يتم توفير التمويل اللازم لنشر بواسطة شركة Cerus، الشركة المصنعة لجهاز الدم INTERCEPT.

<ol>
	<li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Guide to the preparation, use and quality assurance of blood components. , (2010).</li><li>Van Rhenen, D. J., Vermeij, J., Mayaudon, V., 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=Functional+characteristics+of+S-59+photochemically+treated+platelet+concentrates+derived+from+buffy+coats.">Functional characteristics of S-59 photochemically treated platelet concentrates derived from buffy coats.</a> Vox Sang. 79, 206-214 (2000).</li><li>Wollowitz, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeting+DNA+and+RNA+in+pathogens:+mode+of+action+of+amotosalen+HCl.">Targeting DNA and RNA in pathogens: mode of action of amotosalen HCl.</a> Transfus. Med. Rev. 31, 11-16 (2004).</li><li>Irsch, J., Lin, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathogen+Inactivation+of+Platelet+and+Plasma+Blood+Components+for+Transfusion+Using+the+INTERCEPT+Blood+SystemTM.">Pathogen Inactivation of Platelet and Plasma Blood Components for Transfusion Using the INTERCEPT Blood SystemTM.</a> Transfus. Med. Hemother. 38, (2011).</li><li>Lin, L., Dikeman, R., Molini, B., 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=Photochemical+treatment+of+platelet+concentrates+with+amotosalen+and+long-wavelength+ultraviolet+light+inactivates+a+broad+spectrum+of+pathogenic+bacteria.">Photochemical treatment of platelet concentrates with amotosalen and long-wavelength ultraviolet light inactivates a broad spectrum of pathogenic bacteria.</a> Transfusion. 44, 1496-1504 (2004).</li><li>Lin, L., Hanson, C., Alter, H., 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=Inactivation+of+viruses+in+platelet+concentrates+by+photochemical+treatment+with+amotosalen+and+long-wavelength+ultraviolet+light.">Inactivation of viruses in platelet concentrates by photochemical treatment with amotosalen and long-wavelength ultraviolet light.</a> Transfusion. 45, 580-590 (2005).</li><li>Allain, J. P., Cianco, C., Blajchman, 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=Protecting+the+blood+supply+from+emerging+pathogens:+the+role+of+pathogen+inactivation.">Protecting the blood supply from emerging pathogens: the role of pathogen inactivation.</a> Transfus. Med. Rev. 19, 110-126 (2005).</li><li>Stramer, S., Hollinger, F., Katz, L., 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=Emerging+infectious+disease+agents+and+their+potential+threat+to+transfusion+safety.">Emerging infectious disease agents and their potential threat to transfusion safety.</a> Transfusion. 49, 1S-29S (2009).</li><li>Nussbaumer, W., Allesdorfer, D., Grabmer, 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=Prevention+of+transfusion+of+platelet+components+contaminated+with+low+levels+of+bacteria:+a+comparison+of+bacteria+culture+and+pathogen+inactivation+methods.">Prevention of transfusion of platelet components contaminated with low levels of bacteria: a comparison of bacteria culture and pathogen inactivation methods.</a> Transfusion. 47, 1125-1133 (2007).</li><li>Schlenke, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protection+against+Transfusion-Associated+Graft-versus-Host+Disease+in+Blood+transfusion:+Is+Gamma-Irradiation+the+Only+Answer?+Transfus.">Protection against Transfusion-Associated Graft-versus-Host Disease in Blood transfusion: Is Gamma-Irradiation the Only Answer? Transfus.</a> Med. Hemother. 31, 24-31 (2004).</li><li>Lin, L., Corash, L., Osselear, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protection+Against+TA-GVHD+Due+to+Platelet+Transfusion+By+Using+Pathogen+Inactivation+with+the+INTERCEPT+Blood+SystemTM+-+Gamma+Irradiation+is+Not+the+Only+Answer.">Protection Against TA-GVHD Due to Platelet Transfusion By Using Pathogen Inactivation with the INTERCEPT Blood SystemTM - Gamma Irradiation is Not the Only Answer.</a> Haematologica. 95 (Extra 1), 230-237 (2010).</li><li>Corash, L., Lin, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+processes+for+inactivation+of+leukocytes+to+prevent+transfusion-associated+graft-verus-host+disease.">Novel processes for inactivation of leukocytes to prevent transfusion-associated graft-verus-host disease.</a> Bone Marrow Transplant. 33, 1-7 (2004).</li></ol>

ويرعى الإنتاج وحرية الوصول إلى هذه المقالة عن طريق Cerus.

الصفائح الدموية معطف الشهباء تتطلب عدة خطوات المعالجة التي تؤدي إلى ما بعد جمع التكاليف، بما في ذلك وقت الموظفين، والمواد الاستهلاكية، والمعدات، والنفايات، والتي يجب أن يؤخذ في الاعتبار التكلفة الإجمالية للإنتاج وحدات الصفائح الدموية. تحسين العائد الصفائح الدموية من كل معطف الشهباء (عن طريق الاستفادة المثلى من حجم الغلاف الأبيض والهيماتوكريت) يسمح للإنتاج وحدة جرعة مضاعفة معطف الشهباء الصفائح الدموية من بين مجموعة من المعاطف بافي سبعة. مطلوب عدد من المعاطف بافي عندما يتم تنفيذ هذا التحسين، لإنتاج عدد محدد من الجرعات الصفائح الدموية قد خفضها، وبالتالي تحسين الانتفاع العام من المعاطف بافي والتمكين من إنتاج الصفائح الدموية إضافية تركز (أجهزة الكمبيوتر). يتم تخفيض وقت الموظفين وكذلك تكاليف استهلاك والمعدات مثل أجهزة تجميع، الصفائح الدموية حل المضافة، رقائق اتصال العقيمة، ومجموعات التصفية، وإجراءات الطرد المركزي بنسبة تصل إلى 50٪. بالإضافة إلى تحسين إنتاج الصفائح الدموية وتقليل التكاليف المرتبطة بها، ثييمكن تجميع ليالي طريقة توليد التركيز الصفائح الدموية (PC) التي تلبي متطلبات مدخلات للاستخدام مع مجموعة التخزين المزدوج INTERCEPT معالجة الحاويات، مما يسمح لنا لتوفير الحماية المعززة لنقل المستفيدين لدينا أيضا. جهاز الدم INTERCEPT لتعطيل الممرض يستخدم amotosalen والأشعة فوق البنفسجية A الخفيفة (UVA) لDNA عبر الرابط تساهميا وRNA، ومنع تكرار الحمض النووي ومسببات الأمراض غير قادر على تقديم تسبب المرض .. 3 ويثبط نشاط فعال طيف واسع من الجراثيم بما في ذلك الفيروسات والبكتيريا ، والطفيليات، وتلويث الخلايا المانحة الدم البيضاء 4-6 INTERCEPT يوفر بديلا للنموذج اختبار لمسببات الأمراض الناشئة الحالي، الذي ينطوي على تأخير كبير تاريخيا في حين تم تطوير اختبار جديد ويتطلب استثمارات كبيرة في نهاية المطاف المالية لتنفيذ عند اختبار تصبح متوفرة. 7،8 ويمكن أيضا أن توفر بديلا للmeasu السلامة زائدة عن الحاجةالدقة مثل الكشف عن البكتيريا 9 و أشعة جاما. 10-12 وبالإضافة إلى ذلك، يسمح لنا INTERCEPT لعلاج ضعف الصفائح الدموية وحدات الجرعة التي تفيد كفاءة الإنتاج لدينا وتساعدنا على التمسك ميزانياتنا. يمكن تكييفها INTERCEPT وضعف معطف الشهباء طريقة الجرعة إلى مجموعة متنوعة من مهام سير العمل مركز الدم. كأمثلة، ويمكن زيادة مجموعات الدم الكامل إلى 500 مل، ويمكن أيضا أن يتم تخزين مجموعات بين عشية وضحاها في 22 ° C قبل الانفصال. وبالإضافة إلى ذلك، بديل بافي طريقة تجميع معطف من الممكن (على سبيل المثال طريقة الأخطبوط بدلا من أسلوب القطار) و / أو جهاز آلي (مثل TACSI) يمكن استخدامها لفصل الثاني والطرد المركزي لتعليق الصفائح الدموية من الخلايا الحمراء المتبقية . إذا كانت هناك حاجة محتوى الصفائح الدموية المتزايدة، العديد من التقنيات المتاحة بما في ذلك اختيار المسبق للمانحين يعتمد على عدد الصفائح الدموية، بين عشية وضحاها من الحضانة الدم الكامل، adjustment من إعدادات الطرد المركزي، أو استخدام مجموعة من المعاطف بافي 8 في بدلا من 7. بسبب الحد من تجميع مجموعات المتاحة حاليا وقدرة أجهزة الطرد المركزي الدلاء، ينبغي أن الحجم الكلي للتجمع معطف الشهباء تكون حوالي 600 مل. وقد تم تحسين عملية الطرد المركزي والإعدادات المبينة في هذا البروتوكول لانتاج المنتجات التي تلبي المعايير INTERCEPT لمجموعة من المعاطف بافي 7. يجب أن يتم تعديل الإعدادات الطرد المركزي إذا تم تعديل عدد من المعاطف بافي في حوض السباحة. في بعض البلدان، أقل جرعة الصفائح الدموية أعلى ومتطلبات QC هي أكثر صرامة من في السويد. على هذا النحو، قد لا تكون نتائجنا قابلة للتطبيق عالميا. في هذه البلدان، فإن جرعة مضاعفة PC قبل العلاج INTERCEPT بحاجة لاحتواء أكبر عدد من الصفائح الدموية من أجل تلبية المتطلبات المحلية بعد العلاج INTERCEPT والانقسام. لأن محتوى الأقصى الصفائح الدموية والعلاج FO حجمص 11 INTERCEPT هي 7x10 الصفائح الدموية وML 420 على التوالي، فإن نسبة الوحدات التي تتجاوز متطلبات تجهيز INTERCEPT أو الصفائح الدموية تحتوي على محتوى غير كاف لأن تنقسم الى اثنين الجرعات العلاجية، وقد تختلف تلك الرسوم تبعا لعوامل مثل المتطلبات المحلية للجرعة الصفائح الدموية، ومعايير مراقبة الجودة، النتائج بافي الأمثل معطف، والاستقرار الإنتاج. إذا كان PC جرعة مضاعفة تجاوز متطلبات تجهيز INTERCEPT، قد يكون ذلك ضبطها يدويا لتلبية متطلبات وتعامل في وقت لاحق. في تلك المناسبات القليلة التي تجمع معطف 7-بافي ينتج الصفائح الدموية كافية لمنتج جرعة مضاعفة بعد INTERCEPT، لا ننتخب لأداء العلاج INTERCEPT. بدلا من ذلك، قد يختار مراكز الدم الأخرى لعلاج PC مع مجموعة جرعة واحدة تجهيز INTERCEPT (أي مجموعة الحجم الكبير) وتخزين PC في جرعة واحدة أكبر العلاجية، وبالتالي تعطيل أداء الممرض على جميع الوحدات. من أجل ضمان نلتقي لدينامتطلبات QC والممرض. تعطيل والعديد من وحداتنا الصفائح الدموية ممكن، نختار المعاطف بافي استنادا إلى التهم المانحة الصفائح الدموية، والتي عند تجميعها، سيؤدي إلى محتوى أدنى من الصفائح الدموية الصفائح الدموية 5.6x10 11 في حوض السباحة وهذا يضمن محتوى الصفائح الدموية كافية لإنتاج جرعتين العلاجية بعد العلاج INTERCEPT. التحقق من صحة يدل على أن لدينا يمكن بكامل هيئته 7 معطف الشهباء الدم المستمدة حدات مجمعة بنجاح وتعامل مع عملية INTERCEPT لالصفائح الدموية، مما يؤدي إلى 2 المنتجات الممرض الصفائح الدموية المعطل التي تلبي معايير القبول لتصنيع (متطلبات السويدية والمبادئ التوجيهية الأوروبي) ودعم المرضى تتطلب عمليات نقل الصفائح الدموية وفقا لإرشادات الممارسة السريرية ومستوى أساليب التسريب الصفائح الدموية في السويد.

<table border="1">
	<tbody>
		<tr>
			<td>Material / Equipment</td>
			<td>Company</td>
			<td>Catalogue Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>Whole blood donation, primary separation, and platelet production</td>
		</tr>
		<tr>
			<td>Blood collection pack</td>
			<td>Fenwal</td>
			<td>R6485</td>
			<td>Top/Bottom set</td>
		</tr>
		<tr>
			<td>Automated component extractor</td>
			<td>Fenwal</td>
			<td>Optipress-II</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Blood mixer and balance system</td>
			<td>Baxter</td>
			<td>Easymix V3</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Platelet leukocyte filtration set</td>
			<td>Fenwal</td>
			<td>K4R7042</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Centrifuge</td>
			<td>Hettich</td>
			<td>Roto Silenta 63 RS</td>
			<td>Version 5.5</td>
		</tr>
		<tr>
			<td>Platelet additive solution - SSP+</td>
			<td>MacoPharma</td>
			<td>SSP2030U</td>
			<td>300 ml</td>
		</tr>
		<tr>
			<td>Sterile tubing welder</td>
			<td>Terumo</td>
			<td>T-SCD</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>INTERCEPT treatment &amp; storage</td>
		</tr>
		<tr>
			<td>INTERCEPT processing set</td>
			<td>Cerus</td>
			<td>INT2503</td>
			<td>Dual Storage (DS) set</td>
		</tr>
		<tr>
			<td>INTERCEPT Illuminator</td>
			<td>Cerus</td>
			<td>INT100</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>PC sample pack</td>
			<td>Fenwal</td>
			<td>FTX 1122</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Incubator</td>
			<td>Helmer</td>
			<td>PC2200/PC3200</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Agitator</td>
			<td>Helmer</td>
			<td>PF48H/PF96H</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>Evaluation of in vitro Platelet Function</td>
		</tr>
		<tr>
			<td>Blood gas analyzer</td>
			<td>Radiometer</td>
			<td>ABL 735</td>
			<td>Used for pH, blood gases, and lactate measurement</td>
		</tr>
		<tr>
			<td>Chemistry system</td>
			<td>Ortho Clinical Diagnostic</td>
			<td>Vitros 5.1</td>
			<td>Used for glucose measurement</td>
		</tr>
		<tr>
			<td>Hematology analyzer</td>
			<td>Boule Medical AB</td>
			<td>Medonic CA620-Cellguard</td>
			<td>Used for platelet count measurement</td>
		</tr>
		<tr>
			<td>Flow cytometer</td>
			<td>BD</td>
			<td>FACSCanto</td>
			<td>Used for white blood cell measurement</td>
		</tr>
	</tbody>
</table>

preparation and pathogen inactivation of double dose buffy coat platelet products using the intercept blood system

وتواجه مراكز الدم مع العديد من التحديات بما في ذلك إنتاج محصول تعظيم المنتج من التبرعات بالدم التي يتلقونها وكذلك ضمان أعلى مستوى ممكن من السلامة للمرضى نقل الدم، بما في ذلك الحماية من الأمراض المنقولة نقل. يجب أن يتحقق هذا بطريقة مسؤولة من الناحية المالية مما يقلل نفقات التشغيل بما في ذلك المواد الاستهلاكية والمعدات، والنفايات، وتكاليف الموظفين، وغيرها. عدة طرق متاحة لإنتاج الصفائح الدموية لنقل مركزات. واحدة من الاكثر شيوعا هو الأسلوب معطف الشهباء التي أعدت وحدة واحدة الصفائح الدموية العلاجية (≥ 2،0 X10 11 الصفائح الدموية لكل وحدة أو لكل اللوائح المحلية) عن طريق تجميع طبقة الغلاف الأبيض من التبرعات تصل إلى ستة الدم الكامل. إلا في الآونة الأخيرة تم إجراء لإنتاج &quot;المزدوجة جرعة&quot; الصفائح الدموية بأكملها مشتقة المتقدمة. المقدمة هنا هي طريقة جديدة لإعدادجرعة مزدوجة الصفائح الدموية في الدم كله المستمدة من مركزات برك من المعاطف بافي 7 و علاج لاحقا وحدات جرعة مزدوجة مع نظام الدم INTERCEPT لتعطيل الممرض. وقد وضعت INTERCEPT لتعطيل الفيروسات والبكتيريا، والطفيليات، والخلايا البيضاء التي تلوث المانحة قد تكون موجودة في الدم المتبرع به. الإقران مع أسلوب اعتراض جرعة مضاعفة معطف الشهباء من خلال الاستفادة من معالجة INTERCEPT مع مجموعة حاويات التخزين المزدوجة (&quot;مجموعة DS&quot;)، ويتيح مراكز الدم لعلاج كل من وحداتهم جرعة مضاعفة في واحد الممرض تعطيل مجموعة المعالجة، وبالتالي تحقيق أقصى قدر من سلامة المرضى مع تقليل التكاليف. ضعف معطف الشهباء الأسلوب يتطلب جرعة أقل المعاطف بافي ويقلل من استخدام المواد الاستهلاكية بنسبة تصل إلى 50٪ (على سبيل المثال مجموعات تجمع، ومجموعات التصفية، الصفائح الدموية حل المضافة، ورقائق اتصال العقيمة) مقابل إعداد ومعالجة وحدات جرعة واحدة معطف الشهباء الصفائح الدموية . وفورات في التكاليف الأخرى تشمل نفايات أقل، أقل maintenan المعداتم، ومتطلبات خفض استهلاك الطاقة، وانخفاض أفراد الوقت، وبتكلفة أقل مقارنة مع مجموعة تقنية فصل مكونات الدم.

Watch this Scientific Journal Video about Preparation and Pathogen Inactivation of Double Dose Buffy Coat Platelet Products using the INTERCEPT Blood System at JoVE.com

Preparation and Pathogen Inactivation of Double Dose Buffy Coat Platelet Products using the INTERCEPT Blood System

توضح هذه المقالة العملية التي يستخدمها مستشفى جامعة أوريبرو لإنتاج الصفائح الدموية مزدوجة معطف الشهباء جرعة يركز المحضرة من التبرعات بالدم الكامل وتعامل مع نظام الدم INTERCEPT لتعطيل الممرض. ال<em&gt; في المختبر</emيتم تقييم&gt; جودة الوحدات النهائية على الصفائح الدموية 7 أيام من التخزين.

إعداد والتعطيل من العوامل الممرضة جرعة مزدوجة منتجات صفائح الدم معطف بافي باستخدام نظام الدم INTERCEPT

Blood centers are faced with many challenges including maximizing production yield from the blood product donations they receive as well as ensuring the ...

preparation-pathogen-inactivation-double-dose-buffy-coat-platelet

Research

JoVE Journal

Medicine

28.5K Views.  Örebro University Hospital. توضح هذه المقالة العملية التي يستخدمها مستشفى جامعة أوريبرو لإنتاج الصفائح الدموية مزدوجة معطف الشهباء جرعة يركز المحضرة من التبرعات بالدم الكامل وتعامل مع نظام الدم INTERCEPT لتعطيل الممرض. ال في المختبر جودة الوحدات النهائية على الصفائح الدموي?...

Video: Preparation and Pathogen Inactivation of Double Dose Buffy Coat Platelet Products using the INTERCEPT Blood System

Minimal Erythema Dose (MED) Testing

Ultraviolet radiation (UV) therapy is sometimes used as a treatment for various common skin conditions, including psoriasis, acne, and eczema. The dosage of UV light is prescribed according to an individual's skin sensitivity. Thus, to establish the proper dosage of UV light to administer to a patient, the patient is sometimes screened to determine a minimal erythema dose (MED), which is the amount of UV radiation that will produce minimal erythema (sunburn or redness caused by engorgement of capillaries) of an individual's skin within a few hours following exposure. This article describes how to conduct minimal erythema dose (MED) testing. There is currently no easy way to determine an appropriate UV dose for clinical or research purposes without conducting formal MED testing, requiring observation hours after testing, or informal trial and error testing with the risks of under- or over-dosing. However, some alternative methods are discussed.

Minimal Erythema Dose MED Testing

This article describes how to conduct minimal erythema dose (MED) testing in order to determine the lowest dose of ultraviolet radiation that will cause erythema (burning) when administered to an individual.

الجرعة الحمامي الحد الأدنى (MED) اختبار

Ultraviolet radiation (UV) therapy is  sometimes used as a treatment for various common skin conditions, including  psoriasis, acne, and eczema. The ...

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow (ischemia) 1. Middle cerebral artery (MCA) delivers a great majority of blood to the lateral surface of the cortex 2, is the most common site of human stroke 3, and ischemia within its territory can result in extensive dysfunction or death 1,4,5. Survivors of ischemic stroke often suffer loss or disruption of motor capabilities, sensory deficits, and infarct. In an effort to capture these key characteristics of stroke, and thereby develop effective treatment, a great deal of emphasis is placed upon animal models of ischemia in MCA.
Here we present a method of permanently occluding a cortical surface blood vessel. We will present this method using an example of a relevant vessel occlusion that models the most common type, location, and outcome of human stroke, permanent middle cerebral artery occlusion (pMCAO). In this model, we surgically expose MCA in the adult rat and subsequently occlude via double ligature and transection of the vessel. This pMCAO blocks the proximal cortical branch of MCA, causing ischemia in all of MCA cortical territory, a large portion of the cortex. This method of occlusion can also be used to occlude more distal portions of cortical vessels in order to achieve more focal ischemia targeting a smaller region of cortex. The primary disadvantages of pMCAO are that the surgical procedure is somewhat invasive as a small craniotomy is required to access MCA, though this results in minimal tissue damage. The primary advantages of this model, however, are: the site of occlusion is well defined, the degree of blood flow reduction is consistent, functional and neurological impairment occurs rapidly, infarct size is consistent, and the high rate of survival allows for long-term chronic assessment.

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

We describe a highly reproducible method for the permanent occlusion of a rodent major cerebral blood vessel. This technique can be accomplished with very little peripheral damage, minimal blood loss, a high rate of long-term survival, and consistent infarct volume commensurate with the human clinical population.

الدائم انسداد الأوعية الدماغية<em&gt; عبر</em&gt; ضمد مزدوجة وعانى من قطع

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow ...

Drug Repurposing Hypothesis Generation Using the "RE:fine Drugs" System

The promise of drug repurposing is that existing drugs may be used for new disease indications in order to curb the high costs and time for approval. The goal of computational methods for drug repurposing is to enable solutions for safer, cheaper and faster drug discovery. Towards this end, we developed a novel method that integrates genetic and clinical phenotype data from large-scale GWAS and PheWAS studies with detailed drug information on the concept of transitive Drug-Gene-Disease triads. We created "RE:fine Drugs," a freely available, interactive dashboard that automates gene, disease and drug-based searches to identify drug repurposing candidates. This web-based tool supports a user-friendly interface that includes an array of advanced search and export options. Results can be prioritized in a variety of ways, including but not limited to, biomedical literature support, strength and statistical significance of GWAS and/or PheWAS associations, disease indications and molecular drug targets. Here we provide a protocol that illustrates the functionalities available in the "RE:fine Drugs" system and explores the different advanced options through a case study.

Drug Repurposing Hypothesis Generation Using the &quot;RE:fine Drugs&quot; System

Here we describe a protocol using the web-based drug repurposing hypothesis generation tool: "RE:fine Drugs." This protocol can be modified to a user's preferences at the level of the query type (gene, drug or disease) and/or the range of available advanced options.

المخدرات تطويعها لأغراض أخرى فرضية الجيل باستخدام &quot;RE: المخدرات بخير&quot; النظام

The promise of drug repurposing is that existing drugs may be used for new disease indications in order to curb the high costs and time for approval. The ...

Turbidimetry on Human Washed Platelets: The Effect of the Pannexin1-inhibitor Brilliant Blue FCF on Collagen-induced Aggregation

Turbidimetry is a laboratory technique that is applied to measure the aggregation of platelets suspended in either plasma (platelet-rich plasma, PRP) or in buffer (washed platelets), by the use of one or a combination of agonists. The use of washed platelets separated from their plasma environment and in the absence of anticoagulants allows for studying intrinsic platelet properties. Among the large panel of agonists, arachidonic acid (AA), adenosine di-phosphate (ADP), thrombin and collagen are the most frequently used. The aggregation response is quantified by measuring the relative optical density (OD) over time of platelet suspension under continuous stirring. Platelets in homogeneous suspension limit the passage of light after the addition of an agonist, platelet shape change occurs producing a small transitory increase in OD. Following this initial activation step, platelet clots form gradually, allowing the passage of light through the suspension as a result of decreased OD. The aggregation process is ultimately expressed as a percentage, compared to the OD of platelet-poor plasma or buffer. Rigorous calibration is thus essential at the beginning of each experiment. As a general rule: calibration to 0% is set by measuring the OD of a non-stimulated platelet suspension while measuring the OD of the suspension medium containing no platelets represents a value of 100%. Platelet aggregation is generally visualized as a real-time aggregation curve. Turbidimetry is one of the most commonly used laboratory techniques for the investigation of platelet function and is considered as the historical gold standard and used for the development of new pharmaceutical agents aimed at inhibiting platelet aggregation. Here, we describe detailed protocols for 1) preparation of human washed platelets and 2) turbidimetric analysis of collagen-induced aggregation of human washed platelets pretreated with the food dye Brilliant Blue FCF that was recently identified as an inhibitor of Pannexin1 (Panx1) channels.

We describe a straightforward method for the isolation of washed platelets from human blood followed by agonist-induced platelet aggregation measurements by turbidimetry. As an example we apply this method for studying the aggregation response of human platelets to collagen after a pre-incubation with the Pannexin1 channel inhibitor Brilliant Blue FCF.

قياس العكر غسلها لحقوق الإنسان الصفائح الدموية: تأثير من Pannexin1 المانع الزرقاء الرائعة FCF على تجميع الناجمة الكولاجين،

Turbidimetry is a laboratory technique that is applied to measure the aggregation of platelets suspended in either plasma (platelet-rich plasma, PRP) or ...

Routine Screening Method for Microparticles in Platelet Transfusions

Platelet inventory management based on screening microparticle content in platelet concentrates is a new quality improvement initiative for hospital blood banks. Cells fragment off microparticles (MP) when they are stressed. Blood and blood components may contain cellular fragments from a variety of cells, most notably from activated platelets. When performing their roles as innate immune cells and major players in coagulation and hemostasis, platelets change shape and generate microparticles. With dynamic light scattering (DLS)-based microparticle detection, it is possible to differentiate activated (high microparticle) from non-activated (low microparticle) platelets in transfusions, and optimize the use of this scarce blood product. Previous research suggests that providing non-activated platelets for prophylactic use in hematology-oncology patients could reduce their risk of becoming refractory and improve patient care. The goal of this screening method is to routinely differentiate activated from non-activated platelets. The method described here outlines the steps to be performed for routine platelet inventory management in a hospital blood bank: obtaining a sample from a platelet transfusion, loading the sample into the capillary for DLS measurement, performing the DLS test to identify microparticles, and using the reported microparticle content to identify activated platelets.

Platelet inventory management based on screening microparticle content in platelet concentrates is a new quality improvement initiative in hospital blood banks. The goal is to differentiate activated from non-activated platelets to optimize the use of platelets. Providing non-activated platelets to hematology-oncology patients might reduce their high risk to become refractory.

روتين فحص الطريقة المجهرية الدقيقة في عمليات نقل الدم الصفيحات

Platelet inventory management based on screening microparticle content in platelet concentrates is a new quality improvement initiative for hospital blood ...

Preparation, Procedures and Evaluation of Platelet-Rich Plasma Injection in the Treatment of Knee Osteoarthritis

Knee osteoarthritis (KOA) is one of the most frequently encountered diseases in the orthopedic department. Existing non-surgical treatments have a limited effect on the repair of cartilage and on bone regeneration. Platelet-rich plasma (PRP) is an autologous bioactive substance that can repair cartilage injury and accelerate bone regeneration effectively. However, reporting of PRP preparation protocols in clinical studies is highly inconsistent, with the majority of studies providing insufficient information to allow the protocol to be reproduced. We describe a repeatable method of preparing PRP visually, the treatment of KOA using PRP intra-articular injection, and methods of evaluating the outcome. PRP was prepared using manual double centrifugation. The PRP layer was extracted from peripheral blood and used for knee joint cavity injection. Evaluations included assessments of blood platelet concentrations and clinical outcomes. Preparation of PRP by manual centrifugation requires less apparatus and is less costly than plasma filtration or centrifugation using equipment. The centrifugation time of our double centrifugation method was 6 and 5 minutes for the respective centrifugations at forces of 800 and 1400 x g, respectively, to allow for the consistent preparation of standardized PRP. However, a manual method is susceptible to operator error, and PRP batch preparation is not available. Intra-articular injection of PRP proved to be an effective treatment for knee osteoarthritis. The entire treatment procedure took less than 30 minutes, the blood platelet concentration of PRP could be standardized, and treatment was proven to be effective when evaluated by follow-up.

Knee osteoarthritis is frequently seen in the orthopedic department. We introduce in detail the entire knee osteoarthritis treatment process with platelet-rich plasma injection, including preparation, procedures, and evaluation.

إعداد الإجراءات وتقييم لحقن البلازما الغنية بالصفائح الدموية في علاج هشاشة العظام في الركبة

Knee osteoarthritis (KOA) is one of the most frequently encountered diseases in the orthopedic department. Existing non-surgical treatments have a limited ...

An In Vivo Method for Evaluating the Gut-Blood Barrier and Liver Metabolism of Microbiota Products

The gut-blood barrier (GBB) controls the passage of nutrients, bacterial metabolites and drugs from intestinal lumen to the bloodstream. The GBB integrity is disturbed in gastrointestinal, cardiovascular and metabolic diseases, which may result in easier access of biologically active compounds, such as gut bacterial metabolites, to the bloodstream. Thus, the permeability of the GBB may be a marker of both intestinal and extraintestinal diseases. Furthermore, the increased penetration of bacterial metabolites may affect the functioning of the entire organism.
Commonly used methods for studying the GBB permeability are performed ex vivo. The accuracy of those methods is limited, because the functioning of the GBB depends on intestinal blood flow. On the other hand, commonly used in vivo methods may be biased by liver and kidney performance, as those methods are based on evaluation of urine or/and peripheral blood concentrations of exogenous markers. Here, we present a direct measurement of GBB permeability in rats using an in vivo method based on portal blood sampling, which preserves intestinal blood flow and is virtually not affected by the liver and kidney function. 
Polyurethane catheters are inserted into the portal vein and inferior vena cava just above the hepatic veins confluence. Blood is sampled at baseline and after administration of a selected marker into a desired part of the gastrointestinal tract. Here, we present several applications of the method including (1) evaluation of the colon permeability to TMA, a gut bacterial metabolite, (2) evaluation of liver clearance of TMA, and (3) evaluation of a gut-portal blood-liver-peripheral blood pathway of gut bacteria-derived short-chain fatty acids. Furthermore, the protocol may also be used for tracking intestinal absorption and liver metabolism of drugs or for measurements of portal blood pressure.

An In Vivo Method for Evaluating the Gut-Blood Barrier and Liver Metabolism of Microbiota Products

The access of nutrients, microbiota metabolites and medicines to the circulation is controlled by the gut-blood barrier (GBB). We describe a direct method for measuring the GBB permeability in vivo, which, in contrast to commonly used indirect methods, is virtually not affected by liver and kidney functions.

أسلوب في فيفو لتقييم حاجز القناة الهضمية-الدم والايض الكبد من منتجات الحجمية

The gut-blood barrier (GBB) controls the passage of nutrients, bacterial metabolites and drugs from intestinal lumen to the bloodstream. The GBB integrity ...

Platelet-based Detection of Nitric Oxide in Blood by Measuring VASP Phosphorylation

Platelets are the blood components responsible for proper blood clotting. Their function is highly regulated by various pathways. One of the most potent vasoactive agents, nitric oxide (NO), can also act as a powerful inhibitor of platelet aggregation. Direct NO detection in blood is very challenging due to its high reactivity with cell-free hemoglobin that limits NO half-life to the millisecond range. Currently, NO changes after interventions are only estimated based on measured changes of nitrite and nitrate (members of the nitrate-nitrite-NO metabolic pathway). However precise, these measurements are rather difficult to interpret vis a vis actual NO changes, due to the naturally high baseline nitrite and nitrate levels that are several orders of magnitude higher than expected changes of NO itself. Therefore, the development of direct and simple methods that would allow one to detect NO directly is long overdue. This protocol addresses a potential use of platelets as a highly sensitive NO sensor in blood. It describes initial platelet rich plasma (PRP) and washed platelet preparations and the use of nitrite and deoxygenated red blood cells as NO generators. Phosphorylation of VASP at serine 239 (P-VASPSer239) is used to detect the presence of NO. The fact that VASP protein is highly expressed in platelets and that it is rapidly phosphorylated when NO is present leads to a unique opportunity to use this pathway to directly detect NO presence in blood.

Here, we present a protocol to address the potential use of platelets as a highly sensitive nitric oxide sensor in blood. It describes initial platelet preparation and the use of nitrite and deoxygenated red blood cells as nitric oxide generators.

الكشف على أساس الصفائح الدموية من أكسيد النيتريك في الدم عن طريق قياس الفسفرة بسب

Platelets are the blood components responsible for proper blood clotting. Their function is highly regulated by various pathways. One of the most potent ...

Measuring the Effects of Bacteria and Chemicals on the Intestinal Permeability of Caenorhabditis elegans

In living organisms, intestinal hyperpermeability is a serious symptom that leads to many inflammatory bowel diseases (IBDs). Caenorhabditis elegans is a nonmammalian animal model that is widely used as an assay system due to its short lifespan, transparency, cost-effectiveness, and lack of animal ethics issues. In this study, a method was developed to investigate the effects of different bacteria and 3,3'-diindolylmethane (DIM) on the intestinal permeability of C. elegans with a high-throughput image analysis system. The worms were infected with different gut bacteria or cotreated with DIM for 48 h and fed with fluorescein isothiocyanate (FITC)-dextran overnight. Then, the intestinal permeability was examined by comparing the fluorescence images and the fluorescence intensity inside the worm bodies. This method may also have the potential to identify probiotic and pathogenic intestinal bacteria that affect intestinal permeability in the animal model and is effective for examining the effects of harmful or health-promoting chemicals on intestinal permeability and intestinal health. However, this protocol also has some considerable limitations at the genetic level, especially for determining which genes are altered to control illness, because this method is mostly used for phenotypic determination. In addition, this method is limited to determining exactly which pathogenic substrates cause inflammation or increase the permeability of the worms' intestines during infection. Therefore, further in-depth studies, including investigation of the molecular genetic mechanism using mutant bacteria and nematodes as well as chemical component analysis of bacteria, are required to fully evaluate the function of bacteria and chemicals in determining intestinal permeability.

Measuring the Effects of Bacteria and Chemicals on the Intestinal Permeability of Caenorhabditis elegans

This protocol describes how to measure intestinal permeability of Caenorhabditis elegans. This method is helpful for basic biological research on intestinal health related to the interaction between intestinal bacteria and their host and for screening to identify probiotic and chemical agents to cure leaky gut syndrome and inflammatory bowel diseases.

قياس اثار البكتيريا والمواد الكيميائية علي نفاذيه المعوية من اليجان

In living organisms, intestinal hyperpermeability is a serious symptom that leads to many inflammatory bowel diseases (IBDs). Caenorhabditis elegans is a ...

Laminectomy for the Removal of Thoracic Ossification of the Ligamentum Flavum (TOLF) Using Ultrasonic and Conventional Osteotomes

Thoracic ossification of the ligamentum flavum (TOLF) is a common cause of progressive thoracic myelopathy. TOLF is typically treated with surgical decompression. A variety of surgical techniques, including laminoplasty, laminectomy, and lamina fenestration, are used for the effective treatment of TOLF. However, traditional methods are associated with a substantial risk of perioperative complications, including dural laceration and/or iatrogenic spinal cord injury. Therefore, it is important to develop an efficient and secure surgical technique for TOLF. Herein, we describe a method for laminectomyperformed at the thoracic spine using an ultrasonic osteotome combined with a conventional osteotome. This technique can reduce intraoperative complications. This is a relatively safe and easy-to-learn method that should be recommended for the treatment of TOLF.

Laminectomy for the Removal of Thoracic Ossification of the Ligamentum Flavum TOLF Using Ultrasonic and Conventional Osteotomes

We present a methodology for using an ultrasonic osteotome combined with a conventional osteotome for laminectomy in thoracic ossification of the ligamentum flavum. This is a relatively safe and easy-to-learn method that avoids the perioperative complications associated with the conventional method.

استئصال الصفيحة الفقرية لإزالة التعظم الصدري للرباط الفلافوم (TOLF) باستخدام عظم العظم بالموجات فوق الصوتية والتقليدية

Thoracic ossification of the ligamentum flavum (TOLF) is a common cause of progressive thoracic myelopathy. TOLF is typically treated with surgical ...