Name: transurethral induction of mouse urinary tract infection
Uploaded: 2010-08-05T17:30:00
Description: Watch this Scientific Journal Video about Transurethral Induction of Mouse Urinary Tract Infection at JoVE.com

Yazarlar, uzman teknik yardım için Shelly Xie ve Jennifer Payne, kabul etmek istiyorum. Bu çalışma Pediatrik Üroloji Derneği ve Stanford Pediatrik Araştırma Fonu hibe yoluyla mümkün olmuştur.

Üretral kateterler I. Hazırlık <ol><li> Numune alınacak her bir fare mesane ve böbrek için, sırasıyla, bir cryovial tüp içinde 5-6 paslanmaz çelik boncuk (mesane, 1.6 mm boyutlarında veya 0,9-2,0 mm karışımı) veya zirkonyum oksit boncuk (böbrek, 0.5 mm boyutlarında) yerleştirin. Boncuk içeren tüplere Otoklav. </li><li> Ortam sıcaklığına kadar soğumasını tüpleri sonra, her paslanmaz çelik boncuk içeren tüp ve her zirkonyum oksit boncuk içeren tüp steril PBS 400 mikrolitre 200 mikrolitr...

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	<li>Schaeffer, A. J., Schwan, W. R., Hultgren, S. J., Duncan, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relationship+of+type+1+pilus+expression+in+Escherichia+coli+to+ascending+urinary+tract+infections+in+mice.">Relationship of type 1 pilus expression in Escherichia coli to ascending urinary tract infections in mice.</a> Infect Immun. 55, 373-380 (1987).</li><li>Rouschop, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Urothelial+CD44+facilitates+Escherichia+coli+infection+of+the+murine+urinary+tract.">Urothelial CD44 facilitates Escherichia coli infection of the murine urinary tract.</a> J Immunol. 177, 7225-7232 (2006).</li><li>Malaviya, R., Ikeda, T., Abraham, S. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contribution+of+mast+cells+to+bacterial+clearance+and+their+proliferation+during+experimental+cystitis+induced+by+type+1+fimbriated+E.+coli.">Contribution of mast cells to bacterial clearance and their proliferation during experimental cystitis induced by type 1 fimbriated E. coli.</a> Immunol LettM. 91, 103-111 (2004).</li><li>Lane, M. C., Alteri, C. J., Smith, S. N., Mobley, H. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+flagella+is+coincident+with+uropathogenic+Escherichia+coli+ascension+to+the+upper+urinary+tract.">Expression of flagella is coincident with uropathogenic Escherichia coli ascension to the upper urinary tract.</a> Proc Natl Acad Sci U S A. 104, 16669-16674 (2007).</li><li>Hvidberg, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+long-term+ascending+urinary+tract+infection+mouse+model+for+antibiotic+treatment+studies.">Development of a long-term ascending urinary tract infection mouse model for antibiotic treatment studies.</a> Antimicrob Agents Chemother. 44, 156-163 (2000).</li><li>Hopkins, W. J., Hall, J. A., Conway, B. P., Uehling, D. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+urinary+tract+infection+by+intraurethral+inoculation+with+Escherichia+coli:+refining+the+murine+model.">Induction of urinary tract infection by intraurethral inoculation with Escherichia coli: refining the murine model.</a> J Infect Dis. 171, 462-465 (1995).</li><li>Hopkins, W. J., Gendron-Fitzpatrick, A., Balish, E., Uehling, D. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Time+course+and+host+responses+to+Escherichia+coli+urinary+tract+infection+in+genetically+distinct+mouse+strains.">Time course and host responses to Escherichia coli urinary tract infection in genetically distinct mouse strains.</a> Infect Immun. 66, 2798-2802 (1998).</li><li>Gunther, N. W. t., Lockatell, V., Johnson, D. E., Mobley, H. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+dynamics+of+type+1+fimbria+regulation+in+uropathogenic+Escherichia+coli+during+experimental+urinary+tract+infection.">In vivo dynamics of type 1 fimbria regulation in uropathogenic Escherichia coli during experimental urinary tract infection.</a> Infect Immun. 69, 2838-2846 (2001).</li><li>Bishop, B. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cyclic+AMP-regulated+exocytosis+of+Escherichia+coli+from+infected+bladder+epithelial+cells.">Cyclic AMP-regulated exocytosis of Escherichia coli from infected bladder epithelial cells.</a> Nat Med. 13, 625-630 (2007).</li><li>Thumbikat, P., Waltenbaugh, C., Schaeffer, A. J., Klumpp, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antigen-specific+responses+accelerate+bacterial+clearance+in+the+bladder.">Antigen-specific responses accelerate bacterial clearance in the bladder.</a> J Immunol. 176, 3080-3086 (2006).</li><li>Hagberg, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ascending,+unobstructed+urinary+tract+infection+in+mice+caused+by+pyelonephritogenic+Escherichia+coli+of+human+origin.">Ascending, unobstructed urinary tract infection in mice caused by pyelonephritogenic Escherichia coli of human origin.</a> Infect Immun. 40, 273-283 (1983).</li><li>Hopkins, W. J., Hall, J. A., Conway, B. P., Uehling, D. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+urinary+tract+infection+by+intraurethral+inoculation+with+Escherichia+coli:+refining+the+murine+model.">Induction of urinary tract infection by intraurethral inoculation with Escherichia coli: refining the murine model.</a> J Infect Dis. 171, 462-465 (1995).</li><li>Hung, C. S., Dodson, K. W., Hultgren, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+murine+model+of+urinary+tract+infection.">A murine model of urinary tract infection.</a> Nat Protoc. 4, 1230-1243 (2009).</li></ol>

İndüksiyon fare idrar yolu enfeksiyonu, bakteri transüretral aşılama köklü ama teknik olarak zor bir işlemdir 11. Hung ve Hultgren fare idrar yolu enfeksiyonu 13 transüretral teknikler son zamanlarda mükemmel bir inceleme yayınladı. Bu çalışmada, biz bu deneysel yaklaşım daha ince göstermek için çalışırlar. Yeni başlayanlar, mesane ortaya çıkarmak için bir orta hat laparotomi yapmak, Hindistan mürekkep (PBS içinde% 1) bir çözelti enjekte v...

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		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Cryovial tubes, 2.0 ml polypropylene (no caps)</td>
<td>&nbsp;</td>
<td>E&amp;K Scientific</td>
<td>640201</td>
<td>&nbsp;</td>
<tr>
<td>Caps for cryovial tubes, with 'O' ring</td>
<td>&nbsp;</td>
<td>E&amp;K Scientific</td>
<td>440011-N</td>
<td>&nbsp;</td>
<tr>
<td>Bullet Blender</td>
<td>&nbsp;</td>
<td> Next Advance</td>
<td>BBX24</td>
<td>&nbsp;</td>
<tr>
<td>Zirconium Oxide Beads (0.5mm)</td>
<td>&nbsp;</td>
<td>Next Advance</td>
<td>ZROB05</td>
<td>&nbsp;</td>
<tr>
<td>Stainless Steel Beads (1.6mm)</td>
<td>&nbsp;</td>
<td>Next Advance</td>
<td>SSB16</td>
<td>&nbsp;</td>
<tr>
<td>Stainless Steel Bead Blend (0.9-2.0mm)</td>
<td>&nbsp;</td>
<td>Next Advance</td>
<td>SSB14B</td>
<td>&nbsp;</td>
<tr>
<td>CBA/J Mice, 8-12 week old female</td>
<td>&nbsp;</td>
<td>Jackson Laboratories</td>
<td>000656</td>
<td>&nbsp;</td>
<tr>
<td>Phosphate Buffered Saline (NaCl 137mM, KCl 2.7mM, Na2HPO4 4.3mM, KH2PO4 1.4mM)</td>
<td>&nbsp;</td>
<td>EMD</td>
<td>EM-SX0420-13 (NaCl) EM-PX1405-1 (KCl) EM1.06585.0 (Na2HPO4) EM-5108-1 (KH2PO4)</td>
<td>&nbsp;</td>
<tr>
<td>LB agar</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>LB broth</td>
<td>&nbsp;</td>
<td>EMD</td>
<td>1.10285.0500</td>
<td>&nbsp;</td>
<tr>
<td>Isoflurane (Aerrane)</td>
<td>&nbsp;</td>
<td>Baxter</td>
<td>NDC 10019-773-60</td>
<td>&nbsp;</td>
<tr>
<td>Inoculating loops</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>22-363-601</td>
<td>&nbsp;</td>
<tr>
<td>Eosin-methylene blue (EMB) agar powder</td>
<td>&nbsp;</td>
<td>Himedia Laboratories</td>
<td>MM022-500G</td>
<td>&nbsp;</td>
<tr>
<td>MacConkey agar powder</td>
<td>&nbsp;</td>
<td>Difco</td>
<td>212123</td>
<td>&nbsp;</td>
<tr>
<td>LB agar powder</td>
<td>&nbsp;</td>
<td>Difco</td>
<td>244520</td>
<td>&nbsp;</td>
<tr>
<td>Intramedic non-radiopaque polyethylene tubing, PE10 [inner dimension 0.28 mm (0.011 inch); outer dimension 0.61 mm (0.024 inch)]</td>
<td>&nbsp;</td>
<td>BD Biosciences</td>
<td>427401</td>
<td>&nbsp;</td>
<tr>
<td>30 gauge needles, 1/2 inch</td>
<td>&nbsp;</td>
<td>BD Biosciences</td>
<td>305106</td>
<td>&nbsp;</td>
<tr>
<td>1 ml tuberculin Slip-Tip syringe</td>
<td>&nbsp;</td>
<td>BD Biosciences</td>
<td>309602</td>
<td>&nbsp;</td>
<tr>
<td>3 ml Luer-Lock syringe</td>
<td>&nbsp;</td>
<td>BD Biosciences</td>
<td>309585</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

transurethral induction of mouse urinary tract infection

Ilgi üropatojenik bakteri suşlarının agar yetiştirilmektedir. Genellikle, üropatojenik<em&gt; E. coli</em&gt; (UPEC) ve diğer suşların 37 gece Luria-Bertani agar (LB) yetiştirilebilir ° C, havadaki. UPEC suşları LB agar sarımsı-beyaz saydam koloniler olarak büyür. Uygun koloni morfolojisi onayı takiben, tek koloniler, daha sonra suyu olarak yetiştirilebilir toplanır. LB suyu en üropatojenik bakteri suşları için kullanılabilir. İki seri, bir gecede LB suyu kültürleri, tip I pili, üropatojenik bakteriler için iyi tanımlanmış bir virülans faktörü ifade artırmak için istihdam edilebilirler. Broth kültürler istenen konsantrasyona fosfat tamponlu salin (PBS) seyreltilir. Sekiz-12 haftalık dişi farelerde izofluran anestezi altında yerleştirilir ve transüretral 30 göstergesi şırınga boru kaplı polietilen kullanılarak bakteri inoküle. Ampirik / fare her bakteri suşu kombinasyonu için tespit edilmelidir tipik inocula, 10<sup&gt; 6</sup&gt; 10<sup&gt; 8</sup&gt; 10 ila 50 mikrolitre PBS kob başına fare. İstenen enfeksiyon döneminden sonra (birkaç hafta gün), idrar örnekleri ve mesane ve her ikisi de böbrek hasat edilir. Her organ, kıyılmış PBS içinde yer ve Mavi Bullet Homojenizatör homojenize. Idrar ve doku homojenatlarında seri PBS içinde seyreltilmesi ve uygun bir agar kültüre. Ertesi gün, koloni oluşturan birim sayılır.

Watch this Scientific Journal Video about Transurethral Induction of Mouse Urinary Tract Infection at JoVE.com

Transurethral Induction of Mouse Urinary Tract Infection

Bu video transüretral fare idrar yolu enfeksiyonlarına neden ve enfeksiyonların sonucunda dereceye ölçmek için yöntemler gösterecektir.

Transüretral İndüksiyon Fare İdrar Yolu Enfeksiyonu

Uropathogenic bacterial strains of interest are grown on agar. Generally, uropathogenic E. coli (UPEC) and other strains can be grown overnight on ...

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Deneysel Otoimmün hipofizitis İndüksiyon Fare Hipofiz İmmunogen hazırlanması

Autoimmune hypophysitis is a chronic inflammation of the pituitary gland caused or accompanied by autoimmunity1.  It has traditionally been considered a ...

Induction of Experimental Autoimmune Hypophysitis in SJL Mice

SJL Farelerde Deneysel Otoimmün hipofizitis İndüksiyon

Autoimmune hypophysitis can be reproduced experimentally by the injection of pituitary proteins mixed with an adjuvant into susceptible mice1. Mouse ...

RNA Isolation of Pseudomonas aeruginosa Colonizing the Murine Gastrointestinal Tract

RNA Isolation of Pseudomonas aeruginosa Colonizing the Murine Gastrointestinal Tract

RNA İzolasyonu Pseudomonas aeruginosa Murin Gastrointestinal Yolu koloniler

Pseudomonas aeruginosa (PA) infections result in significant morbidity and mortality in hosts with compromised immune systems, such as patients with ...

Isolation of Lymphocytes from Mouse Genital Tract Mucosa

Fare Genital Sistemlerinin Mukozanin gelen Lenfosit İzolasyonu

Mucosal surfaces, including in the gastrointestinal, urogenital, and respiratory tracts, provide portals of entry for pathogens, such as viruses and ...

Urinary Tract Infection in a Small Animal Model: Transurethral Catheterization of Male and Female Mice

İdrar yolu enfeksiyonu küçük bir hayvan modeli: erkek ve dişi farelerin transüretral Kateterizasyon

Urinary tract infections (UTI) are extremely common worldwide, incurring significant morbidity and healthcare-associated expenses. Small animal models, ...

Transurethral Instillation Procedure in Adult Male Mouse

Yetişkin erkek fare yordamda transüretral korumak

Transurethral instillation can be used to deliver different solutions with active ingredients (e.g., drugs, chemicals, bacteria, and viruses) locally into ...

Mouse Models Of Helicobacter Infection And Gastric Pathologies

Mouse Models Of Helicobacter Infection And Gastric Pathologies

Fare modelleri Helicobacter enfeksiyonu ve gastrik patolojiler

Helicobacter pylori is a gastric pathogen that is present in half of the global population and is a significant cause of morbidity and mortality in ...

Isolation of Dendritic Cells from the Human Female Reproductive Tract for Phenotypical and Functional Studies

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The characterization of the human dendritic cells (DCs) resident in mucosal tissues is challenging due to the difficulty in obtaining samples, and the low ...

Isolation of Single Intracellular Bacterial Communities Generated from a Murine Model of Urinary Tract Infection for Downstream Single-cell Analysis

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In this article, we outline a procedure used to isolate individual intracellular bacterial communities from a mouse that has been experimentally infected ...

Induction of Mouse Lung Injury by Endotracheal Injection of Bleomycin

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Pulmonary fibrosis is a hallmark of several human lung diseases with a different etiology. Since current therapies are rather limited, mouse models ...