Suportado pelo NIH HL-10342.

1. Preparando-se para o Procedimento Deve-se primeiro obter e preparar os seguintes itens: <ol><li> A cânula. Para intubação de 20-35 ratinhos g, usamos uma polegada de 1 ou 1,5 de comprimento, 20 gauge cateter IV (BD Insylte, Sparks, MD ou Jelco Optiva, Carlsbad, CA). Um novo cateter estéril pode ser usada para cada rato, mas cateteres também pode ser reutilizado depois da esterilização por imersão em etanol a 70% durante a noite. Embora nem faringe nem traqueia do mouse é estéreis procedimentos adequados de limpeza, incluindo a utilização de luvas estéreis e instrumentos, devem ser seguidas. </li><li> O cabo de fibra ótica. Usamos ≈ 70 cm de 0,5 mm de cabo óptico de Edmund Optics, mas o comprimento não é crítica. É importante para garantir que a fibra tem a sua aresta suavizada, uma vez que após o corte do cabo de comprimento com uma lâmina de barbear, a borda é deixada relativamente acentuada, e não é preciso muito esforço para perfurar a parede traqueal. No entanto, is muito fácil de suavizar esta borda, segurando a fibra de cerca de 2 cm da extremidade e, em seguida, fazendo pequenos círculos durante alguns segundos, com as extremidades da ponta de tocar um pedaço de papel de lixa 1000 grit (ver demonstração no vídeo e na Figura 1 de Macdonald et al. 10). A outra extremidade é introduzido através de uma rolha de borracha. Isto é mais facilmente conseguido, em primeiro lugar empurrando uma agulha de calibre 18 através da tampa, a inserção da fibra óptica através do furo da agulha, depois de retirar a agulha. A rolha de borracha é ligada a uma fonte de halogénio de 150 watt de luz (por exemplo, NCL-150, Volpi EUA, ou qualquer outra fonte de luz, ou, ainda menos do que 150 Watts). É importante certificar-se de usar uma tampa de borracha de silicone (ou material de calor resistente), uma vez que a borracha comum ou de cortiça pode queimar quando localizado tão perto da fonte quente luz. </li></ol> 2. Realizar a intubação <ol><li> Ver Figuras 1 e 2. Insira cabo de fibra óptica through um pequeno pedaço de tubo de borracha de silicone (≈ 0,8 milímetros ID x 4 mm de diâmetro externo, Cole-Palmer, EW-96410-13). Amarre este tubo de borracha bastante apertado, enquanto ainda permitindo que o cabo de fibra óptica a ser ajustado. Inserir o tubo de silicone firmemente na extremidade da cânula Luer corrige a posição do cabo de fibra óptica no interior da cânula. Ajustar a posição do cabo de fibra óptica de modo que ela se estende através da cânula ≈ 4 mm à frente da ponta da cânula. </li><li> Colocar o rato anestesiado num suporte vertical, suspenso pelos seus incisivos superiores (Figura 3). A maioria dos investigadores encontrar a melhor visualização com o lado ventral do rato enfrentam-se. Muito gentilmente puxar a língua para fora e segure com o polegar eo indicador. O dedo médio é colocada entre o pescoço e o suporte de plástico. Tração da língua com o dedo indicador e polegar é utilizado para abrir a boca, e para endireitar o caminho de intubação, o ângulo da cabeça é ajustada com o dedo médio por trás do pescoçomostrado na Figura 3. </li><li> Utilizando o cabo de fibra óptica como uma fonte de luz e do introdutor, empurrá-lo através das cordas vocais visualizadas. Se os cabos não são visíveis, puxe mais sobre a língua usando o dedo médio como apoio. Quando inserido, o avanço da cânula ≈ 5 mm mais. Então, não sendo muito cuidadoso para mover a cânula, retire o cabo de fibra óptica. Deite-se o rato para baixo e segurar a cânula com um pedaço de fita adesiva e apoiar o cubo da cânula sobre um pedaço de plastilina (argila de modelagem), como mostrado na Figura 4. </li><li> O processo no passo 3 não pode ser facilmente ensinado ou mesmo demonstrada, uma vez que é uma operação de solo. No entanto, por ajustes sutis da tração sobre a língua e com o apoio atrás da cabeça quase todos que tentar isso em breve encontrar o caminho certo para posicionar o mouse para visualizar as cordas vocais. </li></ol>

<ol>
	<li>Brown, R. H., Walters, D. M., Greenberg, R. S., Mitzner, W. <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+endotracheal+intubation+and+pulmonary+functional+assessment+for+repeated+studies+in+mice.">A method of endotracheal intubation and pulmonary functional assessment for repeated studies in mice.</a> J. Appl. Physiol. 87, 2362-2365 (1999).</li><li>Walters, D. M., Wills-Karp, M., Mitzner, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+cellular+profile+and+lung+function+with+repeated+bronchoalveolar+lavage+in+individual+mice.">Assessment of cellular profile and lung function with repeated bronchoalveolar lavage in individual mice.</a> Physiol. Genomics. 2, 29-36 (2000).</li><li>Rivera, B., Miller, S., Brown, E., Price, 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+novel+method+for+endotracheal+intubation+of+mice+and+rats+used+in+imaging+studies.">A novel method for endotracheal intubation of mice and rats used in imaging studies.</a> Contemporary Topics in Laboratory Animal Science / American Association for Laboratory Animal Science. 44, 52-55 (2005).</li><li>Hamacher, 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=Microscopic+wire+guide-based+orotracheal+mouse+intubation:+description,+evaluation+and+comparison+with+transillumination.">Microscopic wire guide-based orotracheal mouse intubation: description, evaluation and comparison with transillumination.</a> Laboratory Animals. 42, 222-230 (2008).</li><li>Spoelstra, E. N., 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=A+novel+and+simple+method+for+endotracheal+intubation+of+mice.">A novel and simple method for endotracheal intubation of mice.</a> Laboratory Animals. 41, 128-135 (2007).</li><li>Zhao, X., 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=A+technique+for+retrograde+intubation+in+mice.">A technique for retrograde intubation in mice.</a> Lab Animal. 35, 39-42 (2006).</li><li>Vergari, A., 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=A+new+method+of+orotracheal+intubation+in+mice.">A new method of orotracheal intubation in mice.</a> European Review for Medical and Pharmacological Sciences. 8, 103-106 (2004).</li><li>Vergari, A., Polito, A., Musumeci, M., Palazzesi, S., Marano, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Video-assisted+orotracheal+intubation+in+mice.">Video-assisted orotracheal intubation in mice.</a> Laboratory Animals. 37, 204-206 (2003).</li><li>Hastings, R. H., Summers-Torres, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+Laryngoscopy+in+Mice.">Direct Laryngoscopy in Mice.</a> Contemporary Topics in Laboratory Animal Science / American Association for Laboratory Animal Science. 38, 33-35 (1999).</li><li>MacDonald, K. D., Chang, H. Y., Mitzner, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+improved+simple+method+of+mouse+lung+intubation.">An improved simple method of mouse lung intubation.</a> J. Appl. Physiol. 106, 984-987 (2009).</li><li>Ewart, S. L., Levitt, R. C., Mitzner, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Respiratory+system+mechanics+in+mice+measured+by+end-inflation+occlusion.">Respiratory system mechanics in mice measured by end-inflation occlusion.</a> Journal of Applied Physiology. 79, 560-566 (1995).</li><li>MacDonald, K. D., McKenzie, K. R., Mitzner, W., Zeitlin, P. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lung+Mechanics+in+Heterozygous+CF+Mice+after+Repeated+LPS+Dosing.">Lung Mechanics in Heterozygous CF Mice after Repeated LPS Dosing.</a> Am. J. Respir. Crit. Care Med. 175 (4), A930 (2007).</li></ol>

Nenhum dos autores tem quaisquer conflitos de interesse de divulgar.

O procedimento aqui descrito tem várias vantagens. Primeiro, o aparelho é simples e relativamente barata .. A fabricação do aparelho não necessita de quaisquer ferramentas especiais ou equipamento dispendioso. A utilização de um cateter que apresenta também é a fonte de luz quer dizer que nunca perde-se a abertura traqueal como o introdutor se aproxima da abertura traqueal. O uso de um introdutor de 0,5 mm, também serve para minimizar o trauma que pode ocorrer com uma inserção inicial de uma cânula maior. N...

Em 1999, Brown et al. Publicado um artigo científico que descreve um método para intubação de pulmão de rato 1. Tal técnica tem utilidade considerável em fazer função de repetição pulmonar ou lavado bronco-alveolar em camundongos individuais em estudos longitudinais 2. Desde que o papel original, houve vários outros documentos que descrevem abordagens diferentes para intubação rato 3-9. Embora todos estes métodos podem ser utilizados com sucesso, eles normalmente requerem formação considerável ou custo. Um dos principais problemas para intubação tal é que, como as abordagens cânula de intubação se aproxima a inserção traquéia pendente, a cânula própria bloqueia a luz e, portanto, a visualização de onde ele precisa ir. Assim, a inserção se torna cego no momento mais crítico. Neste artigo vamos mostrar como simples e barata eliminar este problema de visualização, garantindo assim a intubação bem sucedida com relativamente pouco treinamento ouexperiência.

<table border="1"><tbody><tr><td> Material </td><td> Companhia </td><td> Catalogo </td><td> Comentário </td></tr><tr><td> Cânula de intubação </td><td> BD Insylte, Sparks, MD ou Jelco Optiva, Carlsbad, CA </td><td></td><td> 1-pol de comprimento, 20-gauge cateter intravenoso (IV) </td></tr><tr><td> Cabo de fibra óptica </td><td> Edmund Optics, Barrington, NJ </td><td> # NT02-542 </td><td> Cerca de 2 pés de comprimento de 0,5 mm de fibra óptica (Comunicação fibra plástica grau). A aresta da extremidade da fibra, que é inserido na traqueia deve ser suavemente arredondada, segurando a fibra ≈ 2 cm da extremidade e, em seguida, fazendo pequenos círculos ao arrastar a ponta por alguns segundos em 1.000 lixa de esmeril. </td></tr><tr><td> Fonte de Luz </td><td> Volpi </td><td> NCL-150 </td><td> Apesar de usar um 150-W fonte de luz halógena, qualquer fonte de luz equivalente, mesmo com potência muito mais baixapode ser usado. </td></tr><tr><td> Rolha de borracha de silicone </td><td></td><td></td><td> Uma rolha de borracha de silicone n º 1 se ajusta a fonte de luz usada nesta demonstração. Diferentes fontes de luz podem requerer um tamanho diferente. </td></tr><tr><td> Tubo de borracha de silicone pequena </td><td> Cole-Palmer </td><td> EW-96410-13 </td><td> Um pedaço ≈ 1,5 cm de tubo de borracha de silicone (0,8 de diâmetro interno de 4 mm de diâmetro exterior) </td></tr><tr><td> Pé de apoio angular </td><td></td><td></td><td> A nossa é constucted de plexiglass, mas qualquer material ao qual um fio ou fio pode ser afixada para segurar o rato a um ângulo quase vertical pode ser utilizado. </td></tr></tbody></table>

a simple method of mouse lung intubation

Um procedimento simples para entubar ratos para medições de função pulmonar teria várias vantagens em estudos longitudinais com um número limitado de animais ou caro. Uma das razões que isso não é feito rotineiramente mais é que é relativamente difícil, apesar de haver vários estudos publicados que descrevem formas de o conseguir. Neste artigo é demonstrar um procedimento que elimina um dos principais obstáculos associados com este intubação, que de visualizar a traqueia durante todo o tempo de intubação. A abordagem utiliza uma fonte de luz de 0,5 milímetros de fibra óptica, que serve como um introdutor para dirigir a cânula de intubação para dentro da traqueia do rato. Mostra-se que é possível usar este procedimento para medir a mecânica pulmonar em ratos individuais ao longo de um curso de tempo de pelo menos várias semanas. A técnica pode ser configurado com despesa relativamente pequena e experiência, e pode ser realizado rotineiramente com relativamente pouco treinamento. Isto deve makÊ possível para qualquer laboratório para realizar rotineiramente este intubação, permitindo assim estudos longitudinais em ratinhos individuais, minimizando assim o número de ratinhos necessários e aumentar o poder estatístico usando cada rato como seu próprio controlo.

Como uma avaliação do método, utilizou-se quatro 20 semanas de idade BALB / c fêmeas com peso médio (± SEM) de 27,7 ± 0,40 g. Eles foram estudados em cinco semanas consecutivas, em que a resistência pulmonar foi medida utilizando um sistema tal como foi previamente descrito 11. Cada rato foi anestesiado com cetamina (100 mg / g de peso corporal) e xilazina (15 mg / g de peso corporal) em solução salina por injecção IP. Eles foram então entubados, como descrito acima. Se houver alguma dúvida sobr...

Watch this Scientific Journal Video about A Simple Method of Mouse Lung Intubation at JoVE.com

A Simple Method of Mouse Lung Intubation

Este artigo descreve um método striaghforward e eficiente de entubar ratos para medições de função pulmonar ou instilação pulmonar, que permite que os ratos a se recuperar e ser estudado em momentos posteriores. O procedimento envolve uma fonte de luz de fibra óptica de baixo custo que ilumina diretamente a traquéia.

Um método simples de intubação Lung Rato

A  simple procedure to intubate mice for pulmonary function measurements would  have several advantages in longitudinal studies with limited numbers or  ...

Research

JoVE Journal

Medicine

50.5K visualizações.  Johns Hopkins Bloomberg School of Public Health. Este artigo descreve um método striaghforward e eficiente de entubar ratos para medições de função pulmonar ou instilação pulmonar, que permite que os ratos a se recuperar e ser estudado em momentos posteriores. O procedimento envolve uma fonte de luz de fibra óptica de baixo custo que ilumina diretamente a traquéia.

Vídeo: A Simple Method of Mouse Lung Intubation

A Model of Disturbed Flow-Induced Atherosclerosis in Mouse Carotid Artery by Partial Ligation and a Simple Method of RNA Isolation from Carotid Endothelium

Despite the well-known close association, direct evidence linking disturbed flow to atherogenesis has been lacking. We have recently used a modified version of carotid partial ligation methods [1,2] to show that it acutely induces low and oscillatory flow conditions, two key characteristics of disturbed flow, in the mouse common carotid artery. Using this model, we have provided direct evidence that disturbed flow indeed leads to rapid and robust atherosclerosis development in Apolipoprotein E knockout mouse [3]. We also developed a method of endothelial RNA preparation with high purity from the mouse carotid intima [3]. Using this mouse model and method, we found that partial ligation causes endothelial dysfunction in a week, followed by robust and rapid atheroma formation in two weeks in a hyperlipidemic mouse model along with features of complex lesion formation such as intraplaque neovascularization by four weeks. This rapid in vivo model and the endothelial RNA preparation method could be used to determine molecular mechanisms underlying flow-dependent regulation of vascular biology and diseases. Also, it could be used to test various therapeutic interventions targeting endothelial dysfunction and atherosclerosis in considerably reduced study duration.

This describes a partial carotid ligation surgery, which causes disturbed flow conditions and subsequent atherosclerosis development (in two weeks) with intraplaque neo-vascularization (in four weeks) in the mouse common carotid artery. We also describe a novel method of RNA isolation from the carotid intima, providing high purity endothelial RNA.

Um modelo de Disturbed Aterosclerose Flow-induzido em rato artéria carótida pela ligadura parcial e um método simples de isolamento de RNA a partir do endotélio Carótidas

Despite the well-known close association, direct evidence linking disturbed flow to atherogenesis has been lacking. We have recently used a modified ...

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 ...

A Simple Guide Screw Method for Intracranial Xenograft Studies in Mice

The grafting of human tumor cells into the brain of immunosuppressed mice is an established method for the study of brain cancers including glioblastoma (glioma) and medulloblastoma. The widely used stereotactic approach only allows for the injection of a single animal at a time, is labor intensive and requires highly specialized equipment. The guide screw method, initially developed by Lal et al.,1 was developed to eliminate cumbersome stereotactic procedures. We now describe a modified guide screw approach that is rapid and exceptionally safe; both of which are critical ethical considerations. Notably, our procedure now incorporates an infusion pump that allows up to 10 animals to be simultaneously injected with tumor cells.
To demonstrate the utility of this procedure, we established human U87MG glioma cells as intracranial xenografts in mice, which were then treated with AMG102; a fully human antibody directed to HGF/scatter factor currently undergoing clinical evaluation2-5. Systemic injection of AMG102 significantly prolonged the survival of all mice with intracranial U87MG xenografts and resulted in a number of complete cures. 
This study demonstrates that the guide screw method is an inexpensive, highly reproducible approach for establishing intracranial xenografts. Furthermore, it provides a relevant physiological model for validating novel therapeutic strategies for the treatment of brain cancers.

In order to evaluate novel therapeutic paradigms for the treatment of glioma, physiological relevant models are essential. We utilize an implantable guide screw procedure for establishment of intracranial xenograft models that is more rapid and safer than stereotactic approaches.

Um método simples parafuso Guia para intracraniana Estudos Xenoenxerto em Ratos

The grafting of human tumor cells into the brain of immunosuppressed mice is an established method for the study of brain cancers including glioblastoma ...

Noninvasive Intratracheal Intubation to Study the Pathology and Physiology of Mouse Lung

The use of a model that mimics the condition of lung diseases in humans is critical for studying the pathophysiology and/or etiology of a particular disease and for developing therapeutic intervention. With the increasing availability of knockout and transgenic derivatives, together with a vast amount of genetic information, mice provide one of the best models to study the molecular mechanisms underlying the pathology and physiology of lung diseases. Inhalation, intranasal instillation, intratracheal instillation, and intratracheal intubation are the most widely used techniques by a number of investigators to administer materials of interest to mouse lungs. There are pros and cons for each technique depending on the goals of a study. Here a noninvasive intratracheal intubation method that can directly deliver exogenous materials to mouse lungs is presented. This technique was applied to administer bleomycin to mouse lungs as a model to study pulmonary fibrosis.

The use of a model that mimics the condition of lung diseases in humans is critical for studying the pathophysiology and/or etiology of a particular disease and for developing therapeutic intervention. Here a noninvasive intratracheal intubation method that can directly deliver exogenous materials to mouse lungs is presented.

Não invasiva Intratraqueal intubação para estudar a Patologia e Fisiologia do Rato Lung

The use of a model that mimics the condition of lung diseases in humans is critical for studying the pathophysiology and/or etiology of a particular ...

Implantation of Fibrin Gel on Mouse Lung to Study Lung-specific Angiogenesis

Recent significant advances in stem cell research and bioengineering techniques have made great progress in utilizing biomaterials to regenerate and repair damage in simple tissues in the orthopedic and periodontal fields. However, attempts to regenerate the structures and functions of more complex three-dimensional (3D) organs such as lungs have not been very successful because the biological processes of organ regeneration have not been well explored. It is becoming clear that angiogenesis, the formation of new blood vessels, plays key roles in organ regeneration. Newly formed vasculatures not only deliver oxygen, nutrients and various cell components that are required for organ regeneration but also provide instructive signals to the regenerating local tissues. Therefore, to successfully regenerate lungs in an adult, it is necessary to recapitulate the lung-specific microenvironments in which angiogenesis drives regeneration of local lung tissues. Although conventional in vivo angiogenesis assays, such as subcutaneous implantation of extracellular matrix (ECM)-rich hydrogels (e.g., fibrin or collagen gels or Matrigel - ECM protein mixture secreted by Engelbreth-Holm-Swarm mouse sarcoma cells), are extensively utilized to explore the general mechanisms of angiogenesis, lung-specific angiogenesis has not been well characterized because methods for orthotopic implantation of biomaterials in the lung have not been well established. The goal of this protocol is to introduce a unique method to implant fibrin gel on the lung surface of living adult mouse, allowing for the successful recapitulation of host lung-derived angiogenesis inside the gel. This approach enables researchers to explore the mechanisms by which the lung-specific microenvironment controls angiogenesis and alveolar regeneration in both normal and pathological conditions. Since implanted biomaterials release and supply physical and chemical signals to adjacent lung tissues, implantation of these biomaterials on diseased lung can potentially normalize the adjacent diseased tissues, enabling researchers to develop new therapeutic approaches for various types of lung diseases.

Recapitulation of the organ-specific microenvironment, which stimulates local angiogenesis, is indispensable for successful regeneration of damaged tissues. This report demonstrates a novel method to implant fibrin gels on the lung surface of living mouse in order to explore how the lung-specific microenvironment modulates angiogenesis and alveolar regeneration in adult mouse.

Implantação de fibrina Gel no tapete do pulmão para o Estudo de angiogênese Lung-specific

Recent significant advances in stem cell research and bioengineering techniques have made great progress in utilizing biomaterials to regenerate and ...

Intubation-mediated Intratracheal (IMIT) Instillation: A Noninvasive, Lung-specific Delivery System

Respiratory disease studies typically involve the use of murine models as surrogate systems. However, there are significant physiologic differences between the murine and human respiratory systems, especially in their upper respiratory tracts (URT). In some models, these differences in the murine nasal cavity can have a significant impact on disease progression and presentation in the lower respiratory tract (LRT) when using intranasal instillation techniques, potentially limiting the usefulness of the mouse model to study these diseases. For these reasons, it would be advantageous to develop a technique to instill bacteria directly into the mouse lungs in order to study LRT disease in the absence of involvement of the URT. We have termed this lung specific delivery technique intubation-mediated intratracheal (IMIT) instillation. This noninvasive technique minimizes the potential for instillation into the bloodstream, which can occur during more invasive traditional surgical intratracheal infection approaches, and limits the possibility of incidental digestive tract delivery. IMIT is a two-step process in which mice are first intubated, with an intermediate step to ensure correct catheter placement into the trachea, followed by insertion of a blunt needle into the catheter to mediate direct delivery of bacteria into the lung. This approach facilitates a &#62;98% efficacy of delivery into the lungs with excellent distribution of reagent throughout the lung. Thus, IMIT represents a novel approach to study LRT disease and therapeutic delivery directly into the lung, improving upon the ability to use mice as surrogates to study human respiratory disease. Furthermore, the accuracy and reproducibility of this delivery system also makes it amenable to Good Laboratory Practice Standards (GLPS), as well as delivery of a wide range of reagents which require high efficiency delivery to the lung.

Intubation-mediated Intratracheal IMIT Instillation: A Noninvasive, Lung-specific Delivery System

Intubation-mediated intratracheal (IMIT) instillation of reagents is an excellent, noninvasive method for studying respiratory disease, as well as a method for instilling therapeutic reagents directly into the lung. It is a rapid and highly reproducible method which is suitable for preclinical testing.

Intratraqueal (IMIT) instilação mediada por intubação: A não-invasiva, sistema de entrega Lung-specific

Respiratory disease studies typically involve the use of murine models as surrogate systems. However, there are significant physiologic differences ...

Mouse Pneumonectomy Model of Compensatory Lung Growth

In humans, disrupted repair and remodeling of injured lung contributes to a host of acute and chronic lung disorders which may ultimately lead to disability or death. Injury-based animal models of lung repair and regeneration are limited by injury-specific responses making it difficult to differentiate changes related to the injury response and injury resolution from changes related to lung repair and lung regeneration. However, use of animal models to identify these repair and regeneration signaling pathways is critical to the development of new therapies aimed at improving pulmonary function following lung injury. The mouse pneumonectomy model utilizes compensatory lung growth to isolate those repair and regeneration signals in order to more clearly define mechanisms of alveolar re-septation. Here, we describe our technique for performing mouse pneumonectomy and sham pneumonectomy. This technique may be utilized in conjunction with lineage tracing or other transgenic mouse models to define molecular and cellular mechanism of lung repair and regeneration.

Mouse pneumonectomy is a commonly employed model of compensatory lung growth. This procedure can be used in conjunction with lineage tracing or transgenic mouse models to elucidate underlying mechanisms.

Rato Pneumonectomia modelo de crescimento compensatório pulmonar

In humans, disrupted repair and remodeling of injured lung contributes to a host of acute and chronic lung disorders which may ultimately lead to ...

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. As this technology expands and improves, the ability to potentially evaluate and improve the quality of substandard lungs prior to transplant is a critical need. In order to more rigorously evaluate these approaches, a reproducible animal model needs to be established that would allow for testing of improved techniques and management of the donated lungs as well as to the lung-transplant recipient. In addition, an EVLP animal model of associated pathologies, e.g., ventilation induced lung injury (VILI), would provide a novel method to evaluate treatments for these pathologies. Here, we describe the development of a rat EVLP lung program and refinements to this method that allow for a reproducible model for future expansion. We also describe the application of this EVLP system to model VILI in rat lungs. The goal is to provide the research community with key information and &#8220;pearls of wisdom&#8221;/techniques that arose from trial and error and are critical to establishing an EVLP system that is robust and reproducible.

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. Here, we describe the development of a rat EVLP program and refinements that allow for a reproducible model for future expansion.

Método de Isolado<em&gt; Ex Vivo</em&gt; Perfusão pulmonar em um modelo de rato: Lições aprendidas com o desenvolvimento de um Programa PPEV Rat

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed ...

A Simple Device to Rapidly Prepare Whole Mounts of the Mouse Intestine

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe the use of a simple device to cut and &#8216;roll&#8217; mouse intestines to rapidly prepare whole mount preparations of superior and uniform quality to that which can be achieved by hand. The device comprises a base that holds 4 stainless steel rods and a top, which acts a cutting guide. The rods are inserted into the lumen of the small intestine [divided into thirds] and the colon. The rods and samples are then placed over a piece of filter paper or card into the holding slots in the base of the device. The top of the device is then positioned and serves as a cutting guide. The two angled sections in the center of the top piece are used to guide a knife or scalpel and cut the intestines longitudinally on the top of the rods. Once the intestinal sections have been cut, the top is removed and the card,&#160;tissue and rods gently removed from the device and placed on the bench. The rods are then gently rolled sideways to flatten and stick the intestinal segments onto the underlying piece of filter paper or card. The final preparation can then be examined or fixed and stored for later analysis. The preparations are invaluable for the study of intestinal changes in normal or genetically modified mouse models. The preparations have been used for the study and quantification of the effects of inflammation (colitis), damage, pre-cancerous lesions (aberrant crypt foci (ACFs) and mucin depleted foci (MDFs)) and polyps or tumors.

The use of a simple device to cut and &#8216;roll&#8217; mouse intestines to rapidly prepare whole mount preparations is described.

Um dispositivo simples de preparar rapidamente inteiras Mounts do mouse Intestino

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe ...

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 ...