Açıklanan tüm prosedürler uyarınca, araştırma ve tüm hayvan protokolleri Oregon Sağlık ve Bilim Üniversitesi Kurumsal Hayvan Bakım ve Kullanım Kurulu tarafından kabul edildi hayvanların bakımı ve kullanımı için Sağlık kılavuzların National Institutes yapılmaktadır. 1. Cerrahi Hazırlık <ol><li> Fare tartılır. Açıklanan prosedür g. 20 ile 25 arasında tartım C57BL / 6 fareler üzerinde yapılır Anestezi indüksiyon kutusunda izofluran% 3-4 kullanarak bir neden olduğu ve daha sonra hava / oksijen karışımı içinde% 1.5-2.5 izofluran kullanılarak korunur. </li><li> Gözü kayganlaştırır ve ısıtma yastığı hayvan yatar pozisyonda. 4 ekstremitelerde bant kullanarak hareketsiz. Göğüs kompresyonları sırasında tam göğüs duvarı gezi izin mümkün olduğu kadar nötr bir pozisyonda arka-pençeleri bantlanmış olabilir, ancak forepaws, göğüs duvarına yakın olarak güvenli olmalıdır. </li><li> Rektal ısı probu yerleştirin ve yağlayın. Sıcaklık 37.0 korumak için ayarlanmış bir elektronik sıcaklık kontrolörü (Digi-Sense Cole Parmer, Vernon Hills, IL), ° C&#39;ye bağlı bir ısıtma yastığı ve lamba kullanılarak kontrol Hayvan içinde bir sıcaklık gradyanı akış durumuna sırasında gelişebilir mümkün olduğundan, sıcaklık ölçülür ve ilgi organ yakın kontrol edilmesi önemlidir. </li><li> 2,5 cm 22 ga teflon kateter (Insyte-W, BD, Franklin NJ) ve kesme distal ucu açılı bir uygulayıcıya (IN Frova Introducer, Cook Medical, Bloomington) kullanarak trakea entübe. Trakeal entübasyon diğer yöntemler kabul edilebilir, ancak, boyun, intravenöz kateter yerleştirme için cerrahi maruz optimize biraz uzatılmış konumda, uygun pozisyonu sağlar açılı uygulayıcıya kullanımı. Endotrakeal kateter dikiş kesici bir döngü ile güvenli ve göğüs kompresyonları sırasında baş hareketsiz hafif bir gerilim ile sağlanır. </li><li> 140 mcL, 150 soluk / dakika olarak ayarlanmış bir kemirgen ventilatör ile mekanik fare havalandırın. Bu protokol 20-25 g hayvanlar her zaman olduğu gibi bu ağırlığa göre ayarlanabilir değildir. Anestezi üzeri-derinleştirmek etmeyin. Bu bozar resüsitasyon olarak ekspirasyon sonu basınç uygulamak için ventilatör etmeyin. </li><li> Steril teknik ve operasyon mikroskobu kullanarak, juguler ven bir ön-kızarmış PE-10 kateter yerleştirin. Iki tarafında kullanılan, ancak operatöre yakın yan göğüs kompresyonları veya diğer manipülasyonlar sırasında kateterin yerinden çıkması olasılığını azaltır olabilir. % 0.5 &#39;lik bupivakain 0.1 ml, ameliyat sonrası ağrı kontrolü için, yara kenarları içine infitrated. </li><li> PE-10 kateter siyanoakrilat cerrahi yapıştırıcı ile cilt kapatılması sabitleyin. </li><li> Subkutan EKG elektrotları yerleştirin ve izleme cihazı bağlayın. Sinyal kendisi sinyal yolu ve maksimizasyonu dikkat canlandırma başarısı için kritik öneme sahiptir. Tüm teller işletim yüzeye güvenli olduğundan emin olun, sinyal geçişleri en aza indirmek ve sinyal yolu içinde izolatörler (içi boş iğne veya telli hava gibi) en aza indirmek. Nonstranded kurşun tel bağlı Katı iğne satın alınan veya laboratuarda yapılabilir. Bağlantı kurulduktan sonra, monitörde EKG sinyali iyileştirir. </li></ol> 2. Kardiyak Arrest <ol><li> 37.5 ° C rektal sıcaklığı 36.5 ° C olarak tanımlanan, fare normotermik olduğundan emin olun Oda sıcaklığında 0,5 M potasyum klorür intravenöz 40 mcL yönetme ve izleme EKG&#39;de izoelektrik gözlemlemek. Aralığında 20-25 gr ağırlığı için Doz ayarlaması gerekli değildir Tutuklama zamanlayıcı başlatın. </li><li> Ventilatör ayırın. Anestezik buhar durdurun. Isıtma yastığı ve EKG izleme müdahale elektronik gürültü üreten herhangi bir diğer ekipman kapatın. Fare üzerinde yalıtkan bir battaniye koyun. </li><li> Kayıt sıcaklığı kardiyak arrest sırasında her dakika. Gerekirse, bir ısıtma lambası normotermik aralığı çekirdek sıcaklığına getirmek için kullanılıyor olabilir. Kardiyak arrest sırasında, resüsitasyon (örneğin, epinefrin şırınga) malzeme ve teçhizat hazırlamak. Bir kontrol listesi, hayatta kalmak için gerekli kesintisiz resüsitasyon sağlamak için yararlı olabilir. </li><li> Kardiyak arrest 7 dakika, 30 saniye sonra, ventilatör yeniden ve tidal volüm öncesi tutuklama ayarda tutulması, 180 solunum / dk oranını artırmak. </li><li> 8 dakika, 300 BPM göğüs kompresyonları başlatın. EKG Hareket artifakı yargıç CPR oranı kullanılabilir. Göğüs kompresyonları parmağı, 5 mm üstünde kılıç şeklinde bir süreç ve biraz orta hatta sol ile teslim edilmelidir. Göğüs ön-arka mesafe 1/3-1/2 sıkıştırılmış olmalı ve tam bir geri tepme kompresyonları arasında izin verilmelidir. Parmak konumlandırma ve optimal sıkıştırma basıncı, kesinlikle çok kritiktir. Bu modelin sağkalım elde etmek için hemen hemen her zaman optimal CPR kaynaklanmaktadır. </li><li> CPR ilk 30 saniye içinde, 15 mg / ml seyreltilmiş epinefrin 0.5 mL infüze. Toplam epinefrin doz 8-12 mg. Spontan dolaşımı (ROSC) dönmek için EKG dikkatlice gözlemleyin. Dar karmaşık QRS kompleksleri sıkıştırma eserler arasında görülür. ROSC genellikle 90 saniye ve CPR başladıktan sonra 2 dakika arasında gerçekleşir. ROSC 3 dakika meydana değilse Resüsitasyon terk edilmektedir. Sık sık prematüre ventriküler erken vurular ve EKG ekseni değişiklikleri ROSC sonra ilk 2 dakika içinde gözlenen ve hemen hemen her zaman 2 dakika sürekli sinüs taşikardisi çözmek. </li><li> Resüsitasyon ve adrenalin dozu toplam zamanı kaydedin. Kayıt sıcaklığı ROSC sonra 10 dakika için her dakika. </li><li> EKG yol açar, genellikle 12-15 dakika sonra ROSC içinde, spontan solunumu başladığında silinebilir. </li><li> Spontan solunum hızı&gt; 60/min trakea ekstübe. </li><li> 37 ayarlanan sıcaklık kontrollü bir yüzey üzerinde bir kurtarma kafeste fare ° C ilk 2 saat sonrası darlık, yiyecek ve su için kolay erişim sağlar. Kafes sonra fare anesteziden tamamen iyileşti ve aktif olduğu zaman standart ameliyat sonrası konut koşulları taşınmış olabilir. </li></ol> 3. Exsanguination / Perfüzyon-Fiksasyon ve Böbrek Hasat <ol><li> CA / CPR 24 saat sonra,% 3-4 izofluran ile anestezi neden ve hayvan güvenli formalin ile kullanım için uygun bir davlumbaz içinde cerrahi bir yüzey üzerine yatar pozisyonda .. </li><li> Anestezik buhar konsantrasyonu% 5 artırarak yavaş yavaş anestezi derinleştirmek ve spontan solunumun durması kanıtladığı gibi fare derin anestezi sağlamak. </li><li> Kapaklı torakotomi yapın ve standart tekniklere göre kalp sol ventrikül apikal ponksiyon ile fare asıl bulbus olfaktoryusları alındı. </li><li> Aynı iğne yoluyla, yavaş infüzyon ile% 0.9 salin yönetmek. Nick makas ile sağ atriyal apendiks, perfüzyon / exsanguination acclerate için. Laparotomi yapın ve böbrekler gözlemlemek. Beyazlatılmış zaman, tuzlu fiksasyon için% 4 formalin değişmiş olabilir. Böbrekler sonra tübüler epitelyal nekroz ölçümü için Fluoro-yeşim B parafin ve lekeli gömülü. </li></ol> 4. Temsilcisi Sonuçlar: CA indüklenir, ortalama arter basıncı (MAP) ve sıfıra yakın bölgesel böbrek kan akımı (RRBCF) damla ve resüsitasyon başlayana kadar stabil (Şekil 1). 24 saat CA / CPR, böbrek fonksiyon serum endeksleri (kan üre azot, BUN ve kreatinin) anlamlı sham çalışan hayvanlar için göreceli olarak yüksek sonra. AST / ALT, derin bir tüm vücut iskemi (Şekil 2) kanıt sunan da yükselir. Nötrofil-jelatinaz ilişkili lipocalin (NGAL), renal iskemik yaralanma duyarlı bir göstergesi CA / CPR (Şekil 3) upregüle 24 saat sonra ağır. Son olarak, fotomikrografı iskemik renal hasar (leke (Şekil 4, paneli) ve yeşim floro-B ile boyanmış geniş medüller tübüler epitelyum hücre ölümü tübüler epitel ve hematoksilen ve eosin luminal dolum incelmesi ile yamalı, medüller nekroz tipik göstermektedir Şekil 4, panel b). <img src="/files/ftp_upload/3116/3116fig1.jpg" alt="Şekil 1" /> Şekil 1. Kardiyak arrest anında perfüzyon basınç kaybına neden olur, kardiyak arrest dönemi boyunca bölgesel böbrek kortikal kan akımı (RRCBF) yakın tamamen durdurulması ile sonuçlanan femoral arterden ölçülen arter basıncı (MAP), ortalama (gölgeli burada temsil alan). Göğüs kompresyonu ve epinefrin döner Resüsitasyon normal MAP ve RRCBF resüsitasyon sonrası dönemde giderek yükselen 14 ila izni ile yayımlanmaktadır. <img src="/files/ftp_upload/3116/3116fig2.jpg" alt="Şekil 2" /> Şekil 2 - 24 saat sonrası darlık, kan üre azotu (BUN), serum kreatinin ve tübüler hücre ölümü ölçüde CA / CPR gibi sahte prosedürü ile tedavi edilen hayvanlar ile karşılaştırıldığında geçirmiş olan hayvanların önemli ölçüde yükselir. CA / CPR, burada büyük karaciğer fonksiyon enzimlerinde yükseklik ile kanıtlandığı bir pan-organizma iskemik hakaret, neden alanin amino transferaz (ALT) ve plasebo tedavi edilen hayvanlar ile karşılaştırıldığında CA / CPR farelerde (AST), aspartat aminotransferaz. <img src="/files/ftp_upload/3116/3116fig3.jpg" alt="Şekil 3" /> Şekil 3 Western Blot nötrofil jelatinaz ilişkili lipocalin (NGAL), renal iskemik yaralanma duyarlı bir göstergesi poliklonal antikor kullanılarak gerçekleştirildi. İdrar numuneleri hemen önce (&quot;ön&quot;) ve 24 saat sonra (&quot;24&quot;) CA / 4 hayvanlarda CPR (a, b, c ve d yukarıda etiketli) elde edilmiştir. NGAL kitlesel fare upregüleCA / CPR sonra idrar 14 ila izni ile yayımlanmaktadır. <img src="/files/ftp_upload/3116/3116fig4.jpg" alt="Şekil 4" /> Şekil 4 A) Hematoksilen ve eosin böbrek dokusunun 24 CA / CPR sonra kısa bir eksen hiler bölümünde leke. Tübüler takarak medüller ve Kortikomedüller tübüller yamalı ama net bir zarar yoktur. Oklar Kortikomedüller kavşakta şişmiş, pyknotic çekirdekleri ile hasarlı tübüller üzerine gelin. B) Flouro yeşim B CA / CPR sonra, aynı hayvan aynı bölgede 24 saat leke. Fluoro-yeşim B lekeleri nekrotik hücreler, parlak yeşil, yamalı Kortikomedüller tübüler nekroz gösteren. Oklar Kortikomedüller kavşakta Parlak lekeli hasar tübüller üzerine gelin. Bu bulgular, AKI geliştiren insanlarda böbrek biyopsi bulguları ile büyük ölçüde benzer ve diğer hayvan modellerinde AKI tarafından üretilen farklı.

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Çıkar çatışması ilan etti.

Fare kardiyak arrest ve kardiyopulmoner resüsitasyon normotermik model AKI, tüm vücut hipoperfüzyon, en sık görülen klinik nedeni fizyopatolojisi ve morfolojisi çoğaltır bir model değerlendirme birden fazla yollar sunar. Hipotez testleri, genetik manipülasyon teknikleri ve iyi anlaşılmalı ve karakterize bir anatomisi ve fizyolojisi laboratuvar faresi bir yelpazesine erişim destekli olabilir. Burada açıklandığı gibi, deneyimli ellerde sağkalım% 80&#39;dir. Cerrahi hazı...

<table border="1"><tbody><tr><th> Reaktifin Adı </th><th> Şirket </th><th> Katalog numarası </th></tr><tr><td> Potasyum Klorür </td><td> Abbott Medikal </td><td> 06653-5 </td></tr><tr><td> İsofluranın </td><td> Abbott Medikal </td><td> 05.260-05 (ve diğerleri) </td></tr><tr><td> Epinefrin HCl </td><td> Çoklu </td><td> Çoklu </td></tr><tr><td> Digi-Sense sıcaklık kontrolörü </td><td> Cole-Palmer </td><td> EW-89000-00 </td></tr><tr><td> Angiocath IV Cath </td><td> VWR </td><td> 381134 </td></tr><tr><td> Frova açılı havayolu introduser </td><td> Aşçı </td><td> G27282 </td></tr><tr><td> Fareler için MicroVent Vantilatör </td><td> Harvard Apparatus </td><td> 733591 </td></tr></tbody></table>

normothermic cardiac arrest and cardiopulmonary resuscitation: a mouse model of ischemia-reperfusion injury

Akut Böbrek Sakatlık (AKI) yüksek mortalite 1-4 ve tüm vücut hipoperfüzyon tarafından en sık neden olduğu ortak bir kritik hastalık, oldukça ölümcül bir komplikasyon. Kemirgen modelleri tüm vücut hipoperfüzyon 5,6 Başarılı üreme zorluklarla dolu olmuştur. fokal iskemi istihdam 7-9,9,10 modeller art arda klinik ortamda ve genetik manipülasyon tam araç seti tüm vücut hipoperfüzyon eksikliği erişim için izin büyük hayvan modellerinde çevirmek olmayan sonuçları göstermiştir fare mümkün. 11,12 Ancak, son yıllarda, kardiyak arrest ve kardiyopulmoner resüsitasyon bir fare modeli modeli AKI adapte edilebileceği ortaya çıkmıştır. 13 Bu model güvenilir klinik AKI görülen fizyolojik, fonksiyonel, anatomik ve histolojik sonuçlar üretir hızla tekrarlanabilir ve genetik manipülatif teknikleri, büyük hayvanlar için düşük maliyetli akraba ve kullanım kolaylığı erişimi de dahil olmak üzere tüm cerrahi bir fare modelinin önemli avantajlar sunuyor. Grubumuz bu modelin kullanımı ile AKI, organ-spesifik sonuçlar bir sayısını değerlendirmek için geniş bir deneyime sahiptir. 14,15

Watch this Scientific Journal Video about Normothermic Cardiac Arrest and Cardiopulmonary Resuscitation: A Mouse Model of Ischemia-Reperfusion Injury at JoVE.com

Normothermic Cardiac Arrest and Cardiopulmonary Resuscitation: A Mouse Model of Ischemia-Reperfusion Injury

Perioperatif ve yoğun bakım ile ilgili akut böbrek hasarı için güçlü bir model sundu. Neredeyse, klinik AKI, histolojik ve fonksiyonel değişiklikleri çoğaltmak mümkündür kardiyak arrest ile indüklenen tüm vücut hipoperfüzyon kullanma.

Normotermik Kardiyak Arrest ve Kardiyopulmoner Resusitasyon: İskemi-Reperfüzyon Hasarı Fare Modeli

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused ...

normothermic-cardiac-arrest-cardiopulmonary-resuscitation-mouse-model

Research

JoVE Journal

Medicine

17.9K Görüntüleme.  Oregon Health & Sciences University. Perioperatif ve yoğun bakım ile ilgili akut böbrek hasarı için güçlü bir model sundu. Neredeyse, klinik AKI, histolojik ve fonksiyonel değişiklikleri çoğaltmak mümkündür kardiyak arrest ile indüklenen tüm vücut hipoperfüzyon kullanma.

Video: Normothermic Cardiac Arrest and Cardiopulmonary Resuscitation: A Mouse Model of Ischemia-Reperfusion Injury

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 ...

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Induction and Assessment of Ischemia-reperfusion Injury in Langendorff-perfused Rat Hearts

The biochemical events surrounding ischemia reperfusion injury in the acute setting are of great importance to furthering novel treatment options for myocardial infarction and cardiac complications of thoracic surgery. The ability of certain drugs to precondition the myocardium against ischemia reperfusion injury has led to multiple clinical trials, with little success. The isolated heart model allows acute observation of the functional effects of ischemia reperfusion injury in real time, including the effects of various pharmacological interventions administered at any time-point before or within the ischemia-reperfusion injury window. Since brief periods of ischemia can precondition the heart against ischemic injury, in situ aortic cannulation is performed to allow for functional assessment of non-preconditioned myocardium. A saline filled balloon is placed into the left ventricle to allow for real-time measurement of pressure generation. Ischemic injury is simulated by the cessation of perfusion buffer flow, followed by reperfusion. The duration of both ischemia and reperfusion can be modulated to examine biochemical events at any given time-point. Although the Langendorff isolated heart model does not allow for the consideration of systemic events affecting ischemia and reperfusion, it is an excellent model for the examination of acute functional and biochemical events within the window of ischemia reperfusion injury as well as the effect of pharmacological intervention on cardiac pre- and postconditioning. The goal of this protocol is to demonstrate how to perform in situ aortic cannulation and heart excision followed by ischemia/reperfusion injury in the Langendorff model.

The isolated rat heart is an enduring model for ischemia reperfusion injury. Here, we describe the process of harvesting the beating heart from a rat via in situ aortic cannulation, Langendorff perfusion of the heart, simulated ischemia-reperfusion injury, and infarct staining to confirm the extent of ischemic insult.

Langendorff-perfüze sıçan Hearts indüksiyon ve İskemi-reperfüzyon Hasarı Değerlendirilmesi

The biochemical events surrounding ischemia reperfusion injury in the acute setting are of great importance to furthering novel treatment options for ...

Murine Model of Intestinal Ischemia-reperfusion Injury

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, hypotension, necrotizing enterocolitis, bowel transplantation, trauma and chronic inflammation. Intestinal ischemia-reperfusion (IR) injury is a consequence of acute mesenteric ischemia, caused by inadequate blood flow through the mesenteric vessels, resulting in intestinal damage. Reperfusion following ischemia can further exacerbate damage of the intestine. The mechanisms of IR injury are complex and poorly understood. Therefore, experimental small animal models are critical for understanding the pathophysiology of IR injury and the development of novel therapies.
Here we describe a mouse model of acute intestinal IR injury that provides reproducible injury of the small intestine without mortality. This is achieved by inducing ischemia in the region of the distal ileum by temporally occluding the peripheral and terminal collateral branches of the superior mesenteric artery for 60 min using microvascular clips. Reperfusion for 1 hr, or 2 hr after injury results in reproducible injury of the intestine examined by histological analysis. Proper position of the microvascular clips is critical for the procedure. Therefore the video clip provides a detailed visual step-by-step description of this technique. This model of intestinal IR injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

Here we describe the detailed procedure of intestinal ischemia-reperfusion in mice which results in reproducible injury without mortality to encourage the standardization of this technique across the field. This model of intestinal ischemia-reperfusion injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

Bağırsak İskemi-reperfüzyon Hasarı Murin Modeli

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, ...

A Mouse Model of Retinal Ischemia-Reperfusion Injury Through Elevation of Intraocular Pressure

Retinal ischemia-reperfusion (I/R) is a pathophysiological process contributing to cellular damage in multiple ocular conditions, including glaucoma, diabetic retinopathy, and retinal vascular occlusions. Rodent models of I/R injury are providing significant insights into mechanisms and treatment strategies for human I/R injury, especially with regard to neurodegenerative damage in the retinal neurovascular unit. Presented here is a protocol for inducing retinal I/R injury in mice through elevation of intraocular pressure (IOP). In this protocol, the ocular anterior chamber is cannulated with a needle, through which flows the drip of an elevated saline reservoir. Using this drip to raise IOP above systolic arterial blood pressure, a practitioner temporarily halts inner retinal blood flow (ischemia). When circulation is reinstated (reperfusion) by removal of the cannula, severe cellular damage ensues, resulting ultimately in retinal neurodegeneration. Recent studies demonstrate inflammation, vascular permeability, and capillary degeneration as additional elements of this model. Compared to alternative retinal I/R methodologies, such as retinal arterial ligation, retinal I/R injury by elevated IOP offers advantages in its&#160;anatomical specificity, experimental tractability, and technical accessibility, presenting itself as a valuable tool for examining neuronal pathogenesis and therapy in the retinal neurovascular unit.

This article describes a procedure for inducing retinal ischemia-reperfusion injury by elevated intraocular pressure in mice. Retinal ischemia-reperfusion injury by elevated intraocular pressure serves to model human pathologies characterized by compromised oxygen and nutrient delivery in the retina, enabling researchers to examine potential cellular mechanisms and treatments for human diseases of the retinal neurovascular unit.

Göz İçi Basıncı Yükselmesi ile Retina İskemi-Reperfüzyon Hasarı Fare Modeli

Retinal ischemia-reperfusion (I/R) is a pathophysiological process contributing to cellular damage in multiple ocular conditions, including glaucoma, ...

Ultrasonographic Assessment During Cardiopulmonary Resuscitation

The US-CAB (Ultrasound, Circulation/Airway/Breathing) protocol integrates several sonographic techniques into a structured assessment of the circulation, airway, and breathing status of a patient during cardiopulmonary resuscitation (CPR) in an advanced life support-compliant manner. US-C provides a subxiphoid view of the heart, to look for potentially reversible causes of disease, such as pericardial effusion, pulmonary embolism, hypovolemia, and acute coronary thrombosis. Sonographic cardiac activity during CPR not only helps differentiate pseudo-pulseless electrical activity (PEA) from true PEA but also represents a higher chance of the return of spontaneous circulation (ROSC) and survival. Evaluation of the inferior vena cava (IVC) shows the fluid status of the patient and indicates the best methods to use for fluid resuscitation. If aortic dissection is suspected, a subxiphoid view of the aorta is suggested for identifying an intimal flap. Once intubation is done, tracheal ultrasound (US-A) at the suprasternal notch helps differentiate endotracheal intubation (one air-mucosal interface with one comet-tail) from esophageal intubation (double tract sign). Immediately following US-A, bilateral lung US (US-B) should be done to confirm proper bilateral ventilation using the lung sliding sign. In addition, US-C can be serially followed to see the dynamic changes in the cardiac chambers and IVC, or any cardiac contraction suggestive of ROSC. US-B can also detect coexisting lung or pleural pathologies without interfering with the performance of CPR. The main concern when implementing this method is maintaining high-quality CPR without delays in chest compressions when performing US-CAB. Rigorous training and continued practice are key to minimize any interruptions during resuscitation.

Here, we present a US-CAB (Ultrasound, Circulation/Airway/Breathing) protocol for use during cardiopulmonary resuscitation (CPR). US-C evaluates the subxiphoid view of the heart and inferior vena cava. After intubation, tracheal US (US-A) and lung US (US-B) help confirm endotracheal intubation and proper ventilation.

Kardiyopulmoner Resüsitasyon Sırasında Ultrasonografik Değerlendirme

The US-CAB (Ultrasound, Circulation/Airway/Breathing) protocol integrates several sonographic techniques into a structured assessment of the circulation, ...

A Pre-clinical Rat Model for the Study of Ischemia-reperfusion Injury in Reconstructive Microsurgery

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many areas of biomedical research due to its cost-effectiveness and its translation to humans. This protocol describes a method to create a preclinical free skin flap model in rats with ischemia-reperfusion injury. The described 3 cm x 6 cm rat free skin flap model is easily obtained after the placement of several vascular ligatures and the section of the vascular pedicle. Then, 8 h after the ischemic insult and completion of the microsurgical anastomosis, the free skin flap develops the tissue damage. These ischemia-reperfusion injury-related damages can be studied in this model, making it a suitable model for evaluating therapeutic agents to address this pathophysiological process. Furthermore, two main monitoring techniques are described in the protocol for the assessment of this animal model: transit-time ultrasound technology and laser speckle contrast analysis.

Here, we describe a pre-clinical animal model for studying the pathophysiology of ischemia-reperfusion injury in reconstructive microsurgery. This free skin flap model based on the superficial caudal epigastric vessels in the rat may also allow for the evaluation of different therapies and compounds to counteract ischemia-reperfusion injury-related damage.

Rekonstrüktif Mikrocerrahide İskemi-Reperfüzyon Hasarı Nın İnceleştirilmesinde Klinik Öncesi Sıçan Modeli

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many ...

Delayed Intramyocardial Delivery of Stem Cells after Ischemia Reperfusion Injury in a Murine Model

There is significant interest in the use of stem cells (SCs) for the recovery of cardiac function in individuals with myocardial injuries. Most commonly, cardiac stem cell therapy is studied by delivering SCs concurrently with the induction of myocardial injury. However, this approach presents two significant limitations: the early hostile pro-inflammatory ischemic environment may affect the survival of transplanted SCs, and it does not represent the subacute infarction scenario where SCs will likely be used. Here we describe a two-part series of surgical procedures for the induction of ischemia-reperfusion injury and delivery of mesenchymal stem cells (MSCs). This method of stem cell administration may allow for the longer viability and retention around damaged tissue by circumventing the initial immune response. A model of ischemia reperfusion injury was induced in mice accompanied by the delivery of mesenchymal stem cells (3.0 x 105), stably expressing the reporter gene firefly luciferase under the constitutively expressed CMV promoter, intramyocardially 7 days later. The animals were imaged via ultrasound and bioluminescent imaging for confirmation of injury and injection of cells, respectively. Importantly, there was no added complication rate when performing this two-procedure approach for SC delivery. This method of stem cell administration, collectively with the utilization of state-of-the-art reporter genes, may allow for the in vivo study of viability and retention of transplanted SCs in a situation of chronic ischemia commonly seen clinically, while also circumventing the initial pro-inflammatory response. In summary, we established a protocol for the delayed delivery of stem cells into the myocardium, which can be used as a potential new approach in promoting regeneration of the damaged tissue.

Stems cells are continuously investigated as potential treatments for individuals with myocardial damage, however, their decreased viability and retention within injured tissue can impact their long-term efficacy. In this manuscript we describe an alternative method for stem cell delivery in a murine model of ischemia reperfusion injury.

Bir Murine Modelinde İskemi Reperfüzyon Yaralanması Sonrası Kök Hücrelerin Gecikmiş İntramiyokardiyal Teslimi

There is significant interest in the use of stem cells (SCs) for the recovery of cardiac function in individuals with myocardial injuries. Most commonly, ...

A Rat Lung Transplantation Model of Warm Ischemia/Reperfusion Injury: Optimizations to Improve Outcomes

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods for choosing appropriate cuff sizes for the pulmonary vein (PV), pulmonary artery (PA), or bronchus (Br) are varied, thus making the determination of proper cuff size during rat lung transplantation an exercise of trial and error. By standardizing the cuffing technique to use the smallest effective cuff appropriate for the size of the vessel or bronchus, one can make the transplantation procedure safer, faster, and more successful. Since diameters of the PV, PA, and Br are related to the body weight of the rat, we present a strategy to choosing an appropriate size using a weight-based guide. Since lung volume is also related to body weight, we recommend that this relationship should also be considered when choosing the proper volume of air for donor lung inflation during warm ischemia as well as for the proper volume of PBS to be instilled during bronchoalveolar lavage (BAL) fluid collection. We also describe methods for 4th intercostal space dissection, wound closure, and sample collection from both the native and transplanted lobes.

Here, we present optimizations to a rat lung transplantation model that serve to improve outcomes. We provide a size guide for cuffs based on body weight, a measurement strategy to ascertain the 4th intercostal space, and methods of wound closure and BAL (bronchoalveolar lavage) fluid and tissue collection.

Sıcak iskemi / reperfüzyon hasarının bir sıçan akciğer transplantasyon modeli: sonuçları iyileştirmek için optimizasyonlar

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods ...

An Effective Mouse Model of Unilateral Renal Ischemia-Reperfusion Injury

Ischemia-reperfusion injury (IRI) is the leading cause of acute renal failure and is a significant contributor to delayed graft function. Animal models are the only available resources that mimic the complexities of the IRI-associated damage encountered in vivo. This paper describes an effective mouse model of unilateral renal IRI that delivers highly reproducible data. Ischemia is induced by occluding the right renal pedicle for 30 min followed by reperfusion. In addition to the surgical procedure, a sequential overview of the expected physiological and histopathological changes following renal IRI will be provided by comparing data from seven different reperfusion times (4 h, 8 h, 16 h, 1 day, 2 days, 4 days, and 7 days). Critical data for planning experiments ahead, such as mean surgical time, average anesthetic consumption, and body weight changes over time, will be shared. This work will help researchers implement a reliable renal IRI model and select the appropriate reperfusion time that aligns with their intended investigative goals.

Renal ischemia-reperfusion injury is associated with high morbidity and mortality in hospitalized patients. Here, we present a simple and effective mouse model of unilateral renal ischemia-reperfusion injury and provide a sequential overview of representative pathological changes observed in the kidney.

Tek Taraflı İskemi-Reperfüzyon Hasarının Etkili Bir Fare Modeli

Ischemia-reperfusion injury (IRI) is the leading cause of acute renal failure and is a significant contributor to delayed graft function. Animal models ...

A Piglet Perinatal Asphyxia Model to Study Cardiac Injury and Hemodynamics after Cardiac Arrest, Resuscitation, and the Return of Spontaneous Circulation

Neonatal piglets have been extensively used as translational models for perinatal asphyxia. In 2007, we adapted a well-established piglet asphyxia model by introducing cardiac arrest. This enabled us to study the impact of severe asphyxia on key outcomes, including the time taken for the return of spontaneous circulation (ROSC), as well as the effect of chest compressions according to alternative protocols for cardiopulmonary resuscitation. Due to the anatomical and physiological similarities between piglets and human neonates, piglets serve as good models in studies of cardiopulmonary resuscitation and hemodynamic monitoring. In fact, this cardiac arrest model has provided evidence for guideline development through research on resuscitation protocols, pathophysiology, biomarkers, and novel methods for hemodynamic monitoring. Notably, the incidental finding that a substantial fraction of piglets have pulseless electrical activity (PEA) during cardiac arrest may increase the applicability of the model (i.e., it may be used to study pathophysiology extending beyond the perinatal period). However, the model generation is technically challenging and requires various skill sets, dedicated personnel, and a fine balance of the measures, including the surgical protocols and the use of sedatives/analgesics, to ensure a reasonable rate of survival. In this paper, the protocol is described in detail, as well as experiences with adaptations to the protocol over the years.

This piglet model involves surgical instrumentation, asphyxiation until the cardiac arrest, resuscitation, and post-resuscitation observation. The model allows for multiple sampling per animal, and by using continuous invasive arterial blood pressure, ECG, and non-invasive cardiac output monitoring, it provides knowledge about hemodynamics and cardiac pathophysiology in perinatal asphyxia and neonatal cardiopulmonary resuscitation.

Kardiyak Arrest, Resüsitasyon ve Spontan Dolaşımın Geri Dönüşü Sonrası Kardiyak Hasar ve Hemodinamiği İncelemek İçin Bir Domuz Yavrusu Perinatal Asfiksi Modeli

Neonatal piglets have been extensively used as translational models for perinatal asphyxia. In 2007, we adapted a well-established piglet asphyxia model ...