Profiling of generic anti-phosphopeptide antibodies and kinases with peptide microarrays using radioactive and fluorescence-based assays

Kinases represent one of the largest enzyme families and key regulatory proteins in the cell. Only a small subset of these enzymes has been characterised so far. We have prepared different types of phosphopeptide and peptide microarrays displaying peptides deduced from annotated human phosphorylation sites and cytoplasmic domains of all annotated human membrane proteins. This approach was enabled by fully-automated high throughput micro-scale synthesis of peptides by the SPOT technology combined with chemo-selective immobilisation on modified glass slides. The phosphopeptide microarrays displaying 2923 peptides in total have been used for the characterisation of commercially available generic anti-phosphopeptide antibodies. This enabled us to detect Abl kinase activity on a microarray with anti-phosphotyrosine antibodies yielding results comparable to those obtained from a radioactive assay. More than 13 000 peptides deposited on six glass slides were used to profile casein kinase 2 (CK2) using a radioactive assay, since no generic antibody for the reliable detection of serine or threonine phosphorylation could be identified. All previously identified substrates were detected in the microarray experiment. In order to confirm whether substrates on the microarray are substrates in solution phase assays, more than 700 peptides were synthesised and tested with CK2 in a solution phase assay. All substrates identified in the solution phase assay were also detected on the microarray.     

光学非球面凹面零件仿形加工方法的修正

从圆锥体靠模仿形法加工非球面光学零件的基本原理出发,指出加工凸非球面时无原理误差,但加工凹非球面时,只能用圆弧包络法加工。用实心圆锥体靠模时,必然存在误差。解决方法是减小工具的曲率半径,或增加一维数控进行补偿,或使用空心圆锥体仿形。实际应用中前两种方法只能减小误差,不能消除。而采用凹模仿形时,则不存在原理误差。     

复杂多层结构的二次离子质谱定量深度分析

讨论了用ＣＡＭＥＣＡＩＭＳ－４ｆ型二次离子质谱仪对ＡｌｘＧａ１－ｘＡｓ复杂多层结构定量深度分析的方法。采用ＣｓＭ＋技术，完成了无外部参考物质ＡｌｘＧａ１－ｘＡｓ基体成分定量分析，提出并实验验证了一种新的变溅射速率深度校准方法，详细讨论了复杂基体中对杂质进行定量分析时二次离子类型的选择，还尝试了在缺乏足够参考物质时杂质含量的实验估算。实现了二次离子质谱（ＳＩＭＳ）对复杂多层结构的定量分析，同时得到了主成分和杂质的定量深度分布，并保持了ＳＩＭＳ的优良深度分辨本领。     

基于多模态融合技术的用户画像方法

针对当前用户画像工作中各模态信息不能被充分利用的问题,提出一种跨模态学习思想,设计一种基于多模态融合的用户画像模型。首先利用Stacking集成方法,融合多种跨模态学习联合表示网络,对相应的模型组合进行学习,然后引入注意力机制,使得模型能够学习不同模态的表示对预测结果的贡献差异性。改进后的模型具有精心设计的网络结构和目标函数,能够生成一个由特征级融合和决策级融合组成的联合特征表示,从而可以合并不同模态的相关特征。在真实数据集上的实验结果表明,所提模型优于当前最好的基线方法。     

Reversed‐phase liquid chromatography with electrospray mass detection and 1H and 13C NMR characterization of new process‐related impurities,including forced degradants of Efavirenz: Related substances correlated to the synthetic pathway

In this study, a stability‐indicating reversed‐phase liquid chromatographic electrospray mass spectrometric method was developed and validated for the determination of process‐related impurities and forced degradants of Efavirenz in bulk drugs. Efavirenz was subjected to acid, alkaline hydrolysis, H₂O₂ oxidation, photolysis, and thermal stress. Significant degradation was observed during alkaline hydrolysis, and the degradants were isolated on a mass‐based purification system and characterized by high‐resolution mass spectrometry, positive electrospray ionization tandem mass spectrometry, and ¹H and ¹³C NMR spectroscopy. Accurate mass measurement and NMR spectroscopy revealed the possible structure of process‐related impurities and degradant under stress conditions. The acceptable separation was accomplished on Waters bondapak C₁₈ column (250 mm × 4.6 mm; 5 μm), using 5 mM ammonium acetate and acetonitrile as a mobile phase in a gradient elution mode at a flow rate of 1.0 mL/min. The eluents were monitored by diode array detector at 247 nm and quantitation limits were obtained in the range of 0.1–2.5 μg/mL for Efavirenz, degradants, and process‐related impurities. The liquid chromatography method was validated with respect to accuracy, precision, linearity, robustness, and limits of detection and quantification as per International Conference on Harmonization guidelines.     

Opto-thermal Moisture Content and Moisture Depth Profile Measurements in Organic Materials

Opto-thermal transient emission radiometry(OTTER) is a infrared remote sensing technique, which has been successfully used in in vivo skin moisture content and skin moisture depth profiling measurements. In present paper, we extend this moisture content measurement capability to analyze the moisture content of fruit (tomato. grape, etc. ) skins, and to study the relationship between fruits ripening process and their sur-face moisture and moisture depth profiles.     

Extended specificity studies of mRNA assays used to infer human organ tissues and body fluids

Messenger RNA (mRNA) profiling is a technique increasingly applied for the forensic identification of body fluids and skin. More recently, an mRNA‐based organ typing assay was developed which allows for the inference of brain, lung, liver, skeletal muscle, heart, kidney, and skin tissue. When applying this organ typing system in forensic casework for the presence of animal, rather than human, tissue is an alternative scenario to be proposed, for instance that bullets carry cell material from a hunting event. Even though mRNA profiling systems are commonly in silico designed to be primate specific, physical testing against other animal species is generally limited. In this study, human specificity of the organ tissue inferring system was assessed against organ tissue RNAs of various animals. Results confirm human specificity of the system, especially when utilizing interpretation rules considering multiple markers per cell type. Besides, we cross‐tested our organ and body fluid mRNA assays against the target types covered by the other assay. Marker expression in the nontarget organ tissues and body fluids was observed to a limited extent, which emphasizes the importance of involving the case‐specific context of the forensic samples in deciding which mRNA profiling assay to use and when for interpreting results.     

Experimental shift allowance in the deconvolution of SIMS depth profiles

We study the deconvolution of the secondary ion mass spectrometry (SIMS) depth profiles of silicon and gallium arsenide structures with doped thin layers. Special attention is paid to allowance for the instrumental shift of experimental SIMS depth profiles. This effect is taken into account by using Hofmann's mixing‐roughness‐information depth model to determine the depth resolution function. The ill‐posed inverse problem is solved in the Fourier space using the Tikhonov regularization method. The proposed deconvolution algorithm has been tested on various simulated and real structures. It is shown that the algorithm can improve the SIMS depth profiling relevancy and depth resolution. The implemented shift allowance method avoids significant systematic errors of determination of the near‐surface delta‐doped layer position. Copyright © 2013 John Wiley & Sons, Ltd.     

XPS in‐depth composition study on commercial monocrystalline silicon solar cells

From large‐scale production, two monocrystalline silicon solar cells of different quality, i.e. I_SC = 3.0 A (good cell) and I_SC = 1.6 A (bad cell), have been studied by XPS combined with 4 keV Ar⁺ depth profiling. Depth profiling was carried out through the anti‐reflection coating (TiO₂), the passivation layer (SiO₂) and up into the phosphorus‐doped silicon bulk. At the solar cell surface the elemental composition is similar for both cells, although the bad one presents slightly more carbon, phosphorus and lead but less silver than the good one. During profiling, carbon and silver could be followed by XPS. It was found that the carbon content is distinguishably higher in the bad cell than in the good one. Furthermore, it was found that silver atoms have not diffused in the same way in both cells. Only the good cell presents silver atoms up into the silicon bulk. Copyright © 2003 John Wiley & Sons, Ltd.