#39;endoscopio in un bronco fusto principale. </li><li> Confermare il posizionamento del tubo endotracheale nella trachea appena sopra la carena dal controllo visivo attraverso l&#39;endoscopio. Rimuovere l&#39;endoscopio, lasciando il tubo endotracheale sul posto. </li><li> Confermare la posizione endotracheale del tubo misurando fine di CO 2 di marea utilizzando capnografia nel paziente in respiro spontaneo. </li><li> Indurre l&#39;anestesia generale, ad esempio per via endovenosa di Propofol 2 peso mg / kg / corpo. Avviare la ventilazione dei pazienti secondo le norme vigenti. </li></ol> NOTA: Il set di Enk atomizzatore non è attualmente disponibile da Cook Medical.

<ol>
	<li>Simmons, S. T., Schleich, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Airway+regional+anaesthesia+for+awake+fibreoptic+intubation.">Airway regional anaesthesia for awake fibreoptic intubation.</a> Reg Anesth Pain Med. 27, 180-192 (2002).</li><li>Benumof, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Management+of+the+difficult+adult+airway.+With+special+emphasis+on+awake+tracheal+intubation.">Management of the difficult adult airway. With special emphasis on awake tracheal intubation.</a> Anesthesiology. 75, 1087-1110 (1991).</li><li>Piepho, T., Thierbach, A. R., G&#246;bler, S. M., Maybaur, M. O., Werner, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+two+different+techniques+of+fiberoptic+intubation.">Comparison of two different techniques of fiberoptic intubation.</a> Eur J Anaesthesiol. 26, 328-332 (2009).</li><li>Pirlich, N., Lohse, J. A., Schmidtmann, I., Didion, N., Piepho, T., Noppens, R. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparison+of+the+atomizer+with+boluses+of+topical+anaesthesia+for+awake+fibreoptic+intubation.">A comparison of the atomizer with boluses of topical anaesthesia for awake fibreoptic intubation.</a> Anaesthesia. 71, 814-822 (2016).</li><li>Malcharek, M. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+the+Enk+Fibreoptic+Atomizer+with+translaryngeal+injection+for+topical+anaesthesia+for+awake+fibreoptic+intubation+in+patients+at+risk+of+secondary+cervical+injury:+a+randomised+controlled+trial.">Comparison of the Enk Fibreoptic Atomizer with translaryngeal injection for topical anaesthesia for awake fibreoptic intubation in patients at risk of secondary cervical injury: a randomised controlled trial.</a> Eur J Anaesthesiol. 32, 615-623 (2015).</li><li>Williams, K. A., Barker, G. L., Harwood, R. J., Woodall, N. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Combined+nebulization+and+spray-as-you-go+topical+anaesthesia+of+the+airway.">Combined nebulization and spray-as-you-go topical anaesthesia of the airway.</a> Br J Anaesth. 95, 549-553 (2005).</li><li>O'Hare, D. O., Harwood, R., Woodall, N., Barker, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+method+of+administering+topical+anaesthesia+for+flexible+airway+endoscopy.">A method of administering topical anaesthesia for flexible airway endoscopy.</a> Anaesthesia. 55, 616 (2000).</li><li>Long, T. R., Wass, C. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+alternative+to+transtracheal+injection+for+fiberoptic+intubation+in+awake+patients:+A+novel+noninvasive+technique+using+a+standard+multiorifice+epidural+catheter+through+the+bronchoscope+suction+port.">An alternative to transtracheal injection for fiberoptic intubation in awake patients: A novel noninvasive technique using a standard multiorifice epidural catheter through the bronchoscope suction port.</a> Anaesthesiology. 101, 1253 (2004).</li><li>Xue, F. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spray-as-you-go+airway+topical+anesthesia+in+patients+with+a+difficult+airway:+a+randomized,+double-blind+comparison+of+2%+and+4%+lidocaine.">Spray-as-you-go airway topical anesthesia in patients with a difficult airway: a randomized, double-blind comparison of 2% and 4% lidocaine.</a> Anesth Analg. 108, 536-543 (2009).</li><li>Hershey, M. D., Hannenberg, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gastric+distension+and+rupture+from+oxygen+insufflation+during+fiberoptic+intubation.">Gastric distension and rupture from oxygen insufflation during fiberoptic intubation.</a> Anesthesiology. 85, 1479-1480 (1996).</li><li>Ho, C. M., Yin, I. W., Tsou, K. F., Chow, L. H., Tsai, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gastric+rupture+after+awake+fiberoptic+intubation+in+a+patient+with+laryngeal+carcinoma.">Gastric rupture after awake fiberoptic intubation in a patient with laryngeal carcinoma.</a> Br J Anaesth. 94, 856-858 (2005).</li><li>Chapman, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gastric+rupture+and+pneumoperitoneum+caused+by+oxygen+insufflation+via+a+fibreoptic+bronchoscope.">Gastric rupture and pneumoperitoneum caused by oxygen insufflation via a fibreoptic bronchoscope.</a> Anesth Analg. 106, 1592 (2008).</li><li>Ovassapian, A., Mesnick, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oxygen+insufflation+through+the+fiberscope+to+assist+intubation+is+not+recommended.">Oxygen insufflation through the fiberscope to assist intubation is not recommended.</a> Anesthesiology. 87, 183 (1997).</li></ol>

Gli autori non hanno nulla da rivelare. Il set-Enk atomizzatore non è attualmente disponibile da Cook Medical. Secondo l&#39;inventore (comunicazione personale) un dispositivo analogo sarà disponibile in futuro.

Qui mostriamo anestesia topica utilizzando un atomizzatore con alto flusso di ossigeno per l&#39;intubazione da sveglio-endoscopica. Il primo studio per valutare la tecnica atomizzatore confrontato con boli classici di anestesia topica tramite il canale di lavoro dell&#39;endoscopio intubazione flessibile 5. I pazienti nel gruppo atomizzatore valutato il loro livello di comfort migliori rispetto a quelli del gruppo di controllo e sperimentato un minor numero di colpi di tosse e gli episodi di tosse distinti. ...

anestesia topica delle vie aeree per l&#39;intubazione da sveglio è generalmente consigliato per migliorare il comfort del paziente, che contribuirà a rendere la procedura di un successo. La tecnica spray-as-you-go con alto flusso di ossigeno è un concetto semplice e sicura che sta trovando un alto livello di accettazione tra i pazienti. L&#39;atomizzazione dell&#39;anestetico locale ossigeno ad una portata di 10 L / min risultati in meno stimolazione della tosse e gag reflex. Di fronte a un feedback negativo da diversi pazienti, abbiamo deciso di affinare la tecnica istituzionale spray-as-you-go standard. Questa applicazione bolo classica di anestetico locale consisteva di due iniezioni, ciascuna di 5 ml di lidocaina 2% più 5 mL di aria in una siringa da 10 ml, prima nelle corde vocali e poi, dopo un tempo di attesa di 2 min, nella trachea. I pazienti si sono lamentati per la sensazione di essere sommerso da Splash del bolo che colpisce la loro mucosa delle vie aeree. La tecnica spray-as-you-goè comunemente usato in tutto il mondo 1. Questa tecnica comporta l&#39;applicazione di anestetico locale attraverso il canale di lavoro di un endoscopio flessibile intubazione mentre avanza la punta attraverso le vie aeree superiori e inferiori con l&#39;obiettivo di intubazione della trachea del paziente. Ci sono diversi approcci nella pratica e diverse descrizioni dei metodi nella letteratura. Le prime esperienze con lo spray-as-you-go-tecnica in combinazione con l&#39;applicazione di un flusso di ossigeno costante attraverso il canale di lavoro è stato pubblicato nel 1990 2. Il fuoco qui era in effetti dei flussi di ossigeno, come la pulizia della lente disperdendo secrezioni di muco e sangue lontano dall&#39;obiettivo ed elevando la consegna di ossigeno inspiratorio. Il vantaggio del flusso di ossigeno nel favorire atomizzazione di anestetico locale è stato mostrato in precedenza dal Piepho e colleghi 3. Hanno descritto una tecnica di vaporizzazione, dove la fornitura di 3 L / min di ossigeno ha provocato meno tosse dai pazienti. Supponendo che unflusso di 10 L / min sarebbe nebulizzare l&#39;agente locale in particelle più sottili che evocano ancora meno tosse, abbiamo testato con successo l&#39;atomizzatore di questo passo fiore più alto. L&#39;atomizzatore è costituito da un piccolo volume,,, tubo di ossigeno pressione certificata flessibile nodo-resistente e un tubo di collegamento. Questi sono collegati ad un raccordo a tre vie sidearm con una piccola apertura di controllo di flusso. La tecnica atomizzatore può essere utilizzato per tutte le situazioni vie aeree. Tuttavia, il dispositivo deve essere usato con cautela distale dal più alto grado di stenosi delle vie aeree causata da condizioni come masse tumorali. La mancanza di riflusso di ossigeno potrebbe causare un aumento della pressione delle vie aeree e aumenta il rischio di barotrauma. Inoltre, l&#39;atomizzatore è uno strumento utile in un contesto di emergenza. La preparazione del set richiede meno di un minuto e l&#39;utente non ha bisogno di addestramento speciale. Di seguito vi presentiamo le istruzioni operative dettagliate per la tecnica spray-as-you-go con il flusso di ossigeno, che illustra la separatafasi di lavoro.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Lidocaine 10% pump spray&#160;</td>
			<td>e.g. AstraZeneca</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Opiod for i.v. administration</td>
			<td></td>
			<td>Any opiod can be used. We prefer either Sufentanil or Fentanyl</td>
		</tr>
		<tr>
			<td>Enk Fiberoptic Atomizer Set&#160;</td>
			<td>Cook Medical</td>
			<td>Catalogue number: C-EFNS-100&#160;</td>
		</tr>
		<tr>
			<td>Lidocaine 2%&#160;</td>
			<td>e.g. AstraZeneca</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Intubation endoscope</td>
			<td>e.g. Karl Storz Endoscope</td>
			<td>We prefer a fibrescope with an outer diamter of 5 mm and a working channel with a diamter of 2 mm&#160;</td>
		</tr>
		<tr>
			<td>Endotracheal tube</td>
			<td>e.g. R&#252;sch</td>
			<td>Size of the endotracheal tube needs to be adjusted to the patient. We prefer a ID 6.5mm for nasal approach and ID 7.0 mm for oral approach</td>
		</tr>
		<tr>
			<td>Anesthetic drug, e.g. Propofol 1%</td>
			<td>e.g. AstraZeneca</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

topical airway anesthesia for awake-endoscopic intubation using the spray-as-you-go technique with high oxygen flow

la volontà di un paziente di cooperare è un presupposto assoluto per il successo intubazione da sveglio della trachea. Mentre farmaci sedazione dei pazienti può mettere a repentaglio la loro respirazione spontanea, anestesia topica delle vie aeree è una tecnica popolare. La tecnica spray-as-you-go rappresenta una delle opportunità più semplici per anestetizzare la mucosa delle vie aeree. L&#39;applicazione di anestetico locale attraverso il canale di lavoro dell&#39;endoscopio flessibile è prassi diffusa per anestesisti e pulmonologists. Non c&#39;è né necessità di dispositivi aggiuntivi né training speciale come un pre-requisito per eseguire questa tecnica. Tuttavia, un problema clinico noto è la tosse e conati di vomito riflesso che può verificarsi quando l&#39;anestetico liquido colpisce la mucosa delle vie aeree e altre strutture sensibili come le corde vocali. Ciò può essere evitato l&#39;uso di ossigeno applicato attraverso il canale di lavoro al fine di appannamento l&#39;anestetico locale in particelle più fini. per di più, Il flusso di ossigeno fornisce un apporto di ossigeno elevato e contribuisce ad una migliore visione, disperdendo secrezioni di muco e sangue dal lente. L&#39;utilizzo di un atomizzatore con un flusso elevato di ossigeno di 10 L / min abbiamo massimizzato questi vantaggi, ha causato meno tosse e aveva i pazienti più soddisfatti e, pertanto, di cooperazione. Possibile, ma molto rare complicazioni di utilizzare il flusso di ossigeno compresa insufflazione gastrica, la rottura organo o barotrauma non si pone. Attribuiamo l&#39;uso senza complicazione del flusso di ossigeno al disegno dell&#39;insieme, che permette il rilascio di flusso e pressione.

<html> Abbiamo eseguito intubazione endoscopica flessibile utilizzando la tecnica di anestesia topica presentata in 48 pazienti. Siamo riusciti a intubazione tutti i pazienti tranne uno, che soffriva di un tumore stenosante della faringe, che ha reso impossibile per far avanzare il tubo. La qualità del comfort intubazione da sveglio è stata valutata dai pazienti, anestesisti e infermieri che utilizzano una scala analogica vis...

Watch this Scientific Journal Video about Topical Airway Anesthesia for Awake-endoscopic Intubation Using the Spray-as-you-go Technique with High Oxygen Flow at JoVE.com

Topical Airway Anesthesia for Awake-endoscopic Intubation Using the Spray-as-you-go Technique with High Oxygen Flow

The topical anesthetic lidocaine was atomized using a high oxygen flow through the working channel of a flexible intubating endoscope to achieve topical airway anesthesia for awake endotracheal intubation. We prefer this modified spray-as-you-go technique for endoscopic intubation to classical bolus application because of higher patient satisfaction and better compliance.

Topico Airway Anestesia per intubazione da sveglio-endoscopica Utilizzando la Spray-as-you-go Tecnica con alto flusso di ossigeno

A patient&#39;s willingness to cooperate is an absolute precondition for successful awake intubation of the trachea. Whilst drug-sedation of patients can ...

topical-airway-anesthesia-for-awake-endoscopic-intubation-using-spray

Research

JoVE Journal

Medicine

16.5K Views.  University Medical Center of the Johannes Gutenberg-University. The topical anesthetic lidocaine was atomized using a high oxygen flow through the working channel of a flexible intubating endoscope to achieve topical airway anesthesia for awake endotracheal intubation. We prefer this modified spray-as-you-go technique for endoscopic intubation to classical bolus application because of higher patient satisfaction and better compliance.

Video: Topical Airway Anesthesia for Awake-endoscopic Intubation Using the Spray-as-you-go Technique with High Oxygen Flow

A Novel Rescue Technique for Difficult Intubation and Difficult Ventilation

We describe a novel non surgical technique to maintain oxygenation and ventilation in a case of difficult intubation and difficult ventilation, which works especially well with poor mask fit.
Can not intubate, can not ventilate" (CICV) is a potentially life threatening situation. In this video we present a simulation of the technique we used in a case of CICV where oxygenation and ventilation were maintained by inserting an endotracheal tube (ETT) nasally down to the level of the naso-pharynx while sealing the mouth and nares for successful positive pressure ventilation.
A 13 year old patient was taken to the operating room for incision and drainage of a neck abcess and direct laryngobronchoscopy. After preoxygenation, anesthesia was induced intravenously. Mask ventilation was found to be extremely difficult because of the swelling of the soft tissue. The face mask could not fit properly on the face due to significant facial swelling as well. A direct laryngoscopy was attempted with no visualization of the larynx. Oxygen saturation was difficult to maintain, with saturations falling to 80%. In order to oxygenate and ventilate the patient, an endotracheal tube was then inserted nasally after nasal spray with nasal decongestant and lubricant. The tube was pushed gently and blindly into the hypopharynx. The mouth and nose of the patient were sealed by hand and positive pressure ventilation was possible with 100% O2 with good oxygen saturation during that period of time. Once the patient was stable and well sedated, a rigid bronchoscope was introduced by the otolaryngologist showing extensive subglottic and epiglottic edema, and a mass effect from the abscess, contributing to the airway compromise. The airway was secured with an ETT tube by the otolaryngologist.This video will show a simulation of the technique on a patient undergoing general anesthesia for dental restorations.

We describe a technique to maintain oxygenation and ventilation using an endotracheal tube inserted nasally to the level of the naso-pharynx while sealing the mouth and nares for successful positive pressure ventilation.

Una nuova tecnica di soccorso per l&#39;intubazione e ventilazione Difficile Difficile

We describe a novel non surgical technique to maintain oxygenation and ventilation in a case of difficult intubation and difficult ventilation, which ...

Ricerca

Medicina

Guidelines for Elective Pediatric Fiberoptic Intubation

Fiberoptic intubation in pediatric patients is often required especially in difficult airways of syndromic patients i.e. Pierre Robin Syndrome. Small babies will desaturate very quickly if ventilation is interrupted mainly to high metabolic rate. We describe guidelines to perform a safe fiberoptic intubation while maintaining spontaneous breathing throughout the procedure. Steps requiring the use of propofol pump, fentanyl, glycopyrrolate, red rubber catheter, metal insuflation hook, afrin, lubricant and lidocaine spray are shown.

We describe guidelines to perform a safe and efficient elective fiberoptic intubation in pediatric patients while maintaining spontaneous ventilation.

Linee guida per elettive intubazione pediatrica a fibre ottiche

Fiberoptic intubation in pediatric patients is often required especially in difficult airways of syndromic patients i.e. Pierre Robin Syndrome. Small ...

High-throughput Flow Cytometry Cell-based Assay to Detect Antibodies to N-Methyl-D-aspartate Receptor or Dopamine-2 Receptor in Human Serum

Over the recent years, antibodies against surface and conformational proteins involved in neurotransmission have been detected in autoimmune CNS diseases in children and adults. These antibodies have been used to guide diagnosis and treatment. Cell-based assays have improved the detection of antibodies in patient serum. They are based on the surface expression of brain antigens on eukaryotic cells, which are then incubated with diluted patient sera followed by fluorochrome-conjugated secondary antibodies. After washing, secondary antibody binding is then analyzed by flow cytometry. Our group has developed a high-throughput flow cytometry live cell-based assay to reliably detect antibodies against specific neurotransmitter receptors. This flow cytometry method is straight forward, quantitative, efficient, and the use of a high-throughput sampler system allows for large patient cohorts to be easily assayed in a short space of time. Additionally, this cell-based assay can be easily adapted to detect antibodies to many different antigenic targets, both from the central nervous system and periphery. Discovering additional novel antibody biomarkers will enable prompt and accurate diagnosis and improve treatment of immune-mediated disorders.

Over the recent years, live cell-based assays have been used successfully to detect antibodies against surface and conformational antigens. Here, we describe a method using high-throughput flow cytometry enabling the analysis of large cohorts of patients. Detection of novel antibodies will improve diagnosis and treatment of immune-mediated disorders.

High-throughput Citometria a Flusso test cellulare per individuare gli anticorpi per N-metil-D-aspartato Receptor o dopamina-2 recettore nel siero umano

Over the recent years, antibodies against surface and conformational proteins involved in neurotransmission have been detected in autoimmune CNS diseases ...

An Improved Method for Rapid Intubation of the Trachea in Mice

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity is a commonly used model to study both acute lung injury and fibrosis, and multiple methods have been developed for administering bleomycin (and other toxic agents) into the lungs. However, many of these approaches, such as transtracheal instillation, have inherent drawbacks, including the need for strong anesthetics and survival surgery. This paper reports a quick, reproducible method of intratracheal intubation that involves mild inhaled anesthesia, visualization of the trachea, and the use of a surrogate spirometer to confirm exposure. As a proof of concept, 8-12 week old C57BL/6 mice were administered either 2.0 U/kg of bleomycin or an equivalent volume of PBS, and both damage and fibrotic endpoints were measured post-exposure. This procedure allows researchers to treat a large cohort of mice in a relatively short period with little expense and minimal post-procedure care.

This article presents a rapid and simple method for administering bleomycin directly into the mouse trachea via intubation. Key advantages of this method are that it is highly reproducible, easy to master, and does not require specialized equipment or lengthy recovery times.

Un metodo migliore per Rapid intubazione della trachea nei topi

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity ...

Use of a Low-flow Digital Anesthesia System for Mice and Rats

A traditional vaporizer depends on flowing gas and atmospheric pressure for passive anesthetic vaporization. Newly developed direct injection vaporizers utilize a syringe pump to directly administer volatile anesthetics into a gas stream. Unlike a traditional vaporizer, it can be used at very low flow rates, making it ideal for use on mice and rats. 
The equipment's capability to use low flow rates could result in a substantial cost savings due to the reduced need for anesthetic agents, compressed gas, and charcoal scavenging filters1. A lower flow rate means less waste of anesthetic gas and likely reduces the risk of anesthetic exposure to laboratory personnel. Thus, the high levels of precision and safety associated with direct injection vaporizers, along with a reduced need for anesthetic agents, compressed gas, and charcoal filters are beneficial for research requiring small animal anesthesia. 
The goal of this protocol is to demonstrate the use of a syringe-driven direct injection vaporizer as part of a digital, low-flow anesthesia system. The direct injection vaporizer is capable of accurately delivering anesthesia at very low flow rates compared to a traditional vaporizer, making it a promising alternative for controlled gas anesthetic delivery to rodents.

Here, we present a protocol to more safely and efficiently administer anesthetic gas to mice using a digital, low flow anesthesia system utilizing a syringe-driven direct injection vaporizer.

L&#39;utilizzo di un basso flusso di sistema per anestesia digitale per topi e ratti

A traditional vaporizer depends on flowing gas and atmospheric pressure for passive anesthetic vaporization. Newly developed direct injection vaporizers ...

Repeated Orotracheal Intubation in Mice

The literature describes several methods for mouse intubation that either require visualization of the glottis through the oral cavity or incision in the ventral neck for direct confirmation of cannula placement in the trachea. The relative difficulty or the tissue trauma induced to the subject by such procedures can be an impediment to an investigator&#8217;s ability to perform longitudinal studies. This article illustrates a technique in which physical manipulation of the mouse following the use of a depilatory to remove hair from the ventral neck permits transcutaneous visualization of the trachea for orotracheal intubation regardless of degree of skin pigmentation. This method is innocuous to the subject and easily achieved with a limited understanding of murine anatomy. This refined approach facilitates repeated intubation, which may be necessary for monitoring progression of disease or instillation of treatments. Using this method may result in a reduction of the number of animals and technical skill required to measure lung function in mouse models of respiratory disease.

The goal of this article is to describe a refined method of intubation of the laboratory mouse. The method is noninvasive and, therefore, ideal for studies that require serial monitoring of respiratory function and/or instillation of treatments into the lung.

Intubazione orotrachea ripetuta nei topi

The literature describes several methods for mouse intubation that either require visualization of the glottis through the oral cavity or incision in the ...

Human Fetal Blood Flow Quantification with Magnetic Resonance Imaging and Motion Compensation

Magnetic resonance imaging (MRI) is an important tool for the clinical assessment of cardiovascular morphology and heart function. It is also the recognized standard-of-care for blood flow quantification based on phase contrast MRI. While such measurement of blood flow has been possible in adults for decades, methods to extend this capability to fetal blood flow have only recently been developed.
Fetal blood flow quantification in major vessels is important for monitoring fetal pathologies such as congenital heart disease (CHD) and fetal growth restriction (FGR). CHD causes alterations in the cardiac structure and vasculature that change the course of blood in the fetus. In FGR, the path of blood flow is altered through the dilation of shunts such that the oxygenated blood supply to the brain is increased. Blood flow quantification enables assessment of the severity of the fetal pathology, which in turn allows for suitable&#160;in utero&#160;patient management and planning for postnatal care.
The primary challenges of applying phase contrast MRI to the human fetus include small blood vessel size, high fetal heart rate, potential MRI data corruption due to maternal respiration, unpredictable fetal movements, and lack of conventional cardiac gating methods to synchronize data acquisition. Here, we describe recent technical developments from our lab that have enabled the quantification of fetal blood flow using phase contrast MRI, including advances in accelerated imaging, motion compensation, and cardiac gating.

Here we present a protocol for measuring fetal blood flow rapidly with MRI and retrospectively performing motion correction and cardiac gating.

Quantificazione del flusso sanguigno fetale umano con risonanza magnetica e compensazione del movimento

Magnetic resonance imaging (MRI) is an important tool for the clinical assessment of cardiovascular morphology and heart function. It is also the ...

Design and Development of a Model to Study the Effect of Supplemental Oxygen on the Cystic Fibrosis Airway Microbiome

Airway microbial communities are thought to play an important role in the progression of cystic fibrosis (CF) and other chronic pulmonary diseases. Microbes have traditionally been classified based on their ability to use or tolerate oxygen. Supplemental oxygen is a common medical therapy administered to people with cystic fibrosis (pwCF); however, existing studies on oxygen and the airway microbiome have focused on how hypoxia (low oxygen) rather than hyperoxia (high oxygen) affects the predominantly aerobic and facultative anaerobic lung microbial communities. To address this critical knowledge gap, this protocol was developed using an artificial sputum medium that mimics the composition of sputum from pwCF. The use of filter sterilization, which yields a transparent medium, allows optical methods to follow the growth of single-celled microbes in suspension cultures. To create hyperoxic conditions, this model system takes advantage of established anaerobic culturing techniques to study hyperoxic conditions; instead of removing oxygen, oxygen is added to cultures by daily sparging of serum bottles with a mixture of compressed oxygen and air. Sputum from 50 pwCF underwent daily sparging for a 72-h period to verify the ability of this model to maintain differential oxygen conditions. Shotgun metagenomic sequencing was performed on cultured and uncultured sputum samples from 11 pwCF to verify the ability of this medium to support the growth of commensal and pathogenic microbes commonly found in cystic fibrosis sputum. Growth curves were obtained from 112 isolates obtained from pwCF to verify the ability of this artificial sputum medium to support the growth of common cystic fibrosis pathogens. We find that this model can culture a wide variety of pathogens and commensals in CF sputum, recovers a community highly similar to uncultured sputum under normoxic conditions, and creates different culture phenotypes under varying oxygen conditions. This new approach might lead to a better understanding of unanticipated effects induced by the use of oxygen in pwCF on airway microbial communities and common respiratory pathogens.

The goal of this protocol is to develop a model system for the effect of hyperoxia on cystic fibrosis airway microbial communities. Artificial sputum medium emulates the composition of sputum, and hyperoxic culture conditions model the effects of supplemental oxygen on lung microbial communities.

Progettazione e sviluppo di un modello per studiare l'effetto dell'ossigeno supplementare sul microbioma delle vie aeree della fibrosi cistica

Airway microbial communities are thought to play an important role in the progression of cystic fibrosis (CF) and other chronic pulmonary diseases. ...

Rapid Viscoelastic Characterization of Airway Mucus Using a Benchtop Rheometer

In muco-obstructive lung diseases (e.g., asthma, chronic obstructive pulmonary disease, cystic fibrosis) and other respiratory conditions (e.g., viral/bacterial infections), mucus biophysical properties are altered by goblet cell hypersecretion, airway dehydration, oxidative stress, and the presence of extracellular DNA. Previous studies showed that sputum viscoelasticity correlated with pulmonary function and that treatments affecting sputum rheology (e.g., mucolytics) can result in remarkable clinical benefits. In general, rheological measurements of non-Newtonian fluids employ elaborate, time-consuming approaches (e.g., parallel/cone-plate rheometers and/or microbead particle tracking) that require extensive training to perform the assay and interpret the data. This study tested the reliability, reproducibility, and sensitivity of Rheomuco, a user-friendly benchtop device that is designed to perform rapid measurements using dynamic oscillation with a shear-strain sweep to provide linear viscoelastic moduli (G&#39;, G&#34;, G*, and tan &#948;) and gel point characteristics (&#947;c&#160;and &#963;c) for clinical samples within 5 min. Device performance was validated using different concentrations of a mucus simulant, 8 MDa polyethylene oxide (PEO), and against traditional bulk rheology measurements. A clinical isolate harvested from an intubated patient with status asthmaticus (SA) was then assessed in triplicate measurements and the coefficient of variation between measurements is &#60;10%. Ex vivo use of a potent mucus reducing agent, TCEP, on SA mucus resulted in a five-fold decrease in elastic modulus and a change toward a more &#34;liquid-like&#34; behavior overall (e.g., higher tan &#948;). Together, these results demonstrate that the tested benchtop rheometer can make reliable measures of mucus viscoelasticity in clinical and research settings. In summary, the described protocol could be used to explore the effects of mucoactive drugs (e.g., rhDNase, N-acetyl cysteine) onsite to adapt treatment on a case-by-case basis, or in preclinical studies of novel compounds.

The viscoelastic properties of mucus play a critical role in mucociliary clearance. However, traditional mucus rheological techniques require complex and time-consuming approaches. This study provides a detailed protocol for the use of a benchtop rheometer that can rapidly and reliably perform viscoelastic measurements.

Caratterizzazione viscoelastica rapida del muco delle vie aeree utilizzando un reometro da banco

In muco-obstructive lung diseases (e.g., asthma, chronic obstructive pulmonary disease, cystic fibrosis) and other respiratory conditions (e.g., ...

Endotracheal Intubation Using a Flexible Intubation Endoscope as a Standardized Model for Safe Airway Management in Swine

Endotracheal intubation is often a basic requirement for translational research in porcine models for various interventions that require a secured airway or high ventilation pressures. Endotracheal intubation is a challenging skill,&#160;requiring a minimum number of successful endotracheal intubations to achieve a high success rate under optimal conditions, which is often unachievable for non-anaesthesiology researchers. Due to the specific porcine airway anatomy, a difficult airway can usually be assumed. The impossibility of establishing a secure airway can result in injury, adverse events, or death of the laboratory animal. Using a prospective, randomized, controlled evaluation approach, it has been shown that fiberoptic-assisted endotracheal intubation takes longer but has a higher first-pass success rate than conventional intubation without causing clinically relevant drops in oxygen saturation. This model presents a standardized regimen for endoscopically guided endotracheal intubation, providing a secured airway, especially for researchers who are inexperienced in the technique of endotracheal intubation via direct laryngoscopy. This procedure is expected to minimize animal suffering and unnecessary animal losses.

The use of pigs in research has increased in recent years. Nevertheless, pigs are characterized by difficult airway anatomy. By demonstrating how to perform endoscopically guided endotracheal intubation, the present protocol aims to further increase laboratory animals' safety to avoid animal suffering and unnecessary death.

Intubazione endotracheale utilizzando un endoscopio per intubazione flessibile come modello standardizzato per la gestione sicura delle vie aeree nei suini

Endotracheal intubation is often a basic requirement for translational research in porcine models for various interventions that require a secured airway ...

Topico Airway Anestesia per intubazione da sveglio-endoscopica Utilizzando la Spray-as-you-go Tecnica con alto flusso di ossigeno

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