Characterization of N-terminal processing of group VIA phospholipase A<Subscript>2</Subscript> and of potential cleavage sites of amyloid precursor protein constructs by automated identification of signature peptides in LC/MS/MS analyses of proteolytic digests

Dysregulation of proteolytic processing of the amyloid precursor protein (APP) contributes to the pathogenesis of Alzheimer's Disease, and the Group VIA phospholipase A(2) (iPLA(2)beta) is the dominant PLA(2) enzyme in the central nervous system and is subject to regulatory proteolytic processing. We have identified novel N-terminal variants of iPLA(2)beta and previously unrecognized proteolysis sites in APP constructs with a C-terminal 6-myc tag by automated identification of signature peptides in LC/MS/MS analyses of proteolytic digests. We have developed a Signature-Discovery (SD) program to characterize protein isoforms by identifying signature peptides that arise from proteolytic processing in vivo. This program analyzes MS/MS data from LC analyses of proteolytic digests of protein mixtures that can include incompletely resolved components in biological samples. This reduces requirements for purification and thereby minimizes artifactual modifications during sample processing. A new algorithm to generate the theoretical signature peptide set and to calculate similarity scores between predicted and observed mass spectra has been tested and optimized with model proteins. The program has been applied to the identification of variants of proteins of biological interest, including APP cleavage products and iPLA(2)beta, and such applications demonstrate the utility of this approach.     

Comparison of one-, two-, and three-dimensional iron phosphates containing ethylenediamine

A new two-dimensional (2d) iron phosphate, (C₂N₂H₁₀)Fe₂O(PO₄)₂, has been synthesized under hydrothermal conditions in the system of FeCl₃-H₃PO₄-C₂N₂H₈-H₂O. The crystal data is: space group P2₁/c, a=10.670(1) Å, b=10.897(1) Å, c=9.918(1) Å, β=105.632(1)°, Z=4. The layered structure consists of double sheet layers, of composition Fe₂O(PO₄)₂, built from FeO₅ trigonal bipyramids and PO₄ tetrahedra. The amine holds the layers together via H-bonding. The study of the magnetic properties reveals two magnetic transitions at 160 and 30 K with spin-glass-like behavior below 160 K. By varying the hydrothermal conditions, three other iron phosphates were synthesized: the one-dimensional (1d) (C₂N₂H₁₀)Fe(HPO₄)₂(OH)·H₂O, the 2d (C₂N₂H₁₀)Fe₂(PO₄)₂(OH)₂, and the three-dimensional (3d) (C₂N₂H₁₀)₂Fe₄O(PO₄)₄·H₂O. The 1d compound can be used as the starting reagent in the synthesis of both the 2d compound and the 3d lipscombite Fe₃(PO₄)₂(OH)₂ due to the similar building blocks in their structures. In the 3d phosphate (C₂N₂H₁₀)₂Fe₄O(PO₄)₄·H₂O, manganese can substitute for half of the iron atoms. Magnetic study shows ordering transitions at about 30 K, however, manganese substitution depresses the magnetic ordering temperature.     

Application of an integrated microchip system with capillary array electrophoresis to optimization of enzymatic reactions

In this work, a combination of complementary metal-oxide semiconductor (CMOS) microchip system with capillary array electrophoresis (CAE) is demonstrated as a system for optimizing conditions for enzymatic reaction. Dimethylacridinone (DDAO)-phosphate substrate and alkaline phosphatase conjugate were selected for the enzymatic reaction, which was applicable to the enzyme-linked immunosorbent assay (ELISA) technique. Laser-induced fluorometry with a miniature semiconductor laser was used to detect the enzymatic products. The speed of the enzymatic reaction between the DDAO-phosphate and the alkaline phosphatase conjugate was investigated as a function of reaction time. The microchip-CAE detection system could determine the pH condition and the concentration of enzyme that are suitable for rapid and low-cost analysis. This result shows the feasibility of using the microchip-CAE system for application to miniaturized screening systems.     

Metastasis-suppressor KAI1/CD82 induces homotypic aggregation of human prostate cancer cells through Src-dependent pathway

To investigate the functional role of KAI1/CD82, a metastasis suppressor for human prostate cancer, in the regulation of homotypic cell adhesion, we transfected KAI1 cDNA into DU 145 human prostate cancer cells and established stable transfectant clones with high KAI1/CD82 expression. The KAI1 transfectant cells exhibited significantly increased homotypic cell aggregation in comparison with the control transfectant cells. This aggregation of the KAI1 transfectants was further enhanced upon exposure to anti-CD82 antibody, suggesting that KAI1/CD82 may be involved in the intracellular signaling for the cell adhesion. Among several signal pathway inhibitors tested, PP1, an inhibitor of Src family kinases, significantly suppressed homotypic aggregation of the KAI1 transfectant cells. Ligation of KAI1/CD82 with anti-CD82 antibody increased endogenous Src kinase activity of the KAI1 transfectant cells. When different types of src expression constructs were retransfected into the KAI1-transfected DU 145 cells, kinase-negative mutant src transfectant cells exhibited much lower homotypic aggregation than the mock cells transfected with an empty vector. Moreover, homotypic aggregation of the mutant src transfectant cells was not enhanced by KAI1/CD82 ligation with anti- CD82 antibody. These results suggest that Src mediates the intracellular signaling pathway of KAI1/CD82 for the induction of homotypic adhesion of human prostate cancer cells.     

Earthshaker: A mobile rescue robot for emergencies and disasters through teleoperation and autonomous navigation

To deal with emergencies and disasters without rescue workers being exposed to dangerous environments, this paper presents a mobile rescue robot, Earthshaker. As a combination of a tracked chassis and a six-degree-of-freedom robotic arm, as well as miscellaneous sensors and controllers, Earthshaker is capable of traversing diverse terrains and fulfilling dexterous manipulation. Specifically, Earthshaker has a unique swing arm—dozer blade structure that can help clear up cumbersome obstacles and stabilize the robot on stairs, a multimodal teleoperation system that can adapt to different transmission conditions, a depth camera-aided robotic arm and gripper that can realize semiautonomous manipulation and a LiDAR aided base that can achieve autonomous navigation in unknown areas. It was these special systems that supported Earthshaker to win the first Advanced Technology & Engineering Challenge (A-TEC) championships, standing out of 40 robots from the world and showing the efficacy of system integration and the advanced control philosophy behind it.     

鄂尔多斯盆地长7储层致密油水平井CO2补充能量与吞吐参数优化

长7储层是鄂尔多斯盆地主要的致密区块,近一半的致密油分布其中。针对长7储层开采渗透率低、开采难度大、依靠弹性开采产能较低、能量衰竭较快等问题,通过数值模拟的方法,构建水平井分段压裂数值模拟模型,首先对比不同吞吐方式对油田产能的影响,优选出CO₂吞吐为首选的补充能量方式。其次对井网形式、井网参数及吞吐参数进行优化研究。最终优选出合适的吞吐方案,达到提高生产效率的目的。结果表明,该储层的最优注采吞吐参数为：周期注入量为2 673 t、注入压力为21 MPa、注入速度为90 t/d、焖井时间为10 d。研究对鄂尔多斯盆地长7储层提高采收率具有一定的参考意义。     

龙门山中北段重磁场特征与深部构造的关系

利用平均重力场与深部地壳测深资料估算了龙门山及邻区莫霍面的深度变化,表明龙门山地区正处于莫霍面由东南向西北的陡降带上。均衡重力异常计算结果显示该区为正异常。对航磁异常的向上延拓反映出茂汶杂岩体是“无根”的。此外还应用角度平均径向能谱法反演了该区深部磁性基岩的顶面与底面深度。结果表明,龙门山地区上地壳是由若干块体拼合而成的,川西拗陷带内的磁性界面呈现出向龙门山地腹下插的格局。     

Symmetric Skorohod topology onn-variable functions and hierarchical Markov properties ofn-parameter processes

Summary We introduce a new Skorohod topology for functions of several variables. Since ann-variable function may be viewed as a one-variable function with values in the set of (n–1)-variable functions, this topology is defined by induction from the classical Skorohod topology for one-variable functions. This allows us to define the notion of completen-parameter symmetric Markov processes: Such processes are, for any 1pn, rawp-parameter Markov processes (in the sense of our previous paper [17]) with values in the space of (n–p)-variable functions. We prove, for these processes and their Bochner subordinates, a maximal inequality which implies the continuity of additive functionals associated with finite energy measures. We finally present several important examples.