Bu çalışma Körlük (RSB) Önlemek için NIH hibe 1R01EY022901, Research Kariyer Geliştirme Ödülü ile desteklenmiştir, CMReeves &amp; MA Reeves Vakfı, Körler E. Matilda Ziegler Vakfı, Tapınak Şövalyeleri Göz Vakfı ve Bölümü&#39;ne sınırsız hibe RSB Utah Üniversitesi Oftalmoloji. Biz el yazması üzerinde tartışmalar ve yorumlar için Spectralis Multi-Modality Görüntüleme Sistemi ve Tao Zhang teknik yardım için Balamurali Ambati teşekkür ederim.

1.. Araçların hazırlanması <ol><li> Prosedür, bir hayvan tesisinde bir prosedür odada gerçekleştirilir. </li><li> Deney başlamadan önce facemasks, saç bone, cerrahi önlük, steril ayak örtüleri, ve eldiven giyin. </li><li> Bir ocak üzerinde bir behere ~ 40 ° C ısı su. </li><li> Görüntüleme sırasında fare vücut ısısını korumak için daha sonra kullanılacak bir ısıtma yastığı üstüne steril bir mavi yastık yerleştirin. Isıtma pedi açın. </li><li> Görüntüleme Sistemi hazırlayın: <ol><li> Toz kapağını çıkartın ve lazer açın. </li><li> 55 ° lens dışarı atın ve makine üzerine monte. </li><li> Bilgisayar ve girdi yeni bir hastanın sayfanın altında görüntüleme için fare bilgiler (örn. genotip, yaş, vb.) Gelen görüntüleme yazılımını açın. &quot;Cihaz türü&quot; altında, Kızılötesi (IR) modunu seçin. </li></ol></li></ol> Not: Bu rapor edildiğini kullanım oBiz (temsilcisi sonuçlarını görmek, 1-4 Şekiller) harici lens kullanmadan IV-ISYA yüksek kaliteli görüntüler elde hiçbir sorun olmasına rağmen f Harici çift asferik lens görüntü kalitesini 17-20 artırabilir. 2. ISYA Kuyruk Ven Enjeksiyon <ol><li> 1% Tropikamid oftalmik solüsyonu ile fare gözleri dilate ve 5 dakika bekleyin. </li><li> Anestezi gerekli (Ketamin / Ksilazin / Acepromazine 65-100/10-20/1-3 mg / kg) miktarını belirlemek için, fare tartılır. </li><li> Bir 32 G iğne ile birlikte steril 1 ml şırınga al. Anestezi (13 mg / ml ketamin, 2.6 mg / ml Xylazine, steril PBS içinde 0.3 mg / ml Acepromazine) ile periton kesesi içine enjekte fare. Fare tamamen uyuşturulduktan kadar bekleyin (~ 5 dk). </li><li> Venin vazodilatasyona neden 40 ° C ılık suyun içine fare kuyruğu yerleştirin. </li><li> Bir 32 G iğne ile bir 1 ml şırınga al. IC istenilen miktarda geri çekG, tipik haliyle, bir 25 g fare (2 mg / kg) için, bir steril tüp içine, bir 0.2 uM şırınga filtresi ile filtre edilmiş, steril, 1 mg / ml ICG, 50 ul. Şırınga içine herhangi bir hava tanıtmak için dikkatli olun. </li><li> Enjekte edilecek bölgeyi sterilize etmek için bir alkollü bez ile silin kuyruk. </li><li> , Yanal kuyruk damarı yukarı doğru olacak şekilde, bir el ile kuyruk tutun. Yukarı bakacak şekilde iğnenin eğim ile, en az bir açı ile damar içine yaklaşık 2 mm iğne enjekte edilir. Ven delmek için dikkatli olun. Biraz şırınga geri çizin ve iğne başarıyla damar içine yerleştirilen olduğunu gösterir iğne hub, içine kan akımının izleri arayın. </li><li> Yavaş yavaş damar içine enjekte ICG. Enjekte ederken minimal direnç olmalıdır. İğneyi çıkarın ve herhangi bir kanamayı durdurmak için ~ 5-10 saniye enjeksiyon yerinde doğrudan bir alkol bez uygulayın. Fare görüntüleme için hazırdır. Erken faz (enjeksiyondan sonra 0-4 dakika) yakalamak amacıyla, bu esse olduğuhızlı görüntü fareyi ntial. </li></ol> Not: Fare gözler kolayca kuru alabilir ve anestezi altında katarakt gelişebilir. Bu işlem sırasında, steril PBS uygulayarak nemli göz tutmak önemlidir. ISYA Kayıttan önce steril pamuklu çubukla aşırı PBS silin. Diğer laboratuarları kornea 17-20 dehidratasyonu önlemek için bir kontakt lens kullandım. 3. ICG Anjiyografi <ol><li> Retina ve koroid tirajları maksimum parlaklık (0-4 dk) kadar görüntüleri koroid dolum erken faz yakalama izin verir ICG enjeksiyondan sonra 30-40 sn almaya başlar. Retinal damar iyi odak ~ 35-45 dioptri ve koroid damar 10-15 diyoptrilerinde görüntülenmiştir de görüntülenmiştir. Not: bir hayvan modelinin ilk inceleme sırasında, olası tüm abno tanımlamak için tüm açılardan (nazal, temporal, dorsal ve ventral) görüntüleri yakalamak için tavsiye edilirvaskülatürde rmalities. Erken dönemde, orta ve büyük koroid arterler ve venler hem de görüntülenmiştir. Bu protokolde kullanılan hayvan modelinde, koroid lezyonlar (örn. polipoidal dilatasyonlarında) erken fazına 1 dk görünmeye başlayabilir. </li><li> Damar üzerindeki görüntü odak ayarlayın. Parlaklık için Kontrol ve sırasıyla, kontrol modülü ve odak düğmesini kullanarak odaklanır. Bu değerler, ayarlanabilir sayısal olarak ve kolayca sabit tutulur. Aşağıdaki tekniği kullanılarak görüntü kalitesi tekrarlanabilir sağlamak için kamera lensi sabiti fare göz mesafe bırakın. Not: cihaz sadece görüntü, biz kamera lensi ve sürekli fare göz arasındaki odak, parlaklığı ve mesafe tutmak için posterior gözün bir kısmı deneyebilirsiniz beri görüntü farklı açılardan tüm posterior göz gibi. Bunun anahtarı vie alanı ile göz yoluyla ICG tarafından yayılan daire şeklindeki lüminesans hizalamak içinKameranın w. Tüm görüntü karanlık alanları vardır kadar bu, sol-sağ yaparak kamera pozisyonu yukarı ve aşağı, ve-ve-out ayarlamalar yapılır. Kameranın görüş alanında parlaklık ve dizilmiş zaman, lens için göz arasındaki mesafe kaliteli görüntüleme için optimal bir mesafede bir sonraki resim grubu hem de için tekrarlanabilir. </li><li> Damar odak sonra, satın alma modülü üzerinde yuvarlak siyah düğmeye basarak görüntü kareleri yakalamak. Yuvarlak siyah düğmesi de en iyi görüntü kalitesi için ICG sinyalini azaltmak veya artırmak için kullanılabilir. </li><li> Görüntü koroid lezyonların uygun görüş açısını ve odaklama derinliği belirler. Bu gözün konumunu, odaklama derinliği, ve tüm zaman-ders ISYA için sabit diğer cihaz ayarları tutmak için önemlidir. Görüntüler tespit modülüne dokunmatik ekran panelinde acquire düğmesine basarak kaydedilir. </li><li> 6-15 dk kıç orta fazda görüntüler eldeer enjeksiyon. Not: koroid ve retina damarları Hem daha az belirgin hale gelir. Koroid damar gibi yaygın floresan görünür. Hiperflöresansı sergileyen koroid lezyonlar normal bir arka plan floresan çevreleyen solma aksine ortaya. </li><li> Enjeksiyondan sonra 17-25 dk geç fazda görüntü kazanır. Not: hiperflöresansı kaybolur. Hem koroid ve retina damarları artık görünür. Optik sinir başı siyah olur. Hiperfloresan koroid lezyonlar solan arka plan ile maksimum kontrast var. </li><li> Görüntülerin satın bitirdikten sonra, fare gözleri açık bir yağ göz jeli uygulayın ve kurtarma için bir ısıtma pedi üzerinde fareyi bırakın. </li><li> Kafeslerinde tutarak alana fareler dönün. Daha fazla analiz için TIFF ya da JPEG dosyaları olarak dışa aktarın. </li></ol> Not: her bir faz zamanlaması kesin değildir. Bulduğumuz adet zamanlamasıch faz enjekte ICG miktarına bağlı olarak değişebilir. Daha ICG her aşamasını uzatmak için eğilimindedir. Bir faz tanımlamak için en iyi yol, yukarıda listelenen her fazın anahtar özelliklerine göre olduğunu.

<ol>
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YF Bu çalışmada kullanılan AMD fare modeli ile ilgili iki patent bekleyen bir mucididir. SK, ZB, ve ADJ ifşa hiçbir şey yok.

Bu çalışmada, HTRA1 transgenik farelerde görüntü koroid lezyonlara ISYA kullanımını göstermiştir. Bizim fare modelinde erken, orta ve ISYA geç fazların özellikleri insan çalışmaları 1 &#39;de de zaman süreci neticesinde. Bu, AMD olarak koroid ile ilgili koşulların patofizyolojik mekanizmaları ve tedavi stratejileri ile ilgili araştırmalar için çok değerli olan insan patolojisi ve hayvan fenotipleri arasında daha iyi karşılaştırma yapmak önemlidir. ...

İndosiyanin yeşili anjiyografi (ISYA) gözün kan damarları ile ilgili görüntü sorunlarına bir tanı testidir. Emisyon spektrumu 770-880 nm ila 835 nm, 1 de tepe emisyon aralıkları sırasında ICG&#39;nin emilme tayfı 790-805 nm arasında değişir. Bu olan spektrum görünür aralığında yer alır, diğer popüler boya sodyum floresan, farklıdır. Kızılötesi spektrumu kolayca sodyum floresein tabanlı floresein anjiografi (FA) tarafından görselleştirme engelleyebilirsiniz bütün bunlar retina pigment epiteli (RPE), serosanguinöz sıvısı ve lipid eksuda yoluyla nüfuz ICG sağlar. ICG koroid damar ve koroid lezyonların 1,2 gelişmiş görüntüleme sağlayan,% 98 proteine ​​bağlı daha az ekstravazasyonu ile sonuçlanan damar bulunmaktadır. ISYA neredeyse RPE posterior koroid damarlarını görselleştirmek için tek seçenektir. Şekil 1 fare gözlerinde görüntüleme damarsal ISYA ve FA karşılaştırmasını gösterir. FA bGörüntünün retinal damar sistemi kuyu değil, koroid damar kullanılan e. Bunun aksine, ISYA retinal ve koroid damar hem de görüntü için kullanılabilir. ISYA birlikte bu çalışmada kullanacağı kızılötesi hassas video kameralar, yüksek çözünürlüklü dijital görüntüleme sistemleri veya tarayıcı laser oftalmoskobun (SLO) ile gerçekleştirilir. Klinikte, ISYA Polipoidal Koroid Vaskülopati (PCV), Retina Anjiomatöz Yayılmasının (RAP), angioid çizgiler, vitelliform maküler distrofi, SSK&#39;sı, hemanjiomun, kanamalar retina dahil koroidal vaskulatürünü kapsayan koriyoretinal bozukluklar bir dizi teşhis tavsiye edilmiştir Arteriyollerde makroanevrizmaları, koroid tümörleri, posterior ve belirli biçimleri 1,3 üveit. FA ve optik koherens tomografi (OCT) ile ISYA kombinasyonu eksudatif yaşa bağlı maküler tanı ve tedavisinde klinisyenler için güçlü araçlar sağlardejenerasyon (AMD) 4-10. ISYA koroid ilgili koşulların teşhis edilmesi için özellikle yararlıdır. Aslında, ISYA PCV, eksudatif AMD 11-13 bir varyantını teşhisi için altın standart olarak kabul edilir. PCV koroid damar 11-13 terminal polipoidal dilatasyon ile damarların dallanma bir ağ ile karakterizedir. PCV sık sık kaçak ve polipoidal bileşenlerinden 11,14,15 gelen kanama ile RPE ve retinanın tekrarlayan serosanguinöz müfrezeleri ile ilişkilidir. Yakın zamanda transgenik fare retinal pigment epitel (RPE) 16, insan HTRA1, çok fonksiyonlu bir serin proteazı ifade ile birinci PCV hayvan modelinin üretilmesini bildirdi. Bu artan HTRA1 PCV karakteristik özelliklerine sahip, örneğin polipoidal lezyonlar uyardığını göstermiştir. İşte bizim HTRA1 fare modeli kullanılarak AMD araştırma kuyruk damar enjeksiyonu zaman ders ISYA kullanımını gösterdi. Bizim verilerimiz öneririzIV-ISYA IP (ya da deri altına (SC)) güncel koroid lezyonları karakterize etmek için 17,18 alanında kullanılan-ISYA üstündür. Hayvan Araştırma Bildirimi Hayvan deneyleri Kurumsal Hayvan Bakımı ve Kullanımı Komitesi (IACUC) tarafından onaylanmış protokollere göre yapılmıştır ve Oftalmik ve Vizyon Araştırma Hayvanların Kullanım ARVO Tablosuna uygun olarak yapıldı.

<table border="0" cellpadding="0" cellspacing="0" width="667">
	<tbody>
		<tr height="50">
			<td height="50" style="height:50px;width:216px;">Spectralis Multi-Modality Imaging System</td>
			<td style="width:152px;">Heidelberg Engineering, Germany</td>
			<td style="width:132px;">SPECTRALIS HRA+OCT</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Tropicamide ophthalmic solution (1%)</td>
			<td style="width:152px;">Bausch &#38; Lomb</td>
			<td style="width:132px;">NDC 24208-585-64</td>
			<td>for dilation of pupils</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">GenTeal Gel</td>
			<td style="width:152px;">Genteal</td>
			<td style="width:132px;">NDC 58768-791-15</td>
			<td>&#160;clear lubricant eye gel&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Ketamine</td>
			<td style="width:152px;">Vedco Inc</td>
			<td style="width:132px;">NDC 50989-996-06</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Xylazine</td>
			<td style="width:152px;">Lloyd Laboratories</td>
			<td style="width:132px;">NADA 139-236</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Acepromazine</td>
			<td style="width:152px;">Vedco Inc</td>
			<td style="width:132px;">NDC 50989-160-11</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">32-G Needle</td>
			<td>Steriject</td>
			<td style="width:132px;">PRE-32013</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">1-ml syringe</td>
			<td style="width:152px;">BD</td>
			<td style="width:132px;">309659</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Indocyanine Green</td>
			<td style="width:152px;">Pfaltz &#38; Bauer</td>
			<td style="width:132px;">I01250</td>
			<td></td>
		</tr>
	</tbody>
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detecting abnormalities in choroidal vasculature in a mouse model of age-related macular degeneration by time-course indocyanine green angiography

Indosiyanin Yeşil Anjiyografi (veya ISYA) koroid ve böyle yaşa bağlı makula dejenerasyonu (AMD) gibi çeşitli göz hastalıkları retina damar anormallikleri teşhis etmek için göz doktorları tarafından yapılan bir tekniktir. ISYA nedeniyle kızıl ötesi spektrum ile pigmentli tabakası içinden nüfuz kabiliyeti gözün posterior koroid damar görüntüye özellikle yararlıdır. ISYA zaman ders erken, orta ve geç evreye ayrılabilir. Üç aşama göz problemleri patoloji hakkında değerli bilgiler sunmaktadır. Intravenöz zaman ders ISYA (IV) enjeksiyonu yaygın koroid sorunlarının tanı ve tedavisi için klinikte kullanılan rağmen, ISYA intraperitoneal enjeksiyon (IP) tarafından genellikle hayvan araştırmalarda kullanılmaktadır. Burada kuyruk ven enjeksiyon ve konfokal tarayıcı laser oftalmoskopla farelerde yüksek çözünürlüklü ISYA zaman ders görüntüler elde etmek için teknik göstermiştir. Biz koroid le görüntü için bu tekniği kullanılıryaşa bağlı makula dejenerasyonu olan bir fare modelinde lezyonlar. IP tarafından fare damarsal ICG tanıtmak için çok daha kolay olmasına rağmen, bizim veriler IP-ISYA tarafından tekrarlanabilir ISYA zaman ders görüntüler elde etmek zor olduğunu göstermektedir. Buna karşılık, kuyruk ven enjeksiyon yoluyla ISYA insan çalışmalarında karşılaştırılabilir yüksek kaliteli ISYA zaman ders görüntü sağlar. Buna ek olarak, pigmentli ISYA fareler üzerinde gerçekleştirilen daha koroid damarlarının daha net resimler verir albino fareler üzerinde gerçekleştirilen gösterdi. Biz o zaman-ders IV-ISYA hayvan modellerinde dayalı AMD araştırma standart bir uygulama haline gelmelidir öneririz.

Biz, CD1 arka plan üzerinde, her ikisi de, HTRA1 transgenik fareler ve kontrol litermatlarında WT ISYA zaman süreci gerçekleştirildi. Albino CD1 plan indosiyanin yeşil anjiyografi (ISYA) görüntüleme (tartışma) kolaylaştırmak için seçildi. Dilatasyon gibi bazı anevrizma HTRA1 fare erken dönemde ortaya çıkmaya başladı (Şekil 2, kırmızı bir ok bir geminin ucunda dilatasyon gösteren ve kırmızı bir daire, bir küme tipi polipoidal lezyon gösterir). Koroid damarlarının sırası...

Watch this Scientific Journal Video about Detecting Abnormalities in Choroidal Vasculature in a Mouse Model of Age-related Macular Degeneration by Time-course Indocyanine Green Angiography at JoVE.com

Detecting Abnormalities in Choroidal Vasculature in a Mouse Model of Age-related Macular Degeneration by Time-course Indocyanine Green Angiography

Kuyruk ven enjeksiyon yoluyla gerçekleştirilebilir Indocyanine Yeşil Anjiyografi (veya ISYA) fare koroidinde anormallikleri karakterize kaliteli ISYA zaman ders görüntüler sağlar.

Zaman kurs Indocyanine Yeşil Anjiyografi ile Yaşa Bağlı Makula Dejenerasyonu Fare Modeli Koroid vaskülatürüne anormallikleri tespit

Indocyanine Green&#160;Angiography (or ICGA) is a technique performed by ophthalmologists to diagnose abnormalities of the choroidal and retinal ...

detecting-abnormalities-choroidal-vasculature-mouse-model-age-related

Research

JoVE Journal

Medicine

13.5K Görüntüleme.  University of Utah Health Sciences Center.  Kuyruk ven enjeksiyon yoluyla gerçekleştirilebilir Indocyanine Yeşil Anjiyografi (veya ISYA) fare koroidinde anormallikleri karakterize kaliteli ISYA zaman ders görüntüler sağlar. 

Video: Detecting Abnormalities in Choroidal Vasculature in a Mouse Model of Age-related Macular Degeneration by Time-course Indocyanine Green Angiography

Noninvasive Assessment of Cardiac Abnormalities in Experimental Autoimmune Myocarditis by Magnetic Resonance Microscopy Imaging in the Mouse

Myocarditis is an inflammation of the myocardium, but only ~10% of those affected show clinical manifestations of the disease. To study the immune events of myocardial injuries, various mouse models of myocarditis have been widely used. This study involved experimental autoimmune myocarditis (EAM) induced with cardiac myosin heavy chain (Myhc)-&#945; 334-352 in A/J mice; the affected animals develop lymphocytic myocarditis but with no apparent clinical signs. In this model, the utility of magnetic resonance microscopy (MRM) as a non-invasive modality to determine the cardiac structural and functional changes in animals immunized with Myhc-&#945; 334-352 is shown. EAM and healthy mice were imaged using a 9.4 T (400 MHz) 89 mm vertical core bore scanner equipped with a 4 cm millipede radio-frequency imaging probe and 100 G/cm triple axis gradients. Cardiac images were acquired from anesthetized animals using a gradient-echo-based cine pulse sequence, and the animals were monitored by respiration and pulse oximetry. The analysis revealed an increase in the thickness of the ventricular wall in EAM mice, with a corresponding decrease in the interior diameter of ventricles, when compared with healthy mice. The data suggest that morphological and functional changes in the inflamed hearts can be non-invasively monitored by MRM in live animals. In conclusion, MRM offers an advantage of assessing the progression and regression of myocardial injuries in diseases caused by infectious agents, as well as response to therapies.

This study demonstrates the successful establishment of magnetic resonance microscopy imaging as a non-invasive tool to assess the cardiac abnormalities in mice affected with autoimmune myocarditis. The data indicate that the technique can be used to monitor the disease-progression in live animals.

Fare Manyetik Rezonans Görüntüleme Mikroskopi tarafından Deneysel Otoimmün Miyokardit Kardiyak anormallikler noninvaziv değerlendirilmesi

Myocarditis is an inflammation of the myocardium, but only ~10% of those affected show clinical manifestations of the disease. To study the immune events ...

Tıp

A Mouse Model for Laser-induced Choroidal Neovascularization

The mouse laser-induced choroidal neovascularization (CNV) model has been a crucial mainstay model for neovascular age-related macular degeneration (AMD) research. By administering targeted laser injury to the RPE and Bruch&#8217;s membrane, the procedure induces angiogenesis, modeling the hallmark pathology observed in neovascular AMD. 
First developed in non-human primates, the laser-induced CNV model has come to be implemented into many other species, the most recent of which being the mouse. Mouse experiments are advantageously more cost-effective, experiments can be executed on a much faster timeline, and they allow the use of various transgenic models. The miniature size of the mouse eye, however, poses a particular challenge when performing the procedure. Manipulation of the eye to visualize the retina requires practice of fine dexterity skills as well as simultaneous hand-eye-foot coordination to operate the laser. However, once mastered, the model can be applied to study many aspects of neovascular AMD such as molecular mechanisms, the effect of genetic manipulations, and drug treatment effects. 
The laser-induced CNV model, though useful, is not a perfect model of the disease. The wild-type mouse eye is otherwise healthy, and the chorio-retinal environment does not mimic the pathologic changes in human AMD. Furthermore, injury-induced angiogenesis does not reflect the same pathways as angiogenesis occurring in an age-related and chronic disease state as in AMD.
Despite its shortcomings, the laser-induced CNV model is one of the best methods currently available to study the debilitating pathology of neovascular AMD. Its implementation has led to a deeper understanding of the pathogenesis of AMD, as well as contributing to the development of many of the AMD therapies currently available.

Here, we present the mouse laser-induced choroidal neovascularization (CNV) protocol, an experimental model that re-creates the vascular hallmarks of neovascular age-related macular degeneration (AMD). Once mastered, it can reliably and effectively induce CNV as a model system to test various experimental measures.

Lazer kaynaklı Koroid Neovaskülarizasyonlarında A Fare Modeli

The mouse laser-induced choroidal neovascularization (CNV) model has been a crucial mainstay model for neovascular age-related macular degeneration (AMD) ...

The Monoiodoacetate Model of Osteoarthritis Pain in the Mouse

A major symptom of patients with osteoarthritis (OA) is pain that&#160;is triggered by peripheral as well as central changes within the pain pathways. The current treatments for OA pain such as NSAIDS or opiates are neither sufficiently effective nor devoid of detrimental side effects. Animal models of OA are being developed to improve our understanding of OA-related pain mechanisms and define novel pharmacological targets for therapy. Currently available models of OA in rodents include surgical and chemical interventions into one knee joint. The monoiodoacetate (MIA) model has become a standard for modelling joint disruption in OA in both rats and mice. The model, which is easier to perform in the rat, involves injection of MIA into a knee joint that induces rapid pain-like responses in the ipsilateral limb, the level of which can be controlled by injection of different doses. Intra-articular injection of MIA disrupts chondrocyte glycolysis by inhibiting glyceraldehyde-3-phosphatase dehydrogenase and results in chondrocyte death, neovascularization, subchondral bone necrosis and collapse, as well as inflammation. The morphological changes of the articular cartilage and bone disruption are reflective of some aspects of patient pathology. Along with joint damage, MIA injection induces referred mechanical sensitivity in the ipsilateral hind paw and weight bearing deficits that are measurable and quantifiable. These behavioral changes resemble some of the symptoms reported by the patient population, thereby validating the MIA injection in the knee as a useful and relevant pre-clinical model of OA pain.
The aim of this article is to describe the methodology of intra-articular injections of MIA and the behavioral recordings of the associated development of hypersensitivity with a mind to highlight the necessary steps to give consistent and reliable recordings.

Osteoarthritis (OA), or degenerative joint disease, is a debilitating condition associated with pain that remains only partially controlled by available analgesics. Animal models are being developed to improve our understanding of OA-related pain mechanisms. Here we describe the methodology for the monoiodoacetate model of OA pain in the mouse.

Fare Osteoartrit Ağrı Monoiodoacetate Modeli

A major symptom of patients with osteoarthritis (OA) is pain that&#160;is triggered by peripheral as well as central changes within the pain pathways. The ...

Detecting Anastasis In Vivo by CaspaseTracker Biosensor

Anastasis (Greek for &#8220;rising to life&#8221;) is a recently discovered cell recovery phenomenon whereby dying cells can reverse late-stage cell death processes that are generally assumed to be intrinsically irreversible. Promoting anastasis could in principle rescue or preserve injured cells that are difficult to replace such as cardiomyocytes or neurons, thereby facilitating tissue recovery. Conversely, suppressing anastasis in cancer cells, undergoing apoptosis after anti-cancer therapies, may ensure cancer cell death and reduce the chances of recurrence. However, these studies have been hampered by the lack of tools for tracking the fate of cells that undergo anastasis in live animals. The challenge is to identify the cells that have reversed the cell death process despite their morphologically normal appearance after recovery. To overcome this difficulty, we have developed Drosophila and mammalian CaspaseTracker biosensor systems that can identify and permanently track the anastatic cells in vitro or in vivo. Here, we present in vivo protocols for the generation and use of the CaspaseTracker dual biosensor system to detect and track anastasis in Drosophila melanogaster after transient exposure to cell death stimuli. While conventional biosensors and protocols can label cells actively undergoing apoptotic cell death, the CaspaseTracker biosensor can permanently label cells that have recovered after caspase activation - a hallmark of late-stage apoptosis, and simultaneously identify active apoptotic processes. This biosensor can also track the recovery of the cells that attempted other forms of cell death that directly or indirectly involved caspase activity. Therefore, this protocol enables us to continuously track the fate of these cells and their progeny, facilitating future studies of the biological functions, molecular mechanisms, physiological and pathological consequences, and therapeutic implications of anastasis. We also discuss the appropriate controls to distinguish cells that undergo anastasis from those that display non-apoptotic caspase activity in vivo.

Detecting Anastasis In Vivo by CaspaseTracker Biosensor

Anastasis is technically challenging to detect in vivo because the cells that have reversed the cell death process can be morphologically indistinguishable from normal healthy cells. Here we describe protocols for detecting and tracking cells that undergo anastasis in live animals by using our newly developed in vivo CaspaseTracker biosensor system.

Anastasis Vivo içinde CaspaseTracker Biyoalgılayıcı tarafından tespit

Anastasis (Greek for &#8220;rising to life&#8221;) is a recently discovered cell recovery phenomenon whereby dying cells can reverse late-stage cell death ...

Regenerative Therapy by Suprachoroidal Cell Autograft in Dry Age-related Macular Degeneration: Preliminary In Vivo Report

This study is aimed at examining whether a suprachoroidal graft of autologous cells can improve best corrected visual acuity (BCVA) and responses to microperimetry (MY) in eyes affected by dry Age-related Macular Degeneration (AMD) over time through the production and secretion of growth factors (GFs) on surrounding tissue. Patients were randomly assigned to each study group. All patients were diagnosed with dry AMD and with BCVA equal to or greater than 1 logarithm of the minimum angle of resolution (logMAR). A suprachoroidal autologous graft by Limoli Retinal Restoration Technique (LRRT) was carried out on group A, which included 11 eyes from 11 patients. The technique was performed by implanting adipocytes, adipose-derived stem cells obtained from the stromal vascular fraction, and platelets from platelet-rich plasma in the suprachoroidal space. Conversely, group B, including 14 eyes of 14 patients, was used as a control group. For each patient, diagnosis was verified by confocal scanning laser ophthalmoscope and spectral domain-optical coherence tomography (SD-OCT). In group A, BCVA improved by 0.581 to 0.504 at 90 days and to 0.376 logMAR at 180 days (+32.20%) postoperatively. Furthermore, MY test increased by 11.44 dB to 12.59 dB at 180 days. The different cell types grafted behind the choroid were able to ensure constant GF secretion in the choroidal flow. Consequently, the results indicate that visual acuity (VA) in the grafted group can increase more than in the control group after six months.

Regenerative Therapy by Suprachoroidal Cell Autograft in Dry Age-related Macular Degeneration: Preliminary In Vivo Report

The goal of this study is to assess whether the suprachoroidal graft of adipose-derived stem cells included in the stromal vascular fraction and platelets derived from platelet-rich plasma by the Limoli Retinal Restoration Technique can improve visual acuity and retinal sensitivity responses in eyes affected by dry age-related macular degeneration.

Rejeneratif tedavisi ile Suprachoroidal hücre otogrefti içinde Kuru yaşa bağlı makula dejenerasyonu: ön Vivo içinde rapor

This study is aimed at examining whether a suprachoroidal graft of autologous cells can improve best corrected visual acuity (BCVA) and responses to ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

İzometrik Contractility ölçüm tel Myography kullanarak fare mezenterik arter

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

A Neonatal BALB/c Mouse Model of Necrotizing Enterocolitis

Necrotizing enterocolitis (NEC) is the most severe gastrointestinal (GI) disease that often occurs in premature infants, especially very low birth weight infants, with high mortality and unclear pathogenesis. The cause of NEC may be related to inflammatory immune regulatory system abnormalities. An NEC animal model is an indispensable tool for NEC disease immune research. NEC animal models usually use C57BL/6J neonatal mice; BALB/c neonatal mice are rarely used. Related studies have shown that when mice are infected, Th2 cell differentiation is predominant in BALB/c mice compared to C57BL/6J mice. Studies have suggested that the occurrence and development of NEC are associated with an increase in T helper type 2 (Th2) cells and are generally accompanied by infection. Therefore, this study used neonatal BALB/c mice to induce an NEC model with similar clinical characteristics and intestinal pathological changes as those observed in children with NEC. Further study is warranted to determine whether this animal model could be used to study Th2 cell responses in NEC.

Necrotizing enterocolitis (NEC) is the most severe gastrointestinal (GI) disease that often occurs in premature infants, especially very low birth weight infants, with high mortality and unclear pathogenesis. The cause of NEC may be related to inflammatory immune regulatory system abnormalities. An NEC animal model is an indispensable tool for NEC disease immune research. NEC animal models usually use C57BL/6J neonatal mice; BALB/c neonatal mice are rarely used. Related studies have shown that when mice are infected, Th2 cell differentiation is predominant in BALB/c mice compared to C57BL/6J mice. Studies have suggested that the occurrence and development of NEC are associated with an increase in T helper type 2 (Th2) cells and are generally accompanied by infection. Therefore, this study used neonatal BALB/c mice to induce an NEC model with similar clinical characteristics and intestinal pathological changes as those observed in children with NEC. Further study is warranted to determine whether this animal model could be used to study Th2 cell responses in NEC.

Nekrotizan Enterokolit'in Yenidoğan BALB/c Fare Modeli

Necrotizing enterocolitis (NEC) is the most severe gastrointestinal (GI) disease that often occurs in premature infants, especially very low birth weight ...

Robotic Left Hepatectomy using Indocyanine Green Fluorescence Imaging for an Intrahepatic Complex Biliary Cyst

Biliary cysts (BC) are rare congenital dilatations of intra- and extrahepatic parts of the biliary tract and bear a significant risk of carcinogenesis. Surgery is the cornerstone treatment for patients with BC. While total BC excision and Roux-Y hepaticojejunostomy is the treatment method of the choice in patients with extrahepatic BC (i.e., Todani I-IV), patients with intrahepatic BC (i.e., Todani V) benefit the most from a surgical liver resection. In recent years, minimally invasive liver surgery (MILS) including robotic MILS has gained more acceptance as a feasible, safe, and effective procedure for the treatment of both benign and malignant indications. Robotic major MILS is still considered technically demanding and a detailed description of the technical approach during robotic major MILS has only been limitedly discussed in the literature. The current article describes the main steps for a robotic left hepatectomy in a patient with a large BC Todani Type V. The patient is in French position with 5 trocars placed (4 robotic, 1 laparoscopic assistant). After mobilizing the left hemiliver, the left and right hepatic artery are dissected carefully followed by a cholecystectomy. Intraoperative ultrasound is performed to confirm localization and margins of the BC. The Left hepatic artery and left portal vein are isolated, clipped, and divided. Indocyanine green (ICG) fluorescence imaging is used regularly during the entire procedure to visualize and confirm biliary tract anatomy and the BC. Parenchymal transection is performed with robotic cautery hook for the superficial part and robotic cautery spatula, bipolar cautery, and vessel sealer for the deeper parenchyma. The postoperative course was uncomplicated. A robotic left hepatectomy is technically demanding, yet a feasible and safe procedure. ICG-fluorescence imaging aids in delineating the BC and bile duct anatomy. Further, comparative studies are needed to confirm clinical benefits of robotic MILS for benign and malignant indications.

Robotic liver surgery has gained more acceptance as a feasible, safe, and effective procedure for the treatment of both benign and malignant indications. However, robotic left hepatectomy is still technically demanding. We describe our surgical technique of a robotic left hepatectomy using indocyanine green fluorescence imaging for a large biliary cyst.

İntrahepatik Kompleks Biliyer Kist için İndosiyanin Yeşil Floresan Görüntüleme Kullanılarak Robotik Sol Hepatektomi

Biliary cysts (BC) are rare congenital dilatations of intra- and extrahepatic parts of the biliary tract and bear a significant risk of carcinogenesis. ...

Fluorescent Laparoscopic Central Hepatectomy for Liver Cancer Using Indocyanine Green Negative Staining

Laparoscopic hepatectomy is an important treatment method for liver cancer. In the past, the resection boundary was usually determined by intraoperative ultrasound, important vascular structures, and surgeon experience. With the development of anatomical hepatectomy, visual surgery technology has gradually been applied to this type of surgery, particularly indocyanine green (ICG)-guided anatomical hepatectomy. As ICG can be specifically ingested by hepatocytes and used for fluorescence tracing, negative staining techniques have been applied according to different tumor positions. Under ICG fluorescent guidance, the surface boundary and deep resection plane can be more accurately displayed during liver resection. Thus, the tumor-bearing liver segment can be anatomically removed, which helps to avoid damage to important vessels and reduce ischemia or congestion of the remaining liver tissue. Finally, the incidence of postoperative biliary fistula and liver dysfunction is reduced; therefore, a better prognosis is obtained after the resection of liver cancer. Centrally located liver cancer is usually defined as a tumor located at segments 4, 5, or 8 that requires resection of the middle section of the liver. These are among the most difficult hepatectomies to perform because of the large surgical wounds and multiple vessel transections. Based on the specific tumor location, we formulated the required resection ranges by designing personalized fluorescent staining strategies. By completing anatomical resection based on the portal territory, this work aims to achieve the best therapeutic effect.

The present protocol describes fluorescent negative staining in laparoscopic central hepatectomy. This technique can make hepatectomy more accurate and precise.

İndosiyanin Yeşili Negatif Boyama Kullanılarak Karaciğer Kanseri için Floresan Laparoskopik Santral Hepatektomi

Laparoscopic hepatectomy is an important treatment method for liver cancer. In the past, the resection boundary was usually determined by intraoperative ...

Yaşa Bağlı Makula Dejenerasyonunda Kantitatif Otofloresansın Araştırılması

A Workflow to Quantitatively Determine Age-Related Macular Degeneration Lesion-Specific Variations in Fundus Autofluorescence

Fundus autofluorescence (FAF) imaging allows the noninvasive mapping of intrinsic fluorophores of the ocular fundus, particularly the retinal pigment epithelium (RPE), now quantifiable with the advent of confocal scanning laser ophthalmoscopy-based quantitative autofluorescence (QAF). QAF has been shown to be generally decreased at the posterior pole in age-related macular degeneration (AMD). The relationship between QAF and various AMD lesions (drusen, subretinal drusenoid deposits) is still unclear.
This paper describes a workflow to determine lesion-specific QAF in AMD. A multimodal in vivo imaging approach is used, including but not limited to spectral domain optical coherence tomography (SD-OCT) macular volume scanning and QAF. Using customized FIJI plug-ins, the corresponding QAF image is aligned with the near-infrared image from the SD-OCT scan (characteristic landmarks; i.e., vessel bifurcations). The foveola and the edge of the optic nerve head are marked in the OCT images (and transferred to the registered QAF image) for accurate positioning of the analysis grids.
AMD-specific lesions can then be marked on individual OCT BScans or the QAF image itself. Normative QAF maps are created to account for the varying mean and standard deviation of QAF values throughout the fundus (QAF images from a representative AMD group were averaged to build normative standard retinal QAF AMD maps). The plug-ins record the X and Y coordinates, z-score (a numerical measurement that describes the QAF value in relation to the mean of AF maps in terms of standard deviation from the mean), mean intensity value, standard deviation, and number of pixels marked. The tools also determine z-scores from the border zone of marked lesions.&#160;This workflow and the analysis tools will improve the understanding of the pathophysiology and clinical AF image interpretation in AMD.

Yazar Spotlight: QAF İş Akışı ile Yaşa Bağlı Makula Dejenerasyonu Patofizyolojisini Anlama 

This research describes a workflow to determine and compare autofluorescence levels from individual regions of interest (e.g., drusen and subretinal drusenoid deposits in age-related macular degeneration [AMD]) while accounting for varying autofluorescence levels throughout the fundus.

Fundus Otofloresansında Yaşa Bağlı Makula Dejenerasyonu Lezyonuna Özgü Varyasyonları Kantitatif Olarak Belirlemek İçin Bir İş Akışı

Fundus autofluorescence (FAF) imaging allows the noninvasive mapping of intrinsic fluorophores of the ocular fundus, particularly the retinal pigment ...