Name: protocols for vaginal inoculation and sample collection in the experimental mouse model of candida vaginitis
Uploaded: 2011-12-08T12:00:00
Description: Watch this Scientific Journal Video about Protocols for Vaginal Inoculation and Sample Collection in the Experimental Mouse Model of Candida vaginitis at JoVE.com

Bu çalışma, R01 AI32556 (NIAID, Ulusal Sağlık Enstitüsü) tarafından desteklenmiştir. Bu çalışma ayrıca, Louisiana Aşı Merkezi ve Güney Louisiana Louisiana Regents Kurulu tarafından desteklenen Enfeksiyon Hastalıkları Araştırma Enstitüsü tarafından desteklenmiştir.

1. Candida albicans ile Vajinal aşılama <ol><li> Aşılama Üç gün önce, karın ortaya çıkarmak için hayvan frenleyici, alt karın bölgesinde subkütan 0.1-0.5 mg β-östradiol içeren susam yağı enjekte 100 ul. Enjeksiyon yerinden sızıntı en aza indirmek için cilde yan 5 ila 10 mm iğne ilerletin. Alt karın östrojen subkutan uygulanması, genital sistem yakınlığı nedeniyle bu model en uygunudur. Etkili dozlarda fare suşlar, yaşları ya da östrojen türevleri göre değişebilir...

<ol>
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Candida vajinit, deneysel bir fare model kurulmuş ve tarihsel olarak son birkaç yıldır kullanılan 3,4,11,13,16,17,19,21,24 antifungal tedaviler test etmek için Candida mukozal konak yanıtının yanı sıra çalışma olmuştur 25,37. Protokoller, burada verimli ve daha az emek-yoğun yöntemleri dahil, ve Candida vajinit en optimize edilmiş bir model sistemleri bugüne kadar açıklanan gibi görünen sundu. Bu teknikler hızlı kantifikasyon vajinal ö...

<table border="1"><tr><td> Reaktifi Adı </td><td> Şirket </td><td> Katalog numarası </td><td> Yorumlar </td></tr><tr><td> Kadın CBA / J fareler </td><td> Charles River Laboratuvarları </td><td> 01C38 </td><td> 5-6 hafta </td></tr><tr><td> Candida albicans (3153A) </td><td> Patojenik Mantarlar, Birleşik Krallık Ulusal Koleksiyonu </td><td> NCPF3153 </td><td></td></tr><tr><td> Susam yağı </td><td> Sigma-Aldrich </td><td> S3547 </td><td> Kullanmadan önce ön sterilize edilmek üzere ihtiyaç duymaz </td></tr><tr><td> Β-östradiol 17-valerat </td><td> Sigma-Aldrich </td><td> E1631 </td><td> Susam yağı 0.1-0.5mg </td></tr><tr><td> Phytone pepton </td><td> Becton Dickinson </td><td> 211906 </td><td> % 0.1 glukoz ile Ek </td></tr><tr><td> Tripan mavi çözümü </td><td> Sigma-Aldrich </td><td> T8154 </td><td></td></tr><tr><td> Sabouraud dekstroz agar </td><td> Becton Dickinson </td><td> 211584 </td><td></td></tr><tr><td> Kollajenaz tip IV </td><td> Sigma-Aldrich </td><td> C5138 </td><td> % 0.25 </td></tr><tr><td> Dispase </td><td> Invitrogen </td><td> 17105-041 </td><td> 1.7 U / ml </td></tr><tr><td> Tel örgü ekranlar </td><td> TWP </td><td> 060X060S0065W36T </td><td> No 60 mesh, paslanmaz </td></tr><tr><td> Hanks dengeli tuz çözeltisi </td><td> Invitrogen </td><td> 24020-117 </td><td></td></tr><tr><td> CytoPrep fiksatif </td><td> Fisher Scientific </td><td> 12-570-10 </td><td> Smear slaytlar korur </td></tr><tr><td> Leke Papanicolaou EA-65 </td><td> Merck Kimyasallar </td><td> 7054X-85 </td><td></td></tr><tr><td> Leke Papanicolaou RG-6 </td><td> Merck Kimyasallar </td><td> 7052X-85 </td><td></td></tr><tr><td> Harris &#39;Şap hematoksilen </td><td> Merck Kimyasallar </td><td> 638A-85 </td><td></td></tr><tr><td> İsofluranın </td><td> Baxter Healthcare </td><td> NDC 10019-773-60 </td><td> Izofluran buharlaştırıcı ya da bir damla sistemi el anestezi kamara kapalı </td></tr></table>

protocols for vaginal inoculation and sample collection in the experimental mouse model of candida vaginitis

Vulvovajinal kandidiazis (VVC), Candida türlerinin neden olduğu, aksi takdirde sağlıklı kadınların yaklaşık% 75&#39;i üreme yıl boyunca 18,32-34 etkileyen alt dişi genital sistemin bir mantar enfeksiyonu. Predispozan faktörler, antibiyotik kullanımı, kontrolsüz diyabet ve rahatsızlık üreme hormon düzeyleri gebeliğe bağlı, oral kontraseptif veya hormon replasman tedavileri 33,34 içerir . Tekrarlayan VVC (RVVC) yılda üç veya daha fazla bölüm olarak tanımlanan, hiçbir predispozan faktör 33 ile% 8 kadın için ayrı bir 5 etkiler. VVC deneysel bir fare modeli kurulmuş ve patogenezi ve Candida 3,4,11,16,17,19,21,25,37 mukozal konak yanıtı çalışma için kullanılan olmuştur. Bu model aynı zamanda, in vivo 13,24 potansiyel antifungal tedaviler test etmek için işe alındı ​​. Modeli hayvanlar opt pseudoestrus bir devlet muhafaza gerektirirolmuş bölgeler Candida kolonizasyonu / infeksiyonu 6,14,23. Bu koşullar altında, haftalar veya aylar için inoküle hayvanlar vajinal mantar yükünün saptanabilir olacaktır. Geçmişte yapılan çalışmalar, immünolojik ve fizyolojik özellikleri 3,16,21 göre hayvan modeli ve insan enfeksiyonu arasındaki son derece yüksek bir paralellik göstermektedir. Farklılıklar Ancak, normal vajinal flora olarak Candida eksikliği ve farelerde nötr bir vajinal pH içerir. Burada, bir dizi vajinal aşılama, vajinal örneklerin hızlı toplanması, vajinal mantar yükünün değerlendirilmesi ve hücresel çıkarma / izolasyonu için doku hazırlıkları fare vajinit modeli temel yöntemler göstermektedir. Bu vajinal lavaj sıvısı, mantar yükünü, ve drenaj lenf nodu lökosit verim bileşenlerinin temsilcisi sonuçları takip edilmektedir. Anesteziklerin kullanımı ile lavaj örnekleri boyuna değerlendirilmesi için aynı fareler üzerinde birden fazla zaman noktalarında toplananenfeksiyon / kolonizasyon. Ayrıca, tanımlanmış ana koşullar altında immünolojik çalışmalar sağlayan bu model, enfeksiyon başlatmak için herhangi bir immünosupresif ajanlar gerektirir. Son olarak, model ve her iki tekniğin burada potansiyel düşük kadın genital sistem (viral, parazitik, bakteriyel) ve ilgili lokal veya sistemik konak savunmasını diğer bulaşıcı hastalıkları incelemek için metodolojilerin kullanımı doğurabilecek tanıttı.

Watch this Scientific Journal Video about Protocols for Vaginal Inoculation and Sample Collection in the Experimental Mouse Model of Candida vaginitis at JoVE.com

Protocols for Vaginal Inoculation and Sample Collection in the Experimental Mouse Model of Candida vaginitis

Değerlendirilmesinde kullanılacak kilit teknikleri<em&gt; Candida</em&gt; Vajinit deneysel bir hayvan modeli anlatılmaktadır. Yöntemleri, lomber lenf düğümleri drenaj, vajinal örneklerin ve lenfositler hızlı toplama sağlayacaktır. Bu teknikler, kadın alt genital sistem diğer hastalıkların fare modelleri yol açabilir.

Deneysel Fare Modeli Vajinal Aşılama ve Örnek Toplama Protokolleri Candida Vajinit

Vulvovaginal candidiasis (VVC), caused by Candida species, is a fungal infection of the lower female genital tract that affects approximately 75% of ...

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Protocols for Vaginal Inoculation and Sample Collection in the Experimental Mouse Model of Candida vaginitis

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