Mevcut çalışmalar Amerika Birleşik Devletleri Ulusal Sağlık Enstitülerinden hibe HL092188-R01, R01-DK083385 ve R01HL098294 ve Anestezi Eğitim ve Araştırma Vakfı tarafından desteklenmiştir (FAER) HKE ve DFG (Deutsche Forschungsgemeinschaft) Araştırma Bursu (için ArtırdıZAGREP GR2121/1- 1) ve AG Amerikan Kalp Derneği Grant.

Genel notlar. Tüm işlemler dik bir mikroskop (Leica) altında ve cerrahi koagülatör kullanılarak yapılmalıdır. Deneysel ve plasebo gruplarında fareler sonuçların karşılaştırılabilir sağlamak için yaş ve ağırlık uyumlu olmalıdır. Sıcaklık, kan basıncı, anestezi ve sıvı yönetiminin istikrarlı ve tüm deney sırasında takip edilmelidir. 1. Anestezi ve Cerrahi <ol><li> C57BL / 6 fareleri 10 ila 16 haftalık bir yaş aralığında olmalı ve cinsiyet açısından ayrı gruplar halinde olmalıdır. </li><li> Sodyum pentobarbital (70 mg / kg vücut ağırlığı ip) ile anestezi farelerde ve yaklaşık ile anestezi bakımı. 10 mg / kg / saat sodyum pentobarbital. Aşırı Dozlama önemli ölçüde daha düşük kan basıncı ve böylece sonuçlar değiştirebilir. </li><li> Bir rektal termometre probu ile sıcaklık kontrollü ısıtmalı masa (RT, Effenberg, Münih, Almanya) Yer fareler bir termal geribildirim bağlı37 de vücut sıcaklığını koruyacak şekilde kontrol ° C. </li><li> Çıkarılabilir bant ile tablosu bağlı üst ve alt ekstremitelerin ile cerrahi tabloda bir yatar konumda yerinde fareler,. </li><li> Yaşam deneyleri durumunda saç kırpma, iyodoforlar (örneğin Betadine), otoklavlanmış aletlerin steril işleme ve maskeleri cerrah için steril eldiven kullanarak cilt dezenfeksiyonu dahil olmak üzere uygun aseptik teknikler gereklidir. </li></ol> 2. Portal Triad Tıkanıklığı <ol><li> Deri kesme ve böylece fare saç alerji riskini önleyerek fare saç teneffüs önlemek için mineral yağ ile kesi alanı kapsamaktadır. Kolayca karaciğer maruz böylece kadar üst sonuna kadar linea alba orta hat laparotomi insizyonu gerçekleştirin. Mümkünse kanamayı önlemek için bir koagülasyon elektrodu (Erbe, ICC50, Tübingen, Almanya) ile kesi yapar. </li><li> Kuyruk Direc olduğu bir konumda fare ile ısıtma masa yerleştirincerraha ted. Bu pozisyon üçlü yerelleştirilmesine ve dikiş yerleştirmek için en iyi ayarı sağlar. </li><li> Portal triadı ortaya çıkarmak için ıslak bir pamuk uçlu çubuk kullanarak sağ tarafta bağırsak çekilin. </li><li> Biraz diyaframı doğru medyan ve sol lob ve bağırsak doğru sağ lob kaydırır. Yani, sağ / sol medyan lobların çatallanma yukarıda portal triadı net bir görünüm verecektir. </li><li> Dikkatle portal triadı incelemek ve mülayim forseps ile yavaşça aşağı sağ lob tutun. Yani, sol portal triadı güzel bir görünüm verecektir. </li><li> Soldan sağa doğru hepatik arter, hepatik ven ve safra kanalı dahil olmak üzere sol portal triadı altında bir iğne tutucu üzerinden; sonra görsel sütür (Ethicon, Norderstedt, Almanya 7/0 naylon sütür) ile yer iğne tespit Farenin. </li><li> Dikiş üçlü altında yakından olmalıdır. Daha derin bir dikiş altında bulunan vena vaca aşağı yaralayabilirüçlü. Bu nedenle dikiş koymadan önce vena kava tanımlamak için forseps ile karaciğerin sağ lobu basılı tutun. </li><li> Bir forseps ile sağ tarafta iğne ucu almak ve karaciğer zarar vermeden karaciğer alanı dışında hafifçe çekin. </li><li> Sütür bir Eppendorf tüp (su ile dolu yaklaşık 3 g) her bir ucuna takın ve her iki dikiş dikiş sahipleri üzerinde biter yerleştirin. Sütür doğru üçlü karışmaz böylece distalde dikiş dikiş sahibi yerleştirin. </li><li> Ağırlıklar uygulayarak üçlü karaciğer sol ve median lob kan akımı kesintiye neden hemen tıkalı olacak. Başarılı tıkanıklığı kolaylıkla iskemik lobların soluk soluklaşmanın görsel muayene ile teyit edilebilir. Asılı ağırlıkları kutuplar salgılanır ve karaciğer reperfüzyon zaman aksine, renk değişikliği hemen kaybolur. Bu kısmi karaciğer iskemi modeli bloo koruyarak mezenter tıkanıklığı önlerkaraciğer sağ lob d akışı. </li><li> Karın boşluğuna bağırsak değiştirin. 37.0 su ile ıslatılmış ıslak bir bez ile karaciğer ve bağırsak sıcak tutun ° C Ameliyat bittikten sonra piyasada bulunan gıda ambalajlarında ile farelerin kapsayacak vücut ısısını dengede tutmak için ek olarak. </li><li> Deneysel protokol başına gerektiği gibi iskemi gerçekleştirin. Tanımlanmış bir iskemi zamanı bırakılmasından sonra ortanca ve karaciğer sol lob ağırlıkları ve reperfüzyon hemen başlayacaktır. Bu teknik, farelerde tersine çevrilebilir iskemi üreten sağlar. Başka iskemi kez yapılacak ise dikkatle sütür çıkarın. </li><li> 15 dakika önce ve doğrudan reperfüzyon sonrası karın boşluğuna yaklaşık 400 ul sodyum klorür ekleyin. Reperfüzyon süresince sıvı kaybı telafi etmek için 100 ul&#39;lik bir miktarda her 30 dakikada bir sıvı ikame devam etmektedir. </li><li> Sağkalım deneyler için cerrahi yara sürekli kas duvarının sütür ve cilt wi kullanarak kapalı4/0 sütür th. </li></ol> 3. Karaciğer Perfüzyon değişiklikler Aşağıdaki sonuç parametreleri karaciğer hasarı aşağıdaki hepatik iskemi belirlemek için tavsiye edilir: Karaciğer enzimleri (örneğin ALT, AST) ve karaciğer histolojisi (H &amp; E boyama) aşağıdaki karaciğer iskemi ve reperfüzyon 3,9 belirleyin. <img alt="Şekil 1" src="/files/ftp_upload/2550/2550fig1.jpg" /> Şekil 1 karaciğer iskemi için Asılı ağırlık sisteminin Model:. A asılı ağırlık sistemi, sol ve orta lob 3 karaciğer iskemi ikna etmek için kurulmuştur. (A) Bu nedenle kaudat ve sağ lob hafifçe sol lob ayrıldı. Sağ lob açıkça sağ, medyan ve sol lob çatallanma yukarıda portal triadı görüntülemek için hafifçe kaymış oldu. Bir 8.0 naylon sütür sol karaciğer üçlü altına alınmıştır. (B) Her sütür sonunda küçük bir ağırlığı (az 3 g Eppendorf tusu ile doldurulur bes) eklenmiş ve uygulanmıştır. Karaciğer iskemi kırmızıdan soluk için sol ve orta lob bir renk değişimi ile görünür. Bu deneysel tasarım kilo yük uygulamadan ve bırakmadan kesin tıkanma ve sol hepatik üçlünün reperfüzyon sağlar. <img alt="Şekil 2" src="/files/ftp_upload/2550/2550fig2.jpg" /> Şekil 2. Alanin (ALT) ve aspartat (AST) farelerde aminotransferaz farklı karaciğer iskemi süresinin etkisi. Kısmi portal triad iskemi belirtilen (- 60 dakika iskemi 0) olarak oluşturuldu. Reperfüzyon 2 saat sonra, ALT (A) ve AST (B) ölçüldü. 7 fareler / group - Sonuçlar ortalama ± SE 6 olarak ifade edilir. * P 0 dakika iskemi (sham kontrol) ile karşılaştırıldığında &lt;0.05.

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Çıkar çatışması ilan etti.

Bu çalışmada sol hepatik portal triad oklüzyonu Asılı ağırlık sistemi kullanılarak bir fare modelinde iskemi yapan bir teknik anlatılmaktadır. Bunu öğrenmek için kolay bir yöntem olmasına rağmen şu tuzaklar dikkate alınmalıdır. Başlangıçta bir çok yaygın bir hata portal triadı ve vena kava içeren derin bir dikiş olduğunu. Solunum ve kalp hızı artan direniş ve farelerin erken ölümle sonuçlanan kalp kan dönen azalma nedeniyle 15 dakika içinde yavaşlayacaktır. Vena kava dahil önlemenin en iyi yolu bir forseps ile sağ lobu basılı tutarak dikiş önce vena kava yerelleştirilmesine olduğunu. Bir başka önemli konu ağırlıkları da sağ lob kan akışını durdurur sağ ve sol portal yargılanan çok strese yol açabilir ağır olmamasıdır. Bu nedenle Eppendorf şişe içindeki su miktarı fare ağırlığını ve s üzere ayarlanması gerekmektediruture tutucular dikiş yerinden distal yerleştirilmelidir. Karşısında ağırlıkları düşük kan basıncı için ise tamamen durmuş değil. Her ikisi de hataları kolayca sırasıyla medyan ve sol lob sağ lob veya yetersiz renk değişikliği soluk ilave bir renk değişimi tarafından algılanabilen edilir. Bu nedenle ağırlıklarının uygulanması takiben hepatik renk değişimi her 10 dakikada bir kontrol edilmelidir. Başka bir yaygın cerrahi komplikasyon iğne ile karaciğer yaralanmasına neden olduğunu. Bu nedenle sağ lob iğne ucu rahatlıkla görülebilen ve iğne tutucu ile alınabilir böylece mülayim bir forseps ile basılı tutulmalıdır. Başlangıçta bu zorlukları göz önüne alınırsa hepatik iskemi için Asılı ağırlık sistemi portal üçlünün sıkma ile ilişkili karaciğer hasarının değişkenliği en aza indirerek iskemi nedeniyle yüksek oranda tekrarlanabilir yaralanma sağlar. Asılı ağırlık sistemi kullanarak, portal triadı sadece bütün boyunca bir kez manipüle edilircerrahi, karaciğer lobunda önemli ölçüde daha az hasara neden olur. Buna ek olarak, herhangi bir hepatik veya bağırsak konjesyon, bu teknik ile ortaya çıkar. Daha fazla doku travması özellikle iskemik önkoşullama sırasında olduğu gibi birden fazla iskemi döngüleri sırasında kelepçe tekrar uygulama ile, damar kelepçeler kaldırarak ve değiştirilmesi ile oluşur. Bir mengene başvurmamız tarafından karaciğer lobların manipülasyon nedeniyle doku travması önlerken, bizim gözlemlerimize göre, karaciğer dokusunda uzak bir konumda ağırlık asılı kullanımı güvenilir tıkanıklığı avantajı sağlar. Birlikte ele alındığında, bu çalışma, germe ile ilişkili değişkenlik ve sınırlamaları en aza indirerek, iskemi sırasında portal triad oklüzyonu Asılı ağırlık sisteminin fizibilite sağlar. Böylece bu teknik hepatik iskemi okullar diğer araştırmacılar için ilgi çekici olabilir.

<table border="1"><tbody><tr><td> Reaktif Adı </td><td> Şirket </td><td> Katalog numarası </td><td> Yorumlar </td></tr><tr><td> Sodyum pentobarbital (Fatal Plus) </td><td> Vortech İlaç Ls, Ltd </td><td> VPL 9372 </td><td> Tuzlu su içinde 4 mg / ml </td></tr><tr><td> Dikiş, Prolene 8.0 </td><td> Ethicon, ABD </td><td> M8739 </td><td> yeniden </td></tr><tr><td> mikroskop diseksiyon </td><td> Leica </td><td> n / a </td><td> cömert çalışma mesafesi dikkate </td></tr><tr><td> Isıtma Tablo </td><td> Rt, Effenberger, Almanya </td><td> n / a </td><td> Sadece ve tek sağlayıcı </td></tr><tr><td> Cerrahi aletler </td><td> TEFE, Dumont </td><td></td><td> Forseps, iğne tutucu </td></tr><tr><td> Koagülatör </td><td> Erbe, ICC50, Tübingen, Almanya </td><td> n / a </td><td> Coagulators Farklı sağlayıcılar </td></tr></tbody></table> Tablo 1. Özel reaktifleri ve ekipmanları.

use of a hanging-weight system for liver ischemia in mice

Iskemiye bağlı akut karaciğer hasarı, çeşitli klinik prosedürler örneğin karaciğer nakli, karaciğer tümörü rezeksiyonu veya travma onarım sırasında oluşabilir ve yüksek ölüm oranı 1-2 olan karaciğer yetmezliğine neden olabilir. Hepatik iskemi Bu nedenle fare çalışmaları farmakolojik ve genetik çalışmalar 3-9 ile tanışma imkanı sağlayarak önemli bir araştırma alanı haline gelmiştir. Özellikle, bir genin doku spesifik silinmesi ile koşullu fareler (CRE, Flox sistemi) özellikle dokularda 10-13 proteinlerin rolünü içine bir anlayış sağlamak. Çünkü elle farelerde portalı üçlü sıkıştırma ile ilgili teknik zorluk, daha önce 3 tarif edilmiştir portal triad oklüzyonu Asılı ağırlık sistemi kullanılarak sistematik bir değerlendirme yapıldı. Asılı ağırlık sistemi kullanılarak ayrıca bu yana, karaciğer lobunda herhangi bir zarar vermeden portal triadı sol kolu etrafında bir sütür yerleştirinmevcut en iyi kelepçeler yüzünden gemilere karaciğer dokusu yakın konumu karaciğer dokusunda hasara neden olabilir. Ayrıca, hepatik üçlünün sağ kolu hala böylece perfüze hiçbir bağırsak tıkanıklığı korunur karaciğer sağ lob kan akışını olarak bu teknik ile ortaya çıkar. Ayrıca, portal triadı sadece bütün cerrahi işlem boyunca bir kez manipüle edilir. Bunun sonucu olarak, prosedürler kolayca yapılabilir, iskemi ve reperfüzyon kısa süreleri ile, ön şartlandırma gibi. Alanin (ALT) ve aspartat (AST) aminotransferaz serum 3,9 olarak ölçülen farklı iskemi ve reperfüzyon kez yaparak bu modeli sistematik değerlendirmesi karaciğer hasarı ile karaciğer iskemi süresinin yakın bir ilişki tespit edilmedi. Hepatik iskemi ve aralıklı reperfüzyon için Asılı ağırlık sistemi kullanılırken birlikte değerlendirildiğinde, bu çalışmaların yüksek oranda tekrarlanabilir karaciğer hasarı onaylayın. Böylece, bu tekniğin diğer araştırmacılar ilgi için faydalı olabilirfarelerde karaciğer iskemi çalışmalarda ed. Bu nedenle, video klibi, bu tekniğin ayrıntılı adım adım açıklamalar sağlar.

Watch this Scientific Journal Video about Use of a Hanging-weight System for Liver Ischemia in Mice at JoVE.com

Use of a Hanging-weight System for Liver Ischemia in Mice

Biz portal triad oklüzyonu Asılı ağırlık sisteminin yeni bir fare modelini kurdu. Bu teknik, sıçan karaciğer modellerinde iskemi gelecekteki araştırmalar için yararlı olabilir.

Farelerde Karaciğer İskemi için Asılı ağırlık Sistem Kullanımı

Acute liver injury due to ischemia can occur during several clinical procedures e.g. liver transplantation, hepatic tumor resection or trauma repair and ...

use-of-a-hanging-weight-system-for-liver-ischemia-in-mice

Research

JoVE Journal

Medicine

15.9K Görüntüleme.  University of Colorado, Denver. Biz portal triad oklüzyonu Asılı ağırlık sisteminin yeni bir fare modelini kurdu. Bu teknik, sıçan karaciğer modellerinde iskemi gelecekteki araştırmalar için yararlı olabilir.

Video: Use of a Hanging-weight System for Liver Ischemia in Mice

Use of a Hanging Weight System for Coronary Artery Occlusion in Mice

Murine studies of acute injury are an area of intense investigation, as knockout mice for different genes are becoming increasingly available 1-38. Cardioprotection by ischemic preconditioning (IP) remains an area of intense investigation. To further elucidate its molecular basis, the use of knockout mouse studies is particularly important 7, 14, 30, 39. Despite the fact that previous studies have already successfully performed cardiac ischemia and reperfusion in mice, this model is technically very challenging. Particularly, visual identification of the coronary artery, placement of the suture around the vessel and coronary occlusion by tying off the vessel with a supported knot is technically difficult. In addition, re-opening the knot for intermittent reperfusion of the coronary artery during IP without causing surgical trauma adds additional challenge. Moreover, if the knot is not tied down strong enough, inadvertent reperfusion due to imperfect occlusion of the coronary may affect the results. In fact, this can easily occur due to the movement of the beating heart.
Based on potential problems associated with using a knotted coronary occlusion system, we adopted a previously published model of chronic cardiomyopathy based on a hanging weight system for intermittent coronary artery occlusion during IP 39. In fact, coronary artery occlusion can thus be achieved without having to occlude the coronary by a knot. Moreover, reperfusion of the vessel can be easily achieved by supporting the hanging weights which are in a remote localization from cardiac tissues.
We tested this system systematically, including variation of ischemia and reperfusion times, preconditioning regiments, body temperature and genetic backgrounds39. In addition to infarct staining, we tested cardiac troponin I (cTnI)
as a marker of myocardial infarction in this model. In fact, plasma levels of cTnI correlated with infarct sizes (R2=0.8). Finally, we could show in several studies that this technique yields highly reproducible infarct sizes during murine IP and myocardial infarction6, 8, 30, 40, 41. Therefore, this technique may be helpful for researchers who pursue molecular mechanisms involved in cardioprotection by IP using a genetic approach in mice with targeted gene deletion. Further studies on cardiac IP using transgenic mice may consider this technique.

A murine model for myocardial ischemia and ischemic preconditioning is an important tool study cardioprotective mechanisms in vivo. Here, we report an easy applicable in situ model for cardiac IP using a hanging-weight system for coronary artery occlusion.

Asma Ağırlık Sistemi Fare Koroner Arter Tıkanıklığı için kullanın

Murine studies of acute injury are an area of intense investigation, as knockout mice for different genes are becoming increasingly available 1-38. ...

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Use of a Hanging-weight System for Isolated Renal Artery Occlusion

In hospitalized patients, over 50% of cases of acute kidney injury (AKI) are caused by renal ischemia 1-3. A recent study of hospitalized patients revealed that only a mild increase in serum creatinine levels (0.3 to 0.4 mg/dl) is associated with a 70% greater risk of death than in persons without any increase 1. Along these lines, surgical procedures requiring cross-clamping of the aorta and renal vessels are associated with a renal failure rates of up to 30% 4. Similarly, AKI after cardiac surgery occurs in over 10% of patients under normal circumstances and is associated with dramatic increases in mortality. AKI are also common complications after liver transplantation. At least 8-17% of patients end up requiring renal replacement therapy 5. Moreover, delayed graft function due to tubule cell injury during kidney transplantation is frequently related to ischemia-associated AKI 6. Moreover, AKI occurs in approximately 20% of patients suffering from sepsis 6.The occurrence of AKI is associated with dramatic increases of morbidity and mortality 1. Therapeutic approaches are very limited and the majority of interventional trials in AKI have failed in humans. Therefore, additional therapeutic modalities to prevent renal injury from ischemia are urgently needed 3, 7-9. 
To elucidate mechanisms of renal injury due to ischemia and possible therapeutic strategies murine models are intensively required 7-13. Mouse models provide the possibility of utilizing different genetic models including gene-targeted mice and tissue specific gene-targeted mice (cre-flox system). However, murine renal ischemia is technically challenging and experimental details significantly influence results. We performed a systematic evaluation of a novel model for isolated renal artery occlusion in mice, which specifically avoids the use of clamping or suturing the renal pedicle 14. This model requires a nephrectomy of the right kidney since ischemia can be only performed in one kidney due to the experimental setting. In fact, by using a hanging-weight system, the renal artery is only instrumented once throughout the surgical procedure. In addition, no venous or urethral obstruction occurs with this technique. We could demonstrate time-dose-dependent and highly reproducible renal injury with ischemia by measuring serum creatinine. Moreover, when comparing this new model with conventional clamping of the whole pedicle, renal protection by ischemic preconditioning is more profound and more reliable. Therefore his new technique might be useful for other researchers who are working in the field of acute kidney injury.

A precise murine model for acute kidney injury (AKI) due to ischemia is an important tool to investigate acute kidney injury and possibly find therapeutic tools to treat renal injury. The hanging weight system offers a tool for immediate and reliable renal artery occlusion and reperfusion without causing renal congestion.

İzole Renal Arter Tıkanıklığı Asılı ağırlık Sistemi

In hospitalized patients, over 50% of cases of acute kidney injury (AKI) are caused by renal ischemia 1-3. A recent study of hospitalized patients ...

Use of Animal Model of Sepsis to Evaluate Novel Herbal Therapies

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several techniques, including infusion of exogenous bacterial toxin (endotoxemia) or bacteria (bacteremia), as well as surgical perforation of the cecum by cecal ligation and puncture (CLP)1-3. CLP allows bacteria spillage and fecal contamination of the peritoneal cavity, mimicking the human clinical disease of perforated appendicitis or diverticulitis. The severity of sepsis, as reflected by the eventual mortality rates, can be controlled surgically by varying the size of the needle used for cecal puncture2. In animals, CLP induces similar, biphasic hemodynamic cardiovascular, metabolic, and immunological responses as observed during the clinical course of human sepsis3. Thus, the CLP model is considered as one of the most clinically relevant models for experimental sepsis1-3.
Various animal models have been used to elucidate the intricate mechanisms underlying the pathogenesis of experimental sepsis. The lethal consequence of sepsis is attributable partly to an excessive accumulation of early cytokines (such as TNF, IL-1 and IFN-&gamma;)4-6 and late proinflammatory mediators (e.g., HMGB1)7. Compared with early proinflammatory cytokines, late-acting mediators have a wider therapeutic window for clinical applications. For instance, delayed administration of HMGB1-neutralizing antibodies beginning 24 hours after CLP, still rescued mice from lethality8,9, establishing HMGB1 as a late mediator of lethal sepsis. The discovery of HMGB1 as a late-acting mediator has initiated a new field of investigation for the development of sepsis therapies using Traditional Chinese Herbal Medicine. In this paper, we describe a procedure of CLP-induced sepsis, and its usage in screening herbal medicine for HMGB1-targeting therapies.

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection, and can be simulated by a surgical technique termed cecal ligation and puncture (CLP). Here we describe a method to use CLP-induced animal model to screen medicinal herbs for therapeutic agents.

Roman Bitkisel Tedaviler Değerlendirmek için Sepsis Hayvan Modeli kullanımı

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several ...

Intraoperative Detection of Subtle Endometriosis: A Novel Paradigm for Detection and Treatment of Pelvic Pain Associated with the Loss of Peritoneal Integrity

Endometriosis is a common disease affecting 40 to 70% of reproductive-aged women with chronic pelvic pain (CPP) and/or infertility. The purpose of this study was to demonstrate the use of a blue dye (methylene blue) to stain peritoneal surfaces during laparoscopy (L/S) to detect the loss of peritoneal integrity in patients with pelvic pain and suspected endometriosis. Forty women with CPP and 5 women without pain were evaluated in this pilot study. During L/S, concentrated dye was sprayed onto peritoneal surfaces, then aspirated and rinsed with Lactated Ringers solution. Areas of localized dye uptake were evaluated for the presence of visible endometriotic lesions. Areas of intense peritoneal staining were resected and some fixed in 2.5% buffered gluteraldehyde and examined by scanning (SEM) electron microscopy. Blue dye uptake was more common in women with endometriosis and chronic pelvic pain than controls (85% vs. 40%). Resection of the blue stained areas revealed endometriosis by SEM and loss of peritoneal cell-cell contact compared to normal, non-staining peritoneum. Affected peritoneum was associated with visible endometriotic implants in most but not all patients. Subjective pain relief was reported in 80% of subjects. Based on scanning electron microscopy, we conclude that endometrial cells extend well beyond visible implants of endometriosis and appear to disrupt the underlying mesothelium. Subtle lesions of endometriosis could therefore cause pelvic pain by disruption of peritoneal integrity, allowing menstrual or ovulatory blood and associated pain factors access to underlying sensory nerves. Complete resection of affected peritoneum may provide a better long-term treatment for endometriosis and CPP. This simple technique appears to improve detection of subtle or near invisible endometriosis in women with CPP and minimal visual findings at L/S and may serve to elevate diagnostic accuracy for endometriosis at laparoscopy.

Loss of peritoneal integrity provides a new paradigm to understand and treat chronic pelvic pain in women with mild forms of endometriosis and can be easily detected using intraoperative instillation of dye at the time of laparoscopy.

Hafif Endometriozis İntraoperatif Algılama: Periton Bütünlüğü Kaybı ile İlişkili Pelvik Ağrı Tespiti ve Tedavisi İçin Yeni Bir Paradigma

Endometriosis is a  common disease affecting 40 to 70% of reproductive-aged women with chronic  pelvic pain (CPP) and/or infertility. The purpose of this ...

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study model. As the use of mouse models are often a vital step in preclinical drug discovery and drug development1-8, it is necessary to design a system to introduce drugs into mice in a uniform, reproducible manner. Ideally, the system should permit the collection of blood samples at regular intervals over a set time course. The ability to measure drug concentrations by mass-spectrometry, has allowed investigators to follow the changes in plasma drug levels over time in individual mice1, 9, 10. In this study, paclitaxel was introduced into transgenic mice as a continuous arterial infusion over three hours, while blood samples were simultaneously taken by retro-orbital bleeds at set time points. Carotid artery infusions are a potential alternative to jugular vein infusions, when factors such as mammary tumors or other obstructions make jugular infusions impractical. Using this technique, paclitaxel concentrations in plasma and tissue achieved similar levels as compared to jugular infusion. In this tutorial, we will demonstrate how to successfully catheterize the carotid artery by preparing an optimized catheter for the individual mouse model, then show how to insert and secure the catheter into the mouse carotid artery, thread the end of the catheter out through the back of the mouse&#8217;s neck, and hook the mouse to a pump to deliver a controlled rate of drug influx. Multiple low volume retro-orbital bleeds allow for analysis of plasma drug concentrations over time.

This method was developed with the goal of delivering a steady drug solution via the carotid artery, to assess the pharmacokinetics of novel drugs in mouse models.

Farelerde Taksanlar Farmakokinetik ve Farmakodinamik Analiz Karotis Arter İnfüzyonlar

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

Photothrombosis-induced Focal Ischemia as a Model of Spinal Cord Injury in Mice

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) below the site of injury. The secondary ischemia that develops following the initial mechanical insult is a serious complication of the SCI and severely impairs the function and viability of surviving neuronal and non-neuronal cells in the SC. In addition, ischemia is also responsible for the growth of lesion during chronic phase of injury and interferes with the cellular repair and healing processes. Thus there is a need to develop a spinal cord ischemia model for studying the mechanisms of ischemia-induced pathology. Focal ischemia induced by photothrombosis (PT) is a minimally invasive and very well established procedure used to investigate the pathology of ischemia-induced cell death in the brain. Here, we describe the use of PT to induce an ischemic lesion in the spinal cord of mice. Following retro-orbital sinus injection of Rose Bengal, the posterior spinal vein and other capillaries on the dorsal surface of SC were irradiated with a green light resulting in the formation of a thrombus and thus ischemia in the affected region. Results from histology and immunochemistry studies show that PT-induced ischemia caused spinal cord infarction, loss of neurons and reactive gliosis. Using this technique a highly reproducible and relatively easy model of SCI in mice can be achieved that would serve the purpose of scientific investigations into the mechanisms of ischemia induced cell death as well as the efficacy of neuroprotective drugs. This model will also allow exploration of the pathological changes that occur following SCI in live mice like axonal degeneration and regeneration, neuronal and astrocytic Ca2+ signaling using two-photon microscopy.

Photothrombosis is a minimally invasive and highly reproducible procedure to induce focal ischemia in the spinal cord and serves as a model of spinal cord injury in mice.

Farelerde Omurilik Yaralanma Modeli Olarak Odak iskemisi Fototromboz kaynaklı

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) ...

Use of a Low-flow Digital Anesthesia System for Mice and Rats

A traditional vaporizer depends on flowing gas and atmospheric pressure for passive anesthetic vaporization. Newly developed direct injection vaporizers utilize a syringe pump to directly administer volatile anesthetics into a gas stream. Unlike a traditional vaporizer, it can be used at very low flow rates, making it ideal for use on mice and rats. 
The equipment's capability to use low flow rates could result in a substantial cost savings due to the reduced need for anesthetic agents, compressed gas, and charcoal scavenging filters1. A lower flow rate means less waste of anesthetic gas and likely reduces the risk of anesthetic exposure to laboratory personnel. Thus, the high levels of precision and safety associated with direct injection vaporizers, along with a reduced need for anesthetic agents, compressed gas, and charcoal filters are beneficial for research requiring small animal anesthesia. 
The goal of this protocol is to demonstrate the use of a syringe-driven direct injection vaporizer as part of a digital, low-flow anesthesia system. The direct injection vaporizer is capable of accurately delivering anesthesia at very low flow rates compared to a traditional vaporizer, making it a promising alternative for controlled gas anesthetic delivery to rodents.

Here, we present a protocol to more safely and efficiently administer anesthetic gas to mice using a digital, low flow anesthesia system utilizing a syringe-driven direct injection vaporizer.

Fare ve Sıçanlarda için düşük akış Dijital Anestezi Sistemi Kullanımı

A traditional vaporizer depends on flowing gas and atmospheric pressure for passive anesthetic vaporization. Newly developed direct injection vaporizers ...

Confirmation of Myocardial Ischemia and Reperfusion Injury in Mice Using Surface Pad Electrocardiography

Many animal models have been established for the study of myocardial remodeling and heart failure due to its status as the number one cause of mortality worldwide. In humans, a pathologic occlusion forms in a coronary artery and reperfusion of that occluded artery is considered essential to maintain viability of the myocardium at risk. Although essential for myocardial recovery, reperfusion of the ischemic myocardium creates its own tissue injury. The physiologic response and healing of an ischemia/reperfusion injury is different from a chronic occlusion injury. Myocardial ischemia/reperfusion injury is gaining recognition as a clinically relevant model for myocardial infarction studies. For this reason, parallel animal models of ischemia/reperfusion are vital in advancing the knowledge base regarding myocardial injury. Typically, ischemia of the mouse heart after left anterior descending (LAD) coronary artery occlusion is confirmed by visible pallor of the myocardium below the occlusion (ligature). However, this offers only a subjective way of confirming correct or consistent ligature placement, as there are multiple major arteries that could cause pallor in different myocardial regions. A method of recording electrocardiographic changes to assess correct ligature placement and resultant ischemia as well as reperfusion, to supplement observed myocardial pallor, would help yield consistent infarct sizes in mouse models. In turn, this would help decrease the number of mice used. Additionally, electrocardiographic changes can continue to be recorded non-invasively in a time-dependent fashion after the surgery. This article will demonstrate a method of electrocardiographically confirming myocardial ischemia and reperfusion in real time.

During murine myocardial ischemia/reperfusion surgery, correct placement of the occluding ligature is typically confirmed by visible observation of myocardial pallor. Herein, a method of electrocardiographically confirming ischemia and reperfusion, to supplement observed myocardial pallor, is demonstrated in male C57Bl/6 mice.

Yüzey Pad Elektrokardiyografi kullanılması Farelerde Miyokard İskemi ve Reperfüzyon Hasarı Onayı

Many animal models have been established for the study of myocardial remodeling and heart failure due to its status as the number one cause of mortality ...

Ultrasound-guided Intracardiac Injection of Human Mesenchymal Stem Cells to Increase Homing to the Intestine for Use in Murine Models of Experimental Inflammatory Bowel Diseases

Crohn&#39;s disease (CD) is a common chronic inflammatory disease of the small and large intestines. Murine and human mesenchymal stem cells (MSCs) have immunosuppressive potential and have been shown to suppress inflammation in mouse models of intestinal inflammation, even though the route of administration can limit their homing and effectiveness 1,3,4,5. Local application of MSCs to colonic injury models has shown greater efficacy at ameliorating inflammation in the colon. However, there is paucity of data on techniques to enhance the localization of human bone marrow-derived MSCs (hMSCs) to the small intestine, the site of inflammation in the SAMP-1/YitFc (SAMP) model of experimental Crohn&#39;s disease. This work describes a novel technique for the ultrasound-guided intracardiac injection of hMSCs in SAMP mice, a well-characterized spontaneous model of chronic intestinal inflammation. Sex- and age-matched, inflammation-free AKR/J (AKR) mice were used as controls. To analyze the biodistribution and the localization, hMSCs were transduced with a lentivirus containing a triple reporter. The triple reporter consisted of firefly luciferase (fl), for bioluminescent imaging; monomeric red fluorescent protein (mrfp), for cell sorting; and truncated herpes simplex virus thymidine kinase (ttk), for positron emission tomography (PET) imaging. The results of this study show that 24 h after the intracardiac administration, hMSCs localize in the small intestine of SAMP mice as opposed to inflammation-free AKR mice. This novel, ultrasound-guided injection of hMSCs in the left ventricle of SAMP mice ensures a high success rate of cell delivery, allowing for the rapid recovery of mice with minimal morbidity and mortality. This technique could be a useful method for the enhanced localization of MSCs in other models of small-intestinal inflammation, such as TNF&#916;RE6. Future studies will determine if the increased localization of hMSCs by intra-arterial delivery can lead to increased therapeutic efficacy.

Murine studies in models of colonic inflammation have demonstrated that a small percentage (1 - 5%) of mesenchymal stem cells (MSC) injected intravenously or intraperitoneally home to the inflamed colon1,2. This study shows that ultrasound-guided intracardiac injections of MSCs result in increased localization to the intestine.

Ultrason destekli İntrakardiyak enjeksiyon bağırsak için giriş için deneysel iltihabi bağırsak hastalıkları fare modellerinde kullanım artırmak için insan Mezenkimal Kök hücre

Crohn&#39;s disease (CD) is a common chronic inflammatory disease of the small and large intestines. Murine and human mesenchymal stem cells (MSCs) have ...

Murine Precision-Cut Liver Slices as an Ex Vivo Model of Liver Biology

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic fatty liver disease, metabolic liver disease, and liver cancer) requires experimental manipulation of animal models and in vitro cell cultures. Both techniques have limitations, such as the requirement of large numbers of animals for in vivo manipulation. However, in vitro cell cultures do not reproduce the structure and function of the multicellular hepatic environment. The use of precision-cut liver slices is a technique in which uniform slices of viable mouse liver are maintained in laboratory tissue culture for experimental manipulation. This technique occupies an experimental niche that exists between animal studies and in vitro cell culture methods. The presented protocol describes a straightforward and reliable method to isolate and culture precision-cut liver slices from mice. As an application of this technique, ex vivo liver slices are treated with bile acids to simulate cholestatic liver injury and ultimately assess the mechanisms of hepatic fibrogenesis.

This protocol provides a simple and reliable method for the production of viable precision-cut liver slices from mice. The ex vivo tissue samples can be maintained under laboratory tissue culture conditions for multiple days, providing a flexible model to examine liver pathobiology.

Karaciğer Biyolojisi Ex Vivo Modeli olarak Murine Precision-Cut Karaciğer Dilimleri

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic ...