The authors have no acknowledgements.

NOT: Tüm hayvan prosedürleri Cincinnati Üniversitesi Kurumsal Hayvan Bakım ve Kullanım Komitesi (IACUC) ve (NIH Yayın No Ulusal Sağlık Enstitüleri Laboratuar Hayvanlarının Bakımı ve Kullanımı Kılavuzu uyarınca onayı ile yapıldı . 85-23) 1996 Revize. 1. Euthanize ve Fare sterilize <ol><li> Izofluran bir supratheraputic dozunu içeren bir açılır kutu içinde fare yerleştirin ve etkisi nefes sağlar. Fare ötenazi sonra, kutudan çıkarın. </li><li> Cervically ötenazi ikinci aracı olarak yerinden. </li><li> % 70 etanol ile hayvan temizleyerek fare dış yüzeyini sterilize edin. </li></ol> 2. Kimlik ve Üç Farklı Yağ Depoları izolasyonu <ol><li> Kahverengi Yağ Dokusu (BAT) İzolasyon: <ol><li> Emin kürk epidermal ve dermal laye ile kesme yardımcı olmak için alkol temizliği ıslak olduğundan emin olunrs. </li><li> Masanın karşı sırtını bir yatar pozisyonda fare yerleştirin. </li><li> Sadece forseps, asansör ile diyafram altında cilt tut ve makasla kesilirken. </li><li> Periton maruz fare çevresi enine kesin. </li><li> Farenin üst yarısını deglove ile kelebek şeklinde BAT deposu ortaya koymaktadır. Bir yandan alt uzantıları ve karın tutun ve baş yukarı doğru çekerek cildi. </li><li> Masaya eğilimli konumlandırılmış böylece fareyi yönlendirin. Saçlı maruz depo kontamine dikkatli olun. </li><li> Temiz cerrahi aletler ve eldiven taze çifti değişiklik. </li><li> Skapula ve ilgili depo bulun. Dikkatle kelebek üstünde herhangi bir yüzeysel beyaz adipoz kaldırmak ve daha sonra interskapular kahverengi yağ kelebek teşrih. Yakından kahverengi yağ ile ilişkili kas önlemek için dikkatli olun. NOT: Bir diseksiyon mikroskop kullanımı w kaldırılması için tavsiye edilirHite yağ, aynı zamanda, scapulalar kahverengi adipoz ayrılmasında. </li><li> Yağ depo çıkarın ve 2 ml&#39;lik bir mikrosantrifüj tüpüne aktarılır. </li><li> RNA ya da bir protein ekstrakte edilir edilecekse, -80 ° C&#39;de, sıvı azotun ve mağaza içine daldırma ile doku dondurma. Bozulmasını önlemek için bir kerede örnek dondurun. Kültürleme Eğer, DMEF-12 doku kapak ve tüm örnekler kültür (ek bilgi) için tahsil edilene kadar buz üzerinde ayarlayın. </li></ol></li><li> Subkutan Yağ Dokusu (SQ) izolasyonu, Beyaz Yağ Dokusu Depo (WAT): <ol><li> Yağ depoları arasında değil çapraz kontaminasyona olarak eldiven taze çifti koyun. </li><li> Inguinal, fare alt yarısını de-gloving&#39;i üçgen SQ depoları Reveal. Bir yandan üst uzantıları ve toraks tutun ve diğer elinizle ayak aşağı doğru çekerek cildi. </li><li> Saçlı maruz depo kontamine dikkatli olurken bir yatar pozisyonda fare yönlendirin. </li><li> Temiz Surgicaeldiven taze çifti l aletleri ve değişim. </li><li> Dikkatle SQ yağ üçgenleri incelemek. Yağ, meme bezleri komşu, kas ile örnek kontamine veya herhangi damarları keserek ve kanla kontamine örnek için dikkatli olun. NOT: Sınırlar açıkça tanımlanmış değilse, bir diseksiyon mikroskop kullanılması tavsiye edilir. </li><li> Yağ depoları çıkarın ve 2 ml mikrosantrifüj tüpler transfer. </li><li> RNA ya da bir protein ekstrakte edilir edilecekse, -80 ° C&#39;de, sıvı azotun ve mağaza içine daldırma ile doku dondurma. Bozulmasını önlemek için bir kerede örnek dondurun. </li><li> Boyama için, düzeltmek veya OCT embed. Kültürleme Eğer, DMEF-12 doku kapak ve tüm örnekler kültür (ek bilgi) için tahsil edilene kadar buz üzerinde ayarlayın. </li></ol></li><li> Gonadal Fat bir Visseral Yağ Dokusu (KDV) ve Beyaz Yağ Dokusu (WAT) Depo İzolasyonu: <ol><li> Yağ depoları arasında değil çapraz kontaminasyona olarak eldiven taze çifti koyun. </li><li> Sci ilessors doğrudan diyafram altında, enine periton kesti. Koronal karın organları açığa rektum ortasına kadar diyaframdan periton kesin. </li><li> Testisler veya yumurtalıkların bulun ve kadınlarda epididimal erkeklerde adipoz veya gonadal adipoz olarak bilinen ekli beyaz yağ dokusu, tespit. </li><li> Eldiven taze çifti Temiz cerrahi aletler ve değişim </li><li> Dikkatle testis, epididim ve vas deferensler epididimal yağ depolarını hem teşrih. Ya da kadın, eğer dikkatle yumurtalıkların hem gonadal yağ yastıkları incelemek. </li><li> Yağ depoları çıkarın ve 2 ml mikrosantrifüj tüpler aktarabilirsiniz. </li><li> RNA ya da bir protein ekstrakte edilir edilecekse, -80 ° C&#39;de, sıvı azotun ve mağaza içine daldırma ile doku dondurma. Bozulmasını önlemek için bir kerede örnek dondurun. </li><li> Boyama için, düzeltmek veya OCT embed. Kültürleme Eğer, DMEF-12 doku kapak ve tüm örnekler kültür (ek bilgi) için tahsil edilene kadar buz üzerinde ayarlayın. </li&gt; </ol></li></ol> Perivasküler Yağ Doku 3. izolasyonu (PVAT) <ol><li> Kalp İzolasyonu: <ol><li> Dışa genişletilmiş üst ve alt uzantıları ile bir yatar pozisyonda fare yatıyordu. </li><li> Cerrahi bant kullanarak Güvenli uzantıları. </li><li> Yukarıda listelenen fareyi yerleştirdikten sonra, forseps ile kılıç şeklinde süreci kaldırarak gerilim yaratmak. Göğüs boşluğunun alt kısmını açığa diyafram aracılığıyla yatay kesin. </li><li> Kılıç şeklinde süreci kaldırarak, gerginlik korurken, sadece sternum tarafına, kafasına doğru superiorda göğüs kafesi kesti. </li><li> Klavikula sadece alt göğüs kafesi Pick up ve her iki yönde aksillada doğru dışarı klavikula alt uzunluğu boyunca kesilmiş. göğüs boşluğu ve içeriği (kalp, akciğerler, vb) şimdi açıkça görülebilir olmalıdır. </li><li> Steril ga kullanarak gereksiz kan ve sıvı göğüs boşluğu temizleyinUZE sıvıyı emmek için. Toplama organları veya gemiler (ek bilgi) bol emin olunuz. </li><li> Alan sıvı temizlendikten sonra, kalbin daha iyi poz sağlayan, akciğerler kaldırmak için bronşlar ve takılarak gemilerin kesti. </li></ol></li><li> Yerelleştirme ve Eksizyon Aort ve Perivasküler Yağ Dokusu (PVAT): <ol><li> Karaciğer, mide, dalak, pankreas, bağırsak ve kolon: iyi aort alt bölümlerini tanımlamak için aşağıdaki organları çıkarın. </li><li> İlk mide ve yemek borusu belirlenmesi ile başlar. Vücuttan mideyi boşaltmak için gastro-özofageal kavşakta yemek borusu kesin. </li><li> Sonraki, bağırsakları ve çevredeki mezentere belirlemek. O aort böbrek kısmına çok yakından yatıyor ardından &quot;bağırsak çalıştırın.&quot;, Mezenterden ile yüzeysel Cut </li><li> Kolon mümkün olduğunca rektum kadar yakın özgür kesin. Böylece, fare mide, bağırsak ve kolon kurtararak. </li><li> Tekrartutturma mezenteri ve damarları aracılığıyla keserek mide, bağırsak, kolon, pankreas ve dalak hareket ettirin. pankreas ve dalak mide, bağırsak ve kolon ücretsiz gelmelidir. </li><li> Tüm lobları kaldırmak, mezentere hepatik venler ile kesme ve takarak karaciğer çıkarın. </li><li> Visseral tabaka kesip ve yağ böbrekleri çevreliyor. Aortun farklı kesimleri için coğrafi işaretler olarak hizmet etmek in vivo aorta bağlı böbrekleri bırakın. </li><li> Steril 1x PBS ile bölgeyi durulayın ve bir steril gazlı bez ile emilim bütün sıvıyı çıkarın. </li><li> Mikro-makas ve mikro-forseps kullanarak, omurga dorsal eki ve yemek borusu onun ventral eki aort ayırın. </li><li> Aşağıdaki ve iliak bölgede çatallanma kalbinde kökenli inen aorta aorta uzunluğunu ayırarak aort izole edin. </li><li> Belirlemek ve subklavian damarları izole. Bu gemiler yalıtmakAort kök boyundan daha iyi kalp aort kavşak ve kök maruz. </li><li> Sonra timus çıkarın kalbin hareketini sağlayan, brakiyosefalik arter, sol karotis arter ve sol subklavian arter kesti. </li><li> Bir diseksiyon mikroskobu yardımıyla, aort çevreleyen perivasküler adipoz dokusu (PVAT) katmanı görüntüleyin. </li><li> Sıkıştırmayın veya PVAT sıkmak, yavaşça mikro-forseps ile aortadan PVAT şekilde çekmeyin büyük özen. Yavaşça diyafram nerede sadece üstün torakal bölgede başlayan mikro-makas ile aorta PVAT eki kesti. NOT: A diseksiyon mikroskop tavsiye edilir. </li><li> Aort geminin iliak çatallanma sadece üstün bulunan İnfrarenal aort bölgesinde, terbiye, bu sürecini aortun tüm uzunluğu tekrarlayın. </li><li> Aortik ark PVAT isteniyorsa, daha az c PVAT kaldırmak için aynı yöntem kullanılırkemer urvature. </li><li> 2 ml mikrosantrifüj tüpü (lar) içine PVAT numuneler. </li><li> RNA ya da bir protein ekstrakte edilir edilecekse, -80 ° C&#39;de, sıvı azotun ve mağaza içine daldırma ile doku dondurma. Bozulmasını önlemek için bir kerede örnek dondurun. Kültürleme Eğer, DMEF-12 doku kapak ve tüm örnekler kültür (ek bilgi) için tahsil edilene kadar buz üzerinde ayarlayın. NOT: Kapsamlı bir yağ depo analizi ilgilenen varsa ek yağ depoları dikkate şunlardır: retroperitoneal, mezenterik, omentum, perikard ve popliteal. </li></ol></li></ol>

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The authors declare that they have no competing financial interests or other conflicts of interest.

Obezite morbidite büyük bir ev sahibi ve adipoz tam olarak anlaşılmış değildir oynadığı rolün tam bir anlayış yol açabilir; adipoz alanında nedenle devam araştırma gereklidir. Hayvan modelleri, özellikle fare modelleri hastalıklar ve potansiyel ilaç tedavilerinin test ilerlemesinde ilk araştırma için idealdir. Bu modelleri kullanarak, kesin izolasyon ve yağ depolarında eksizyonu yağ etkilenen hastalıkların patolojisi çalışmada son derece önemli ve gerekli bir araçtır. <p class="jove...

Yağ nedeniyle 20. yüzyılın son birkaç on yıl içinde obezite en dramatik artış, medya spot kayda değer bir görünüm getirmişti. Obezite anda yetişkinlerin üçte biri ve Amerika Birleşik Devletleri&#39;nde (ABD) 1 çocuk ve ergenlerin% 17 daha fazla etkiler. Tüm etnik grupları Kapsayan, obezite salgını çevreleyen istatistiksel araştırma Meksikalı Amerikalılar (% 40.4), tüm İspanyollar (% 39.1), ve olmayan kıyasla İspanyol olmayan siyahlar obezite (% 49.5) en yüksek yaşa göre düzeltilmiş oranına sahip olduğunu göstermiştir Hispanik beyazlar (% 34.3) 2. Obezitenin ekonomik etkisi de sağlık sistemi için büyüyen bir husustur. 2012 yılında, 2005 yılında ABD&#39;de obezite için bakım yıllık tıbbi maliyeti 190.200.000.000 $, genel tıbbi harcama bütçesinin yaklaşık% 21 olduğu tahmin edilmiştir. Ne yazık ki, çocukluk çağı obezitesi tek başına doğrudan tıbbi maliyetler 14 milyar dolar sorumlu olduğu tahmin edilmiştir. İstatistiksel olarak, bu ortalama tıbbi maliyeti olduğu tespit edildiobezite ile bireylerin bu morbidite 3-5 olmayanlara göre daha yüksek 2741 $ bir yıl oldu. Tip 2 diyabet, dislipidemi, kardiyovasküler hastalıklar, kanser, kas iskelet bozuklukları ve kronik inflamasyon: Obezite gibi durumların çeşitli için önemli bir risk faktörüdür. Obezite derinden metabolik sendrom ve diğer kronik hastalıkların 6-8 patogenezi bağlıdır. Obezite ve diğer yandaş arasında böyle sert ve olumsuz bağlantıları ile, bilimsel araştırma daha güncel salgın ve adipoz oynadığı çeşitli ve önemli rolleri anlamak için dikkat odaklanmıştır. Tarihsel, yağ dokusu önemsiz kabul edildi ve basit bir dolgu dokusu gibi sadece görülüyordu. Metabolizma, hormon regülasyonu, inflamasyon, koruma ve izolasyon 9: Şu anda, yağ vücudun işlevi birçok temel rol oynamaya gösterilmiştir. Yağ dokusu öncelikle oluşuradipositler de perisitler, endotel hücreleri, monositler, makrofajlar ve pluripotent kök hücrelerin 8 içerir. Yağ dokusu farklı depolarda vücuda dağılır. ana depolar deri altından, kas içinden, deri altından bulunan ve içgüdüsel 10 edilebilir. Yağ depolar bozuklukları 8 obeziteye bir depo belirli duyarlılık göstermiştir ve ilgili olan depo belirli metabolik profillerin sahip olduğu gösterilmiştir. Geleneksel, adipoz doku iki ana gruba sınıflandırılmıştır: beyaz yağ dokusu (WAT) ve kahverengi adipoz doku (BAT); Son literatür gösterir, ancak üçüncü bir grup varlıkları brite veya bej adipoz 11 vaftiz. Adipoz doku farklı renkler, morfolojileri, metabolik fonksiyonları, biyokimyasal özellikleri ve ifade 10 genetik modellerine sahip olduğu gösterilmiştir. WAT içinde Adipositler tek, büyük lipit damlacık ve mitokondri değişken miktarlarda bulunmaktadır. WATbaskın olarak vücudun deri altına ve visseral yerlerde bulunur. Birincil enerji depolama ve organ koruma alanı olarak WAT çalışır. BAT Adipositler bir multiloküler morfoloji ve bol mitokondri var. BAT öncelikle boyun ve göğüs büyük kan damarlarının yanı sıra, skapula 12 yer almaktadır. Thermogenesis 7 düzenleyen enerji sokmaksızın davranışları BAT öncelikle çalışır. Brite veya bej yağ BAT benzer bir morfoloji ve ifade paylaşmaya gösterilmiştir fakat beyaz adipositlerden 11 kaynaklandığı tespit edilmiştir. Bu yazıda anlatılan cerrahi yöntem gibi faktörler farklı etkilerini analiz etmek kapasitesine sahip araştırmacılar sunar: çevre, ilaç ve genetik, adipoz var; de yararlı veya zararlı rol adipoz hastalığı ve genel sağlık oynar gibi. Aynı zamanda, bir yol sağlayarak tespit ve adipoz dokuda farklı izole etmek içindaha izin depoları arasındaki biyokimyasal ilişkiler ve farklılıklar anlamak. Bu vücut içindeki konumu, işlev ve yağ arasındaki ilişkinin belirlenmesinde yardımcı olabilir. Bu tarif edilen yöntem, in vitro çalışmalar için brüt görselleştirme, gen ekspresyon analizi, protein ifadesi analizi, histolojik ve birincil hücre hatlarının izolasyonu için araçlar sağlayarak gerçekleştirir. Şu anda farklı yağ depoları, yanı sıra anatomik yerlerde metabolik davranışı içgörü sağlayan birçok makaleleri vardır; ama özellikle, lokalize belirlemek ve bu depoları izole etmek için nasıl bir derinlemesine bir yöntem vermeyin. Bu durum bunların cerrahi, bir veya iki depo 13-14 bir izolat için tasarlanan diğerleri yöntemlerine göre diseksiyon ve kontaminasyon en az bir miktar ile birden fazla depo izolasyonu sağlayan bir hassas bir teknik sağlamaktadır. Bu protokolün amacı sağlamaktırBirden fazla anatomik bölgelerden yağ depoları farklı tiplerinin belirlenmesi ve izolasyonu için kesin bir metot.

<table border="0" cellpadding="0" cellspacing="0" width="730">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:243px;">Name of Reagent/ Equipment</td>
			<td style="width:175px;">Company</td>
			<td style="width:125px;">Catalog Number</td>
			<td style="width:188px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">isoflurane</td>
			<td>Med-Vet International&#160;</td>
			<td>#RXISO-250</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:243px;">70% Ethanol</td>
			<td style="width:175px;">Fisher</td>
			<td style="width:125px;">07-678-001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:243px;">DMEF-12</td>
			<td style="width:175px;">Sigma Aldrich</td>
			<td style="width:125px;">D-6421</td>
			<td>Warm in waterbath before putting on tissue</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:243px;">2 mL microcentrifuge tubes</td>
			<td style="width:175px;">Midsci</td>
			<td style="width:125px;">MCT-200-C-S</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:243px;">Phosphate buffered saline</td>
			<td style="width:175px;">Sigma Aldrich</td>
			<td style="width:125px;">P5368-10PAK</td>
			<td></td>
		</tr>
	</tbody>
</table>

localization, identification, and excision of murine adipose depots

Obesity has increased dramatically in the last few decades and affects over one third of the adult US population. The economic effect of obesity in 2005 reached a staggering sum of $190.2 billion in direct medical costs alone. Obesity is a major risk factor for a wide host of diseases. Historically, little was known regarding adipose and its major and essential functions in the body. Brown and white adipose are the two main types of adipose but current literature has identified a new type of fat called brite or beige adipose. Research has shown that adipose depots have specific metabolic profiles and certain depots allow for a propensity for obesity and other related disorders. The goal of this protocol is to provide researchers the capacity to identify and excise adipose depots that will allow for the analysis of different factorial effects on adipose; as well as the beneficial or detrimental role adipose plays in disease and overall health. Isolation and excision of adipose depots allows investigators to look at gross morphological changes as well as histological changes. The adipose isolated can also be used for molecular studies to evaluate transcriptional and translational change or for in vitro experimentation to discover targets of interest and mechanisms of action. This technique is superior to other published techniques due to the design allowing for isolation of multiple depots with simplicity and minimal contamination.

Kimlik ve kasık deri altı yağ, interskapular kahverengi adipoz, viseral adipoz epididim (Şekil 1), yanı sıra aort arkı perivasküler adipoz, torakal aort adipoz, böbreküstü aort adipoz ve infrarenal aort adipoz lokalizasyonu (Şekil 2) kullanılarak başarıyla gerçekleştirilmiştir açıklanan cerrahi yöntem. BAT ve WAT örnekler arasında histolojik muayene ve farklılaşma pozitif boyanan (Şekil 3) hematoksilen ve eozin (H &amp; E) ile değerlendirildi....

Watch this Scientific Journal Video about Localization, Identification, and Excision of Murine Adipose Depots at JoVE.com

Localization, Identification, and Excision of Murine Adipose Depots

Nedeniyle obezite ve diğer yandaş arasındaki şiddetli ve negatif bağlantı, rol adipoz araştırma hastalığında oynadığı ve genel sağlık garantilidir. Biz izolasyon ve yerinde kullanarak adipoz çalışması için izin yağ depolarında eksizyonu ve in vitro yöntemler bir protokol mevcut.

Yerelleştirme, Kimlik ve Eksizyon Kemirgen Yağ depolarının

Obesity has increased dramatically in the last few decades and affects over one third of the adult US population. The economic effect of obesity in 2005 ...

localization-identification-and-excision-of-murine-adipose-depots

Research

JoVE Journal

Medicine

40.8K Görüntüleme.  University of Cincinnati College of Medicine.  Nedeniyle obezite ve diğer yandaş arasındaki şiddetli ve negatif bağlantı, rol adipoz araştırma hastalığında oynadığı ve genel sağlık garantilidir. Biz izolasyon ve yerinde kullanarak adipoz çalışması için izin yağ depolarında eksizyonu ve in vitro yöntemler bir protokol mevcut. 

Video: Localization, Identification, and Excision of Murine Adipose Depots

A Simplified Technique for In situ Excision of Cornea and Evisceration of Retinal Tissue from Human Ocular Globe

Enucleation is the process of retrieving the ocular globe from a cadaveric donor leaving the rest of the globe undisturbed. Excision refers to the retrieval of ocular tissues, especially cornea, by cutting it separate from the ocular globe. Evisceration is the process of removing the internal organs referred here as retina. The ocular globe consists of the cornea, the sclera, the vitreous body, the lens, the iris, the retina, the choroid, muscles etc (Suppl. Figure 1). When a patient is suffering from corneal damage, the cornea needs to be removed and a healthy one must be transplanted by keratoplastic surgeries. Genetic disorders or defects in retinal function can compromise vision. Human ocular globes can be used for various surgical procedures such as eye banking, transplantation of human cornea or sclera and research on ocular tissues. However, there is little information available on human corneal and retinal excision, probably due to the limited accessibility to human tissues. Most of the studies describing similar procedures are performed on animal models. Research scientists rely on the availability of properly dissected and well-conserved ocular tissues in order to extend the knowledge on human eye development, homeostasis and function. As we receive high amount of ocular globes out of which approximately 40% (Table 1) of them are used for research purposes, we are able to perform huge amount of experiments on these tissues, defining techniques to excise and preserve them regularly.
The cornea is an avascular tissue which enables the transmission of light onto the retina and for this purpose should always maintain a good degree of transparency. Within the cornea, the limbus region, which is a reservoir of the stem cells, helps the reconstruction of epithelial cells and restricts the overgrowth of the conjunctiva maintaining corneal transparency and clarity. The size and thickness of the cornea are critical for clear vision, as changes in either of them could lead to distracted, unclear vision. The cornea comprises of 5 layers; a) epithelium, b) Bowman's layer, c) stroma, d) Descemet's membrane and e) endothelium. All layers should function properly to ensure clear vision4,5,6. The choroid is the intermediate tunic between the sclera and retina, bounded on the interior by the Bruch's membrane and is responsible for blood flow in the eye. The choroid also helps to regulate the temperature and supplies nourishment to the outer layers of the retina5,6. The retina is a layer of nervous tissue that covers the back of the ocular globe (Suppl. Figure 1) and consists of two parts: a photoreceptive part and a non-receptive part. The retina helps to receive the light from the cornea and lens and converts it into the chemical energy eventually transmitted to the brain with help of the optic nerve5,6.
The aim of this paper is to provide a protocol for the dissection of corneal and retinal tissues from human ocular globes. Avoiding cross-contamination with adjacent tissues and preserving RNA integrity is of fundamental importance as such tissues are indispensable for research purposes aimed at (i) characterizing the transcriptome of the ocular tissues, (ii) isolating stem cells for regenerative medicine projects, and (iii) evaluating histological differences between tissues from normal/affected subjects. In this paper we describe the technique we currently use to remove the cornea, the choroid and retinal tissues from an ocular globe. Here we provide a detailed protocol for the dissection of the human ocular globe and the excision of corneal and retinal tissues. The accompanying video will help researchers to learn an appropriate technique for the retrieval of precious human tissues which are difficult to find regularly.

A Simplified Technique for In situ Excision of Cornea and Evisceration of Retinal Tissue from Human Ocular Globe

The paper describes a simplified technique to excise corneal and to eviscerate retinal tissues from the ocular globe of human cadaveric donors. The technique described here will help to excise good quality tissues to be used for transplantation, surgical or research purposes without damaging other tissues of the ocular globe.

Için Basitleştirilmiş Tekniği<em&gt; Yerinde</emİnsan Oküler Globe gelen Kornea ve Retina Doku Eviserasyon of&gt; eksizyonu

Enucleation is the process of retrieving the ocular globe from a cadaveric donor leaving the rest of the globe undisturbed. Excision refers to the ...

Tıp

Isolation and Excision of Murine Aorta; A Versatile Technique in the Study of Cardiovascular Disease

Cardiovascular disease is a broad term describing disease of the heart and/or blood vessels. The main blood vessel supplying the body with oxygenated blood is the aorta. The aorta may become affected in diseases such as atherosclerosis and aneurysm. Researchers investigating these diseases would benefit from direct observation of the aorta to characterize disease progression as well as to evaluate efficacy of potential therapeutics. The goal of this protocol is to describe proper isolation and excision of the aorta to aid investigators researching cardiovascular disease. Isolation and excision of the aorta allows investigators to look at gross morphometric changes as wells as allowing them to preserve and stain the tissue to look at histologic changes if desired. The aorta may be used for molecular studies to evaluate protein and gene expression to discover targets of interest and mechanisms of action. This technique is superior to imaging modalities as they have inherent limitations in technology and cost. Additionally, primary isolated cells from a freshly isolated and excised aorta can allowing researchers to perform further in situ and in vitro assays. The isolation and excision of the aorta has the limitation of having to sacrifice the animal however, in this case the benefits outweigh the harm as it is the most versatile technique in the study of aortic disease.

Pathology of the aorta can lead to severe morbidity and mortality, therefore research of disease progression and potential therapies is warranted. Here, we present a protocol to isolate and excise the murine aorta to aid researchers in their investigation of cardiovascular disease.

İzolasyon ve Kemirgen Aort eksizyonu; Kardiyovasküler Hastalığı Çalışmasında Çok Yönlü Tekniği

Cardiovascular disease is a broad term describing disease of the heart and/or blood vessels. The main blood vessel supplying the body with oxygenated ...

Human Brown Adipose Tissue Depots Automatically Segmented by Positron Emission Tomography/Computed Tomography and Registered Magnetic Resonance Images

Reliably differentiating brown adipose tissue (BAT) from other tissues using a non-invasive imaging method is an important step toward studying BAT in humans. Detecting BAT is typically confirmed by the uptake of the injected radioactive tracer 18F-Fluorodeoxyglucose (18F-FDG) into adipose tissue depots, as measured by positron emission tomography/computed tomography (PET-CT) scans after exposing the subject to cold stimulus. Fat-water separated magnetic resonance imaging (MRI) has the ability to distinguish BAT without the use of a radioactive tracer. To date, MRI of BAT in adult humans has not been co-registered with cold-activated PET-CT. Therefore, this protocol uses 18F-FDG PET-CT scans to automatically generate a BAT mask, which is then applied to co-registered MRI scans of the same subject. This approach enables measurement of quantitative MRI properties of BAT without manual segmentation. BAT masks are created from two PET-CT scans: after exposure for 2 hr to either thermoneutral (TN) (24 &#176;C) or cold-activated (CA) (17 &#176;C) conditions. The TN and CA PET-CT scans are registered, and the PET standardized uptake and CT Hounsfield values are used to create a mask containing only BAT. CA and TN MRI scans are also acquired on the same subject and registered to the PET-CT scans in order to establish quantitative MRI properties within the automatically defined BAT mask. An advantage of this approach is that the segmentation is completely automated and is based on widely accepted methods for identification of activated BAT (PET-CT). The quantitative MRI properties of BAT established using this protocol can serve as the basis for an MRI-only BAT examination that avoids the radiation associated with PET-CT.

The method presented here uses 18F-Fluorodeoxyglucose (18F-FDG) positron emission tomography/computed tomography (PET-CT) and fat-water separated magnetic resonance imaging (MRI), each scanned following 2 hr exposure to thermoneutral (24 &#176;C) and cold conditions (17 &#176;C) in order to map brown adipose tissue (BAT) in adult human subjects.

İnsan Brown Yağ Dokusu Depolar otomatik Pozitron Emisyon Tomografi / Bilgisayarlı Tomografi tarafından Parçalı ve Manyetik Rezonans Görüntüler Kayıtlı

Reliably differentiating brown adipose tissue (BAT) from other tissues using a non-invasive imaging method is an important step toward studying BAT in ...

Encapsulation Thermogenic Preadipocytes for Transplantation into Adipose Tissue Depots

Cell encapsulation was developed to entrap viable cells within semi-permeable membranes. The engrafted encapsulated cells can exchange low molecular weight metabolites in tissues of the treated host to achieve long-term survival. The semipermeable membrane allows engrafted encapsulated cells to avoid rejection by the immune system. The encapsulation procedure was designed to enable a controlled release of bioactive compounds, such as insulin, other hormones, and cytokines. Here we describe a method for encapsulation of catabolic cells, which consume lipids for heat production and energy dissipation (thermogenesis) in the intra-abdominal adipose tissue of obese mice. Encapsulation of thermogenic catabolic cells may be potentially applicable to the prevention and treatment of obesity and type 2 diabetes. Another potential application of catabolic cells may include detoxification from alcohols or other toxic metabolites and environmental pollutants.

Here, we present a protocol for encapsulation of catabolic cells, which consume lipids for heat production in intra-abdominal adipose tissue and increase energy dissipation in obese mice.

Yağ Dokusu Depolarında içine Nakli Enkapsülasyon termojenik Preadipositler

Cell encapsulation was developed to entrap viable cells within semi-permeable membranes. The engrafted encapsulated cells can exchange low molecular ...

Assessment of Human Adipose Tissue Microvascular Function Using Videomicroscopy

While obesity is closely linked to the development of metabolic and cardiovascular disease, little is known about mechanisms that govern these processes. It is hypothesized that pro-atherogenic mediators released from fat tissues particularly in association with central/visceral adiposity may promote pathogenic vascular changes locally and systemically, and the notion that cardiovascular disease may be the consequence of adipose tissue dysfunction continues to evolve. Here, we describe a unique method of videomicroscopy that involves analysis of vasodilator and vasoconstrictor responses of intact small human arterioles removed from the adipose depot of living human subjects. Videomicroscopy is used to examine functional properties of isolated microvessels in response to pharmacological or physiological stimuli using a pressured system that mimics in vivo conditions. The technique is a useful approach to gain understanding of the pathophysiology and molecular mechanisms that contribute to vascular dysfunction locally within the adipose tissue milieu. Moreover, abnormalities in the adipose tissue microvasculature have also been linked with systemic diseases. We applied this technique to examine depot-specific vascular responses in obese subjects. We assessed endothelium-dependent vasodilation to both increased flow and acetylcholine in adipose arterioles (50 - 350 &#181;m internal diameter, 2 - 3 mm in length) isolated from two different adipose depots during bariatric surgery from the same individual. We demonstrated that arterioles from visceral fat exhibit impaired endothelium-dependent vasodilation compared to vessels isolated from the subcutaneous depot. The findings suggest that the visceral microenvironment is associated with vascular endothelial dysfunction which may be relevant to clinical observation linking increased visceral adiposity to systemic disease mechanisms. The videomicroscopy technique can be used to examine vascular phenotypes from different fat depots as well as compare findings across individuals with different degrees of obesity and metabolic dysfunction. The method can also be used to examine vascular responses longitudinally in response to clinical interventions.

Videomicroscopy systems are used to examine functional properties of isolated adipose tissue arterioles in response to physiological and pharmacological stimuli. This technique can be used to examine microvascular phenotypes in different adipose tissue domains in obese humans.

İnsan yağ dokusu mikrovasküler işlevini kullanarak Videomicroscopy değerlendirilmesi

While obesity is closely linked to the development of metabolic and cardiovascular disease, little is known about mechanisms that govern these processes. ...

Vessel-sparing Excision and Primary Anastomosis

Urethroplasty is considered to be the standard treatment for urethral strictures since it provides excellent long-term success rates. For isolated short bulbar or posterior urethral strictures, urethroplasty by excision and primary anastomosis (EPA) is recommended. As EPA only requires the excision of the narrowed segment and the surrounding spongiofibrosis, a full-thickness transection of the corpus spongiosum, as performed in the traditional transecting EPA (tEPA), is usually unnecessary. Jordan et al. introduced the idea of a vessel-sparing approach in 2007, aiming to reduce surgical trauma, especially to the dual arterial blood supply of the urethra, and, thus, potentially reducing the risk of postoperative erectile dysfunction or glans ischemia. This approach could also be beneficial for subsequent urethral interventions such as redo urethroplasty using a free graft, in which a well-vascularized graft bed is imperative. Nevertheless, these potential benefits are only assumptions as prospective studies comparing the functional outcome of both techniques with validated questionnaires are currently lacking. Moreover, vessel-sparing EPA (vsEPA) should at least be able to provide similar surgical outcomes as tEPA. The aim of this paper is to give an elaborate, step-by-step overview of how to manage patients with isolated short bulbar or posterior urethral strictures with vsEPA. The main objective of this manuscript is to outline the surgical technique and to report the representative surgical outcome. A total of 117 patients were managed according to the described protocol. The analysis was performed on the entire patient cohort and on the bulbar (n = 91) and posterior (n = 26) vsEPA group separately. Success rates were 93.4% and 88.5% for the bulbar and posterior vsEPA, respectively. To conclude, vsEPA, as outlined in the protocol, provides excellent success rates with low complication rates for isolated short bulbar and posterior urethral strictures.

Here, we present an elaborate and efficient protocol to treat isolated short bulbar or posterior urethral strictures with vessel-sparing excision and primary anastomosis.

Gemi tutumlu eksizyon ve birincil anastomoz

Urethroplasty is considered to be the standard treatment for urethral strictures since it provides excellent long-term success rates. For isolated short ...

Electromagnetic Navigation Transthoracic Nodule Localization for Minimally Invasive Thoracic Surgery

The increased use of chest computed tomography (CT) has led to an increased detection of pulmonary nodules requiring diagnostic evaluation and/or excision. Many of these nodules are identified and excised via minimally invasive thoracic surgery; however, subcentimeter and subsolid nodules are frequently difficult to identify intra-operatively. This can be mitigated by the use of electromagnetic transthoracic needle localization. This protocol delineates the step-by-step process of electromagnetic localization from the pre-operative period to the postoperative period and is an adaptation of the electromagnetically guided percutaneous biopsy previously described by Arias et al. Pre-operative steps include obtaining a same day CT followed by the generation of a three-dimensional virtual map of the lung. From this map, the target lesion(s) and an entry site are chosen. In the operating room, the virtual reconstruction of the lung is then calibrated with the patient and the electromagnetic navigation platform. The patient is then sedated, intubated, and placed in the lateral decubitus position. Using a sterile technique and visualization from multiple views, the needle is inserted into the chest wall at the prechosen skin entry site and driven down to the target lesion. Dye is then injected into the lesion and, then, continuously during needle withdrawal, creating a tract for visualization intra-operatively. This method has many potential benefits when compared to the CT-guided localization, including a decreased radiation exposure and decreased time between the dye injection and the surgery. Dye diffusion from the pathway occurs over time, thereby limiting intra-operative nodule identification. By decreasing the time to surgery, there is a decrease in wait time for the patient, and less time for dye diffusion to occur, resulting in an improvement in nodule localization. When compared to electromagnetic bronchoscopy, airway architecture is no longer a limitation as the target nodule is accessed via a transparenchymal approach. Details of this procedure are described in a step-by-step fashion.

Presented here is a protocol for lung nodule localization using dye marking via electromagnetically navigated transthoracic needle access. The technique described here can be accomplished in the peri-operative period to optimize nodule localization and to successful resection when performing minimally invasive thoracic surgery.

Minimal İnvaziv Göğüs Cerrahisinde Elektromanyetik Navigasyon Transtorasik Nodül Lokalizasyonu

The increased use of chest computed tomography (CT) has led to an increased detection of pulmonary nodules requiring diagnostic evaluation and/or ...

Murine Appendectomy Model of Chronic Colitis Associated Colorectal Cancer by Precise Localization of Caecal Patch

The human appendix has been recently implicated to play important biological roles in the pathogenesis of various complex diseases, such as colorectal cancer, inflammatory bowel disease, and Parkinson&#8217;s disease. To study the function of the appendix, a gut disease-associated murine appendectomy model has been established and its step-by-step protocol is described here. This report introduces a facile protocol for caecal patch removal in mice followed by the chemical induction of chronic colitis-associated colorectal cancer using a combination of dextran sulfate sodium (DSS) and azoxymethane (AOM). IgA specific cells and IgA concentration were significantly reduced upon removal of the caecal patch in male C57BL/6 mice&#160;compared to those in the sham group. Simultaneously administering 2% DSS and AOM resulted in nearly 80% mice survival in both sham and appendectomy groups without significant body weight loss. Histological results confirmed colonic inflammation and different degrees of adenocarcinoma. This model can be used for the study of the functional role of the appendix in maintaining gut microbiota homeostasis and pathogenesis of gut colitis and malignancies, as well as for the potential development of drug targeting therapies.

The presented protocol describes a facile surgical removal of the appendix (caecal patch) in a mouse followed by the induction of inflammatory bowel disease-associated colorectal cancer. This murine appendectomy model enables investigation of the biological role of the appendix in the pathogenesis of human gastrointestinal disease.

Kaekal Patch Kesin Lokalizasyonu ile Kronik Kolit İlişkili Kolorektal Kanser Murine Apendektomi Modeli

The human appendix has been recently implicated to play important biological roles in the pathogenesis of various complex diseases, such as colorectal ...

Inguinal Subcutaneous White Adipose Tissue (ISWAT) Transplantation Model of Murine Islets

Pancreatic islet transplantation is a well-established therapeutic treatment for type 1 diabetes. The kidney capsule is the most commonly used site for islet transplantation in rodent models. However, the tight kidney capsule limits the transplantation of sufficient islets in large animals and humans. The inguinal subcutaneous white adipose tissue (ISWAT), a new subcutaneous space, was found to be a potentially valuable site for islet transplantation. This site has better blood supply than other subcutaneous spaces. Moreover, the ISWAT accommodates a larger islet mass than the kidney capsule, and transplantation into it is simple. This manuscript describes the procedure of mouse islet isolation and transplantation in the ISWAT site of syngeneic diabetic mouse recipients. Using this protocol, murine pancreatic islets were isolated by standard collagenase digestion and a basement membrane matrix hydrogel was used for fixing the purified islets in the ISWAT site. The blood glucose levels of the recipient mice were monitored for more than 100 days. Islet grafts were retrieved at day 100 after transplantation for histological analysis. The protocol for islet transplantation in the ISWAT site described in this manuscript is simple and effective.

Inguinal Subcutaneous White Adipose Tissue ISWAT Transplantation Model of Murine Islets

In this protocol, a method of murine islet isolation and transplantation into the inguinal subcutaneous white adipose tissue is described. Isolated syngeneic murine islets are transplanted into a murine recipient using a basement membrane hydrogel. The blood glucose level of the recipients is monitored, and histology analysis of the islet grafts is performed.

Inguinal Subkutan Beyaz Yağ Dokusu (ISWAT) Transplantasyon Modeli Murine Adacıkları

Pancreatic islet transplantation is a well-established therapeutic treatment for type 1 diabetes. The kidney capsule is the most commonly used site for ...

PCL Kist Eksizyonu için Artroskopik Çift Posteromedial Yaklaşım

Arthroscopic Excision of Posterior Cruciate Ligament Cysts Using a Double Posteromedial Approach

Cruciate ligament cysts of the knee are a rare condition. Posterior cruciate ligament cysts of the knee are less common than anterior cruciate ligament cysts. In patients with asymptomatic isolated cruciate ligament cysts of the knee, conservative treatment is recommended. Symptomatic cruciate ligament cysts of the knee are mostly manifested as knee hyperflexion pain, straightening pain, knee discomfort after standing for a long time or walking for a long time, etc., which seriously affects the quality of life, surgical treatment can be performed. The surgical treatments can be divided into ultrasound-guided cyst puncture and fluid extraction procedure and arthroscopic cystectomy. Cysts are mostly lobulated with a multi-layer cyst wall, cyst fluid extraction does not remove the cyst wall completely but simply extracts cyst fluid, leading to a high recurrence rate. Arthroscopic surgery can completely remove the cyst wall with little trauma, a low recurrence rate, and fast postoperative recovery, so arthroscopic resection is the most common and preferred method of treatment. Since posterior cruciate ligament cysts mostly occur posterior to the ligament, we remove the cyst wall by adding a double posteromedial approach to the knee joint, and the cyst wall is removed under direct vision, which is simple to operate, the cyst wall is completely cleared, the trauma is small, the postoperative recovery is fast, and there is no recurrence. Here, 8 posterior cruciate ligament cysts were removed with complete postoperative symptom relief, no surgical complications, and no recurrence at 1-year follow-up.

Yazar Gündemi: Arka Çapraz Bağ Kistlerinin Tedavisinde Çift Posteromedial Yaklaşım

We present a surgical approach to treat posterior cruciate ligament cysts by an arthroscopic double posteromedial approach.

Arka Çapraz Bağ Kistlerinin Çift Posteromedial Yaklaşımla Artroskopik Eksizyonu

Cruciate ligament cysts of the knee are a rare condition. Posterior cruciate ligament cysts of the knee are less common than anterior cruciate ligament ...