Supportato da NIH HL-10342.

1. Preparazione per la procedura di Si deve prima ottenere e preparare i seguenti elementi: <ol><li> La cannula. Per l&#39;intubazione di 20-35 g di topi, usiamo un 1 o 1,5 pollici di lunghezza, 20 calibro catetere IV (BD Insylte, Sparks, MD o Jelco Optiva, Carlsbad, CA). Un nuovo catetere sterile può essere utilizzato per ciascun topo, cateteri ma può anche essere riutilizzato dopo la sterilizzazione immergendo in etanolo al 70% durante la notte. Sebbene né la faringe, né trachea del mouse è sterili procedure di pulizia adeguate, compreso l&#39;uso di guanti sterili e strumenti, devono essere seguite. </li><li> Il cavo a fibre ottiche. Usiamo ≈ 70 cm di cavo da 0,5 mm ottico da Edmund Optics, ma la lunghezza non è critica. È importante assicurarsi che la fibra ha il suo bordo smussato, in quanto dopo il taglio del cavo di lunghezza con un rasoio, il bordo rimane relativamente forte, e non ci vuole molto sforzo per perforare la parete tracheale. Tuttavia, is molto facile per lisciare questo bordo tenendo la fibra di circa 2 cm dalla fine e poi facendo piccoli cerchi per pochi secondi con i bordi della punta toccare un pezzo di carta 1000 grana abrasiva (vedi dimostrazione nel video e alla figura 1 Macdonald, et al. 10). L&#39;altra estremità è inserita attraverso un tappo di gomma. Questo è più facilmente realizzabile dal primo spingendo un ago calibro 18 attraverso il tappo, inserire la fibra ottica attraverso il foro dell&#39;ago, quindi estrarre l&#39;ago. Il tappo di gomma è collegato ad una sorgente alogena da 150 watt di luce (ad esempio NCL-150, Volpi USA, o qualsiasi altra sorgente luminosa o, addirittura inferiore a 150 watt). E &#39;importante assicurarsi di utilizzare un tappo in gomma di silicone (o materiale resistente al calore), in quanto normale gomma o sughero può bruciare quando si trova così vicino alla fonte di luce calda. </li></ol> 2. Esecuzione della intubazione <ol><li> Vedere le figure 1 e 2. Inserire il cavo in fibra ottica tttraverso un breve pezzo di tubo in gomma siliconica (≈ 0.8 mm ID x 4 mm, Cole-Palmer, EW-96410-13). Tie questo tubo di gomma abbastanza stretto, pur consentendo al cavo in fibra ottica da regolare. Inserimento del tubo al silicone comodamente alla fine Luer della cannula fissa la posizione del cavo in fibra ottica è all&#39;interno della cannula. Regolare la posizione del cavo a fibre ottiche in modo che si estende attraverso la cannula 4 mm ≈ davanti alla punta della cannula. </li><li> Posizionare il mouse anestetizzato su un supporto verticale, sospesa dai suoi incisivi superiori (Figura 3). La maggior parte dei ricercatori a trovare la migliore visualizzazione con il lato ventrale del mouse affrontare se stessi. Molto delicatamente tirare fuori la lingua e tenere premuto con il pollice e l&#39;indice. Il dito medio è posto tra il collo e il supporto plastico. Trazione sulla linguetta con il dito indice e pollice viene utilizzato per aprire la bocca, e per raddrizzare il percorso intubazione, l&#39;angolo della testa viene regolata con il dito medio dietro il collomostrato in Figura 3. </li><li> Utilizzando il cavo in fibra ottica da una sorgente di luce e introduttore, spingerlo attraverso le corde vocali visualizzate. Se i cavi non sono visibili, delicatamente tirare con forza sulla lingua con il dito medio come supporto. Una volta inserito, far avanzare la cannula ≈ 5 mm più. Quindi, non essendo molto attento a spostare la cannula, ritirare il cavo in fibra ottica. Giacciono il mouse e fissare la cannula con un pezzo di nastro e sostenere il mozzo cannula su un pezzo di plastilina (modellazione argilla), come mostrato in Figura 4. </li><li> La procedura del passo 3 non può essere facilmente insegnato o anche dimostrato, poiché è un&#39;operazione da solo. Tuttavia, regolazioni fini della trazione sulla lingua e il sostegno dietro la testa quasi tutti coloro che provare questo presto trovare il modo giusto per posizionare il mouse per visualizzare le corde vocali. </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>

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La procedura qui descritta presenta diversi vantaggi. Prima l&#39;apparecchiatura è semplice e relativamente poco costoso .. La fabbricazione del dispositivo non richiede utensili speciali o attrezzature costose. L&#39;uso di un catetere che presenta anche la sorgente luminosa significa che non si perde di vista dell&#39;apertura tracheale come introduttore avvicina l&#39;apertura tracheale. L&#39;uso di un introduttore 0,5 millimetri serve anche a minimizzare il trauma che potrebbe verificarsi con un inserimento inizi...

Nel 1999, Brown et al. Pubblicato un documento che descrive un metodo per intubazione del polmone mouse 1. Tale tecnica ha utilità notevole facendo funzione polmonare ripetizione o lavaggio broncoalveolare nei topi singoli studi longitudinali 2. Dal momento che la carta originale, ci sono stati diversi altri giornali che hanno descritto i diversi approcci per l&#39;intubazione del mouse 3-9. Mentre tutti questi metodi possono essere utilizzati con successo, di solito richiedono formazione considerevole o costo. Uno dei problemi principali con l&#39;intubazione come è che con l&#39;approssimarsi delle cannula di intubazione si avvicina l&#39;inserimento trachea in attesa, la stessa cannula blocca la luce e quindi la visualizzazione di dove deve andare. Pertanto, l&#39;inserimento diventa cieco nel momento più critico. In questo articolo mostriamo come eliminare semplice ed economico il problema di visualizzazione, assicurando in tal modo l&#39;intubazione di successo con la formazione relativamente poco oesperienza.

<table border="1"><tbody><tr><td> Materiale </td><td> Azienda </td><td> Catalogare # </td><td> Commento </td></tr><tr><td> Cannula per intubazione </td><td> BD Insylte, Sparks, MD o Jelco Optiva, Carlsbad, CA </td><td></td><td> 1-trovi-lunga, calibro 20 per via endovenosa (IV) del catetere </td></tr><tr><td> Cavi in ​​fibra ottica </td><td> Edmund Optics, Barrington, NJ </td><td> # NT02-542 </td><td> Circa 2-ft lunghezza di 0,5 mm in fibra ottica (fibra plastica Comunicazione grado). Il bordo l&#39;estremità della fibra che viene inserito nella trachea deve essere leggermente arrotondato tenendo la fibra ≈ 2 cm dalla fine e poi fare piccoli cerchi mentre si trascina la punta per alcuni secondi su 1.000 grana Emory. </td></tr><tr><td> Fonte di luce </td><td> Volpi </td><td> NCL-150 </td><td> Anche se si usa un 150-W sorgente di luce alogena, qualsiasi fonte di luce equivalente, anche con potenza molto più bassapuò essere utilizzato. </td></tr><tr><td> Tappo di gomma di silicone </td><td></td><td></td><td> Un tappo di gomma siliconica # 1 adatta sorgente luminosa utilizzata in questa dimostrazione. Fonti di luce diverse possono richiedere una dimensione diversa. </td></tr><tr><td> Piccolo tubo di gomma di silicone </td><td> Cole-Palmer </td><td> EW-96410-13 </td><td> Un pezzo ≈ 1.5 cm di tubo di gomma di silicone (0,8 diametro interno di 4 mm di diametro esterno) </td></tr><tr><td> Il piede di supporto angolare </td><td></td><td></td><td> Ours è costruzione di plexiglas, ma qualsiasi materiale a cui un filo o filo può essere apposto tenere il mouse con un angolo quasi verticale può essere utilizzato. </td></tr></tbody></table>

a simple method of mouse lung intubation

Una procedura semplice per intubare i topi per le misure di funzionalità polmonare avrebbe diversi vantaggi in studi longitudinali con un numero limitato di animali o costoso. Uno dei motivi per cui questo non è fatto più di routine è che è relativamente difficile, nonostante sia diversi studi pubblicati che descrivono il modo di realizzarla. In questo lavoro dimostrare una procedura che elimina uno dei principali ostacoli associati a questa intubazione, che di visualizzare la trachea durante l&#39;intero tempo di intubazione. L&#39;approccio utilizza una sorgente di luce a fibre ottiche 0,5 millimetri che serve come un introduttore di dirigere l&#39;intubazione cannula nella trachea mouse. Abbiamo dimostrato che è possibile utilizzare questa procedura per misurare meccanica polmonare in singoli topi in un corso di tempo di almeno parecchie settimane. La tecnica può essere impostato con una spesa relativamente poco e competenze, e può essere realizzato con la formazione di routine relativamente poco. Questo dovrebbe makE &#39;possibile per qualsiasi laboratorio per eseguire routine out questo intubazione, consentendo studi longitudinali in singoli topi, minimizzando il numero di topi necessari e aumentare la potenza statistica utilizzando ogni topo come proprio controllo.

Come una valutazione del metodo, abbiamo utilizzato quattro 20 settimane vecchio maschio topi BALB / c con peso medio (± SEM) di 27,7 ± 0,40 g. Essi sono stati studiati su cinque settimane consecutive, in cui la resistenza polmonare è stata misurata usando un sistema come descritto in precedenza 11. Ogni mouse è stato anestetizzato con ketamina (100 mg / g di peso corporeo) e xilazina (15 mg / g di peso corporeo) in soluzione salina tramite iniezione IP. Sono stati quindi intubati come descritto sopra. Se...

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

A Simple Method of Mouse Lung Intubation

Il presente documento descrive un metodo striaghforward ed efficiente intubazione topi per le misure di funzionalità polmonare o instillazione polmonare, che consente ai topi di recuperare e da studiare in tempi successivi. La procedura prevede una fonte economica di luce a fibre ottiche che illumina direttamente la trachea.

Un metodo semplice di intubazione mouse polmone

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

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JoVE Journal

Medicine

50.5K Views.  Johns Hopkins Bloomberg School of Public Health. Il presente documento descrive un metodo striaghforward ed efficiente intubazione topi per le misure di funzionalità polmonare o instillazione polmonare, che consente ai topi di recuperare e da studiare in tempi successivi. La procedura prevede una fonte economica di luce a fibre ottiche che illumina direttamente la trachea.

Video: 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.

Un modello di Disturbed flusso indotta aterosclerosi carotidea in Arteria mouse da legatura parziale e un semplice metodo di isolamento dell&#39;RNA da Endotelio carotidea

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

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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 ...

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.

Un semplice metodo vite Guida per intracranica Studi dello xenotrapianto in topi

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.

Non invasivo intratracheale intubazione per studiare la Patologia e Fisiologia del mouse polmone

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.

L&#39;impianto di fibrina gel sul mouse polmone per studiare angiogenesi Lung-specifica

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.

Intubazione-mediata intratracheale (IMIT) instillazione: A non invasiva, Lung-specifico sistema di rilascio

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.

Topo Pneumonectomia Modello di crescita compensatoria Lung

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.

Metodo di isolato<em&gt; Ex Vivo</em&gt; Lung Perfusion in un modello di ratto: Lezioni apprese da Sviluppo di un programma EVLP 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.

Un dispositivo semplice da preparare rapidamente intere Monti della Intestino mouse

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.

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 ...