Histidine affinity chromatography-based methodology for the simultaneous isolation of Escherichia coli small and ribosomal RNA

Research on RNA has led to many important biological discoveries and the improvement of therapeutic technologies. In particular, there is a great focus on small RNA and ribosomal RNA owing to their key functions in the cell, which make them excellent therapeutic targets. Although the study of these RNA classes is progressing, some limitations have been found regarding the use of suitable techniques that are able to produce and isolate biologically competent and chemically stable RNA. To address this, we have developed a novel histidine affinity chromatography-based isolation methodology for small and ribosomal RNA molecules. The new procedure involves three main steps: (1) cell lysis with guanidinium buffer, (2) RNA primary isolation with ammonium sulfate precipitation and (3) histidine affinity chromatography to specifically purify small RNA and ribosomal RNA from other Escherichia coli impurities (genomic DNA and proteins). The RNA quality assessment revealed that both RNA species were obtained with a high recovery, integrity and purity. The potential of this method to achieve a reproducible RNA isolation with appropriate quality has been demonstrated and it should have broad application in the structural, biophysical and biomedical investigation of systems involving RNA components.     

Detection of PCR products amplified from DNA of epizootic pathogens using magnetic nanoparticles and SERS

Here we present a novel approach using surface‐enhanced Raman scattering (SERS) spectroscopy for the sequence‐specific detection of DNA utilizing magnetic nanoparticles (MNPs) for the enrichment of the target molecules. To achieve fast and efficient binding of longer DNA strands, e.g. PCR products, the hybridization procedure is performed in solution. To further purify and enrich the DNA strands of interest, MNPs are used for their separation. Following the binding of the target DNA, a dye‐modified, short synthetic ssDNA is hybridized, which serves as label for the SERS detection. The SERS spectra are used to identify the bound molecules. The applicability of this approach was first tested with short synthetic oligonucleotides to evaluate its specificity. Afterward, the system was applied to detect PCR products amplified from DNA of specific agents of epizootic diseases. Sequences of the bacterium Mycoplasma mycoides subspecies mycoides small colony type (MmmSC), causing contagious bovine pleuropneumonia (CBPP) were used as PCR targets. To demonstrate the multiplexing capability of SERS, the simultaneous detection of three different PCR products labeled with three dyes was performed. Copyright © 2010 John Wiley & Sons, Ltd.     

有机小分子空穴传输材料取代基效应的理论研究

在B3LYP/6-31G(d)水平下对N,N′-二苯基-N,N′-二(1-萘基)-1,1′-联苯-4,4′-二胺(以下简称NTD)及其衍生物(以下简称NTDs)的中性态和离子态结构进行优化,得到各物质的重组能并进行比较。结果显示,NTD分子的化学修饰对重组能λ变化趋势的影响是电子诱导效应和共轭效应共同作用的结果：(1)吸电子诱导效应会引起NTD分子重组能的升高;多重取代位的吸电子效应对分子重组能影响具有相互抵消的作用,不具有简单加和性;给电子诱导效应会引起NTD分子重组能的降低;(2)o位、m位的正共轭效应使体系重组能剧烈升高,共轭作用在o位、m位取代时占主导作用;p位的正共轭效应使体系重组能降低;(3)大范围的共轭π键可能引起所取代的苯环电子分布发生根本性变化,导致分子构型随之发生剧烈变化,从而重组能急剧升高。     

Morphological operator for infrared dim small target enhancement using dilation and erosion through structuring element construction

To effectively enhance infrared dim small target, a new morphological operator is proposed by constructing two structuring elements based on the properties of target regions and their surrounding background. By utilizing the two constructed structuring elements, the proposed morphological operator uses dilation and erosion to enhance the target regions and suppresses the surrounding background, which directly achieves infrared small target enhancement. Also, the proposed method is simplified by using flat structuring elements. Experimental results show that the proposed method can effectively enhance infrared dim small target embedded in clutter background.     

Propagation characteristics of flattened Gaussian beams through a misaligned optical system with a misaligned annular aperture

The approximate analytical formula for flattened Gaussian beams through a misaligned optical system with a misaligned annular aperture was derived by the extended Huygens–Fresnel principle. Some numerical simulations are illustrated to the effects on the propagation of flattened Gaussian beams by the misaligned annular aperture. To compare the difference between annular apertured system and circular apertured system, the circular apertured system is also studied. The results show that angle misalignments and lateral displacements of aperture create asymmetrical average intensity distribution at receiving plane z = 500. The effects on intensity distribution by angle misalignments of annular aperture were small. In annular aperture case, the smooth of intensity distribution was worse with escalating obscure ratio ? in near-field; the side-lobes increased and the central lobe decreased with escalating obscure ratio ? in far-field. At receiving plane z = 500: for circular aperture, the side-lobes decreased, even to be neglected, with the increasing of truncation parameter δ; for annular aperture, the side-lobes increased with the increasing of truncation parameter δ. In addition, it is found that the aligned thin lens can fix asymmetry of intensity distribution which was caused by the misaligned annular aperture.     

Investigation on diffraction characteristics of continuous terahertz beam limited by hard-edge apertures

Hard-edge apertures are common devices in optical systems. The light beam would be diffracted while transmitting through an aperture. The diffraction characteristics of terahertz (THz) beam passing through two hard-edge apertures have been simulated based on the Fresnel diffraction integral function, and an approximate parallel beam has been obtained. The effect on the diffraction spot has been compared and analyzed by changing the aperture diameter and distance between the two apertures and the diffraction plane. Then the experimental results were presented, which basically accorded with the numerical results. It can be inferred that the diameter of THz beam diminishes and the divergence angle can be neglected after the restriction of two apertures. The method is preferably available in some experiments which require small beam diameter and divergence angle.     

Energy calibration issues in nuclear resonant vibrational spectroscopy: observing small spectral shifts and making fast calibrations

The conventional energy calibration for nuclear resonant vibrational spectroscopy (NRVS) is usually long. Meanwhile, taking NRVS samples out of the cryostat increases the chance of sample damage, which makes it impossible to carry out an energy calibration during one NRVS measurement. In this study, by manipulating the 14.4 keV beam through the main measurement chamber without moving out the NRVS sample, two alternative calibration procedures have been proposed and established: (i) an in situ calibration procedure, which measures the main NRVS sample at stage A and the calibration sample at stage B simultaneously, and calibrates the energies for observing extremely small spectral shifts; for example, the 0.3 meV energy shift between the 100%‐⁵⁷Fe‐enriched [Fe₄S₄Cl₄]⁼ and 10%‐⁵⁷Fe and 90%‐⁵⁴Fe labeled [Fe₄S₄Cl₄]⁼ has been well resolved; (ii) a quick‐switching energy calibration procedure, which reduces each calibration time from 3–4 h to about 30 min. Although the quick‐switching calibration is not in situ, it is suitable for normal NRVS measurements.     

利用小尺寸电荷耦合器件实现数字全息高分辨成像

为了在降低数字全息显微成像系统成本的同时实现高分辨成像,对像面数字全息显微术的记录与再现过程进行了理论分析,结合系统的点扩散函数,对该系统的横向分辨率进行了分析. 得到了如下结论：像面数字全息显微系统的横向分辨率对电荷耦合器件（CCD）光敏面尺寸变化不敏感;对于常见的CCD 器件,其像元尺寸的变化对该系统的横向分辨率影响甚微. 此外,对像面数字全息显微系统的成像特点进行了分析,结果表明：利用像面数字全息系统可以实现物体信息的完整记录与再现,其成像分辨率及像质优于预放大数字全息系统. 利用搭建的数字全息实验记录系统,从强度及位相两方面对理论分析结果进行了验证,实验结果表明了理论分析的正确性. 关键词： 像面数字全息术 横向分辨率 点扩散函数 小尺寸电荷耦合器件     

高分辨率合成孔径雷达图像的Gamma分布下最大后验概率降斑算法

针对传统的Gamma分布下最大后验概率降斑算法不能有效保留均匀区域的点目标, 不能有效保留弱边缘以及不能有效滤除强边缘区域的斑点等问题, 提出了基于第二类统计量的先验参数估计的高分辨率合成孔径雷达图像Gamma 分布下最大后验概率降斑算法. 使用Mellin卷积和斑点的乘性模型, Gamma先验分布的参数可由观察图像的前两阶对数累积量精确估计.所提算法具有解析的滤波输出, 便于实现.农田和城区的高分辨率合成孔径雷达图像的降斑实验表明, 与传统的Gamma分布下最大后验概率降斑算法相比, 所提算法既能有效保留均匀区域的点目标, 又能有效保留弱边缘, 还能有效滤除强边缘区域的斑点. 关键词： 高分辨率合成孔径雷达图像 Gamma分布下最大后验概率降斑算法 第二类统计量 对数累积量     

目标粗糙对合成孔径激光雷达回波的退相干效应

本文研究了目标表面粗糙对回波信号探测的影响,通过蒙特卡罗方法建立一维和二维目标高斯随机粗糙面模型,并对目标回波信号的探测过程进行仿真,研究目标表面粗糙对中频信号的影响.还进行了粗糙面和光滑面两组光外差探测实验,验证目标粗糙对合成孔径激光雷达回波信号严重的“退相干”效应.同时,采用数字波前分析仪对探测器处本振光和信号光波前分别进行检测,检测结果与仿真一致,证明粗糙面回波相位畸变严重.文中研究结果对设计光源参数、接收系统参数以及评估系统作用距离奠定基础,为合成孔径激光雷达系统设计提供定量参考.