Name: application of an in vitro dna protection assay to visualize stress mediation properties of the dps protein
Uploaded: 2013-05-31T13:00:00
Description: Watch this Scientific Journal Video about Application of an In vitro DNA Protection Assay to Visualize Stress Mediation Properties of the Dps Protein at JoVE.com

威尔弗雷德·哈根，德马蒂诺MICHELA，和Kourosh Honarmand埃布拉希米有益的讨论，我们非常感谢。这项工作是由代尔夫特理工大学的启动资金支持。

1。 DPS表达和纯化获得高纯度蛋白的DNA保护实验是必不可少的第一步。可以在4至5天的Dps蛋白的分离纯化。 <ol><li>变换的蛋白酶缺陷型菌株大肠杆菌大肠杆菌 （如BL21（DE3）pLysS中）已被克隆到其中的Dps的蛋白编码序列的pET载体（如pET17）。 </li><li>连胜转化细胞到的Luria肉汤（LB）琼脂平板，含适当的抗生素（如氨苄青霉素和氯霉素1.1中提供的例子）。过夜孵育板在37°C?...

<ol>
	<li>Almiron, M., Link, A. J., Furlong, D., Kolter, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+DNA-binding+protein+with+regulatory+and+protective+roles+in+starved+Escherichia+coli.">A novel DNA-binding protein with regulatory and protective roles in starved Escherichia coli.</a> Genes Dev. 6, 2646-2654 (1992).</li><li>Chiancone, E., Ceci, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+multifaceted+capacity+of+Dps+proteins+to+combat+bacterial+stress+conditions:+Detoxification+of+iron+and+hydrogen+peroxide+and+DNA+binding.">The multifaceted capacity of Dps proteins to combat bacterial stress conditions: Detoxification of iron and hydrogen peroxide and DNA binding.</a> Biochimica et biophysica acta. 1800 (8), 798-805 (2010).</li><li>Ali Azam, T., Iwata, A., Nishimura, A., Ueda, S., Ishihama, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Growth+phase-dependent+variation+in+protein+composition+of+the+Escherichia+coli+nucleoid.">Growth phase-dependent variation in protein composition of the Escherichia coli nucleoid.</a> J. Bacteriol. 181 (20), 6361-6370 (1999).</li><li>Grainger, D. C., Goldberg, M. D., Lee, D. J., Busby, 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=Selective+repression+by+Fis+and+H-NS+at+the+Escherichia+coli+dps+promoter.">Selective repression by Fis and H-NS at the Escherichia coli dps promoter.</a> Molecular Microbiology. 68 (6), 1366-1377 (2008).</li><li>Martinez, A., Kolter, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protection+of+DNA+during+oxidative+stress+by+the+nonspecific+DNA-binding+protein+Dps.">Protection of DNA during oxidative stress by the nonspecific DNA-binding protein Dps.</a> J. Bacteriol. 179 (16), 5188-5194 (1997).</li><li>Nair, S., Finkel, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dps+protects+cells+against+multiple+stresses+during+stationary+phase.">Dps protects cells against multiple stresses during stationary phase.</a> J. Bacteriol. 186 (13), 4192-4198 (2004).</li><li>Grant, R. A., Filman, D. J., Finkel, S. E., Kolter, R., Hogle, 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=The+crystal+structure+of+Dps,+a+ferritin+homolog+that+binds+and+protects+DNA.">The crystal structure of Dps, a ferritin homolog that binds and protects DNA.</a> Nat. Struct. Biol. 5 (4), 294-303 (1998).</li><li>Zhao, G., Ceci, P., 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=Iron+and+hydrogen+peroxide+detoxification+properties+of+DNA-binding+protein+from+starved+cells.+A+ferritin-like+DNA-binding+protein+of+Escherichia+coli.">Iron and hydrogen peroxide detoxification properties of DNA-binding protein from starved cells. A ferritin-like DNA-binding protein of Escherichia coli.</a> J. Biol. Chem. 277 (31), 27689-27696 (2002).</li><li>Ceci, P., Cellai, 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=DNA+condensation+and+self-aggregation+of+Escherichia+coli+Dps+are+coupled+phenomena+related+to+the+properties+of+the+N-terminus.">DNA condensation and self-aggregation of Escherichia coli Dps are coupled phenomena related to the properties of the N-terminus.</a> Nucleic Acids Res. 32 (19), 5935-5944 (2004).</li><li>Ilari, A., Ceci, P., Ferrari, D., Rossi, G. L., Chiancone, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Iron+incorporation+into+Escherichia+coli+Dps+gives+rise+to+a+ferritin-like+microcrystalline+core.">Iron incorporation into Escherichia coli Dps gives rise to a ferritin-like microcrystalline core.</a> J. Biol. Chem. 277 (40), 37619-37623 (2002).</li><li>Swift, J., Wehbi, W. 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=Design+of+functional+ferritin-like+proteins+with+hydrophobic+cavities.">Design of functional ferritin-like proteins with hydrophobic cavities.</a> Journal of the American Chemical Society. 128 (20), 6611-6619 (2006).</li><li>Ceci, P., Chiancone, E., 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=Synthesis+of+iron+oxide+nanoparticles+in+Listeria+innocua+Dps+(DNA-binding+protein+from+starved+cells):+a+study+with+the+wild-type+protein+and+a+catalytic+centre+mutant.">Synthesis of iron oxide nanoparticles in Listeria innocua Dps (DNA-binding protein from starved cells): a study with the wild-type protein and a catalytic centre mutant.</a> Chemistry. 16 (2), 709-717 (2010).</li><li>Ceci, P., Ilari, A., Falvo, E., Chiancone, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Dps+protein+of+Agrobacterium+tumefaciens+does+not+bind+to+DNA+but+protects+it+toward+oxidative+cleavage:+x-ray+crystal+structure,+iron+binding,+and+hydroxyl-radical+scavenging+properties.">The Dps protein of Agrobacterium tumefaciens does not bind to DNA but protects it toward oxidative cleavage: x-ray crystal structure, iron binding, and hydroxyl-radical scavenging properties.</a> The Journal of Biological Chemistry. 278 (22), 20319-20326 (2003).</li><li>Su, M., Cavallo, S., Stefanini, S., Chiancone, E., Chasteen, N. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+so-called+Listeria+innocua+ferritin+is+a+Dps+protein.+Iron+incorporation,+detoxification,+and+DNA+protection+properties.">The so-called Listeria innocua ferritin is a Dps protein. Iron incorporation, detoxification, and DNA protection properties.</a> Biochemistry. 44 (15), 5572-5578 (2005).</li><li>Kumar, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protective+effects+of+Koelreuteria+paniculata+Laxm.+on+oxidative+stress+and+hydrogen+peroxide-induced+DNA+damage.">Protective effects of Koelreuteria paniculata Laxm. on oxidative stress and hydrogen peroxide-induced DNA damage.</a> Phytopharmacology. 1 (5), 177-189 (2011).</li><li>Stinefelt, B., Leonard, S. S., Blemings, K. P., Shi, X., Klandorf, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Free+radical+scavenging,+DNA+protection,+and+inhibition+of+lipid+peroxidation+mediated+by+uric+acid.">Free radical scavenging, DNA protection, and inhibition of lipid peroxidation mediated by uric acid.</a> Annals of Clinical and Laboratory Science. 35 (1), 37-45 (2005).</li><li>Lopez-Gomollon, S., Sevilla, E., Bes, M. T., Peleato, M. L., Fillat, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+insights+into+the+role+of+Fur+proteins:+FurB+(All2473)+from+Anabaena+protects+DNA+and+increases+cell+survival+under+oxidative+stress.">New insights into the role of Fur proteins: FurB (All2473) from Anabaena protects DNA and increases cell survival under oxidative stress.</a> The Biochemical Journal. 418 (1), 201-207 (2009).</li><li>Ebrahimi, K. H., Hagedoorn, P. L., Hagen, W. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibition+and+stimulation+of+formation+of+the+ferroxidase+center+and+the+iron+core+in+Pyrococcus+furiosus+ferritin.">Inhibition and stimulation of formation of the ferroxidase center and the iron core in Pyrococcus furiosus ferritin.</a> Journal of Biological Inorganic Chemistry. 15 (8), 1243-1253 (2010).</li><li>Smith, F. E., Herbert, J., Gaudin, J., Hennessy, D. J., Reid, G. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Serum+iron+determination+using+ferene+triazine.">Serum iron determination using ferene triazine.</a> Clinical Biochemistry. 17 (5), 306-310 (1984).</li></ol>

在这篇文章中所描述的净化过程中的DPS是非常强大的。一直纯度高（&gt; 99％）;没有其他蛋白质出现在可见光波段的SDS-PAGE凝胶。尽管这样，似乎有部分批次纯化DPS核酸酶活性，证明了部分DNA的降解，当孵育浓度非常高的DPS。这可能表明在低浓度高活性的DNA酶的存在，我们无法通过净化除去。然而，这种DNA降解只观察到的浓度通常不用于在体外测定的 ，所以这通常不存在任何问题。 <...

需氧生物必须不断抗衡，可以破坏它们的DNA，以及其他重要的生物大分子的活性氧。对抗氧化损伤的毒性作用的一个有力的工具是从饥饿细胞的DNA结合蛋白（DPS）。自从被发现于1992年，从饥饿E.大肠杆菌培养1，DPS中已发现300余种细菌和古细菌2。 DPS在固定相的大规模上调使得它表达最高度的拟核相关蛋白的大肠杆菌饥饿条件下3，4的 大肠杆菌 。此外，已被证明的Dps保留在许多不同的应力，包括饥饿，铁浓度高，紫外线照射，热休克，氧化应激5，6细菌的生存能力和DNA完整性。 在结构上，DPS自联营成一个稳定的同源寡聚体复合物的12个单体，W一族组装成一个球形的空壳。 〜4.5 nm的范围内的内部腔体的溶剂可允许通过的小分子7的毛孔的外部访问，和，它可以螯合矿化的金属如铁8。 DPS的保护作用来源于几个生化活动，其中包括非特异性DNA结合，亚铁活动，和铁存储8。 对人体有益的生化活动的DPS，首先需要详细研究其净化。 Dps的净化是一个复杂的过程，Dps的必须不仅从其他蛋白质分离，但也从任何结合的DNA 7。我们已优化的纯化过程中使用许多常用的技术，包括两个离子交换柱和硫酸铵沉淀步骤。几个缓冲区的交流是必要的，高度集中的DPS可以在溶液中沉淀出来低盐条件。一旦DPS蛋白已被纯化，它可能是用于检测直接测量亚铁活性8，DNA-结合的化学计量学9，与铁结合的机制10。纯净DPS也有其他潜在的应用。稳定的中空球状结构的Dps被用作脚手架，用于存储在蛋白质内部空腔11的憎水颗粒，甚至作为一个反应室，以合成新颖的磁性纳米粒子12。 保护能力的DPS调解活性氧自由基造成的损害，可以清楚地直接证明使用DNA保护实验13，14。在这种体外过程中，产生自由基种，铁催化H 2 O 2通过Fenton化学降解。这些自由基直接损伤DNA存在于反应中，可以在高浓度下完全降解。两个关键的DPS活动都可能直接抵消的影响FENT在介导的自由基的产生。 Dps的催化铁的浓度降低通过矿化，在这个过程中消耗的可用过氧化氢。此外，DPS与DNA结合可能屏蔽它身体免受自由基的损害，并凝结成更小的体积与反应表面积少。这两个属性的组合，使得DNA为目的测量保护的Dps活动的适合的保护测定。 的DNA保护分析法是相当灵活的，可用于各种各样的应用程序之外的Dps表征。自由基损伤是一种常见的细胞形式的压力，和许多不同的蛋白质和化学品的使用，抵制它。测定的一般原则，使用作为自由基损伤的标记物的DNA的完整性，可以结合使用几乎任何产生自由基的反应剂或抵消。其中包括，该法已成功地用于确定抗氧化性能K的穿心莲提取物使用在食品行业15，表征尿酸对羟基损害调解16的影响，并获得新的见解毛皮转录调节蛋白17的功能。 尽管在发表的论文中检测的众多用途中，我们发现许多优化和故障排除步骤，这使得第一次不必要的艰苦的过程，许多研究者测定。在这篇文章中，我们提出的协议旨在消除这一屏障，为入境。

<table border="1">
 <tbody>
 <tr>
 <td>Name of Reagent/Material</td>
 <td>Company</td>
 <td>Catalog Number</td>
 </tr>
 <tr>
 <td>BL21(DE3)pLysS competent cells</td>
 <td>Promega</td>
 <td>L1195</td>
 </tr>
 <tr>
 <td>pET-17b DNA</td>
 <td>EMD-Millipore</td>
 <td>69663-3</td>
 </tr>
 <tr>
 <td>LB broth powder</td>
 <td>Sigma-Aldrich</td>
 <td>L3022</td>
 </tr>
 <tr>
 <td>Ampicillin sodium salt</td>
 <td>Sigma-Aldrich</td>
 <td>A0166</td>
 </tr>
 <tr>
 <td>Chloramphenicol</td>
 <td>Sigma-Aldrich</td>
 <td>C0378</td>
 </tr>
 <tr>
 <td>IPTG</td>
 <td>Sigma-Aldrich</td>
 <td>I6758</td>
 </tr>
 <tr>
 <td>HEPES</td>
 <td>BDH</td>
 <td>441476L</td>
 </tr>
 <tr>
 <td>Potassium hydroxide</td>
 <td>Merck</td>
 <td>105033</td>
 </tr>
 <tr>
 <td>Sodium chloride</td>
 <td>VWR</td>
 <td>443824T</td>
 </tr>
 <tr>
 <td>EDTA</td>
 <td>Sigma-Aldrich</td>
 <td>E9884</td>
 </tr>
 <tr>
 <td>Protease Inhibitor Cocktail Set III</td>
 <td>EMD-Millipore</td>
 <td>539134</td>
 </tr>
 <tr>
 <td>DEAE-Sepharose</td>
 <td>Sigma-Aldrich</td>
 <td>DFF100</td>
 </tr>
 <tr>
 <td>Ammonium sulphate</td>
 <td>Sigma-Aldrich</td>
 <td>A4418</td>
 </tr>
 <tr>
 <td>PD-10 Desalting Columns</td>
 <td>GE Healthcare LS</td>
 <td>17-0851-01</td>
 </tr>
 <tr>
 <td>SP-Sepharose</td>
 <td>Sigma-Aldrich</td>
 <td>S1799</td>
 </tr>
 <tr>
 <td>Amicon Ultra Centr. Filter (10K MWCO)</td>
 <td>Millipore</td>
 <td>UFC901024</td>
 </tr>
 <tr>
 <td>Ferrous Sulphate heptahydrate</td>
 <td>Sigma-Aldrich</td>
 <td>F8048</td>
 </tr>
 <tr>
 <td>Hydrogen peroxide solution</td>
 <td>Sigma-Aldrich</td>
 <td>216763</td>
 </tr>
 <tr>
 <td>MOPS</td>
 <td>Calbiochem</td>
 <td>475898</td>
 </tr>
 <tr>
 <td>SDS solution</td>
 <td>Bio-Rad</td>
 <td>161-0418</td>
 </tr>
 <tr>
 <td>Ethidium bromide</td>
 <td>Sigma-Aldrich</td>
 <td>E1510</td>
 </tr>
 <tr>
 <td>Equipment</td>
 <td>Company</td>
 <td>Model</td>
 </tr>
 <tr>
 <td>Static incubator</td>
 <td>Hettich</td>
 <td>INE500</td>
 </tr>
 <tr>
 <td>Shaking Incubator </td>
 <td>New Brunswick Sc.</td>
 <td>Inova 44</td>
 </tr>
 <tr>
 <td>Cooled centrifuge</td>
 <td>Beckman Coulter</td>
 <td>Avanti J-E</td>
 </tr>
 <tr>
 <td>Table-top centrifuge</td>
 <td>Eppendorf</td>
 <td>5424</td>
 </tr>
 <tr>
 <td>Cell disrupter</td>
 <td>Constant Systems Ltd.</td>
 <td>TS2/40/AA/AA</td>
 </tr>
 <tr>
 <td>FPLC Purifier</td>
 <td>General Electric</td>
 <td>AKTA</td>
 </tr>
 <tr>
 <td>Airtight vials</td>
 <td>Cole-Parmer</td>
 <td>EW-08918-85</td>
 </tr>
 <tr>
 <td>Syringe needles</td>
 <td>BD</td>
 <td>305128</td>
 </tr>
 <tr>
 <td>Pipettes</td>
 <td>Eppendorf</td>
 <td>Z683779-1EA, Z683795-1EA</td>
 </tr>
 </tbody>
</table>

application of an in vitro dna protection assay to visualize stress mediation properties of the dps protein

氧化应激是一个不可避免的副产品的有氧生活。地面代谢分子氧气是必不可少的，但它也需要在许多破坏性反应于生物体的部分。通过Fenton化学降解细胞成分产生自由基相结合的有氧代谢和铁，这是生活的另一种重要的化合物，是足够的。 DNA降解无疑是最具破坏性的过程，涉及细胞内的自由基，DNA修复是远从琐碎。在这篇文章中提出的检测提供了一个定量的测量和可视化技术，分子和酶的作用自由基介导的DNA损伤。 DNA保护法是一种简单，快速和强大的工具， 在体外蛋白质或化学品的防护性能表征。它涉及曝光的DNA的损害性的氧化反应，加入不同浓度的化合物还。作为化合物浓度的函数的DNA损伤的减少或增加，然后可视化，使用凝胶电泳。在这篇文章中，我们展示了保护的DNA检测技术，通过测量DNA结合蛋白的保护性饥饿细胞（DPS）。 DPS是一个小型铁，动用300余种细菌，有力地打击环境压力。在这里，我们提出的的DPS净化协议和评估DNA保护DPS优化实验条件。

这里描述的Dps的纯化方法是高度可再现性。 Dps的纯化，根据所描述的协议，使用2升的大肠杆菌大肠杆菌文化为出发点，通常会产生2.5毫升含蛋白质浓度在5和12微米之间的DPS 12。较长的诱导时间（4小时）似乎减少这种可变性。蛋白质纯度为99％以上，始终SDS-PAGE凝胶（ 图1）可见一斑。 DNA污染的水平，一直是可以忽略不计，证明通过琼脂糖凝胶染色DNA和OD 260 / OD ...

Watch this Scientific Journal Video about Application of an In vitro DNA Protection Assay to Visualize Stress Mediation Properties of the Dps Protein at JoVE.com

Application of an In vitro DNA Protection Assay to Visualize Stress Mediation Properties of the Dps Protein

（DPS）从饥饿细胞的DNA结合蛋白起着至关重要的作用打击细菌应力的。本文讨论了净化<em&gt; E。大肠杆菌</em&gt; DPS和一个协议<em&gt;在体外</em&gt;试验示范DPS-介导的保护DNA降解的活性氧物种。

应用的<em在体外</em&gt;保护DNA检测，以可视化应力调解属性的DPS蛋白质

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Long-term Lethal Toxicity Test with the Crustacean Artemia franciscana

致命的甲壳类动物长期毒性试验卤虫franciscana</em

Our research activities target the use of biological methods for the evaluation of environmental quality, with particular reference to saltwater/brackish ...

In vitro Methylation Assay to Study Protein Arginine Methylation

In vitro Methylation Assay to Study Protein Arginine Methylation

在体外甲基化分析研究蛋白质精氨酸甲基化

Protein arginine methylation is one of the most abundant post-translational modifications in the nucleus. Protein arginine methylation can be identified ...

Quantitative Immunofluorescence Assay to Measure the Variation in Protein Levels at Centrosomes

定量免疫检测，以衡量在中心体的变异蛋白水平

Centrosomes are small but important organelles that serve as the poles of mitotic spindle to maintain genomic integrity or assemble primary cilia to ...

Comet Assay as an Indirect Measure of Systemic Oxidative Stress

彗星试验系统性氧化应激的间接测量

Higher eukaryotic organisms cannot live without oxygen; yet, paradoxically, oxygen can be harmful to them. The oxygen molecule is chemically relatively ...

Measuring DNA Damage and Repair in Mouse Splenocytes After Chronic In Vivo Exposure to Very Low Doses of Beta- and Gamma-Radiation

Measuring DNA Damage and Repair in Mouse Splenocytes After Chronic In Vivo Exposure to Very Low Doses of Beta- and Gamma-Radiation

测量DNA损伤与修复小鼠脾细胞慢性后<em&gt;在体内</em&gt;暴露于β和γ辐射剂量很低

Low dose radiation exposure may produce a variety of biological effects that are different in quantity and quality from the effects produced by high ...

Chromatin Immunoprecipitation Assay for the Identification of Arabidopsis Protein-DNA Interactions In Vivo

Chromatin Immunoprecipitation Assay for the Identification of Arabidopsis Protein-DNA Interactions In Vivo

染色质免疫沉淀拟南芥蛋白-DNA相互作用的鉴定<em&gt;在体内</em

Intricate gene regulatory networks orchestrate biological processes and developmental transitions in plants. Selective transcriptional activation and ...

The Assembly and Application of 'Shear Rings': A Novel Endothelial Model for Orbital, Unidirectional and Periodic Fluid Flow and Shear Stress

大会和“剪切环”中的应用：一种新型内皮模型轨道，单向和定期流体流动和剪应力

Deviations from normal levels and patterns of vascular fluid shear play important roles in vascular physiology and pathophysiology by inducing adaptive as ...