نشكر الأستاذ JP شباتز (قسم كيمياء البيوفيزيائية في جامعة هايدلبرغ وإدارة المواد الجديدة والنظم البيولوجية، ومعهد ماكس بلانك للأنظمة الذكية، ومقرها شتوتغارت) لدعمه النوع. كما اعترف الدعم المالي من ماكس بلانك غزلشافت وجمعية الألمانية للبحوث (DFG SFB/TR79 لEAC-A.) إلى حد كبير.

1. تركيب 11-Mercaptoundecanoyl-N-hydroxysuccinimide استر (MU-NHS) <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> إضافة قطرة قطرة محلول من 500 ملغم N-hydroxysuccinimide و 30 ملغ 4 - (dimethylamino) البيريدين في 10 مل الأسيتون (السلطة الفلسطينية) إلى 1 ز حامض 11-mercaptoundecanoic في 40 مل ثنائي كلورو ميثان (السلطة الفلسطينية) في درجة حرارة الغرفة (RT). </li><li style=";text-align:right;direction:rtl"> تهدئة رد الفعل إلى 0 درجة مئوية وإضافة قطرة قطرة 1.1 ز ن، ن &#39;dicyclohexylcarbodiimide في 10 مل ثنائي كلورو ميثان (تحت جو من النيتروجين). الحفاظ على التفاعل في درجة حرارة منخفضة لمدة 1 ساعة ثم يقلب في RT بين عشية وضحاها. </li><li style=";text-align:right;direction:rtl"> تحديد راسب وجففه تحت ضغط منخفض. تنقية المنتج من قبل فلاش اللوني مع البنزين البترول وخلات الإيثيل (سنويا) في نسبة 1:1. </li></ol> 2. إعداد طبقات متجانسة الذهب <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> coverslips الزجاج نظيفة مع مناديل الدقة ويصوتن لهم لمدة 5 دقائق في محلول يحتوي على 1:1 خليط من خلات الإيثيل (السلطة الفلسطينية) والميثانول (السلطة الفلسطينية). Rinsركائز الإلكترونية مع الميثانول ويجف حالا في ظل تدفق النيتروجين. </li><li style=";text-align:right;direction:rtl"> وضع ركائز نظيفة في الجهاز تفل الطلاء. إخلاء القاعة. إزالة طبقة أكسيد الكروم العلوي من الكروم الهدف الاخرق من قبل لمدة 60 ثانية. وضع العينات تحت شعاع المعادن ومعطف لهم مع طبقة الكروم 10 نانومتر، تلتها طلاء بطبقة من الذهب 40 نانومتر. </li></ol> 3. تجميد سطح BMP-2 <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> حل MU-NHS في N، N-dimethlyformamide (DMF) إلى تركيز النهائي من حوالي 1 ملم واحتضان ركائز الذهب في حل MU-NHS في RT لمدة 4 ساعة تحت جو النيتروجين. </li><li style=";text-align:right;direction:rtl"> السطوح يصوتن في DMF لمدة 2 دقيقة، وشطف مع DMF وMeOH وتجف حالا في ظل تدفق النيتروجين. </li><li style=";text-align:right;direction:rtl"> إعداد محلول المخزون من rhBMP-2 في العقيمة 4 ملم حمض الهيدروكلوريك بتركيز 100 ميكروغرام / مل (مأخوذة تخزينها في -80 درجة مئوية). لالتخفيفات العمل (3.5 ميكروغرام / مل)، وتمييع الحل الأسهم في برنامج تلفزيوني / كلوريد الصوديوم (PBS تحتوي على 1 M كلوريد الصوديوم) وdjust لدرجة الحموضة 8.5 مباشرة قبل الاستعمال. </li><li style=";text-align:right;direction:rtl"> احتضان السطوح بين functionalized MU-NHS في حل العاملة rhBMP-2 في 4 درجات مئوية خلال الليل. إزالة طاف الحضانة. السطوح يصوتن في برنامج تلفزيوني / كلوريد الصوديوم لمدة 2 دقيقة ويغسل 3X مع برنامج تلفزيوني العقيمة / كلوريد الصوديوم. </li></ol> 4. توصيف السطح <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> لمنع امتصاص غير محدد من الأجسام المضادة، واحتضان الأسطح مع BSA 5٪ (ث / ت) في حل برنامج تلفزيوني لمدة 1 ساعة على RT، تليها الحضانة مع الأضداد المضادة للBMP-2 (1:100 في 1٪ (ث / ت ) حل BSA / PBS) لمدة 1 ساعة على RT. غسل الأسطح مرتين مع برنامج تلفزيوني ويصوتن لمدة 30 ثانية. </li><li style=";text-align:right;direction:rtl"> احتضان ركائز مع الضد HRP، مترافق الثانوية (1:1،000 في 1٪ (ث / ت) حل BSA / PBS) لمدة 30 دقيقة في RT، ثم يغسل مرتين مع برنامج تلفزيوني. </li><li style=";text-align:right;direction:rtl"> استخدام Ampliflu الأحمر فحص لقياس HRP النشاط الأنزيمي في 570 نانومتر مع قارئ لوحة. </li></ol> 5. تحليل النشاط الحيوي <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> البذور 1 × 10 5 الماوس myoblasts C2C12 لكل بئر في 6 جيدا ررآتش في النمو المتوسط، وتتألف من DMEM الجلوكوز المرتفعة التي تحتوي على البيروفات تستكمل مع FBS 10٪، 1٪ البنسلين / الستربتوميسين واحتضان لهم لمدة 24 ساعة عند 37 درجة مئوية / 5٪ CO 2. </li><li style=";text-align:right;direction:rtl"> تجويع الخلايا في المصل خالية DMEM لمدة 3-5 ساعة قبل البذر على أسطح مزينة يجمد BMP-2. </li><li style=";text-align:right;direction:rtl"> للتحقيق في المدى القصير يشير الاستقراء، استبدال المتوسطة بنسبة 200 ميكرولتر الطازجة DMEM المصل خالية ووضع يجمد BMP-2 فوق أسطح الخلايا. وينبغي التعامل مع السطح مع ملاقط غيض غرامة لتجنب خدش. </li><li style=";text-align:right;direction:rtl"> إزالة بلطف الأسطح ونضح المتوسطة مع ماصة، وغسل الخلايا مرتين مع برنامج تلفزيوني. الشروع في تحليل استجابات الخلايا. </li><li style=";text-align:right;direction:rtl"> لتحليل النشاط البيولوجي على المدى الطويل، وخلايا لوحة على السطوح والثقافة لهم عند 37 درجة مئوية / 5٪ CO 2 في ظروف المصل منخفضة (2٪ FBS) لمدة ستة أيام. </li></ol>

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في هذا البروتوكول وصفنا إعداد الأسطح النشطة بيولوجيا بين functionalized مع rhBMP-2. ويتألف هذا النهج خطوتين: 1) التشكيل الأولي للأحادي الطبقة الذاتي تجميع (SAM) من رابط bifunctional على سطح الذهب، 2) تجميد التساهمية من البروتين rhBMP-2. في الأعمال السابقة، ونحن التحقق من صحة الربط الفعال لل...

عظم بروتين مخلق 2 (BMP-2) هو عضو عامل النمو التحويلي (TGF-β) الأسرة وبمثابة محفز للدي نوفو تكوين العظام وكذلك تنظيم العديد من الأنسجة أثناء التطور الجنيني والكبار التوازن 1-3. كل مونومر من النشطة بيولوجيا homodimeric البروتين BMP-2 يحتوي على &quot;عقدة السيستين&quot; عزر، والتي يتم حفظها للغاية في جميع أفضل الممارسات الإدارية 4. ستة من بقايا السيستين شكل سندات ثاني كبريتيد سبعة داخل الجزيء أن استقرار كل مونومر، في حين تشارك السيستين السابع في dimerization، تشكيل السندات الجزيئات بين اثنين من مونومرات 5،6. يحدد هذا عقدة السيستين حفظا للغاية هيكل ثلاثي الأبعاد للبروتين BMP-2 ويحدد خصائص فريدة من نوعها، مثل مقاومة ضد الحرارة، وdenaturants الحمضية درجة الحموضة 7-9. BMP-2 بربط سيرين / ثريونين كيناز مستقبلات عبر الغشاء، مما يحفز نقل الإشارة <sup&gt; 10-12. اعتمادا على طريقة مستقبلات oligomerization، يتم تنشيط مسارات إشارات مختلفة: شلال يشير صماد مستقلة يؤدي إلى الفوسفاتيز القلوية تحريض عبر P38 إشارات، في حين أن المسار التي تعتمد على صماد تفعيلها من خلال النتائج مستقبلات الفسفرة في صماد إزفاء النووية المعقدة وتفعيل النسخ من الجينات المستهدفة محددة، مثل المانع من التمايز (هوية) 12-14. في العظام، BMP-2 الحث على تمايز الخلايا الجذعية الوسيطة في بانيات، وبالتالي تحفيز الشفاء ودي نوفو تشكيل العظام. حاليا، أعرب recombinantly يتم تطبيق BMP-2 سريريا لتعزيز الشفاء من مواقع كسر. استراتيجية مشتركة في هندسة الأنسجة العظام هو استخدام عوامل النمو عن طريق الحقن، والتي هي أقل الغازية مقارنة مع أنظمة التسليم المحلي. ومع ذلك، فقد أظهرت الدراسات المجراة في والتطبيقات السريرية أن قصيرة العمر النصفي البيولوجي، في الموضع غير محددةالتخلف والتهميش والتخليص السريع للالمحلية BMP-2 قد يؤدي إلى العديد من المشاكل المحلية، خارج الرحم والنظامية 15. وبالتالي، للحصول على العرض الفعال، وانحباس أو تجميد BMP-2 داخل أو على مواد ضرورية للتسليم المحلي والمستدام لها في الموقع المستهدف. ويمكن تحقيق استدامة التسليم مع نهج الاستبقاء غير التساهمية، مثل فخ المادية، والامتزاز أو أيون complexation 16. ومع ذلك، فمن المعروف أن الامتزاز غير محدد من البروتينات على الأسطح يجوز النتائج في تمسخ من الجزيئات 17. لالتساهمية ملزم من عوامل النمو، وقد تم تطوير أنواع مختلفة من الدعم على مدى العقد الماضي. استخدام جزيئات ربط bifunctional التي تستهدف الأمينية أو الكربوكسيل مجموعات من البروتين على سبيل المثال، هو نوع واحد من النهج الذي لا يتطلب بالضرورة تعديل البروتين لتحقيق الشلل فيها. في الواقع، في حين أن التعديل البروتين يوفر ميزة السيطرة على التوجه البروتين،مقدمة من المجالات الصناعية، والعلامات وسلاسل الببتيد في مواقع محددة قد يغير النشاط البيولوجي من عوامل النمو 17. وبالتالي، للتحايل تمسخ بسبب التفاعل مع المواد الداعمة، السطوح يمكن بين functionalized مسبقا، على سبيل المثال، مع أحادي الطبقة الذاتي تجميعها (SAM) من جزيء ربط، تليها اقتران عامل المطلوب 18. وقد استخدمنا نهج قائم على SAM-لشل حركة تساهميا BMP-2 على سطح من خلال استهداف مخلفاتها أمين حرة وأظهرت أن بروتين يجمد يحتفظ كلا من النشاط البيولوجي القصيرة والطويلة الأجل و19. يوفر هذا البروتوكول طريقة بسيطة وفعالة لتقديم BMP-2 إلى الخلايا في المختبر لدراسات عن الآليات التي تحدث في غشاء الخلية وتنظيم الإشارات بين الخلايا المكونة للعظم مسؤولة عن إرسال الإشارات.

<table border="1">
	<tbody>
		<tr>
			<td>Name of Material/ Equipment</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments/Description</td>
		</tr>
		<tr>
			<td>N-hydroxysuccinimide</td>
			<td>Sigma-Aldrich</td>
			<td>130672</td>
			<td></td>
		</tr>
		<tr>
			<td>4-(dimethylamino)pyridin</td>
			<td>Sigma-Aldrich</td>
			<td>522805</td>
			<td></td>
		</tr>
		<tr>
			<td>Acetone</td>
			<td>AppliChem</td>
			<td>A2282</td>
			<td></td>
		</tr>
		<tr>
			<td>11-mercaptoundecanoic acid</td>
			<td>Sigma-Aldrich</td>
			<td>674427</td>
			<td></td>
		</tr>
		<tr>
			<td>Dichlormethane</td>
			<td>Merck</td>
			<td>106050</td>
			<td></td>
		</tr>
		<tr>
			<td>N,N&#39;-dicyclohexylcarbodiimide</td>
			<td>Sigma-Aldrich</td>
			<td>D80002</td>
			<td></td>
		</tr>
		<tr>
			<td>Petroleum benzene</td>
			<td>Merck</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Glass coverslips</td>
			<td>Carl Roth</td>
			<td>M 875</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethylacetate</td>
			<td>AppliChem</td>
			<td>A3550</td>
			<td></td>
		</tr>
		<tr>
			<td>Methanol</td>
			<td>Carl Roth</td>
			<td>4627</td>
			<td></td>
		</tr>
		<tr>
			<td>N,N-dimethylformamide</td>
			<td>Carl Roth</td>
			<td>T921</td>
			<td></td>
		</tr>
		<tr>
			<td>rhBMP-2</td>
			<td>R&#38;D Systems</td>
			<td>355-BM</td>
			<td>Carrier-free; expressed in E.coli</td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>PAA</td>
			<td>H15-002</td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>Carl Roth</td>
			<td>HN00.2</td>
			<td></td>
		</tr>
		<tr>
			<td>Poly(dimethyl siloxane) (PDMS)</td>
			<td>Dow Corning</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sylgard 184 silicone elastomer kit</td>
			<td>Dow Corning</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-rhBMP-2</td>
			<td>Sigma</td>
			<td>B9553</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat anti-mouse IgG-HRP</td>
			<td>Santa Cruz</td>
			<td>sc-2005</td>
			<td>Secondary antibody</td>
		</tr>
		<tr>
			<td>Ampliflu Red assay</td>
			<td>Sigma</td>
			<td>90101</td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbecco&#39;s Modified Eagle Medium (DMEM) (1x), liquid</td>
			<td>Gibco</td>
			<td>41966</td>
			<td>High glucose</td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum (FBS)</td>
			<td>Sigma</td>
			<td>F7524</td>
			<td>Sterile filtered, cell culture tested</td>
		</tr>
		<tr>
			<td>Pen/Strep</td>
			<td>Gibco</td>
			<td>15140</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypsin 0.05% (1x) with EDTA 4Na</td>
			<td>Gibco</td>
			<td>25300</td>
			<td></td>
		</tr>
		<tr>
			<td>Glycine (0.1 M)</td>
			<td>Riedel-de Ha&#235;n</td>
			<td>33226</td>
			<td></td>
		</tr>
		<tr>
			<td>IGEPAL CA-630 (1%)</td>
			<td>Sigma</td>
			<td>I8896</td>
			<td>Lysis buffer (ALP assay)19</td>
		</tr>
		<tr>
			<td>Magnesium chloride (MgCl2)(1 mM)</td>
			<td>Carl Roth</td>
			<td>HNO3.2</td>
			<td></td>
		</tr>
		<tr>
			<td>Zinc chloride (ZnCl2) (1 mM)</td>
			<td>Carl Roth</td>
			<td>3533.1</td>
			<td></td>
		</tr>
		<tr>
			<td>p-nitrophenylphosphate (pNPP)</td>
			<td>Sigma</td>
			<td>S0942</td>
			<td>Phosphatase substrate</td>
		</tr>
		<tr>
			<td>Anti-mysin heavy chain (MHC)</td>
			<td>Developmental Studies Hybridoma Bank, University of Iowa</td>
			<td>MF20</td>
			<td>Monoclonal antibody</td>
		</tr>
		<tr>
			<td>Alexa Fluor 488 Goat anti-mouse IgG</td>
			<td>Invitrogen</td>
			<td>A11001</td>
			<td></td>
		</tr>
		<tr>
			<td>DAPI</td>
			<td>Sigma</td>
			<td>D9542</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Equipment</td>
		</tr>
		<tr>
			<td>Ultrsonic bath (Sonorex Super RK 102H), Frequency 35 kHz</td>
			<td>BANDELIN electronic GmbH &#38; Co. KG</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>MED 020 Sputtercoating system</td>
			<td>BAL-TEC AG</td>
			<td></td>
			<td>Coating conditions 
			Cr: 120 mA, 1.3 x 10-2 mbar, 30 sec 
			Au: 60 mA, 5.0 x 10-2 mbar, 45 sec</td>
		</tr>
		<tr>
			<td>Tecan Infinite M200 Plate reader</td>
			<td>Tecan</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

covalent binding of bmp-2 on surfaces using a self-assembled monolayer approach

بروتين مخلق العظام 2 (BMP-2) هو عامل النمو جزءا لا يتجزأ من المصفوفة خارج الخلية من الأنسجة والعظام. BMP-2 بمثابة الزناد من تمايز الخلايا الوسيطة في بانيات، وبالتالي تحفيز الشفاء وتكوين العظام نوفو دي. وقد أثار استخدام السريرية البشرية المؤتلف BMP-2 (rhBMP-2) بالتزامن مع السقالات الخلافات الأخيرة، استنادا إلى طريقة العرض وكمية ليتم تسليمها. بروتوكول المعروضة هنا يوفر وسيلة بسيطة وفعالة لتقديم BMP-2 للدراسات في المختبر على خلايا. ونحن تصف كيفية تشكيل أحادي الطبقة الذاتي تجميعها يتكون من رابط heterobifunctional، وتظهر الخطوة اللاحقة ملزمة للحصول على تجميد التساهمية من rhBMP-2. مع هذا النهج فمن الممكن لتحقيق العرض المستمر من BMP-2، مع المحافظة على النشاط البيولوجي من البروتين. في الواقع، فإن تجميد سطح BMP-2 يسمح التحقيقات المستهدفة من خلال منع الإعلانات غير محددةorption، مع تقليل كمية عامل النمو، وعلى الأخص، مما يعوق جامحا من على سطح الأرض. كلا الحدثين الإشارات القصيرة والطويلة الأجل والناجمة عن BMP-2 تجري عندما تتعرض الخلايا على الأسطح تقديم يجمد تساهميا rhBMP-2، مما يجعل هذا النهج مناسب لفي المختبر دراسات عن مدى استجابة الخلية لBMP-2 التحفيز.

في الإعداد لدينا، وقد تم اختيار الذهب وسواغ لأنه يوفر نظام غير محددة بيولوجيا ولكن الانضباطي كيميائيا. وعلاوة على ذلك، فإن تطبيق الطبقات الوحيدة الذاتي تجميع يستلزم العديد من الفوائد: صواريخ سام كثف بعفوية عبر بهم &quot;وجها لمجموعات&quot; على المعادن والطبقات الوحيدة ش?...

Watch this Scientific Journal Video about Covalent Binding of BMP-2 on Surfaces Using a Self-assembled Monolayer Approach at JoVE.com

Covalent Binding of BMP-2 on Surfaces Using a Self-assembled Monolayer Approach

نحن تصف طريقة لإنجاز الشلل كفاءة BMP-2 على السطوح. ويستند نهجنا على تشكيل أحادي الطبقة الذاتي تجميعها لتحقيق التساهمية ملزم من BMP-2 عبر مخلفاتها أمين الحرة. هذا الأسلوب هو أداة مفيدة لدراسة الإشارات في غشاء الخلية.

التساهمية تجليد BMP-2 على السطوح باستخدام نهج أحادي الطبقة تجميعها الذاتي

Bone morphogenetic protein 2 (BMP-2) is a growth factor embedded in the extracellular matrix of bone tissue. BMP-2 acts as trigger of mesenchymal cell ...

covalent-binding-bmp-2-on-surfaces-using-self-assembled-monolayer

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Unit 1

Covalent Bonding and Structure

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13.8K Views.  University of Heidelberg.  نحن تصف طريقة لإنجاز الشلل كفاءة BMP-2 على السطوح. ويستند نهجنا على تشكيل أحادي الطبقة الذاتي تجميعها لتحقيق التساهمية ملزم من BMP-2 عبر مخلفاتها أمين الحرة. هذا الأسلوب هو أداة مفيدة لدراسة الإشارات في غشاء الخلية. 

Video: Covalent Binding of BMP-2 on Surfaces Using a Self-assembled Monolayer Approach

Stabilizing Hepatocellular Phenotype Using Optimized Synthetic Surfaces

Currently, one of the major limitations in cell biology is maintaining differentiated cell phenotype. Biological matrices are commonly used for culturing and maintaining primary and pluripotent stem cell derived hepatocytes. While biological matrices are useful, they permit short term culture of hepatocytes, limiting their widespread application. We have attempted to overcome the limitations using a synthetic polymer coating. Polymers represent one of the broadest classes of biomaterials and possess a wide range of mechanical, physical and chemical properties, which can be fine-tuned for purpose. Importantly, such materials can be scaled to quality assured standards and display batch-to-batch consistency. This is essential if cells are to be expanded for high through-put screening in the pharmaceutical testing industry or for cellular based therapy. Polyurethanes (PUs) are one group of materials that have shown promise in cell culture. Our recent progress in optimizing a polyurethane coated surface, for long-term culture of human hepatocytes displaying stable phenotype, is presented and discussed.

This article will focus on developing polymer coated surfaces for long-term, stable culture of stem cell derived human hepatocytes.

استقرار الكبدي النمط الظاهري عن طريق الأسطح الاصطناعية محسن

Currently, one of the major limitations in cell biology is maintaining differentiated cell phenotype. Biological matrices are commonly used for culturing ...

An Inverse Analysis Approach to the Characterization of Chemical Transport in Paints

The ability to directly characterize chemical transport and interactions that occur within a material (i.e., subsurface dynamics) is a vital component in understanding contaminant mass transport and the ability to decontaminate materials. If a material is contaminated, over time, the transport of highly toxic chemicals (such as chemical warfare agent species) out of the material can result in vapor exposure or transfer to the skin, which can result in percutaneous exposure to personnel who interact with the material. Due to the high toxicity of chemical warfare agents, the release of trace chemical quantities is of significant concern. Mapping subsurface concentration distribution and transport characteristics of absorbed agents enables exposure hazards to be assessed in untested conditions. Furthermore, these tools can be used to characterize subsurface reaction dynamics to ultimately design improved decontaminants or decontamination procedures. To achieve this goal, an inverse analysis mass transport modeling approach was developed that utilizes time-resolved mass spectroscopy measurements of vapor emission from contaminated paint coatings as the input parameter for calculation of subsurface concentration profiles. Details are provided on sample preparation, including contaminant and material handling, the application of mass spectrometry for the measurement of emitted contaminant vapor, and the implementation of inverse analysis using a physics-based diffusion model to determine transport properties of live chemical warfare agents including distilled mustard (HD) and the nerve agent VX.

In this paper, a procedure for quantifying the mass transport parameters of chemicals in various materials is presented. This process involves employing an inverse-analysis based diffusion model to vapor emission profiles recorded by real-time, mass spectrometry in high vacuum.

منهج تحليل معكوس لتوصيف النقل الكيميائية في مواد الطلاء

The ability to directly characterize chemical transport and interactions that occur within a material (i.e., subsurface dynamics) is a vital component in ...

Facile Preparation of Internally Self-assembled Lipid Particles Stabilized by Carbon Nanotubes

We present a facile method to prepare nanostructured lipid particles stabilized by carbon nanotubes (CNTs). Single-walled (pristine) and multi-walled (functionalized) CNTs are used as stabilizers to produce Pickering type oil-in-water (O/W) emulsions. Lipids namely, Dimodan U and Phytantriol are used as emulsifiers, which in excess water self-assemble into the bicontinuous cubic Pn3m phase. This highly viscous phase is fragmented into smaller particles using a probe ultrasonicator in presence of conventional surfactant stabilizers or CNTs as done here. Initially, the CNTs (powder form) are dispersed in water followed by further ultrasonication with the molten lipid to form the final emulsion. During this process the CNTs get coated with lipid molecules, which in turn are presumed to surround the lipid droplets to form a particulate emulsion that is stable for months. The average size of CNT-stabilized nanostructured lipid particles is in the submicron range, which compares well with the particles stabilized using conventional surfactants. Small angle X-ray scattering data confirms the retention of the original Pn3m cubic phase in the CNT-stabilized lipid dispersions as compared to the pure lipid phase (bulk state). Blue shift and lowering of the intensities in characteristic G and G&#39; bands of CNTs observed in Raman spectroscopy characterize the interaction between CNT surface and lipid molecules. These results suggest that the interactions between the CNTs and lipids are responsible for their mutual stabilization in aqueous solutions. As the concentrations of CNTs employed for stabilization are very low and lipid molecules are able to functionalize the CNTs, the toxicity of CNTs is expected to be insignificant while their biocompatibility is greatly enhanced. Hence the present approach finds a great potential in various biomedical applications, for instance, for developing hybrid nanocarrier systems for the delivery of multiple functional molecules as in combination therapy or polytherapy.

We report on a smart application of carbon nanotubes for kinetic stabilization of lipid particles that contain self-assembled nanostructures in their cores. The preparation of lipid particles requires rather low concentrations of carbon nanotubes permitting their use in biomedical applications such as drug delivery.

إعداد سطحي من تجميعها الذاتي داخليا الدهن الجسيمات استقرت بواسطة أنابيب الكربون النانوية

We present a facile method to prepare nanostructured lipid particles stabilized by carbon nanotubes (CNTs). Single-walled (pristine) and multi-walled ...

Highly Stable, Functional Hairy Nanoparticles and Biopolymers from Wood Fibers: Towards Sustainable Nanotechnology

Nanoparticles, as one of the key materials in nanotechnology and nanomedicine, have gained significant importance during the past decade. While metal-based nanoparticles are associated with synthetic and environmental hassles, cellulose introduces a green, sustainable alternative for nanoparticle synthesis. Here, we present the chemical synthesis and separation procedures to produce new classes of hairy nanoparticles (bearing both amorphous and crystalline regions) and biopolymers based on wood fibers. Through periodate oxidation of soft wood pulp, the glucose ring of cellulose is opened at the C2-C3 bond to form 2,3-dialdehyde groups. Further heating of the partially oxidized fibers (e.g., T = 80 &#176;C) results in three products, namely fibrous oxidized cellulose, sterically stabilized nanocrystalline cellulose (SNCC), and dissolved dialdehyde modified cellulose (DAMC), which are well separated by intermittent centrifugation and co-solvent addition. The partially oxidized fibers (without heating) were used as a highly reactive intermediate to react with chlorite for converting almost all aldehyde to carboxyl groups. Co-solvent precipitation and centrifugation resulted in electrosterically stabilized nanocrystalline cellulose (ENCC) and dicarboxylated cellulose (DCC). The aldehyde content of SNCC and consequently surface charge of ENCC (carboxyl content) were precisely controlled by controlling the periodate oxidation reaction time, resulting in highly stable nanoparticles bearing more than 7 mmol functional groups per gram of nanoparticles (e.g., as compared to conventional NCC bearing &#60;&#60; 1 mmol functional group/g). Atomic force microscopy (AFM), transmission electron microscopy (TEM), and scanning electron microscopy (SEM) attested to the rod-like morphology. Conductometric titration, Fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR), dynamic light scattering (DLS), electrokinetic-sonic-amplitude (ESA) and acoustic attenuation spectroscopy shed light on the superior properties of these nanomaterials.

Synthesis schemes to prepare highly stable wood fiber-based hairy nanoparticles and functional cellulose-based biopolymers have been detailed.

مستقرة للغاية، وظيفية مشعر النانوية والبوليمرات الحيوية من ألياف الخشب: نحو تقنية النانو المستدام

Nanoparticles, as one of the key materials in nanotechnology and nanomedicine, have gained significant importance during the past decade. While ...

Microfluidic-based Synthesis of Covalent Organic Frameworks (COFs): A Tool for Continuous Production of COF Fibers and Direct Printing on a Surface

Covalent Organic Frameworks (COFs) are a class of porous covalent materials which are frequently synthesized as unprocessable crystalline powders. The first COF was reported in 2005 with much effort centered on the establishment of new synthetic routes for its preparation. To date, most available synthetic methods for COF synthesis are based on bulk mixing under solvothermal conditions. Therefore, there is increasing interest in developing systematic protocols for COF synthesis that provide for fine control over reaction conditions and improve COF processability on surfaces, which is essential for their use in practical applications. Herein, we present a novel microfluidic-based method for COF synthesis where the reaction between two constituent building blocks, 1,3,5-benzenetricarbaldehyde (BTCA) and 1,3,5-tris(4-aminophenyl)benzene (TAPB), takes place under controlled diffusion conditions and at room temperature. Using such an approach yields sponge-like, crystalline fibers of a COF material, hereafter called MF-COF. The mechanical properties of MF-COF and the dynamic nature of the approach allow the continuous production of MF-COF fibers and their direct printing onto surfaces. The general method opens new potential applications requiring advanced printing of 2D or 3D COF structures on flexible or rigid surfaces.

Microfluidic-based Synthesis of Covalent Organic Frameworks COFs: A Tool for Continuous Production of COF Fibers and Direct Printing on a Surface

We present a novel microfluidic-based method for synthesis of covalent organic frameworks (COFs). We demonstrate how this approach can be used to produce continuous COF fibers, and also 2D or 3D COF structures on surfaces.

ميكروفلويديك التوليف القائم على الأطر العضوية التساهمية (كوفس): أداة للإنتاج المستمر للألياف كوف والطباعة المباشرة على سطح

Covalent Organic Frameworks (COFs) are a class of porous covalent materials which are frequently synthesized as unprocessable crystalline powders. The ...

Light-driven Molecular Motors on Surfaces for Single Molecular Imaging

The design and synthesis of a synthetic system that aims for the direct visualization of a synthetic rotary motor at the single molecule level on surfaces are demonstrated. This work requires careful design, considerable synthetic effort, and proper analysis. The rotary motion of the molecular motor in solution is shown by 1H NMR and UV-vis absorption spectroscopy techniques. In addition, the method to graft the motor onto an amine-coated quartz is described. This method helps to gain more insight into molecular machines.

The manuscript describes how to synthesize and graft a molecular motor on surfaces for single molecular imaging.

المحركات الجزيئية يحركها الضوء على الأسطح للتصوير الجزيئي واحد

The design and synthesis of a synthetic system that aims for the direct visualization of a synthetic rotary motor at the single molecule level on surfaces ...

Analysis of Complex Molecules and Their Reactions on Surfaces by Means of Cluster-Induced Desorption/Ionization Mass Spectrometry

Desorption/Ionization Induced by Neutral SO2 Clusters (DINeC) is employed as a very soft and efficient desorption/ionization technique for mass spectrometry (MS) of complex molecules and their reactions on surfaces. DINeC is based on a beam of SO2 clusters impacting on the sample surface at low cluster energy. During cluster-surface impact, some of the surface molecules are desorbed and ionized via dissolvation in the impacting cluster; as a result of this dissolvation-mediated desorption mechanism, low cluster energy is sufficient and the desorption process is extremely soft. Both surface adsorbates and molecules of which the surface is composed of can be analyzed. Clear and fragmentation-free spectra from complex molecules such as peptides and proteins are obtained. DINeC does not require any special sample preparation, in particular no matrix has to be applied. The method yields quantitative information on the composition of the samples; molecules at a surface coverage as low as 0.1 % of a monolayer can be detected. Surface reactions such as H/D exchange or thermal decomposition can be observed in real-time and the kinetics of the reactions can be deduced. Using a pulsed nozzle for cluster beam generation, DINeC can be efficiently combined with ion trap mass spectrometry. The matrix-free and soft nature of the DINeC process in combination with the MSn capabilities of the ion trap allows for very detailed and unambiguous analysis of the chemical composition of complex organic samples and organic adsorbates on surfaces.

Neutral SO2 clusters of low kinetic energy (&#60; 0.8 eV/constituent) are used to desorb complex surface molecules such as peptides or lipids for further analysis by means of mass spectrometry using an ion trap mass spectrometer. No special sample preparation is required, and real-time observation of reactions is possible.

تحليل الجزيئات المعقدة وردود أفعالها على الأسطح عن طريق الامتصاص/التأيّن الطيفي الكتلي الناجم عن الكتلة

Desorption/Ionization Induced by Neutral SO2 Clusters (DINeC) is employed as a very soft and efficient desorption/ionization technique for mass ...

Covalent Attachment of Single Molecules for AFM-based Force Spectroscopy

Atomic force microscopy (AFM)-based single molecule force spectroscopy is an ideal tool for investigating the interactions between a single polymer and surfaces. For a true single molecule experiment, covalent attachment of the probe molecule is essential because only then can hundreds of force-extension traces with one and the same single molecule be obtained. Many traces are in turn necessary to prove that a single molecule alone is probed. Additionally, passivation is crucial for preventing unwanted interactions between the single probe molecule and the AFM cantilever tip as well as between the AFM cantilever tip and the underlying surface. The functionalization protocol presented here is reliable and can easily be applied to a variety of polymers. Characteristic single molecule events (i.e., stretches and plateaus) are detected in the force-extension traces. From these events, physical parameters such as stretching force, desorption force and desorption length can be obtained. This is particularly important for the precise investigation of stimuli-responsive systems at the single molecule level. As exemplary systems poly(ethylene glycol) (PEG), poly(N-isopropylacrylamide) (PNiPAM) and polystyrene (PS) are stretched and desorbed from SiOx (for PEG and PNiPAM) and from hydrophobic self-assembled monolayer surfaces (for PS) in aqueous environment.

Covalent attachment of probe molecules to atomic force microscopy (AFM) cantilever tips is an essential technique for the investigation of their physical properties. This allows us to determine the stretching force, desorption force and length of polymers via AFM-based single molecule force spectroscopy with high reproducibility.

مرفق التكافؤ من جزيئات واحدة للتنظير الطيفي للقوة المستندة إلى AFM

Atomic force microscopy (AFM)-based single molecule force spectroscopy is an ideal tool for investigating the interactions between a single polymer and ...

Synthesis of pH Dependent Pyrazole, Imidazole, and Isoindolone Dipyrrinone Fluorophores using a Claisen-Schmidt Condensation Approach

Methine-bridged conjugated bicyclic aromatic compounds are common constituents of a range of biologically relevant molecules such as porphyrins, dipyrrinones, and pharmaceuticals. Additionally, restricted rotation of these systems often results in highly to moderately fluorescent systems as observed in 3H,5H-dipyrrolo[1,2-c:2',1'-f]pyrimidin-3-ones, xanthoglows, pyrroloindolizinedione analogs, BODIPY analogs, and the phenolic and imidazolinone ring systems of Green Fluorescent Protein (GFP). This manuscript describes an inexpensive and operationally simple method of performing a Claisen-Schmidt condensation to generate a series of fluorescent pH dependent pyrazole/imidazole/isoindolone dipyrrinone analogs. While the methodology illustrates the synthesis of dipyrrinone analogs, it can be translated to produce a wide range of conjugated bicyclic aromatic compounds. The Claisen-Schmidt condensation reaction utilized in this method is limited in scope to nucleophiles and electrophiles that are enolizable under basic conditions (nucleophile component) and non-enolizable aldehydes (electrophile component). Additionally, both the nucleophilic and electrophilic reactants must contain functional groups that will not inadvertently react with hydroxide. Despite these limitations, this methodology offers access to completely novel systems that can be employed as biological or molecular probes.

The Claisen-Schmidt condensation reaction is an important methodology for the generation of methine-bridged conjugated bicyclic aromatic compounds. Through utilizing a base-mediated variant of the aldol reaction, a range of fluorescent and/or biologically relevant molecules can be accessed through a generally inexpensive and operationally simple synthetic approach.

توليف البيرازول المعتمد على درجة الحموضة، إيميدازول، وإيزيندولون ديبيرينون فلوروفوريس باستخدام نهج التكثيف كلايسين شميدت

Methine-bridged conjugated bicyclic aromatic compounds are common constituents of a range of biologically relevant molecules such as porphyrins, ...

Analysis of Organochlorine Pesticides in a Soil Sample by a Modified QuEChERS Approach Using Ammonium Formate

Currently, the QuEChERS method represents the most widely used sample preparation protocol worldwide for analyzing pesticide residues in a broad variety of matrices both in official and non-official laboratories. The QuEChERS method using ammonium formate has previously proven to be advantageous compared to the original and the two official versions. On the one hand, the simple addition of 0.5 g of ammonium formate per gram of sample is sufficient to induce phase separation and achieve good analytical performance. On the other hand, ammonium formate reduces the need for maintenance in routine analyses. Here, a modified QuEChERS method using ammonium formate was applied for the simultaneous analysis of organochlorine pesticide (OCP) residues in agricultural soil. Specifically, 10 g of the sample was hydrated with 10 mL of water and then extracted with 10 mL of acetonitrile. Next, phase separation was carried out using 5 g of ammonium formate. After centrifugation, the supernatant was subjected to a dispersive solid-phase extraction clean-up step with anhydrous magnesium sulfate, primary-secondary amine, and octadecylsilane. Gas chromatography-mass spectrometry was used as the analytical technique. The QuEChERS method using ammonium formate is demonstrated as a successful alternative for extracting OCP residues from a soil sample.

The present protocol describes the utilization of ammonium formate for phase partitioning in QuEChERS, together with gas chromatography-mass spectrometry, to successfully determine organochlorine pesticide residues in a soil sample.

تحليل مبيدات الآفات الكلورية العضوية في عينة التربة بواسطة نهج QuEChERS المعدل باستخدام فورمات الأمونيوم

Currently, the QuEChERS method represents the most widely used sample preparation protocol worldwide for analyzing pesticide residues in a broad variety ...