Intentional formation of a protein corona on nanoparticles: Serum concentration affects protein corona mass,surface charge,and nanoparticle–cell interaction

The protein corona, which immediately is formed after contact of nanoparticles and biological systems, plays a crucial role for the biological fate of nanoparticles. In the here presented study we describe a strategy to control the amount of corona proteins which bind on particle surface and the impact of such a protein corona on particle-cell interactions. For corona formation, polyethyleneimine (PEI) coated magnetic nanoparticles (MNP) were incubated in a medium consisting of fetal calf serum (FCS) and cell culture medium. To modulate the amount of proteins bind to particles, the composition of the incubation medium was varied with regard to the FCS content. The protein corona mass was estimated and the size distribution of the participating proteins was determined by means of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS–PAGE). Additionally, the zeta potential of incubated particles was measured. Human blood–brain barrier-representing cell line HBMEC was used for in vitro incubation experiments. To investigate the consequences of the FCS dependent protein corona formation on the interaction of MNP and cells flow cytometry and laser scanning microscopy were used. Zeta potential as well as SDS–PAGE clearly reveal an increase in the amount of corona proteins on MNP with increasing amount of FCS in incubation medium. For MNP incubated with lower FCS concentrations especially medium-sized proteins of molecular weights between 30 kDa and 100 kDa could be found within the protein corona, whereas for MNP incubated within higher FCS concentrations the fraction of corona proteins of 30 kDa and less increased. The presence of the protein corona reduces the interaction of PEI-coated MNP with HBMEC cells within a 30 min-incubation.     

Directional memory and caged dynamics in cytoskeletal remodelling

We report directional memory of spontaneous nanoscale displacements of an individual bead firmly anchored to the cytoskeleton of a living cell. A novel method of analysis shows that for shorter time intervals cytoskeletal displacements are antipersistent and thus provides direct evidence in a living cell of molecular trapping and caged dynamics. At longer time intervals displacements are persistent. The transition from antipersistence to persistence is indicative of a time-scale for cage rearrangements and is found to depend upon energy release due to ATP hydrolysis and proximity to a glass transition. Anomalous diffusion is known to imply memory, but we show here that memory is attributed to direction rather than step size. As such, these data are the first to provide a molecular-scale physical picture describing the cytoskeletal remodelling process and its rate of progression.     

The Met243 sulfonium ion linkage is responsible for the anomalous magnetic circular dichroism and optical spectral properties of myeloperoxidase

 The heme group of myeloperoxidase shows anomalous optical properties, and the enzyme possesses the unique ability to catalyze the oxidation of chloride. However, the nature of the covalently bound heme macrocycle has been difficult to identify. In this work, the electronic and magnetic properties of the heme groups in oxidized and reduced forms of wild-type and Met243Thr mutant myeloperoxidase and wild-type lactoperoxidase have been investigated using variable-temperature (1.6–273 K) magnetic circular dichroism (MCD) spectroscopy along with parallel optical absorption and electron paramagnetic resonance studies. The results provide assessment of the spin state mixtures of the oxidized and reduced samples at ambient and liquid helium temperatures and show that the anomalous MCD properties of myeloperoxidase, e.g. red-shifted and inverted signs for bands in the high-spin ferric and low-spin ferrous forms compared to other heme peroxidases and heme proteins in general, are a direct consequence of a novel electron-withdrawing sulfonium ion heme linkage involving Met243. Received: 3 May 1999 / Accepted: 9 August 1999     

Central and peripheral contributions to muscle fatigue in humans during sustained maximal effort

The purpose of this study was to estimate the relative contributions of central and peripheral factors to the development of human muscle fatigue. Nine healthy subjects [five male, four female; age = 30 (2) years, mean (SE)] sustained a maximum voluntary isometric contraction (MVC) of the ankle dorsiflexor muscles for 4 min. Fatigue was quantitated as the fall in MVC. Three measures of central activation and one measure of peripheral activation (compound muscle action potential, CMAP) were made using electromyography (EMG) and electrical stimulation. Measures of intramuscular metabolism were made using magnetic resonance spectroscopy. After exercise, MVC and electrically stimulated tetanic contraction (50 Hz, 500 ms) forces were 22.2 (3.7)% and 37.3 (7.1)% of pre-exercise values, respectively. The measures of central activation suggested some central fatigue during exercise: (1) the central activation ratio [MVC/(MVC + superimposed tetanic force)] fell from 0.94 (0.03) to 0.78 (0.09), (2) the MVC/tetanic force ratio fell from 2.3 (0.7) to 1.3 (0.7), and (3) the integral of the EMG (iEMG) signal decreased to 72.6 (9.1)% of the initial value, while the CMAP amplitude was unchanged. Intramuscular pH was associated by regression with the decline in MVC force (and therefore fatigue) and iEMG. The results indicate that central factors, which were not associated with altered peripheral excitability, contributed approximately 20% to the muscle fatigue developed, with the remainder being attributable to intramuscular (i.e., metabolic) factors. The association between pH and iEMG is consistent with proton concentration as a feedback mechanism for central motor drive during maximal effort.     

三种提取RNA的方法对汉滩病毒检测敏感性影响的研究

分别采用标准法、ＧＰ法和ＭＢＰ法抽提汉滩病毒ＲＮＡ,结合套式ＰＣＲ比较这三种方法的敏感性。结果表明,所用引物对这两株病毒具有相同的特异性。这三种提取ＲＮＡ的敏感性差异不大,敏感性从高到低依次为：ＧＰ、ＭＢＰ和标准法,检测汉滩病毒的效价分别为０．０１、０．０１和０．１ＴＣＩＤ５０／２００μｌ;提取ＲＮＡ的时间长短分别为：标准法为２．５ｈ,ＧＰ为１ｈ,ＭＢＰ为０．５ｈ,可见ＭＢＰ用时最少。使用ＧＰ和ＭＢＰ提取ＲＮＡ时,可以在室温下进行,不需要低温离心。所以,ＭＢＰ最适合于实验室和临床病原体的快速检测,特别是对采集的环境样品的快速检测     

磁流体的制备条件对FeO粒径的影响

目的：探讨纳米磁流体的制备方法及制德条件,如反应温度,搅拌速度,ＮａＯｈ溶液的滴加速度对其粒径,磁化强度的影响。方法：用化学共沉淀法制备,扫描电镜和透视电镜观察粒径大小及形状,古埃磁天平测磁化强度。结果：粒子形态为近似球形,最小可达１０ｎｍ,最大为１μｍ,Ｆｅ＾２＋：Ｆｅ＾３＋＝１：２为最佳投料比,投粒比与粒径无关。     

磁共振水成像对脊柱疾病诊断中的应用价值

目的:探讨磁共振水成像技术对脊柱疾病诊断的应用价值。方法:采用快速高级自旋回波(FASE)、重T2WI及脂肪抑制序列对300例病人检查行磁共振椎管水成像(MRmyelography,MRM)。结果:MRM显示正常25例,MRM异常275例,清楚显示原发病变与邻近脊髓腔、脊髓、神经根的相关关系。结论:MRM具有无创伤、无辐射、速度快,不需对比剂,患者易接受的特点。MRM与常规MRI图像结合可获得全面、客观的病变信息,MRM图像可取代X线脊髓造影和CT脊髓造影。     

磁流体肿瘤热疗技术的研究进展

热疗在近年来已经成为肿瘤治疗最为重要的手段之一,但存在一定的局限性。磁流体热疗技术作为新兴发展起来的热疗手段,克服了常规热疗的缺陷,可以辅助治疗,甚至发展成独立治疗手段。本文综述了国内外近年来有关磁流体热疗基础研究及试验领域的最近进展,首先对磁流体特性、常见磁流体材料和交变磁场装置等方面进行了介绍,最后介绍了磁流体肿瘤热疗技术在体外试验、动物试验和临床研究方面的进展状况。虽然磁流体热疗逐步进入临床阶段,但仍存在不足,需要进一步的完善提高治疗效果。     

CT与MRI对肝硬化背景下小肝癌检出率的比较

目的:分析比较CT与MR对肝硬化背景下小肝癌检出情况,探究CT与MR在肝硬化背景下小肝癌的诊断价值。方法:选择2010年6月~2015年6月期间,我院收治确诊为肝硬化背景下小肝癌患者91例为研究对象,病理及临床相关方法确诊102个病灶,其中小肝癌69个和微小肝癌33个,患者均在不同时期或序列下行多排螺旋CT与MRI检查,分析比较两者对小肝癌和微小肝癌的检出率。结果:多排螺旋CT检查发现肝癌小病灶91个,其中66个小肝癌,25个微小肝癌;MRI检查发现95个病灶,小肝癌67个,微小肝癌28个;69个小肝癌病灶,检出率最高的为CT动脉期(92.75%)与LAVA动脉期(92.75%),检出率最低的为CT平扫(76.81%);33个微小肝癌病灶,检出率最高为LAVA动脉期(75.76%),检出率最低的为LAVA平衡期(36.36%);CT平扫、门静脉期、动脉期、平衡期、MRI-IN-PHASE、LAVA平衡期、LAVA平扫对小肝癌的检出率显著高于对微小肝癌的检出率(P0.05);CT对小肝癌的检出率显著高于微小肝癌的检出率(P0.05),MRI对小肝癌与微小肝癌的检出率无显著差异(P0.05);MRI与CT对小肝癌的检出率不存在差异(P0.05),但MRI对微小肝癌的检出率显著高于CT(P0.05)。结论:MRI-LAVA的动脉期序列对小肝癌病灶与微小肝癌病灶的检出率最高;CT与MRI在对小肝癌的检出率不存在差异,但MRI对微小肝癌的检出具有更明显的优势。     

鞍旁海绵状血管瘤的MRI诊断及误诊分析

目的:分析鞍旁海绵状血管瘤MR影像特点及误诊原因,提高对该疾病的诊断及鉴别诊断水平。方法:收集我院经手术病理证实的13例鞍旁海绵状血管瘤,术前均行MRI平扫及增强扫描,5例行3D-ASL检查,分析其影像学资料。结果:9例表现为横向哑铃状,鞍旁大,鞍内小,病灶主体位于颈内动脉外侧,颈内动脉海绵窦段被病灶包绕;1例鞍旁与鞍内病灶大小相似,1例病灶主体位于颈内动脉内侧,2例病灶完全位于颈内动脉外侧;7例垂体显示不清,6例垂体受推移;6例T2W I表现为类似脑脊液的极高信号;仅5例行3D-ASL检查,病灶均呈低灌注。误诊9例,其中4例误诊垂体腺瘤,5例误诊脑膜瘤。结论:横向哑铃状、病灶主体位于颈内动脉外侧及T2W I类似脑脊液的极高信号是鞍旁海绵状血管瘤的典型影像特征。对于不典型病变,借助3D-ASL可以减少误诊,充分掌握MRI影像特征及鉴别诊断的要点,对提高临床术前诊断水平具有重要价值。