Os autores não têm confirmações.

Todos os procedimentos foram aprovados pelo comitê de ética da Associação Médica do Estado Renânia-Palatinado e realizada de acordo com a Declaração de Helsinki. 1. Atomizador 4 <ol><li> Remova o conjunto de pacote. Inspecionar o produto para garantir que não haja danos. NOTA: O dispositivo destina-se a um tempo de uso. Por favor, consulte as instruções de utilização para obter mais informações sobre o armazenamento e fornecimento (ver Materiais e Tabela Equipment). </li></ol> 2. Preparação geral do paciente <ol><li> Administrar pré-medicação, por exemplo, Lorazepam 1 - 2,5 mg à noite antes e / ou 2-4 mg 1-2 h antes da intervenção (com atenção aos contra-indicações). </li><li> Estabelecer parâmetros como oximetria de pulso (SpO2), eletrocardiograma (ECG) e pressão arterial não invasiva (PNI) de monitoramento. NOTA: Monitor de sedação do paciente utilizando padrões como Sedação de Ramsay Escala e Bispectmonitoramento Índice ral. Fechar monitorização do nível de sedação pode ajudar a prevenir potenciais complicações, como depressão respiratória. </li><li> Garantir um acesso intravenoso periférico. </li></ol> 3. Anestesia Tópica das vias aéreas superiores <ol><li> Aplicar uma mistura mL de lidocaína a 2% com 0,25% de fenilefrina em cada narina. </li><li> Aplicar lidocaína 10%, duas vezes por pulverização directamente sobre a mucosa da orofaringe, a pulverização a partir da ponta para a parte posterior da língua. Peça para o paciente gargarejar a lidocaína na boca durante o maior tempo possível. NOTA: Anestesia do dispositivo supraglótico pode ser realizada alternativamente, usando cotonetes ou vias aéreas nasofaríngeas, gotas, gel, abanada e gargarejo, aspiração e pulverização anestésicos locais. </li></ol> 4. Analgo-sedação <ol><li> Comece analgo-sedação uso de medicação iv, por exemplo bolus sufentanil inicial (&lt;60 kg: 5 mg;&gt; 60 kg: 10 mg;&gt; 100 kg: 15 mg). NOTA: Dose sedação induzida pela droga no nível mais baixo possível, a fim de evitar a apneia ou vias aéreas constrição. </li><li> Dê um bolus sufentanil iv extra, por exemplo, 5 mg, como tratamento &#39;resgate&#39; até que uma pontuação de Sedação de Ramsay de 2 (cooperativo, orientado, tranquilo) é alcançada. </li></ol> 5. Preparação Equipamento <ol><li> Montar e verificar o endoscópio de intubação flexível de acordo com as normas institucionais. </li><li> Ligue o tubo de oxigênio do atomizador ao medidor de fluxo de oxigênio diretamente. Cuidado! medidores de fluxo de oxigênio com garrafas umidificador não deve ser utilizado. </li><li> Anexar o tubo de ligação à Luer sobre o canal de trabalho do endoscópio. Definir a taxa de fluxo de oxigênio a 10 L / min, como recomendado no manual de instruções. </li><li> Ligue um seringa de 1 mL contendo a solução de lidocaína a 2% para três vias montagem arma. NOTA: A lidocaína 2% com o seu perfil de eficácia favorável é um um adequadoestético. </li></ol> Técnica 6. Atomizador <ol><li> Feche a abertura de controle de fluxo usando um dedo. Rapidamente injetar a medicação. liberte imediatamente a abertura de controle de fluxo. NOTA: A seringa tem de ser ligado à montagem de três vias braço lateral para se obter um fluxo de oxigénio de baixa contínua através do canal de trabalho. </li><li> Enquanto avança o endoscópio flexível nas vias aéreas, injetar o anestésico local usando o atomizador nos seguintes locais: no interior da narina, em narinas posteriores; epiglote; glote; e cordas vocais. </li><li> Repita essas etapas tantas vezes quanto necessário, enquanto avança o endoscópio ( &quot;spray-as-you-go&quot;). Antes de passar através das cordas vocais, esperar por 2 min para permitir a droga efeito suficiente no local. NOTA: monitorar o nível de sedação do paciente e do topo para cima sedação uso de medicação iv (por exemplo, sufentanil 5 mg), conforme necessário. </li></ol> 7. Flexível endoscópica intubação <ol><li> Avançar o Endoscope cuidadosamente através da glote na traqueia e a posição da ponta do endoscópio apenas acima da carina. Estrada de ferro, um tubo endotraqueal adequado tamanho algemado e lubrificado flexível ao longo do endoscópio na traquéia sob rotação. Nota: Evite avanço acidental da ponta do endoscópio em um brônquio principal. </li><li> Confirmar o posicionamento do tubo endotraqueal na traqueia imediatamente acima da carina por controlo visual através do endoscópio. Remover o endoscópio, deixando o tubo endotraqueal no lugar. </li><li> Confirme a posição traqueal do tubo através da medição final CO maré 2 usando capnografia no paciente respirando espontaneamente. </li><li> Induzir a anestesia geral, por exemplo intravenosa de propofol 2 peso mg / kg / corpo. Comece ventilação dos pacientes de acordo com as normas vigentes. </li></ol> NOTA: O conjunto Enk Atomizador não está disponível a partir de 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>

Os autores não têm nada para revelar. O conjunto Enk-Atomizador não está disponível a partir de Cook Medical. De acordo com o inventor (comunicação pessoal) um dispositivo comparável estarão disponíveis no futuro.

Aqui demonstramos anestesia tópica usando um atomizador com alto fluxo de oxigênio para a intubação acordada-endoscópica. O primeiro estudo a avaliar a técnica de atomizador em comparação com bolos clássicos de anestesia tópica, através do canal de trabalho do endoscópio flexível intubating 5. Os pacientes do grupo atomizador avaliado o seu nível de conforto, melhor do que aqueles no grupo de controle e apresentaram menos tosse e episódios de tosse distintas. Além disso, a técnica de atomizad...

A anestesia tópica das vias aéreas por intubação acordada é geralmente recomendado para melhorar o conforto do paciente, o que vai ajudar a tornar o procedimento um sucesso. A técnica de spray-as-you-go com alto fluxo de oxigênio é um conceito simples e seguro que é encontrar um alto nível de aceitação entre os pacientes. A atomização de anestésico local por oxigénio a uma velocidade de fluxo de 10 l / min Resultados em menos estimulação da tosse e gag reflexo. Diante de feedback negativo a partir de vários pacientes, decidimos para refinar a técnica padrão institucional pulverizador-as-you-go. Esta aplicação de bolus clássica de anestésico local consistiu em duas injecções, cada de 5 mL de lidocaína a 2% mais 5 mL de ar em uma seringa de 10 ml, em primeiro lugar para as cordas vocais e, em seguida, depois de um tempo de espera de 2 minutos, para a traqueia. Os pacientes queixaram-se da sensação de ser abafado pelo respingo do bolo golpeando sua mucosa das vias aéreas. A técnica de spray-as-you-goé comumente usado em todo o mundo 1. Esta técnica envolve a aplicação de anestésico local através do canal de trabalho de um endoscópio flexível intubating enquanto avança a ponta através das vias respiratórias superiores e inferiores com o objetivo de entubar a traqueia do paciente. Existem várias abordagens na prática e várias descrições de métodos na literatura. Experiências iniciais com o spray-as-you-go-técnica em combinação com aplicação de um fluxo de oxigênio constante através do canal de trabalho foram publicados na década de 1990 2. O foco aqui foi sobre os efeitos de fluxo de oxigênio como limpar a lente através da dispersão de secreções muco e sangue longe da lente e elevando a oferta de oxigênio inspiratório. A vantagem de o fluxo de oxigénio em ajudar a atomização de anestésico local foi mostrado anteriormente por Piepho e colegas 3. Eles descreveram uma técnica de vaporização, em que o fornecimento de 3 L / min de oxigénio resultou em menos tosse pelos pacientes. Partindo do pressuposto de que umafluxo de 10 L / min seria atomizar o agente local em partículas mais finas que evocam ainda menos tosse, testou-se com sucesso o atomizador a esta taxa superior flor. O atomizador consiste em um volume pequeno,,, tubo de oxigênio certificada pressão flexível kink-resistente e um tubo de ligação. Estes estão ligados a uma de três vias de montagem braço lateral com uma abertura de controlo de fluxo pequeno. A técnica de atomizador pode ser utilizado para todas as situações das vias aéreas. No entanto, o dispositivo deve ser usado com cautela distal da estenose das vias aéreas de maior grau causada por condições tais como massas tumorais. A falta de refluxo de oxigênio pode causar aumento da pressão das vias aéreas e aumenta o risco de barotrauma. Além disso, o atomizador é uma ferramenta útil em situações de emergência,. A preparação do conjunto precisa de menos de um minuto eo usuário não precisa de treinamento especial. A seguir, apresentamos instruções detalhadas para a técnica de spray-as-you-go com alto fluxo de oxigênio, ilustrando a separadaetapas de trabalho.

<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

a vontade do paciente de cooperar é um pré-requisito absoluto para a intubação acordada sucesso da traqueia. Enquanto drogas sedação dos pacientes pode comprometer a sua respiração espontânea, anestesia tópica das vias aéreas é uma técnica popular. A técnica de spray-as-you-go representa uma das oportunidades mais simples para anestesiar a mucosa das vias aéreas. A aplicação de anestésico local através do canal de trabalho do endoscópio flexível é uma prática generalizada para anestesistas, bem como pneumologistas. Não há nem necessidade de dispositivos adicionais, nem treinamento especial como um pré-requisito para executar esta técnica. No entanto, um problema clínico conhecido é o de tosse e náusea reflexo que pode ocorrer quando o anestésico líquido atinge a mucosa das vias aéreas e outras estruturas sensíveis como as cordas vocais. Isto pode ser evitado pelo uso de oxigénio aplicada através do canal de trabalho, com o objectivo de embaciamento do anestésico local em partículas mais finas. além disso, O fluxo de oxigénio proporciona uma oferta maior de oxigênio e contribui para uma melhor visualização, dispersando secreções muco e sangue longe da lente. A utilização de um atomizador com um alto fluxo de oxigénio de 10 L / min, que maximizada estes benefícios, causou menos tosse e tinha pacientes mais satisfeitos, e, por conseguinte, de cooperação. As possíveis complicações, mas muito raros de usar o fluxo de oxigênio incluindo insuflação gástrica, ruptura de órgãos ou barotrauma não surgiu. Atribuímos o uso livre de complicações de alto fluxo de oxigênio para o desenho do conjunto, que permite o fluxo e pressão release.

<html> Foi realizada acordado intubação endoscópica flexível utilizando a técnica de anestesia tópica apresentado em 48 pacientes. Conseguimos intubating todos os pacientes, exceto um, que sofria de um tumor estenosante da faringe, o que tornou impossível avançar o tubo. A qualidade de conforto intubação acordada foi avaliada pelos pacientes, anestesistas e enfermeiros utilizando uma escala analógica visual de 11 pon...

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.

Tópica das vias aéreas Anestesia para intubação acordada-endoscópica Usando o Spray-as-you-go Técnica com fluxo de oxigênio elevado

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 visualizações.  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.

Vídeo: 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.

A Técnica de Resgate Novel para intubação difícil e Ventilação Difícil

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

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.

Diretrizes para Intubação Pediátrica Eletiva Fiberoptic

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 de Fluxo Ensaio Celular para detectar anticorpos em N-metil-D-aspartato ou dopamina-2 Receptor em Soro Humano

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.

Um método melhorado para rápida de intubação da traqueia em Ratos

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.

O uso de um baixo fluxo Sistema de Anestesia Digital para camundongos e ratos

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.

Intubação Orotraqueal Repetida em Camundongos

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.

Quantificação do Fluxo Sanguíneo Fetal Humano com Ressonância Magnética e Compensação de 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.

Projeto e Desenvolvimento de um Modelo para Estudar o Efeito do Oxigênio Suplementar no Microbioma da Fibrose Cística

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.

Caracterização rápida da Viscoelástica do Muco das Vias Aéreas usando um Reômetro Benchtop

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.

Intubação endotraqueal usando um endoscópio de intubação flexível como modelo padronizado para o manejo seguro das vias aéreas em suínos

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