Name: introducing shear stress in the study of bacterial adhesion
Uploaded: 2011-09-02T12:00:00
Duration: 13 min 28 s
Description: Watch this Scientific Journal Video about Introducing Shear Stress in the Study of Bacterial Adhesion at JoVE.com

लेखकों एमिली Mairey और Emmanuel Donnadieu प्रक्रिया के प्रारंभिक स्थापित करने के लिए धन्यवाद देना चाहूंगा.

<ol>
	<li>Mairey, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cerebral+microcirculation+shear+stress+levels+determine+Neisseria+meningitidis+attachment+sites+along+the+blood-brain+barrier.">Cerebral microcirculation shear stress levels determine Neisseria meningitidis attachment sites along the blood-brain barrier.</a> J Exp Med. 203, 1939-1950 (2006).</li><li>Pries, A. R., Secomb, T. W., Gaehtgens, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design+Principles+of+Vascular+Beds.">Design Principles of Vascular Beds.</a> Circ Res. 77, 1017-1023 (1995).</li><li>Zarbock, A., Ley, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neutrophil+adhesion+and+activation+under+flow.">Neutrophil adhesion and activation under flow.</a> Microcirculation. 16, 31-42 (2009).</li><li>Tissot, O., Pierres, A., Foa, C., Delaage, M., Bongrand, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motion+of+cells+sedimenting+on+a+solid+surface+in+a+laminar+shear+flow.">Motion of cells sedimenting on a solid surface in a laminar shear flow.</a> Biophys J. 61, 204-215 (1992).</li><li>Mikaty, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extracellular+bacterial+pathogen+induces+host+cell+surface+reorganization+to+resist+shear+stress.">Extracellular bacterial pathogen induces host cell surface reorganization to resist shear stress.</a> PLoS Pathog. 5, e1000314-e1000314 (2009).</li><li>Chamot-Rooke, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Posttranslational+modification+of+pili+upon+cell+contact+triggers+N.+meningitidis+dissemination.">Posttranslational modification of pili upon cell contact triggers N. meningitidis dissemination.</a> Science. 331, 778-782 (2011).</li><li>Soyer, M., Dum&#237;nil, G., Christodoulides, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bacterial+adhesion+under+shear+stress.">Bacterial adhesion under shear stress.</a> Neisseria meningitidis: Methods and Protocols (Methods in Molecular Biology). , (2011).</li><li>Abramoff, M. D., Magelhaes, P. J., Ram, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Image+Processing+with+ImageJ.">Image Processing with ImageJ.</a> Biophotonics International. 11, 36-42 (2004).</li><li>Thomas, W. E., Trintchina, E., Forero, M., Vogel, V., Sokurenko, E. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bacterial+adhesion+to+target+cells+enhanced+by+shear+force.">Bacterial adhesion to target cells enhanced by shear force.</a> Cell. 109, 913-923 (2002).</li><li>Tchesnokova, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Shear-enhanced+binding+of+intestinal+colonization+factor+antigen+I+of+enterotoxigenic+Escherichia+coli.">Shear-enhanced binding of intestinal colonization factor antigen I of enterotoxigenic Escherichia coli.</a> Mol Microbiol. 76, 489-502 (2010).</li><li>Konto-Ghiorghi, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dual+role+for+pilus+in+adherence+to+epithelial+cells+and+biofilm+formation+in+Streptococcus+agalactiae.">Dual role for pilus in adherence to epithelial cells and biofilm formation in Streptococcus agalactiae.</a> PLoS Pathog. 5, e1000422-e1000422 (2009).</li></ol>

ब्याज की कोई संघर्ष की घोषणा की.

कतरनी तनाव का महत्व और आम तौर पर जीव विज्ञान में यांत्रिक पहलुओं की तेजी से मान्यता प्राप्त किया जा रहा है है. उदाहरण के लिए अत्यधिक अनुकूलित लिम्फोसाइटों आसंजन और संवहनी दीवार पर रोलिंग की प्रक्रिया में प्रोटीन की selectin परिवार के चिपकने वाला गुण प्रक्रिया में कतरनी तनाव शुरू करने से मान्यता प्राप्त किया गया. ऊपर वर्णित की प्रक्रिया ग्राम नकारात्मक जीवाणुओं नेइसेरिया meningitidis लिए आवेदन किया था लेकिन रोगजनकों की एक विस्तृत श्रृंखला के लिए लागू किया जाना चाहिए . कतरनी तनाव के महत्व को अन्य रोगज़नक़ों और अन्य संक्रमण साइटों के लिए किया गया है भी दिखाया गया है. बैक्टीरियल आसंजन कतरनी तनाव से वातानुकूलित uropathogenic Escherichia कोलाई (UPEC) 9 पर पाया FimH adhesin के अध्ययन के द्वारा वर्णित किया गया है . Selectins करने के लिए इसी प्रकार, FimH और अपने मेजबान सेल रिसेप्टर के बीच बातचीत कतरनी प्रेरित यांत्रिक 9 बलों और enterotoxigenic ई. कोलाई के CfaE adhesin से मजबूत होना दिखाया गया था भी सूचित किया गया है एक कतरनी पर निर्भर के माध्यम से आंतों उपकला कोशिकाओं के आसंजन मध्यस्थता 10 तंत्र. हम सूचना दी, चैम्बर लामिना का प्रवाह परख है कि स्ट्रैपटोकोकस agalactiae pili इस रोगज़नक़ के प्रवाह शर्तों के तहत 11 उपकला कोशिकाओं का पालन करने के लिए आवश्यक थे इस अध्याय में वर्णित प्रोटोकॉल का उपयोग. इस तरह के अध्ययन की पुष्टि करते हैं कि हमारे प्रवाह चैम्बर परख कतरनी तनाव की शर्तों के तहत मेजबान कोशिका रोगज़नक़ बातचीत की जांच के लिए एक उपयोगी उपकरण है.

<table border="1"><tbody><tr><td> अभिकर्मक के नाम </td><td> कंपनी </td><td> सूचीपत्र संख्या </td></tr><tr><td> μ-स्लाइड छठी 0.4 प्रवाह किट </td><td> IBIDI </td><td> 80606 </td></tr><tr><td> सिरिंज 50 मिलीलीटर Plastipak Luer - बंद </td><td> Becton Dickinson </td><td> 300865 </td></tr><tr><td> Tygon टयूबिंग R3603 3.2 x 4.8mm </td><td> फिशर वैज्ञानिक </td><td> R3603 </td></tr><tr><td> पानी निकलने की टोंटी 3 तरह, 2 पुरुष luer महिला luer </td><td> जैव - रेड </td><td> 7328103 </td></tr><tr><td> सिरिंज पंप </td><td> हार्वर्ड उपकरण </td><td> 2000 पीएचडी </td></tr><tr><td> उल्टे माइक्रोस्कोप, Nikon </td><td> Nikon </td><td> ग्रहण टी मैं </td></tr><tr><td> सीसीडी कैमरा </td><td> हमामात्सू </td><td> ORCA 285 सीसीडी या 3-सीसीडी ORCA </td></tr><tr><td> ImageJ सॉफ्टवेयर </td><td> एनआईएच </td><td> फ्रीवेयर ( <a href="http://rsbweb.nih.gov/ij/">http://rsbweb.nih.gov/ij/</a> ) </td></tr></tbody></table>

introducing shear stress in the study of bacterial adhesion

During bacterial infections a sequence of interactions occur between the pathogen and its host. Bacterial adhesion to the host cell surface is often the initial and determining step of the pathogenesis. Although experimentally adhesion is mostly studied in static conditions adhesion actually takes place in the presence of flowing liquid. First encounters between bacteria and their host often occur at the mucosal level, mouth, lung, gut, eye, etc. where mucus flows along the surface of epithelial cells. Later in infection, pathogens occasionally access the blood circulation causing life-threatening illnesses such as septicemia, sepsis and meningitis. A defining feature of these infections is the ability of these pathogens to interact with endothelial cells in presence of circulating blood. The presence of flowing liquid, mucus or blood for instance, determines adhesion because it generates a mechanical force on the pathogen. To characterize the effect of flowing liquid one usually refers to the notion of shear stress, which is the tangential force exerted per unit area by a fluid moving near a stationary wall, expressed in dynes/cm2. Intensities of shear stress vary widely according to the different vessels type, size, organ, location etc. (0-100 dynes/cm2). Circulation in capillaries can reach very low shear stress values and even temporarily stop during periods ranging between a few seconds to several minutes 1. On the other end of the spectrum shear stress in arterioles can reach 100 dynes/cm2 2. The impact of shear stress on different biological processes has been clearly demonstrated as for instance during the interaction of leukocytes with the endothelium 3. To take into account this mechanical parameter in the process of bacterial adhesion we took advantage of an experimental procedure based on the use of a disposable flow chamber 4. Host cells are grown in the flow chamber and fluorescent bacteria are introduced in the flow controlled by a syringe pump. We initially focused our investigations on the bacterial pathogen Neisseria meningitidis, a Gram-negative bacterium responsible for septicemia and meningitis. The procedure described here allowed us to study the impact of shear stress on the ability of the bacteria to: adhere to cells 1, to proliferate on the cell surface 5and to detach to colonize new sites 6 (Figure 1). Complementary technical information can be found in reference 7. Shear stress values presented here were chosen based on our previous experience1 and to represent values found in the literature. The protocol should be applicable to a wide range of pathogens with specific adjustments depending on the objectives of the study.

Watch this Scientific Journal Video about Introducing Shear Stress in the Study of Bacterial Adhesion at JoVE.com

Introducing Shear Stress in the Study of Bacterial Adhesion

संक्रमण की प्रक्रिया के दौरान, एक महत्वपूर्ण कदम मेजबान कोशिकाओं के साथ रोगजनकों के आसंजन है. ज्यादातर मामलों में इस आसंजन कदम यांत्रिक तरल बह द्वारा उत्पन्न तनाव की उपस्थिति में होता है. हम एक तकनीक है कि बैक्टीरियल आसंजन के अध्ययन में एक महत्वपूर्ण पैरामीटर के रूप में कतरनी तनाव परिचय का वर्णन.

बैक्टीरियल आसंजन के अध्ययन में कतरें तनाव का परिचय

During bacterial infections a sequence of interactions occur between the pathogen and its host. Bacterial adhesion to the host cell surface is often the ...

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तरीके बैक्टीरियल Pyomelanin उत्पादन रोकना और ऑक्सीडेटिव तनाव के प्रति संवेदनशीलता में इसी वृद्धि निर्धारित करने के लिए

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