A wide-range, low-cost 150 bp ladder for sizing DNA fragments between 150 and 4500 bp

Agarose gel electrophoresis, a very routine procedure, requires molecular weight standards; these are usually manufactured from plasmid or viral DNA fragments, or more recently, from PCR products of defined sizes. We describe here the preparation of a molecular weight standard from a completely different DNA source - the uniquely organized genome of the beetle Tenebrio molitor. The standard can be used to accurately size DNAs between 150 and 4500 bp, a useful range of sizes for many agarose gel electrophoresis applications, including separation of PCR products and plasmid cloning targets. In addition, it is easy to prepare, inexpensive, and rivals the best of the commercial ladders.     

Direct numerical simulation of wall turbulent flows with microbubbles

The marker‐density‐function (MDF) method has been developed to conduct direct numerical simulation (DNS) for bubbly flows. The method is applied to turbulent bubbly channel flows to elucidate the interaction between bubbles and wall turbulence. The simulation is designed to clarify the structure of the turbulent boundary layer containing microbubbles and the mechanism of frictional drag reduction. It is deduced from the numerical tests that the interaction between bubbles and wall turbulence depends on the Weber and Froude numbers. The reduction of the frictional resistance on the wall is attained and its mechanism is explained from the modulation of the three‐dimensional structure of the turbulent flow. Copyright © 2001 John Wiley & Sons, Ltd.     

银覆盖冠状硅柱阵列基底的制备、SERS特性分析及肿瘤标志物miRNA-106a的检测

以聚苯乙烯(PS)小球为模板,采用金属辅助刻蚀和湿法化学刻蚀技术,制备大面积冠状硅柱阵列,再原位生长银纳米粒子后得到银覆盖冠状硅柱阵列(Ag/Si CPA)基底。实验表明,制备的基底具有优良的表面增强拉曼散射(SERS)特性,电磁增强因子达到1.81×10~6。同时,将制备的罗丹明分子(R6G)标记的DNA发卡探针与基底链接,在与miRNA-106a互补杂交后进行SERS信号检测,获得相应的剂量-响应曲线。结果表明,基于(Ag/Si CPA)基底的SERS特性,开展miRNA-106a的检测,具有特异性好和灵敏度高的优势,检测范围为1 fmol·L~(-1)～100 pmol·L~(-1),检测极限为0.917 fmol·L~(-1)。此外,与实时荧光定量多聚核苷酸链式反应(RT-qPCR)方法相比,不仅检测结果一致,而且基于SERS光谱技术的检测方法具有更高的灵敏度。     

基于液相色谱-串联质谱法的肉类特征肽段鉴别及掺假测定

建立了液相色谱-串联质谱技术鉴别肉类特征肽段及定量检测羊肉中常见外源肉掺假的方法。样品经蛋白质提取、胰蛋白酶水解和固相萃取小柱净化后,利用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱（UPLC-Q/Exactive-HRMS）和Proteinpilot软件,实现蛋白质和多肽的鉴定;再通过基本局部比对搜索工具（BLAST）与Uniprot数据库对比分析,筛选出羊肉、鸭肉、猪肉和鸡肉的20个物种特征性多肽标志物;最后利用高效液相色谱-三重四极杆质谱（UPLC-QqQ-MS）系统对羊肉、鸭肉、猪肉和鸡肉的特征性多肽进行了验证和多反应监测（MRM）定量研究。将鸭肉、猪肉和鸡肉分别按照质量分数为1%、5%、10%、20%、50%的比例掺加到羊肉中,得到鸭肉最低掺假检出限为0.25%、猪肉最低掺假检出限为0.17%、鸡肉最低掺假检出限为0.10%。     

AC susceptibility of magnetic markers in suspension for liquid phase immunoassay

AC susceptibility of magnetic markers in solution was studied for biosensor applications. First, frequency dependence of the susceptibility was measured, and size distribution of the markers was estimated by analyzing the experimental result with the so-called singular value decomposition (SVD) method. The size distribution estimated with the magnetic measurement agreed with that obtained from conventional optical measurement. Next, susceptibility measurement was applied to the liquid-phase immunoassay without bound/free (B/F) separation. We performed the detection of biotin-coated polymer beads in suspension using avidin-coated magnetic markers. Changes of the susceptibility and the size distribution caused by the binding reaction were shown.     

Mass spectrometric characterization of lipid-modified peptides for the analysis of acylated proteins

The analysis of acylated proteins by mass spectrometry (MS) has largely been overshadowed in proteomics by the analysis of glycosylated and phosphorylated proteins; however, lipid modifications on proteins are proving to be of increasing importance in biomedical research. In order to identify the marker ions and/or neutral loss fragments that are produced upon collision-induced dissociation, providing a means to identify the common lipid modifications on proteins, peptides containing an N-terminally myristoylated glycine, a palmitoylated cysteine and a farnesylated cysteine were chemically synthesized. Matrix-assisted laser desorption/ionization time-of-flight time-of-flight (MALDI-TOF-TOF), electrospray ionization quadrupole time-of-flight (ESI Q-TOF), and electrospray ionization hybrid triple-quadrupole/linear ion trap (ESI QqQ(LIT)) mass spectrometers were used for the analysis. The peptide containing the N-terminally myristoylated glycine, upon CID, produced the characteristic fragments a1 (240.4 Th) and b1 (268.4 Th) ions as well as a low-intensity neutral loss of 210 Da (C14H26O). The peptides containing a farnesylated cysteine residue fragmented to produce a marker ion at a m/z of 205 Th (C15H25) as well as other intense farnesyl fragment ions, and a neutral loss of 204 Da (C15H24). The peptides containing a palmitoylated cysteine moiety generated neutral losses of 238 Da (C16H30O) and 272 Da (C16H32OS); however, no marker ions were produced. The neutral losses were more prominent in the MALDI-TOF-TOF spectra, whereas the marker ions were more abundant in the ESI QqQ(LIT) and Q-TOF mass spectra.     

Nonribosomal cyclic peptides: specific markers of fungal infections

Some cyclic peptides and depsipeptides are synthesized in microorganisms by large multienzymes called nonribosomal peptide synthetases. The structures of peptide products originating in this way are complex and diverse and are microorganism-specific. This work proposes the use of fungal cyclic peptides and depsipeptides as extremely specific markers of fungal infections. Since a reliable molecular tool for diagnosing fungal infections at an early stage is still missing, we present mass spectrometry as a new, modern, broadband (with respect to fungal strain) and specific tool for clinical mycologists. More than 40 different fungal species can be rapidly characterized according to specific families of cyclic peptides, and in some cases, a particular fungal strain can be identified on the basis of its cyclopeptide profile. This paper is also aimed at initiating discussion on the biological role of these secondary metabolites, especially of those synthesized by medically important strains. Proven cytotoxic, anti-inflammatory or immunosuppressive activities of some cyclic peptides indicate that these molecules may contribute to the synergistic array of fungal virulence factors and support microbial invasion during fungal infection. In addition to an overview on recent mass spectrometric protocols for cyclic peptide sequencing, the structures of new peptides from Paecilomyces and Pseudallescheria are presented.     

玉米作物多光谱图像精准分割与叶绿素诊断方法研究

为了快速获取大田玉米作物长势信息, 基于多光谱图像开展了大田玉米叶绿素指标的非破坏性诊断研究。应用自主开发的2-CCD多光谱图像感知系统, 在田间采集玉米冠层可见光［Blue(B), Green(G), Red(R);400～700 nm］和近红外(Near-infrared: NIR, 760～1 000 nm)图像, 并使用SPAD同步测量样本叶绿素指标。采集后图像经自适应平滑滤波处理后, 进行图像玉米植株提取。为了选择最优算法实现玉米植株与杂草、土壤背景的分割, 首先比较了最大类间方差(OTSU)分割算法和局部阈值处理分割算法, 选取了基于局部统计的可变阈值处理方法对玉米NIR图像进行初步分割, 进而采用区域标记算法进行精细分割, 分割准确率达95.59%。将分割结果应用于玉米植株可见光图像R, G, B各通道, 从而实现了玉米植株多光谱图像中可见光图像的整体分割。基于分割后R, G, B和NIR四个通道的玉米冠层图像, 提取了各通道图像灰度均值(ANIR, ARed, AGreen和ABlue)并计算了归一化植被指数(NDVI)、比值植被指数(RVI)和绿色归一化植被指数(NDGI)作为光谱特征参数, 建立了玉米冠层叶绿素指标诊断的偏最小二乘法回归模型。结果表明, 建模R2达0.596 0, 预测R2达0.568 5, 该方法通过玉米多光谱图像特征参数评估叶片叶绿素含量, 可为大田玉米长势监测提供支持。     

航空机电作动器健康管理验证系统研究

针对健康管理智能算法与推理模型性能的验证与评估需求,设计了一种航空机电作动器健康管理验证系统;通过分析航空机电作动器的故障机理,提出了电机、控制器、齿轮减速器的数学建模方法和对应的主要失效模式的故障注入方法,设计了验证系统硬件平台、基于组件模式的通用软件运行环境及支持实时显示功能的验证与评估模块;仿真结果表明,该平台能够支持各种健康管理算法与系统模型的加载和调用,并能够实现验证与评估信息的在线显示。     

Spectrophotofluorimetric Study of the Interaction between Trazodone Hydrochloride and Bovine Serum Albumin

Abstract

The binding of trazodone hydrochloride (TZH), an antidepressant drug, to bovine serum albumin (BSA) has been investigated by fluorescence spectroscopic analysis. The fluorescence emission of BSA (λ_em=350 nm) was quenched by TZH while that of this ligand was enhanced (λ_em=443 nm). The spectral behavior was consistent with the static quenching mechanism, and the apparent binding constant, K _a (1.05×10⁴ l mol^?1) as well as binding site number, n (～1), were estimated. Thermodynamic parameters obtained from the measured data at different temperatures showed that the binding of TZH to BSA involved predominantly hydrophobic interactions as well as smaller contributions from electrostatic forces. Phenylbutazone and ibuprofen were utilized as competitive markers for sites I and II, respectively, in the interaction of TZH with BSA. This competitive displacement procedure indicated that the likely binding was site I, i.e., subdomain IIA, and this was supported by the observation that up to 50% of this site marker, phenylbutazone, could be exchanged with TZH whilst only a few percent of ibuprofen were so affected.