The evolution of the device started with somewhat different cutting prototype demonstrated at the Imperial Cancer Research Fund (ICRF) mouse club by Bass Hassan, following this there were several discussions with Rob Rudling and Nikki Mandir in the containment facility at ICRF Clare Hall. Jo Kitau of the mechanical workshop at ICRF constructed the initial versions and Andy Lucas designed and perfected the final model in the CRUK workshop in Oxford.

تمت الموافقة على الإجراءات التجريبية المختلفة والقتل الرحيم إنسانية من قبل لجنة أخلاقية المؤسسات المضيفة والهيئات التنظيمية المناسبة. ويتحقق القتل الرحيم عادة باستخدام CO 2 الخنق تليها خلع عنق الرحم. 1. جهاز <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> آلة الجزء الرئيسي للقاعدة الجهاز من خارج كتلة صلبة عالية الكثافة الراتنج الاسيتيل، كما هو موضح في هذه المخطوطة. في نهاية كل إنشاء 3 مم عالية ارتقى الشرائط، وآلة جزء من هذه 2.5 ملم بساتين لاتخاذ قضبان (انظر الشكل 2 والشكل 3). </li><li style=";text-align:right;direction:rtl"> جعل قضبان من 2.4 مم الفولاذ المقاوم للصدأ مع نهايات مدورة حالا. تم تشكيله الإطار الرئيسي للغطاء من لوحة الدوراليومين مزيج من ألمونيوم و نحاس: ملاحظة. كما يتوفر عدد محدود من أجهزة الانتهاء من الدكتور Goodlad. </li></ol> 2. استخدام الجهاز <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> تشريح خارج الأمعاء الدقيقة والقولون من تجويف البطن باستخدام مقصوأصابع القفاز. إزالة مساريق من خلال عقد مع ملقط المنحنية وبلطف سحبه بعيدا من الأمعاء. </li><li style=";text-align:right;direction:rtl"> شطف الأمعاء مع الفوسفات الباردة مخزنة المالحة (PBS، ودرجة الحموضة 7.4) باستخدام نوع جيلسون ماصة، الذي كان قد خفض في نهاية الأوسع بحيث يمكن أن يصلح على 10 أو 20 مل بالتركيبة حقنة بلاستيكية. </li><li style=";text-align:right;direction:rtl"> وضع الأمعاء الدقيقة (SB) إلى ورقة منشفة ورقية وتقسم إلى ثلاثة أقسام متساوية في الطول (الداني، منتصف، والقاصي - SB1، SB2 وSB3) باستخدام المقص. إزالة الأعور ووضع القولون. </li><li style=";text-align:right;direction:rtl"> جعل العديد من الجروح الصغيرة جدا في قطاعات المعوية مع مقص لتسمح للسائل للهروب ووضع ورقة منشفة ورقية على الأمعاء وبلطف تشغيل الاصبع على التحضيرات للضغط على السائل وصمة عار الجافة. </li><li style=";text-align:right;direction:rtl"> رفع المقاطع وإدخال قضبان الفولاذ المقاوم للصدأ التي تم المبللة مسبقا عن طريق نقع في كوب من الفوسفات مخزنة المالحة. ملاحظة: الأمعاء يمكن أن يكون فيverted إذا رغبت في ذلك (تحولت الداخل الى الخارج). </li><li style=";text-align:right;direction:rtl"> تسمية قطعة من بطاقة أو تصفية قطع الورق إلى حجم لتناسب قاعدة الجهاز. قلم رصاص هو أفضل، لأنه لا يتأثر كل الحلول تقريبا. واقترح تاريخ التشريح، رمز التجريبية ورقم هوية الحيوان. </li><li style=";text-align:right;direction:rtl"> وضع بطاقة أو ورقة مرشح في قاعدة الجهاز. إدراج قضبان والأمعاء في الفتحات الموجودة في قاعدة الجهاز. تأكد من وضع نهاية القريبة من القطاعات بطريقة موحدة. يجب أن تكون نهايات القريبة بالقرب من التسميات البطاقة. </li><li style=";text-align:right;direction:rtl"> ضع قطعة العلوي من الجهاز على قاعدة. استخدام أشرطة الزاوية لتوجيه شفرة مشرط لقطع طولي الأقسام. ملاحظة: كما أنه يساعد إذا كان النسيج هو بلطف وعناية عقدت مع ساكنا لضمان أن لا يتحرك مع السكين. </li><li style=";text-align:right;direction:rtl"> إزالة قطعة العلوي من الجهاز. إزالة بعناية ورقة والأنسجة مرشح (لا يزال على عصيهم). ملاحظة: قطعة من بطاقة شديدة وضعت تحت سن الفيلوتستعرض ورقة ثالثا مساعدة الرفع. </li><li style=";text-align:right;direction:rtl"> الرطب قليلا (مع الإصبع القفاز مغموسة في PBS) قطاعات. لفة قضيب، جنبا إلى جنب لفتح القناة الهضمية وينتشر بشكل مسطح. سوف الأنسجة ثم الانضمام إلى ورقة الترشيح. </li><li style=";text-align:right;direction:rtl"> فحص البصر الاستعدادات لأي الآفات الجسيمة. </li><li style=";text-align:right;direction:rtl"> نقل الأنسجة الالتزام بها ورقة الترشيح إلى حمام ضحل (أو شطيرة مربع) تحتوي على مثبت. ملاحظة: بعد تتمسك الأنسجة إلى ورقة الترشيح أو بطاقة سيبقى تعلق بحزم. وهناك مجموعة واسعة من المثبتات يمكن استخدامها للحفاظ على التحضير. فمن المستحسن أن السائل Carnoy ينبغي أن تستخدم لدراسة انتشار الاسلحة الخلايا المعوية 6، وهو تثبيت المثالي لأفضل الطرق تكاثر الخلايا. ومع ذلك الفورمالين أو يمكن أن تستخدم أيضا العديد من مثبتات أخرى إذا كانت هناك حاجة النهاية الأخرى. </li><li style=";text-align:right;direction:rtl"> بعد تثبيت (عادة 3 ساعات)، ونقل التحضيرات إلى 70٪ من الإيثانول. ويمكن بعد ذلك يتم تخزين العينات حتى المطلوبة. التحضيرات جلدراستها أون الوجه و / أو المستخدمة في إعداد العينات النسيجية لH &amp; E أو غيرها من تقنيات تلطيخ النسيجية، المناعية و / أو التهجين في الموقع. ملاحظة: المطول التثبيت يمكن أن تقلل من القابلية للتفاعل المناعى و / أو التهجين الموضعي للتفاعل في. صناديق البولي بروبلين شطيرة مثالية لتخزين عدد كبير من التحضيرات. </li><li style=";text-align:right;direction:rtl"> إذا لزم الأمر الأنسجة الثابتة من مختلف شرائح المعوية يمكن أن يتم في &#39;القوائم السويسرية وملطخة مجموعة كاملة من وسائل النسيجية 6 بما في ذلك تلك المذكورة أعلاه. لجعل &#39;لفة السويسرية تتم إزالة أجزاء الأمعاء من ورقة الترشيح، وتوالت حول عصا كوكتيل أو مسواك، المضمون مع دبوس الحشري ومعالجتها للشمع التضمين 6. </li><li style=";text-align:right;direction:rtl"> إذا لزم الأمر، وصمة عار على النسيج السائبة مع الميثيلين الأزرق إلى تصور الشاذة سرداب البؤر 7. </li><li style=";text-align:right;direction:rtl"> يسجل الاستعدادات الثابتة فيأوقات الفراغ. </li></ol>

<ol>
	<li>Goodlad, R. A., Alison, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=+Cancer+Handbook."> Cancer Handbook.</a> , 243-262 (2007).</li><li>Ward, J. M., Treuting, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rodent+intestinal+epithelial+carcinogenesis:+pathology+and+preclinical+models.">Rodent intestinal epithelial carcinogenesis: pathology and preclinical models.</a> Toxicol Pathol. 42, 148-161 (2014).</li><li>Sundberg, J. P., Hogenesch, H., Nikitin, A. Y., Treuting, P. M., Ward, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Training+mouse+pathologists:+ten+years+of+workshops+on+the+Pathology+of+Mouse+Models+of+Human+Disease.">Training mouse pathologists: ten years of workshops on the Pathology of Mouse Models of Human Disease.</a> Toxicol Pathol. 40, 823-825 (2012).</li><li>Wasan, H. S., Novelli, M., Bee, J., Bodmer, W. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dietary+fat+influences+on+polyp+phenotype+in+multiple+intestinal+neoplasia+mice.">Dietary fat influences on polyp phenotype in multiple intestinal neoplasia mice.</a> Proc Natl Acade Sci USA. 94, 3308-3313 (1997).</li><li>Rudling, R., Hassan, A. B., Kitau, J., Mandir, N., Goodlad, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+device+to+rapidly+prepare+whole+mounts+of+murine+intestine.">A simple device to rapidly prepare whole mounts of murine intestine.</a> Cell Prolif. 39, 415-420 (2006).</li><li>Alferez, D., Goodlad, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=To+best+measure+cell+proliferation+in+samples+from+the+intestine.">To best measure cell proliferation in samples from the intestine.</a> Cell Prolif. 40, 231-240 (2007).</li><li>Park, H. S., Goodlad, R. A., Wright, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+incidence+of+aberrant+crypt+foci+and+colonic+carcinoma+in+dimethylhydrazine-treated+rats+varies+in+a+site-specific+manner+and+depends+on+tumor+histology.">The incidence of aberrant crypt foci and colonic carcinoma in dimethylhydrazine-treated rats varies in a site-specific manner and depends on tumor histology.</a> Cancer Res. 57, 4507-4510 (1997).</li><li>Park, H. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+epidermal+growth+factor+and+dimethylhydrazine+on+crypt+size,+cell+proliferation,+and+crypt+fission+in+the+rat+colon.+Cell+proliferation+and+crypt+fission+are+controlled+independently.">Effects of epidermal growth factor and dimethylhydrazine on crypt size, cell proliferation, and crypt fission in the rat colon. Cell proliferation and crypt fission are controlled independently.</a> Am J Pathol. 151, 843-852 (1997).</li><li>Femia, A. P., Dolara, P., Caderni, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mucin-depleted+foci+(MDF)+in+the+colon+of+rats+treated+with+azoxymethane+(AOM)+are+useful+biomarkers+for+colon+carcinogenesis.">Mucin-depleted foci (MDF) in the colon of rats treated with azoxymethane (AOM) are useful biomarkers for colon carcinogenesis.</a> Carcinogenesis. 25, 277-281 (2004).</li><li>Ruehl-Fehlert, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Revised+guides+for+organ+sampling+and+trimming+in+rats+and+mice--part+1.">Revised guides for organ sampling and trimming in rats and mice--part 1.</a> Exp Toxicol Pathol. 55, 91-106 (2003).</li><li>Goodlad, R. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibiting+vascular+endothelial+growth+factor+receptor-2+signaling+reduces+tumor+burden+in+the+ApcMin/++mouse+model+of+early+intestinal+cancer.">Inhibiting vascular endothelial growth factor receptor-2 signaling reduces tumor burden in the ApcMin/+ mouse model of early intestinal cancer.</a> Carcinogenesis. 27, 2133-2139 (2006).</li><li>Mandir, N., Englyst, H., Goodlad, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Resistant+carbohydrates+stimulate+cell+proliferation+and+crypt+fission+in+wild-type+mice+and+in+the+Apc(Min/+)+mouse+model+of+intestinal+cancer,+association+with+enhanced+polyp+development.">Resistant carbohydrates stimulate cell proliferation and crypt fission in wild-type mice and in the Apc(Min/+) mouse model of intestinal cancer, association with enhanced polyp development.</a> Br J Nutr. 100, 711-721 (2008).</li></ol>

The authors have nothing to disclose.

الجهاز تسرع كثيرا من عملية بحيث يمكن للمشغلين اثنين تشريح الجثة بالكامل ماوس في غضون 6 دقائق. وسوف يشمل ذلك فحص بصري للآفات جسيمة واضحة وزنها وتحديد الأجهزة الرئيسية. وسوف تسمح أيضا للشركات لشطف، وصمة عار وتزن المعدة، الأعور والأمعاء الدقيقة والقولون واعداد يتصاعد ك...

الماوس والماوس المعدلة وراثيا 1-3 ونموذجا لا تقدر بثمن لدراسة مختلف الأمراض وخاصة سرطان وعلاقته التهاب في الجهاز الهضمي. هذا وغالبا ما يتطلب إعداد يتصاعد كلها من الأمعاء الدقيقة و / أو القولون. إعداد يتصاعد كلها يمكن القيام به مع زوج دقة مقص تعويض ولكن يمكن أن يستغرق حوالي 20 دقيقة في الماوس 4. هذا ليس عملي جدا للدراسات المقارنة، والتي تتطلب قدرا كبيرا من الفئران (ويقول 10 في مجموعة تخضع للتحليل الإحصائي). وبالإضافة إلى ذلك، وطول الوقت الذي يستغرقه يزيد من خطر تدهور الأنسجة. محاولة للتخفيف من حدة هذه المشكلة أدت إلى محاكمة مختلف الوسائل إعداد القطع. واستند الأول في سلسلة من لوحات معدنية، مع بساتين القطع، ولكن حجب لوحات النسيج وكان قطع وبالتالي من الصعب السيطرة عليها. A conventionaل قلم رصاص سداسية قدمت أخيرا إلهام للجهاز القطع، ونحن يمكن بعد ذلك نرى كيف شكل نصف قلم رصاص من شأنه أن يجعل دليلا جيدا لمشرط (الشكل 1). وأدى ذلك إلى تصميم لإطار مع أربعة أدلة قطع 5. كانت فعالية هذا الجهاز دراماتيكية كما الاستعدادات في الشكل (4) تبين الفرق بين إعداد مقص وإعداد الجهاز. شيد قمم الأجهزة الأصلية من عدة قطع من المعدن، ولكن تلك في وقت لاحق، وكان تشكيله النموذج الحالي من قطعة صلبة من الدوراليومين مزيج من ألمونيوم و نحاس. تم تشكيله على قاعدة من أعلى مستويات الجودة الراتنج عالية الكثافة. وقد أثبت الجهاز أن تكون ذات فائدة كبيرة لإعداد يتصاعد كاملة من الماوس الأمعاء لأنها تسرع كثيرا ويحسن من جودة واتساق والاستعدادات، التي لديها مجموعة واسعة من التطبيقات.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Delrin</td>
			<td>Ryan Plastics, Earls Barton, UK</td>
			<td>N/A</td>
			<td>High-density acetal resin similar material would suffice</td>
		</tr>
		<tr>
			<td>Duralumin plate</td>
			<td>Metal Supplies Ltd, Park Road,Dukinfield. SK16 5LP UK</td>
			<td>N/A</td>
			<td>Aluminium allow dating back to 1909 so alterative suppliers are available&#160;</td>
		</tr>
		<tr>
			<td>Finished device(s) ready for use</td>
			<td>Contact Dr Goodlad&#160; r.goodlad@imperial.ac.uk</td>
			<td>N/A</td>
			<td>Contact&#160; r.goodlad@imperial.ac.uk&#160; for supply details</td>
		</tr>
	</tbody>
</table>

a simple device to rapidly prepare whole mounts of the mouse intestine

يمكن إعداد يتصاعد كلها من الأمعاء الدقيقة والقولون الماوس للتحليل الكمي أو لاحقة تكون مضيعة للوقت وصعبا. وصفنا استخدام جهاز بسيط لخفض والأمعاء &quot;لفة&quot; الماوس للتحضير بشكل سريع التحضير جبل كلها ذات جودة عالية وموحدة لتلك التي يمكن تحقيقها عن طريق اليد. ويتألف الجهاز قاعدة أن يحمل 4 قضبان الفولاذ المقاوم للصدأ وأعلى، والذي يعمل دليلا القطع. يتم إدراج قضبان في تجويف الأمعاء الدقيقة [مقسمة إلى ثلثي] والقولون. ثم توضع قضبان والعينات على قطعة من الورق أو بطاقة مرشح في فتحات عقد في قاعدة الجهاز. ثم يتم وضع الجزء العلوي من الجهاز وبمثابة دليل القطع. وتستخدم الأقسام الزاوية اثنين في وسط قطعة كبار لتوجيه سكين أو مشرط وقطع الأمعاء طوليا على الجزء العلوي من قضبان. مرة واحدة وقد تم قطع الأجزاء المعوية، تتم إزالة الجزء العلوي وبطاقة، TISSرق وقضبان إزالة بلطف من الجهاز ووضعها على مقاعد البدلاء. ثم يتم إرجاع قضبان بلطف جانبية للشد وعصا شرائح المعوية على قطعة الكامن وراء ورقة أو بطاقة مرشح. ويمكن بعد ذلك فحص الإعداد النهائي أو ثابتة وتخزينها لتحليلها لاحقا. الاستعدادات لا تقدر بثمن لدراسة التغيرات المعوية في نماذج الفئران العادية أو المعدلة وراثيا. وقد استخدمت الاستعدادات لدراسة وتقدير حجم آثار التهاب (التهاب القولون)، والضرر، والآفات ما قبل السرطانية (الشاذة سرداب البؤر (ACFs) والميوسين المنضب بؤر (MDFs)) واللحمية أو الأورام.

الاورام الحميدة هي سهلة لتحديد إطار stereomicroscope مع مصدر ضوء بارد إلى جانب إلقاء الضوء على الأنسجة (الشكل 4). ومن المفيد للاحتفال البطاقة مع علامات قلم رصاص عندما لوحظ ورم من اجل ان يحرز يمكن مناقشتها في وقت لاحق ويمكن لمشغلي مختلفة التوصل إلى اتفاق. الأنسجة يمكن ...

Watch this Scientific Journal Video about A Simple Device to Rapidly Prepare Whole Mounts of the Mouse Intestine at JoVE.com

A Simple Device to Rapidly Prepare Whole Mounts of the Mouse Intestine

The use of a simple device to cut and &#8216;roll&#8217; mouse intestines to rapidly prepare whole mount preparations is described.

جهاز بسيط للتحضير السريع الجبال الجامعة للماوس الأمعاء

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe ...

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19.5K Views.  National Institutes of Health. The use of a simple device to cut and ‘roll’ mouse intestines to rapidly prepare whole mount preparations is described.

Video: A Simple Device to Rapidly Prepare Whole Mounts of the Mouse Intestine

A Simple Method of Mouse Lung Intubation

A simple procedure to intubate mice for pulmonary function measurements would have several advantages in longitudinal studies with limited numbers or expensive animal. One of the reasons that this is not done more routinely is that it is relatively difficult, despite there being several published studies that describe ways to achieve it. In this paper we demonstrate a procedure that eliminates one of the major hurdles associated with this intubation, that of visualizing the trachea during the entire time of intubation. The approach uses a 0.5 mm fiberoptic light source that serves as an introducer to direct the intubation cannula into the mouse trachea. We show that it is possible to use this procedure to measure lung mechanics in individual mice over a time course of at least several weeks. The technique can be set up with relatively little expense and expertise, and it can be routinely accomplished with relatively little training. This should make it possible for any laboratory to routinely carry out this intubation, thereby allowing longitudinal studies in individual mice, thereby minimizing the number of mice needed and increasing the statistical power by using each mouse as its own control.

This paper describes a striaghforward and efficient method of intubating mice for pulmonary function measurements or pulmonary instillation, that allows the mice to recover and be studied at later times. The procedure involves an inexpensive fiberoptic light source that directly illuminates the trachea.

طريقة بسيطة لتركيب الانبوبة الرئة ماوس

A  simple procedure to intubate mice for pulmonary function measurements would  have several advantages in longitudinal studies with limited numbers or  ...

Isolating and Using Sections of Bovine Mesenteric Artery and Vein as a Bioassay to Test for Vasoactivity in the Small Intestine

Mammalian gastrointestinal systems are constantly exposed to compounds (desirable and undesirable) that can have an effect on blood flow to and from that system. Changes in blood flow to the small intestine can result in effects on the absorptive functions of the organ. Particular interest in toxins liberated from feedstuffs through fermentative and digestive processes has developed in ruminants as an area where productive efficiencies could be improved. The video associated with this article describes an in vitro bioassay developed to screen compounds for vasoactivity in isolated cross-sections of bovine mesenteric artery and vein using a multimyograph. Once the blood vessels are mounted and equilibrated in the myograph, the bioassay itself can be used: as a screening tool to evaluate the contractile response or vasoactivity of compounds of interest; determine the presence of receptor types by pharmacologically targeting receptors with specific agonists; determine the role of a receptor with the presence of one or more antagonists; or determine potential interactions of compounds of interest with antagonists. Through all of this, data are collected real-time, tissue collected from a single animal can be exposed to a large number of different experimental treatments (an in vitro advantage), and represents vasculature on either side of the capillary bed to provide an accurate picture of what could be happening in the afferent and efferent blood supply supporting the small intestine.

The small intestine is frequently exposed to toxins that can influence blood flow and negatively impact nutrient absorption. Using a multimyograph and mesenteric artery and vein isolates, compounds or toxins of interest can be screened for vasoactivity.

عزل وعن طريق أقسام الأبقار الشريان والوريد المساريقي باعتباره الأحيائي لاختبار لVasoactivity في الأمعاء الدقيقة

Mammalian gastrointestinal systems are constantly exposed to compounds (desirable and undesirable) that can have an effect on blood flow to and from that ...

A Simple Mechanical Procedure to Create Limbal Stem Cell Deficiency in Mouse

Limbal stem cell deficiency (LSCD) is a state of malfunction or loss of limbal epithelial stem cells, after which the corneal epithelium is replaced with conjunctiva. Patients suffer from recurrent corneal defects, pain, inflammation, and loss of vision.
Previously, a murine model of LSCD was described and compared to two other models. The goal was to produce a consistent mouse model of LSCD that both mimics the phenotype in humans and lasts long enough to make it possible to study the disease pathophysiology and to evaluate new treatments. Here, the technique is described in more detail.
A motorized tool with a rotating burr has been designed to remove the rust rings from the corneal surface or to smooth the pterygium bed in patients. It is a suitable device to create the desired LSCD model. It is a readily available, easy-to-use tool with a fine tip that makes it appropriate for working on small eyes, as in mice. Its application prevents unnecessary trauma to the eye and it does not result in unwanted injuries, as often is the case with chemical injury models. As opposed to a blunt scraper, it removes the epithelium with the basement membrane. In this protocol, the limbal area was abraded two times, and then the whole corneal epithelium was shaved from limbus to limbus. To avoid stroma injury, care was taken not to brush the corneal surface once the epithelium was already removed.

The following article provides an easy, reproducible technique to effectively create a sustainable mouse model of limbal stem cell deficiency (LSCD). This animal model is useful in testing and comparing the efficacy of treatments for limbal stem cell diseases.

إجراء ميكانيكي بسيط لإنشاء الحوفي نقص الخلايا الجذعية في الفأر

Limbal stem cell deficiency (LSCD) is a state of malfunction or loss of limbal epithelial stem cells, after which the corneal epithelium is replaced with ...

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

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

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

حقن إينتراكاردياك الموجهة بالموجات فوق الصوتية البشرية الخلايا الجذعية الوسيطة لزيادة صاروخ موجه للاستخدام في نماذج مورين من أمراض التهاب الأمعاء التجريبية للامعاء

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

Ileectomy-induced Bile Overaccumulation in Mouse Intestine

Intestinal resection is a common therapeutic approach for human diseases such as obesity, inflammatory bowel disease, Crohn's disease, and colon cancer that often results in severe short bowel syndrome-like adverse effects including bile acid diarrhea, dehydration, electrolyte disturbances, and nutrient malabsorption. Here we introduce a murine ileal resection model, termed ileectomy, to evaluate tissue communication and the maintenance of systemic homeostasis. After ileal resection, circulating blood is permanently devoid of the ileum-specific endocrine hormone fibroblast growth factor 15 (FGF15), which releases its endocrinal inhibition of bile acid synthesis in the liver. In combination with the increased production and abolished reabsorption of bile acids after removing the ileum, mice that underwent surgery suffer from bile salt overaccumulation in the intestine and associated diarrhea, morbidity, and mortality. Novel usage of the surgery model introduced in this study may provide mechanistic and functional insights into ileal control of systemic metabolic regulation in physiology and disease.

Small intestine-dependent bile acid reabsorption and feedback inhibition of hepatic bile acid synthesis is important for systemic homeostasis and health. In this study, we describe a mouse model for ileal resection to evaluate ileectomy-induced bile malabsorption, overaccumulation, and toxicity in mouse intestine.

أوفيراككوموليشن الناجمة عن إيليكتومي الصفراوية في الأمعاء الماوس

Intestinal resection is a common therapeutic approach for human diseases such as obesity, inflammatory bowel disease, Crohn's disease, and colon cancer ...

Mechanisms Underlying Gut Hormone Secretion Using the Isolated Perfused Rat Small Intestine

The gut is the largest endocrine organ of the body, producing more than 15 different peptide hormones that regulate appetite and food intake, digestion, nutrient absorption and distribution, and post-prandial glucose excursions. Understanding the molecular mechanisms that regulate gut hormone secretion is fundamental for understanding and translating gut hormone physiology. Traditionally, the mechanisms underlying gut hormone secretion are either studied in vivo (in experimental animals or humans) or using gut hormone-secreting primary mucosal cell cultures or cell lines. Here, we introduce an isolated perfused rat small intestine as an alternative method for studying gut hormone secretion. The virtues of this model are that it relies on the intact gut, meaning that it recapitulates most of the physiologically important parameters responsible for the secretion in in vivo studies, including mucosal polarization, paracrine relationships and routes of perfusion/stimulus exposure. In addition, and unlike in vivo studies, the isolated perfused rat small intestine allows for almost complete experimental control and direct assessment of secretion. In contrast to in vitro studies, it is possible to study both the magnitude and the dynamics of secretion and to address important questions, such as what stimuli cause secretion of different gut hormones, from which side of the gut (luminal or vascular) is secretion stimulated, and to analyze in detail molecular sensors underlying the secretory response. In addition, the preparation is a powerful model for the study of intestinal absorption and details regarding the dynamics of intestinal absorption including the responsible transporters.

Here, we present a powerful and physiological model to study the molecular mechanisms underlying gut hormone secretion and intestinal absorption &#8212; the isolated perfused rat small intestine.

الآليات الأساسية لإفراز هرمون الأمعاء التي تستخدم عزل Perfused الفئران الأمعاء

The gut is the largest endocrine organ of the body, producing more than 15 different peptide hormones that regulate appetite and food intake, digestion, ...

A Cryoinjury Model to Study Myocardial Infarction in the Mouse

The use of animal models is essential for developing new therapeutic strategies for acute coronary syndrome and its complications. In this article, we demonstrate a murine cryoinjury infarct model that generates precise infarct sizes with high reproducibility and replicability. In brief, after intubation and sternotomy of the animal, the heart is lifted from the thorax. The probe of a handheld liquid nitrogen delivery system is applied onto the myocardial wall to induce cryoinjury. Impaired ventricular function and electrical conduction can be monitored with echocardiography or optical mapping. Transmural myocardial remodeling of the infarcted area is characterized by collagen deposition and loss of cardiomyocytes. Compared to other models (e.g., LAD-ligation), this model utilizes a handheld liquid nitrogen delivery system to generate more uniform infarct sizes.

This article demonstrates a model to study cardiac remodeling after myocardial cryoinjury in mice.

نموذج الإصابة بالتبريد لدراسة احتشاء عضلة القلب في الماوس

The use of animal models is essential for developing new therapeutic strategies for acute coronary syndrome and its complications. In this article, we ...

A Modified Simple Method for Induction of Myocardial Infarction in Mice

Myocardial infarction (MI) represents one of the leading causes of death. MI models are widely used for investigating the pathomechanisms of post-MI remodeling and evaluation of novel therapeutics. Different methods (e.g., isoproterenol treatment, cryoinjury, coronary artery ligation, etc.) have been used to induce MI. Compared with isoproterenol treatment and cryoinjury, coronary artery ligation may better reflect the ischemic response and chronic remodeling after MI. However, traditional methods for coronary ligation in mice are technically challenging. The current study describes a simple and efficient process for induction of MI in mice with readily available materials. The mouse chest skin was cut open under stable anesthesia. The heart was immediately externalized through the intercostal space after blunt separation of the pectoralis major and pectoralis minor. The left anterior descending branch (LAD) was ligated with a 6-0 suture 3 mm from its origin. Following LAD ligation, staining with 2,3,5-Triphenyltetrazolium chloride (TTC) indicated successful induction of MI and temporal changes of post-MI scar size. Meanwhile, survival analysis results showed overt mortality within 7 days after MI, mainly due to cardiac rupture. Moreover, post-MI echocardiographic assessment demonstrated successful induction of contractile dysfunction and ventricular remodeling. Once mastered, an MI model can be established in mice within 2-3 min with readily available materials.

Under adequate anesthesia, the mouse heart was externalized through the intercostal space, and myocardial infarction was successfully induced by ligating the left anterior descending artery (LAD) using materials readily available in most laboratories.

طريقة بسيطة معدلة لتحريض احتشاء عضلة القلب في الفئران

Myocardial infarction (MI) represents one of the leading causes of death. MI models are widely used for investigating the pathomechanisms of post-MI ...

Application of Optical Coherence Tomography to a Mouse Model of Retinopathy

Optical coherence tomography (OCT) offers a noninvasive method for the diagnosis of retinopathy. The OCT machine can capture retinal crosssectional images from which the retinal thickness can be calculated. Although OCT is widely used in clinical practice, its application in basic research is not as prevalent, especially in small animals such as mice. Because of the small size of their eyeballs, it is challenging to conduct fundus imaging examinations in mice. Therefore, a specialized retinal imaging system is required to accommodate OCT imaging on small animals. This article demonstrates a small-animal-specific system for OCT examination procedures and a detailed method for image analysis. The results of retinal OCT examination of very-low-density lipoprotein receptor (Vldlr) knockout mice and C57BL/6J mice are presented. The OCT images of C57BL/6J mice showed retinal layers, while those of Vldlr knockout mice showed subretinal neovascularization and retinal thinning. In summary, OCT examination could facilitate the noninvasive detection and measurement of retinopathy in mouse models.

Here, we describe an in vivo imaging technique using optical coherence tomography to facilitate the diagnosis and quantitative measurement of retinopathy in mice.

تطبيق التصوير المقطعي للتماسك البصري على نموذج فأر لاعتلال الشبكية

Optical coherence tomography (OCT) offers a noninvasive method for the diagnosis of retinopathy. The OCT machine can capture retinal crosssectional images ...

Complete and Partial Resuscitative Endovascular Balloon Occlusion of the Aorta for Hemorrhagic Shock

Resuscitative endovascular balloon occlusion of the aorta (REBOA) devices grew out of a military-civilian partnership to develop new capabilities for hemorrhage control. With the advent of purpose-built devices, REBOA has become increasingly common in civilian trauma and acute care settings. Currently available REBOA catheters were designed as complete aortic occlusion devices. However, the therapeutic window for complete aortic occlusion is time-limited due to ischemia-reperfusion injury. The partial procedure allows blood flow past the level of occlusion while maintaining targeted proximal pressure, which has been shown to reduce distal ischemia and adjunctive resuscitation requirements in preclinical studies with prolonged occlusion times as compared to traditional complete occlusion.
pREBOA-PRO is the first catheter designed to enable partial and complete aortic occlusion and is currently in limited market release at seven Level I trauma centers in North America. This paper will focus on procedural considerations for REBOA, including patient selection criteria and a comparison of complete and partial aortic occlusion in a simulator, along with highlighting critical steps to improve clinical outcomes. Additionally, this paper reviews a contrast-enhanced CT scan from a trauma patient that shows distal perfusion after 2 h of partial aortic occlusion using this newly designed catheter and discusses representative results from the limited market release to highlight the profound effect of technological innovation on outcomes in vascular emergencies.

A commercial catheter was designed to facilitate true partial resuscitative endovascular balloon occlusion of the aorta (REBOA) and address the complications associated with complete aortic occlusion. Initial clinical reports indicate that partial REBOA improves transition to reperfusion, reduction of distal ischemia, and extension of safe occlusion time compared to complete occlusion.

انسداد بالون الأوعية الدموية الانعاشية الكاملة والجزئية للشريان الأورطي للصدمة النزفية

Resuscitative endovascular balloon occlusion of the aorta (REBOA) devices grew out of a military-civilian partnership to develop new capabilities for ...