Selective monitoring of peptidase activities with synthetic polypeptide substrates and polyion-sensitive membrane electrode detection

A novel method to monitor specific peptidase activities in biological samples as complex as undiluted plasma/blood is described. The approach is based on the design of synthetic polypeptide substrates in which di- or triarginine sequences are linked to each other via one or more other amino acids recognized specifically by the peptidase to be determined. Detection of chymotrypsin and renin activities using synthetic substrates P4 (F-R-R-R-F-V-R-R-F-NH2) and P5 (R-R-R-L-L-R-R-L-L-R-R-R), respectively, serves to demonstrate the principles of this new assay system. A polyion-sensitive membrane electrode, prepared by doping polymer films with dinonylnaphthalene-sulfonate (DNNS), is shown to exhibit significant nonequilibrium electromotive force (EMF) responses toward these and other polycationic substrates at microgram/milliliter levels under physiological conditions. The same electrode, however, exhibits much smaller total EMF response toward the shorter fragments of the synthetic peptides generated by peptidase activity; hence, the addition of peptidase to a solution containing the synthetic substrate yields a change in electrode EMF response, the rate of which is proportional to the activity of peptidase present. Other synthetic polycationic peptides as well as natural polycationic peptides (e.g., protamine) that lack specific cleavage sites for chymotrypsin and renin, yet are detected by the DNNS-based membrane electrode, do not elicit any significant change in EMF response in the presence of the peptidases, confirming the feasibility and utility of the proposed bioanalytical method.     

Changes in mitochondrial calcium concentration during the cardiac contraction cycle

OBJECTIVE: The aim was to examine whether mitochondrial Ca2+ fluxes are high enough to change mitochondrial and cytosolic calcium concentration during the contraction cycle. METHODS: Isolated guinea pig ventricular myocytes were stimulated with paired voltage clamp pulses until contractions were maximal (2 mM [Ca2+]o, 36 degrees C). At defined times of diastole or systole, the cells were shock frozen. Electron-probe microanalysis measured the concentration of total calcium in mitochondria (sigma Ca(mito)) and surrounding cytosol (sigma Cac). Other experiments were performed to evaluate DNP sensitive mitochondrial Ca2+ uptake from depolarisation induced [Ca2+]c transients (K5indo-1 fluorescence). RESULTS: At end of diastole, sigma Ca(mito) was 446 mumol.litre-1. During systole, sigma Ca(mito) increased with a 20 ms delay. A peak sigma Ca(mito) of 1050 mumol.litre-1 was measured 40 ms after start of systole, while 95 ms after start of systole sigma Ca(mito) had fallen to 530 mumol.litre-1. From the changes in sigma Ca(mito) the rates of net mitochondrial Ca2+ flux were estimated at 100 nmol.s-1 x mg-1 protein for Ca2+ influx and 36 nmol.s-1 x mg-1 protein for Ca2+ egress. Decay of sigma Ca(mito) was coupled to a rise in sigma Na(mito). sigma Cl(mito) and sigma K(mito) rose and fell in parallel with sigma Ca(mito), suggesting Ca2+ activation of mitochondrial anion and cation channels. Activation of the non-specific permeability can be excluded. Block of mitochondrial Ca2+ uptake with DNP (100 microM) or FCCP (10 microM) increased the amplitude of the [Ca2+]c transients for 1-3 min by about 50%; evaluation of mitochondrial Ca2+ uptake from DNP sensitive difference signals, however, was hampered by sequestration of mitochondrial Ca2+ into the sarcoplasmic reticulum. CONCLUSIONS: Mitochondrial calcium content changes during each individual contraction cycle; a substantial amount of calcium is taken up during the systole and released during later systole and diastole.     

真空熔炼高压气体雾化制粉技术及设备

介绍了气体雾化粉的品种、用途及工业化生产。还介绍了真空熔炼-高压气体雾化制粉所需的设备。     

汛期采金船的缆绳牵拉

本文介绍了采金船渡汛时的停泊方向及采金船在洪水中可能产生拖曳力的估算方法，为了防止采金船汛期发生溜船事故，建议采用缆绳牵拉固船的措施，以保障采金船的汛期安全。     

多功能滚齿机的数字控制系统

本文叙述了以ＳＴＤＰＣ／ＸＴ（或ＰＣ／ＡＴ）机为主要核心构成的滚齿机数字控制系统的结构，软件设计和特点，系统采用混合语言编程，极大地提高了软件设计的灵活性与快速性，也分析了ＣＮＣ系统的加工过程，提出了加工中的软件预处理控制方法，并给出了轴伺服控制的并行算法分析与实现。     

面向大规模定制的虚拟设计研究

综述了大规模定制的范围经济性和面向大规模定制的基本设计方法，并提出了一个虚拟设计框架，该框架改变了传统的设计模式，利用先进的设计方法学和仿真支撑技术为产品开发提供了最大柔性和敏捷性。同时，它提供了一个多学科合作的设计环境，用户直接参与进设计循环中。最后，结合新型剑杆织机的开发，展示了一个能够支持新产品快速成功开发的虚拟设计环境。     

安钢宽板坯连铸机达产实践

分析了安钢宽板坯连铸机在热试车期间出现的生产问题,总结出制约生产主要因素,制定针对性措施,使该连铸机在较短时间内实现达产创效.     

山药

山药是一种常用的中药，营养丰富，具有较高的药疗价值，现对山药的化学成分、药理作用、临床应用的研究进展作一介绍。