<p class="jove_content"&gt; Mevcut çalışmalar, Ulusal Kalp, Akciğer ve Kan Enstitüsü Hibe R01-HL0921, HK Eltzschig R01-DK083385 ve R01HL098294, T. Eckle 1K08HL102267-01, ve Anestezi Eğitim ve Araştırma T. Eckle Hibeler Vakfı tarafından desteklenen HK Eltzschig ve Amerikan Kalp Derneği, T. Eckle ve HK Eltzschig ve M. Koeppen Deutsche Forschungsgemeinschaft (DFG) araştırma bursu Grant.</p

Genel notlar: Tüm işlemler, dik bir diseksiyon mikroskobu (Olympus, STU2 Standı Boom Stand ile Z-Eksen Crank Post SZX10) altında bir cerrahi koagülatör (11) kullanılarak yapılmalıdır. Deneysel gruplar sonuçlarının karşılaştırılabilirliği sağlamak için en iyi yaş ve ağırlığı mümkün olduğunca uyumlu olmalıdır. Sıcaklık, kan basıncı, anestezi ve sıvı yönetimi boyunca sabit olmalıdır. 1. Anestezi ve trakea hazırlık <ol><li> Kullanın ve en az 10 hafta eski C57BL / 6 fareler 22-25 g. bir vücuda sahip. Anestezi neden doz 70 mg / kg vücut ağırlığı ip (6) pentobarbital sodyum kullanarak. Yaklaşık pentobarbital 10 mg / kg / saat sodyum ile anestezi koruyun. Bu kan basıncını önemli ölçüde azaltabilirsiniz doz aşımı ile dikkatli olun. Pentobarbital yeniden dozlama sonra bile plazma düzeylerinde ciddi artışlara yol saat-. </li><li> Sonra anestezi indüksiyonu, bir teyp ve ayak bileklerine kadar bağlanmış bir dikiş kullanarak tabloya bağlı üst ve alt ekstremite ile yatar pozisyonda güvenli fareler. Dişler kullanarak kafa için de aynı işlemi yapın. Yeterli frenleyici başarılı bir entübasyon ve iyi kontrollü bir cerrahi için önemlidir. Ameliyat öncesinde, fare saç alerji riskini azaltmak için, mineral yağ ile fare kaplayın. Vücut ısısı, piyasada bulunan gıda ambalajlarında farelerin kapağı sabit kalmasını sağlamak için. </li><li> 37 vücut ısısını korumak için termal geribildirim denetleyicisine bağlı bir rektal termometre probu ile sıcaklık kontrollü ısıtmalı masa (RT, Effenberg, Münih, Almanya) ° C Yeri fareler </li><li> Trakeanın cerrahi Açığa. Bağ dokusu yer ve iki 3.0 ipek cerrahi sütür (Harvard cihazları, ABD) her 10 cm altında uzun trakea trakea lateral ve dorsal taraf parçalara ayır. Sütür dışında yaklaşık 1 cm olmalıdır. </li><li> McPherson-Vannas Makas (Dünya Hassas Aletler, ABD 8 cm, düz bıçak) kullanarak iki dairesel kıkırdaklar arasındaki gırtlak altında 4 mm - dikkatlice trakea 3 insizyon. Kanamaya neden bu sonuç parametreleri etkileyebilecek, emin olun. </li><li> Trakeal entübasyon künt polietilen kanül (Insyte 22g, Beckton Dickinson, ABD) kullanarak gerçekleştirin. Polietilen kanül ucu trakea içine 85 derecelik bir açıyla yerleştirin. Trakeal lümen böylece Sonra kanüller yatırın. Kanül ucu toraks diyafram kayboldu kadar tüp trakea aşağı yavaşça ilerler. Trakea (bkz. 1.4) dorsal yerleştirilen iki cerrahi ipek sütür ile bu pozisyonda tüp sabitleşmek. </li></ol> 2. Ventilatöre bağlı akciğer hasarı Tekniği <ol><li> Tüpün bir vantilatör bağlayın. Akciğer hasarına neden için Servo 900 C, Siemens (PRM Veteriner, ABD) kullanılarak bir basınç kontrollü ventilasyon tekniği kullanır . Hayvanlar FiO 2 = 1.0 ile 80 nefes / dk 45 mbar, frekans pik inspiratuar basınç ve 0-3 mbar bir pozitif ekspirasyon sonu basınç ile havalandırılmalıdır. Ilham sona erme oranı 1:1 olmalıdır. Servo 900 C, insanlar için vantilatör olarak inşa olduğu gerçeğine rağmen, bir basınç kontrollü ventilatör ortamda kullanılması farelerin havalandırma için mükemmel çalışır. </li><li> Monitör EKG (örneğin, Hewlett Packard, B blingen, Almanya) ile kalp atım hızı. Kalp hızı 400 altına düştüğü olmadığından emin olun. Bir mekanik ventilasyon intratorasik basınç artışı takiben artan pulmoner arter basınçları bir işareti olarak tesis sağ kalp eksen kayması, göreceksiniz. Fare bradikardi oluşursa, sıcaklık ve anestezi dozu / konsantrasyon kontrol edin. Xylacin / Ketamin anestezisi 250 / dk bir kalp neden olur ve bu nedenle tavsiye edilmez. </li><li> Uygun bir sıvı replasmanı uygulayın. Olan bir infüzyon serum fizyolojik 0.1 ml / saat arteriyel ya da venöz kateter yoluyla yeterli venöz dosyalama sağlamak için havalandırma önce yapılmalıdır. Ortalama arter basıncında önemli bir düşüşe yol açabilecek yüksek havalandırma basınçları, kalbe venöz dönüş bozulur nedeniyle. Ayrıca, 500 ul tuzlu bolus cerrahi öncesi ip verilebilir. </li><li> Karotis arter kan basıncı (27) sürekli kayıt için bir kateter yerleştirin. Arter diseksiyon başlamadan önce vücut kol takın. Karotis arter paratrakeal kasların künt diseksiyon ile maruz kalmaktadır. Daha fazla maruz kalma ve herhangi bir doku travması (özellikle vagal sinir) dikkatli kaçınma ardından, iki sütür ve küçük bir kelepçe (37) ile damar içine bir kateter yerleştirilir. Bu arter uzun bir segment gösterecektir. Karotis arter en sonunda bir ligatür yerleştirin. Gerginlik veya sabitleşmek inci elde etmek için dikiş sonuna kadar büyük bir kelepçe takınbant kullanarak tablo e dikin. Arter çevresinde başka bir dikiş yerleştirin ve çok distal ucu arter disseke. Burada, küçük bir kelepçe yerleştirin. Arter içine bir küçük çapraz açılış kesmek için mikro makas kullanın. Ince bir forseps (Dumont, TEFE) ile açılış tutun ve ellerinizi / forseps ile uygun büyüklükte kateter önceden. Ikinci sütür ile düğüm yapın ve arter güvenli. Kelepçe gevşetin ve kateter daha önceden. Kateter birkaç knot ve bant ile sabitleyin. Alternatif olarak, karotis arter kateter arter kan gazı analizi için kan örnekleri toplamak için deney sonunda yer olabilir. </li></ol> 3. Kurtarma doku örneklerinin 3 saat sonra mekanik ventilasyon, örnekler, akciğer hasarı ölçüde değerlendirmek için toplanır. Brochnoalveolar lavaj sıvısı (BAL), arteryel kan ve akciğer dokusunda toplayarak öneririz. <ol><li> Deneyin sonunda BAL sıvı alın. Anestezi derinleşen sonra, 1 ml fosfat ile trakeal tüp yıkayın tuzlu su (PBS) tamponlu. Bağlı enjektör ile kurtarma önce sıvı üç saniye için trakea / akciğer kalmalıdır. BAL, sıvı nitrojen içinde snap-dondurulmuş ve -80 ° C&#39;de daha fazla analiz için saklanır. Kurtarılan hacmi 1 ml önemli ölçüde daha az olabileceğini farkında olun. </li><li> Deney sonunda kan gazı analizi yapın. Bunu yapmak için, bir kesi sağ sternum altında yapılmalıdır. Forseps ve göğüs kafesi boyunca kesi ölçüde ile sternum tutun. Daha sonra, diyaframın kenarlarında kazıma ve kaburga dışarı kesilir. Şimdi alt toraks boşluğuna açık bir görüş vardır. Forseps ile sternum yukarı kaldırın ve tam anterior toraks duvar-koğuşta kadar açık olduğunu, sağ ve sol tarafında uzun kesintiler (mümkün olduğunca lateral) göğüs kafesi açmak. Deneysel hayvan hala mekanik ventilasyon ise bu yapılmalıdır. Sol ventrikül, 27 delinirse ½ G iğne ve arteryel kan analizi i-STAT Sistemi (Abbott, USA) kullanılarak yapılır. Bu sonuç önemli bir karıştırıcı olacağından, arteryel kan gazı analizi yapılmalıdır BAL, elde edilebilir değildir. </li><li> Alternatif Yukarıdaki yöntem, arteryel kan örnekleri karotis arter kateter yoluyla toplanabilir. Ancak BAL elde önce yapılmalıdır. </li><li> Kalp yukarı çekerek ve trakea kesmek Vergi akciğerlerin en blok. Kan emmek için hazır bir doku parçası vardır, cerrahi sitesi görünür. Karın yönünde kalp çekin ve tüm torasik organlar dikkatli bir şekilde harekete geçirmek için, omurga boyunca kesilir. Aort Kes, torasik organlar kaldırmak ve temiz bir cerrahi masanın üzerine koyun. </li><li> Kalp ve büyük damar doku örneği kesip. Hiçbir timus dokusu hala akciğere bağlı olduğundan emin olun. Bireysel boru ve ek dondurucu bir makas ve yer ile akciğerlere ayırın. Daha fazla analiz için -80 ° C&#39;de saklayın. </li></ol> 4. Akciğer hasarı ölçümü Biz akciğer hasarı ölçüde değerlendirmek için aşağıdaki sonuç parametreleri kullanmanızı tavsiye ederim: albümin ELISA (Bethyl Laboratories, USA) ve bariyer disfonksiyonu ölçüde ve inflamatuar miktarını değerlendirmek için bir miyeloperoksidaz (MPO) ELISA (Hycult Biyoteknoloji, ABD) yapın BAL sıvısı hücre. Da akciğer dokusunda MPO ELISA gerçekleştirin oluşturur. Islak-kuru oranı ölçülecek ise BAL sıvısı elde yoktur ve pulmoner dolaşım (3.3) kızarmış değildir. Eksizyon sonrası akciğer ağırlığı ölçün. Sonra akciğerler 48 saat ve akciğer dokusunun yeniden ölçülür için liyofilize. Sonra ıslak-kuru oranı kuru doku mg başına düşen su mg (5) olarak ölçülür. <img src="/files/ftp_upload/2525/2525fig1.jpg" alt="Şekil 1" /> Şekil 1. Protein içeriği VILI. Fareler yanıt BAL pentobarbital ile anestezi, mekanik ventilasyon tesis edildi ve fareler, basınç kontrollü ayarları (45 mbar inspiratuar basınç, son exspiratory pozitif basınç 3 mbar, ilham% 100 kullanarak havalandırılan oksijen konsantrasyonu). 0 1, 2 ve 3 saat sonra, havalandırma BAL, hasat ve protein içeriği bicinchoninic asit tayini (BCA tahlil) kullanarak sayısal oldu. Protein içeriği göreceli değişim havalandırma 0 saat normalize gösterilir (n = 4 grup başına, * p &lt;0.05, kontrole göre gösterir, ortalama ± SEM )

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K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selective+deletion+of+the+A1+adenosine+receptor+abolishes+heart-rate+slowing+effects+of+intravascular+adenosine+in+vivo.">Selective deletion of the A1 adenosine receptor abolishes heart-rate slowing effects of intravascular adenosine in vivo.</a> PLoS One. 4, e6784-e6784 (2009).</li><li>Kohler, D., Eckle, T., Faigle, M., Grenz, A., Mittelbronn, M., Laucher, S., Hart, M. L., Robson, S. C., Muller, C. E., Eltzschig, H. 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C., Rosenberger, P., Kuhlicke, J., Eltzschig, H. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HIF-1-dependent+repression+of+adenosine+kinase+attenuates+hypoxia-induced+vascular+leak.">HIF-1-dependent repression of adenosine kinase attenuates hypoxia-induced vascular leak.</a> Blood. 111, 5571-5580 (2008).</li><li>Morote-Garcia, J. C., Rosenberger, P., Nivillac, N. M., Coe, I. R., Eltzschig, H. 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P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Crucial+role+for+ecto-5'-nucleotidase+(CD73)+in+vascular+leakage+during+hypoxia.">Crucial role for ecto-5'-nucleotidase (CD73) in vascular leakage during hypoxia.</a> J. Exp. Med. 200, 1395-1405 (2004).</li><li>Aherne, C. M., Kewley, E. M., Eltzschig, H. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+resurgence+of+A2B+adenosine+receptor+signaling.">The resurgence of A2B adenosine receptor signaling.</a> Biochim Biophys Acta. , (2010).</li><li>Eckle, T., Koeppen, M., Eltzschig, H. 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Bu çalışmada farelerde ventilatöre bağlı akciğer hasarı yapan bir teknik açıklar. Bu model, yüksek basınçlı havalandırma nedeniyle yüksek tekrarlanabilir yaralanma göstermektedir. Farelerde akut akciğer hasarı okullar Müfettişler bu model yararlanabilir.

<table border="1"><tbody><tr><th> Reaktifin Adı </th><th> Şirket </th><th> Katalog numarası </th><th> Yorumlar (isteğe bağlı) </th></tr><tr><td> Pentobarbital Sodyum (Fatal Plus,) </td><td> Vortech İlaç Ls, Ltd. </td><td> VPL 9372 </td><td> Içinde 4mg/mL serum fizyolojik </td></tr><tr><td> Insyte 22 G </td><td> Beckton Dickinson </td><td> n / a </td><td></td></tr><tr><td> Dikiş, ipek 4.0 </td><td> Harvard Apparatus </td><td> 517698 </td><td></td></tr><tr><td> Dikiş, Prolen 8.0 </td><td> Ethicon, ABD </td><td> M8739 </td><td> yeniden kullanılabilir </td></tr><tr><td> Siemens, 900 ° C </td><td> PRM Veteriner, ABD </td><td> # 336 </td><td> yenilenmiş </td></tr><tr><td> diseksiyon mikroskobu (SZX10) </td><td> Olympus </td><td> n / a </td><td> geniş çalışma mesafesi düşünün </td></tr><tr><td> Isıtma Tablo </td><td> Rt, Effenberger, Almanya </td><td> n / a </td><td> sadece ve tek bir sağlayıcı </td></tr><tr><td> Kan basıncı cihazı </td><td> Cyber ​​Sense, Inc. </td><td> BPM02 </td><td></td></tr><tr><td> Ben STAT </td><td> Abbott </td><td> n / a </td><td></td></tr></tbody></table>

pressure controlled ventilation to induce acute lung injury in mice

Sıçan modellerinde farklı organların sistemleri (1-34) akut yaralanmalar kapsamlı araştırmak için kullanılır. Uzamış mekanik ventilasyon ile ortaya çıkan akut akciğer hasarı (ALI), kritik hastalık morbidite ve mortalite katkıda bulunur ve yeni genetik veya farmakolojik hedefler üzerinde çalışmalar yoğun bir soruşturma alanlarda (1-3, 5, 8, 26, 30, 33 -36). ALİ non-kardiyak pulmoner ödem ve pulmoner gaz değişimi (36) sonraki bozukluğu neden olan hastalık, akut başlangıçlı tarafından tanımlanır. Biz ventilatöre bağlı akciğer hasarı (2) ikna etmek için bir basınç kontrollü ventilasyon ile ALI bir fare modeli geliştirdiler. Bu amaçla, C57BL / 6 fareler anestezi ve mekanik ventilasyon ile ALI indüksiyon bir trakeotomi takip yapılır. 3 saat - Fareler üzerinde 45 mbar 1 inspiratuar tepe basıncı ile basınç kontrollü bir ortamda havalandırılan. Sonuç parametreleri, pulmoner ödem (ıslak-kuru oranı), bronkoalveoler sıvı albümin içerik, bronkoalveoler sıvı ve akciğer dokusunda miyeloperoksidaz içeriği ve pulmoner gaz değişimi (2) değerlendirilir. Bu tekniği kullanarak Yeteri kadar akut akciğer iltihabı neden olur ve farklı tedavi grupları ya da genotipler (1-3, 5) ayırt edebilirsiniz gösterebilirim. Bu nedenle bu tekniğin ALİ gen hedefli bir silme ile farelerde genetik bir yaklaşım kullanarak ilgili moleküler mekanizmaları takip araştırmacılar için yararlı olabilir.

Watch this Scientific Journal Video about Pressure Controlled Ventilation to Induce Acute Lung Injury in Mice at JoVE.com

Pressure Controlled Ventilation to Induce Acute Lung Injury in Mice

Ventilatöre bağlı akciğer hasarı için bir fare modeli akut akciğer hasarı incelemek için önemli bir araçtır<em&gt; In vivo</em&gt;. Burada, kolay uygulanabilir rapor<em&gt; In situ</em&gt; Yüksek basınçlı mekanik ventilasyon akut akciğer yetmezliğine yol kullanarak akut akciğer hasarı modeli.

Farelerde Akut Akciğer Hasarı tetikleyebilecek Basınç Kontrollü Havalandırma

Murine models are extensively used to investigate acute injuries of different organs systems (1-34). Acute lung injury (ALI), which occurs with prolonged ...

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18.1K Görüntüleme.  University of Colorado. Ventilatöre bağlı akciğer hasarı için bir fare modeli akut akciğer hasarı incelemek için önemli bir araçtır In vivo. Burada, kolay uygulanabilir rapor In situ Yüksek basınçlı mekanik ventilasyon akut akciğer yetmezliğine yol kullanarak akut akciğer hasarı modeli.

Video: Pressure Controlled Ventilation to Induce Acute Lung Injury in Mice

Controlled Cervical Laceration Injury in Mice

Use of genetically modified mice enhances our understanding of molecular mechanisms underlying several neurological disorders such as a spinal cord injury (SCI). Freehand manual control used to produce a laceration model of SCI creates inconsistent injuries often associated with a crush or contusion component and, therefore, a novel technique was developed. Our model of cervical laceration SCI has resolved inherent difficulties with the freehand method by incorporating 1) cervical vertebral stabilization by vertebral facet fixation, 2) enhanced spinal cord exposure, and 3) creation of a reproducible laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm in depth without associated contusion. Compared to the standard methods of creating a SCI laceration such as freehand use of a scalpel or scissors, our method has produced a consistent lesion. This method is useful for studies on axonal regeneration of corticospinal, rubrospinal, and dorsal ascending tracts.

A novel technique to create a reproducible in vivo model of cervical spinal cord laceration injury in the mouse is described. This technique is based on spine stabilization by fixation of the cervical facets and laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm.

Farelerde Kontrollü Servikal yırtığı Sakatlık

Use of genetically modified mice enhances our  understanding of molecular mechanisms underlying several neurological disorders  such as a spinal cord ...

Tıp

Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often unreported mortality rates that may confound analyses. Bilateral renal pedicle clamping is commonly used to induce IR-AKI, but differences between effective clamp pressures and/or renal responses to ischemia between kidneys often lead to more variable results. In addition, shorter clamp times are known to induce more variable tubular injury, and while mice undergoing bilateral injury with longer clamp times develop more consistent tubular injury, they often die within the first 3 days after injury due to severe renal insufficiency. To improve post-injury survival and obtain more consistent and predictable results, we have developed two models of unilateral ischemia-reperfusion injury followed by contralateral nephrectomy. Both surgeries are performed using a dorsal approach, reducing surgical stress resulting from ventral laparotomy, commonly used for mouse IR-AKI surgeries. For induction of moderate injury BALB/c mice undergo unilateral clamping of the renal pedicle for 26 min and also undergo simultaneous contralateral nephrectomy. Using this approach, 50-60% of mice develop moderate AKI 24 hr after injury but 90-100% of mice survive. To induce more severe AKI, BALB/c mice undergo renal pedicle clamping for 30 min followed by contralateral nephrectomy 8 days after injury. This allows functional assessment of renal recovery after injury with 90-100% survival. Early post-injury tubular damage as well as post injury fibrosis are highly consistent using this model.

We describe models of moderate and severe ischemia-reperfusion-induced kidney injury in which the mice undergo unilateral renal pedicle clamping followed by simultaneous or delayed contralateral nephrectomy, respectively. These models consistently give rise to renal dysfunction and post-injury fibrosis, but injury severity and survival are dependent on mouse background, age and surgical equipment.

Farelerde Akut Böbrek Hasarı ve Post Yaralanma Fibrozis iskemi-reperfüzyon Model

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often ...

Controlled Cortical Impact Model for Traumatic Brain Injury

Every year over a million Americans suffer a traumatic brain injury (TBI). Combined with the incidence of TBIs worldwide, the physical, emotional, social, and economical effects are staggering. Therefore, further research into the effects of TBI and effective treatments is necessary. The controlled cortical impact (CCI) model induces traumatic brain injuries ranging from mild to severe. This method uses a rigid impactor to deliver mechanical energy to an intact dura exposed following a craniectomy. Impact is made under precise parameters at a set velocity to achieve a pre-determined deformation depth. Although other TBI models, such as weight drop and fluid percussion, exist, CCI is more accurate, easier to control, and most importantly, produces traumatic brain injuries similar to those seen in humans. However, no TBI model is currently able to reproduce pathological changes identical to those seen in human patients. The CCI model allows investigation into the short-term and long-term effects of TBI, such as neuronal death, memory deficits, and cerebral edema, as well as potential therapeutic treatments for TBI.

Traumatic brain injuries (TBIs) remain a serious health problem. Using the controlled cortical impact surgery model, research on the effects of TBI and possible treatment methods may be performed.

Travmatik Beyin Hasarı için Kontrollü Kortikal Etki Modeli

Every year over a million Americans suffer a traumatic brain injury (TBI).  Combined with the incidence of TBIs worldwide, the physical, emotional, ...

Cardiac Catheterization in Mice to Measure the Pressure Volume Relationship: Investigating the Bowditch Effect

Animal models that mimic human cardiac disorders have been created to test potential therapeutic strategies. A key component to evaluating these strategies is to examine their effects on heart function. There are several techniques to measure in vivo cardiac mechanics (e.g., echocardiography, pressure/volume relations, etc.). Compared to echocardiography, real-time left ventricular (LV) pressure/volume analysis via catheterization is more precise and insightful in assessing LV function. Additionally, LV pressure/volume analysis provides the ability to instantaneously record changes during manipulations of contractility (e.g., &#946;-adrenergic stimulation) and pathological insults (e.g., ischemia/reperfusion injury). In addition to the maximum (+dP/dt) and minimum (-dP/dt) rate of pressure change in the LV, an accurate assessment of LV function via several load-independent indexes (e.g., end systolic pressure volume relationship and preload recruitable stroke work) can be attained. Heart rate has a significant effect on LV contractility such that an increase in the heart rate is the primary mechanism to increase cardiac output (i.e., Bowditch effect). Thus, when comparing hemodynamics between experimental groups, it is necessary to have similar heart rates. Furthermore, a hallmark of many cardiomyopathy models is a decrease in contractile reserve (i.e., decreased Bowditch effect). Consequently, vital information can be obtained by determining the effects of increasing heart rate on contractility. Our and others data has demonstrated that the neuronal nitric oxide synthase (NOS1) knockout mouse has decreased contractility. Here we describe the procedure of measuring LV pressure/volume with increasing heart rates using the NOS1 knockout mouse model.

This article describes the measurement of murine left ventricular function via pressure/volume analysis at different heart rates.

Bowditch Efekt incelenmesi: Basınç Hacim İlişkisi Tedbir Fare Kardiyak Kateterizasyon

Animal models that mimic human cardiac disorders have been created to test potential therapeutic strategies. A key component to evaluating these ...

Open Tracheostomy Gastric Acid Aspiration Murine Model of Acute Lung Injury Results in Maximal Acute Nonlethal Lung Injury

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute respiratory distress syndrome (ARDS), characterized by neutrophilic alveolitis, and injury to both alveolar epithelium and vascular endothelium. Clinically, ARDS is defined by acute onset of hypoxemia, bilateral patchy pulmonary infiltrates and non-cardiogenic pulmonary edema. Human studies have provided us with valuable information about the physiological and inflammatory changes in the lung caused by ARDS, which has led to various hypotheses about the underling mechanisms. Unfortunately, difficulties determining the etiology of ARDS, as well as a wide range of pathophysiology have resulted in a lack of critical information that could be useful in developing therapeutic strategies.
Translational animal models are valuable when their pathogenesis and pathophysiology accurately reproduce a concept proven in both in vitro and clinical settings. Although large animal models (e.g., sheep) share characteristics of the anatomy of human trachea-bronchial tree, murine models provide a host of other advantages including: low cost; short reproductive cycle lending itself to greater data acquisition; a well understood immunologic system; and a well characterized genome leading to the availability of a variety of gene deletion and transgenic strains. A robust model of low pH induced ARDS requires a murine ALI that targets mainly the alveolar epithelium, secondarily the vascular endothelium, as well as the small airways leading to the alveoli. Furthermore, a reproducible injury with wide differences between different injurious and non-injurious insults is important.
The murine gastric acid aspiration model presented here using hydrochloric acid employs an open tracheostomy and recreates a pathogenic scenario that reproduces the low pH pneumonitis injury in humans. Additionally, this model can be used to examine interaction of a low pH insult with other pulmonary injurious entities (e.g., food particles, pathogenic bacteria).

This protocol induces acute lung injury in a mouse that has close fidelity to the pathogenesis of acid pneumonitis observed in humans. We generate a maximal acute nonlethal low pH lung injury and account for differences in rodent-human anatomic respiratory structure using an open tracheostomy coupled with circumferential pressure release.

Akut Akciğer Hasarı Açık Trakeostomi Gastrik Asit Aspirasyon Fare Modeli Maksimal Akut Ölümcül olmayan Akciğer Hasarı Sonuçlar

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute ...

Balloon-based Injury to Induce Myointimal Hyperplasia in the Mouse Abdominal Aorta

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine balloon denudation model, which is comparable with established vessel injury models in large animals. The aorta denudation model with balloon catheters mimics the clinical setting and leads to comparable pathobiological and physiological changes. Briefly, after performing a horizontal incision in the aorta abdominalis, a balloon catheter will be inserted into the vessel, inflated, and introduced retrogradely. Inflation of the balloon will lead to intima injury and overdistension of the vessel. After removing the catheter, the aortic incision will be closed with single stiches. The model shown in this article is reproducible, easy to perform, and can be established quickly and reliably. It is especially suitable for evaluating expensive experimental therapeutic agents, which can be applied in an economical fashion. By using different knockout-mouse strains, the impact of different genes on MH development can be assessed.

This article demonstrates a murine model to study the development of myointimal hyperplasia (MH) after aortic balloon injury.

Fare Abdominal aort Myointimal Hiperplazi ikna etmek için balon tabanlı yaralanma

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine ...

A Closed-chest Model to Induce Transverse Aortic Constriction in Mice

Research on cardiac hypertrophy and heart failure is frequently based on pressure overload mouse models induced by TAC. The standard procedure is to perform a partial thoracotomy to visualize the transverse aortic arch. However, the surgical trauma caused by the thoracotomy in open-chest models changes the respiratory physiology as the ribs are dissected and left unattached after chest closure. To prevent this, we established a minimally invasive, closed chest approach via lateral thoracotomy. Herein we approach the aortic arch via the 2nd intercostal space without entering the chest cavities, leaving the mouse with a less traumatic injury to recover from. We perform this operation using standard laboratory settings for open chest TAC procedures with equal survival rates. Apart from maintaining physiological breathing patterns due to the closed chest approach, the mice seem to benefit by showing rapid recovery, as the less invasive technique appears to facilitate a fast healing process and to reduce immune response after trauma.

Here, we present a protocol of Transverse Aortic constriction (TAC) via a lateral thoracotomy. This technique is a minimally invasive, closed chest surgical procedure aiming to simulate pressure overload and heart failure in mice utilizing standard TAC laboratory settings.

Transvers aort daralma farelerde ikna etmek için kapalı-göğüs manken

Research on cardiac hypertrophy and heart failure is frequently based on pressure overload mouse models induced by TAC. The standard procedure is to ...

Real-time Pressure-volume Analysis of Acute Myocardial Infarction in Mice

Acute myocardial infarction can lead to acute heart failure and cardiogenic shock. The evaluation of hemodynamics is critical for the evaluation of any potential therapeutic approach directed against acute left ventricular (LV) dysfunction. Current imaging modalities (e.g., echocardiography and magnetic resonance imaging) have several limitations since data on LV pressure cannot directly be measured. LV catheterization in mice undergoing coronary artery occlusion could serve as a novel method for a real-time evaluation of LV function.
At the beginning of the procedure, mice were anesthetized followed by endotracheal intubation. For LV catheterization, the right carotid artery was exposed via middle-neck incision. The catheter was introduced and placed into the LV cavity. Left thoracotomy was conducted and the left main coronary artery (LCA) was ligated. To induce reperfusion, the suture was released after 45 min. Pressure-volume data was recorded at all times.
Ligation of the LCA caused a decrease in LV systolic function as evidenced by a 30% reduction in stroke volume, LV ejection fraction (EF), and cardiac output. Maximum dP/dt as a parameter for LV contractility was also significantly reduced and diastolic function was severely impaired (minimum dP/dt -40%). Reperfusion over a period of 20 min did not lead to a complete recovery of LV function.
Real-time pressure-volume analysis served as a valid procedure for monitoring cardiac function during acute myocardial infarction in mice. Maintaining stable anesthesia and a standardized surgical approach was crucial to ensure valid results. As the early phase of acute myocardial infarction is critical for morbidity and mortality, the delineated method could be beneficial for preclinical evaluation of new strategies for cardioprotection.

Acute myocardial infarction in mice induces acute but incompletely characterized changes in left ventricular (LV) function. LV catheterization in mice undergoing coronary artery occlusion serves as a novel method for a real-time evaluation of LV function.

Akut miyokard infarktüsünde farelerde gerçek zamanlı basınç-hacim Analizi

Acute myocardial infarction can lead to acute heart failure and cardiogenic shock. The evaluation of hemodynamics is critical for the evaluation of any ...

Lung Fixation under Constant Pressure for Evaluation of Emphysema in Mice

Emphysema is a significant feature of chronic obstructive pulmonary disease (COPD). Studies involving an emphysematous mouse model require optimal lung fixation to produce reliable histological specimens of the lung. Due to the nature of the lung&rsquo;s structural composition, which consists largely of air and tissue, there is a risk that it collapses or deflates during the fixation process. Various lung fixation methods exist, each of which has its own advantages and disadvantages. The lung fixation method presented here utilizes constant pressure to enable optimal tissue evaluation for studies using an emphysematous mouse lung model. The main advantage is that it can fix many lungs with the same condition at one time. Lung specimens are obtained from chronic cigarette smoke-exposed mice. Lung fixation is performed using specialized equipment that enables the production of constant pressure. This constant pressure maintains the lung in a reasonably inflated state. Thus, this method generates a histological specimen of the lung that is suitable to evaluate cigarette smoke-induced mild emphysema.

Presented here is a useful protocol for lung fixation that creates a stable condition for histological evaluate of lung specimens from a mouse model of emphysema. The main advantage of this model is that it can fix many lungs with the same constant pressure without lung collapse or deflation.

Farelerde AmfizemIn Değerlendirilmesi İçin Sabit Basınç Altında Akciğer Fiksasyonu

Emphysema is a significant feature of chronic obstructive pulmonary disease (COPD). Studies involving an emphysematous mouse model require optimal lung ...

Inducing Acute Lung Injury in Mice by Direct Intratracheal Lipopolysaccharide Instillation

Airway administration of lipopolysaccharide (LPS) is a common way to study pulmonary inflammation and acute lung injury (ALI) in small animal models. Various approaches have been described, such as the inhalation of aerosolized LPS as well as nasal or intratracheal instillation. The presented protocol describes a detailed step-by-step procedure to induce ALI in mice by direct intratracheal LPS instillation and perform FACS analysis of blood samples, bronchoalveolar lavage (BAL) fluid, and lung tissue. After intraperitoneal sedation, the trachea is exposed and LPS is administered via a 22 G venous catheter. A robust and reproducible inflammatory reaction with leukocyte invasion, upregulation of proinflammatory cytokines, and disruption of the alveolo-capillary barrier is induced within hours to days, depending on the LPS dosage used. Collection of blood samples, BAL fluid, and lung harvesting, as well as the processing for FACS analysis, are described in detail in the protocol. Although the use of the sterile LPS is not suitable to study pharmacologic interventions in infectious diseases, the described approach offers minimal invasiveness, simple handling, and good reproducibility to answer mechanistic immunological questions. Furthermore, dose titration as well as the use of alternative LPS preparations or mouse strains allow modulation of the clinical effects, which can exhibit different degrees of ALI severity or early vs. late onset of disease symptoms.

Presented here is a step-by-step procedure to induce acute lung injury in mice by direct intratracheal lipopolysaccharide instillation and to perform FACS analysis of blood samples, bronchoalveolar lavage fluid, and lung tissue. Minimal invasiveness, simple handling, good reproducibility, and titration of disease severity are advantages of this approach.

Doğrudan Intratracheal Lipopolysaccharide Instilasyon ile farelerde akut akciğer hasarının İndükleme

Airway administration of lipopolysaccharide (LPS) is a common way to study pulmonary inflammation and acute lung injury (ALI) in small animal models. ...