Purification and renaturation of recombinant human lymphotoxin (tumour necrosis factor beta) expressed in Escherichia coli as inclusion bodies

High level expression of recombinant human tumour necrosis factor β (rh TNF-β) in Escherichia coli results in the formation of two portions of protein, namely soluble active protein and insoluble protein which is inactive and aggregates in the form of inclusion bodies (IBs). In this study, a procedure for purification and renaturation of rh TNF-β from inclusion bodies has been designed and verified experimentally with a product purity of more than 90% and a recovery of about 30%. The procedure includes washing of IBs with specific wash buffer (Triton X-100/EDTA/lysozyme/PMSF), their solubilization with 8 mol dm^?3 alkaline urea, purification with ion-exchange columns, refolding with renaturation buffer and finally concentration and desalination with an ultrafiltration membrane. The characteristics of the renatured protein were identical with those of purified protein from the soluble fraction as demonstrated by (1) SDS-PAGE, (2) cytotoxic activity on mouse L929 cells, (3) N-terminal amino acid sequence, and (4) gel filtration chromatography.     

Application of artificial neural networks to the analysis of two‐dimensional fluorescence spectra in recombinant E coli fermentation processes

A two‐dimensional (2D) spectrofluorometer was used to monitor various fermentation processes with recombinant E coli for the production of 5‐aminolevulinic acid (ALA). The whole fluorescence spectral data obtained during a process were analyzed using artificial neural networks, ie self‐organizing map (SOM) and feedforward backpropagation neural network (BPNN). The SOM‐based classification of the whole spectral data has made it possible to qualitatively associate some process parameters with the normalized weights and variances, and to select some useful combinations of excitation and emission wavelengths. Based on the classified fluorescence spectra a supervised BPNN algorithm was used to predict some of the process parameters. It was also shown that the BPNN models could elucidate some sections of the process's performance, eg forecasting the process's performance. Copyright © 2005 Society of Chemical Industry     

Distribution and Fate of Escherichia coli in Lake Michigan Following Contamination with Urban Stormwater and Combined Sewer Overflows

Escherichia coli distribution and persistence in nearshore Lake Michigan were assessed following heavy rains and sanitary sewer overflow (SSO) and combined sewer overflow (CSO) events over a 5-year period, including an 18-day period following 25.4 cm of rainfall in which intensive studies were conducted following multiple CSO and SSO events. E. coli levels in the Milwaukee estuary and harbor following SSO and CSO events ranged from 10⁴ to nearly 10⁵ CFU/100 mL, which were significantly higher (p ≤ 0.05) than levels following rainfall alone. Sites outside of the breakwall but within the contamination plume (e.g., within 2 km of the harbor) were an order of magnitude lower. Locations 2–5 km from the harbor ranged from below detection limits, of < 1 to 5 CFU/100 mL. E. coli levels corrected for dilution based on specific conductivity measurements were lower than what would be expected for loss due to dilution alone, suggesting a combination of die-off and dilution, were responsible for the rapid disappearance of these organisms outside of the harbor. E. coli and fecal coliforms measured concurrently demonstrated that fecal coliforms could be recovered longer than E. coli in the open waters of the lake. E. coli isolated directly from sewage treatment plant influent were found to have a marked increase in antibiotic resistance traits for ten antibiotics commonly used in the human population compared with isolates from two animal sources of fecal pollution. However, E. coli obtained from sewage impacted water (n = 2,513) and from stormwater impacted water (n = 1,465) collected the previous year when there were no sewage overflows, were found to have no significant difference (p < 0.05) in the frequency of resistance when comparing the two conditions. E. coli survival characteristics and population dynamics are most likely influenced by multiple factors in complex systems such as the watershed/estuarine/lake environments of the Great Lakes.     

TFT有源矩阵α－Si∶H薄膜PECVD淀积工艺研究

分析了α－Ｓｉ∶Ｈ薄膜的质量和厚度对α－Ｓｉ∶ＨＴＦＴ关键性指标的影响，深入、详细地讨论了其ＰＥＣＶＤ淀积工艺，并在实验的基础上确定了最佳淀积工艺参数，从而获得了高性能的７５ｍｍ３７２×２７６像素α－Ｓｉ∶ＨＴＦＴ有源矩阵     

提高E^2PROM中NMOS管源漏穿通电压的实验研究

为了提高Ｅ＾２ＰＲＯＭ中Ｎ管源漏穿通电压，用实验的方法对制造工艺进行了研究。结果表明，高能量注入是提高ＶＰＴ的有效手段，但受到ｐｎ结击穿的限制，只适用于低区短沟Ｎ管；ＤＤＤ工艺大幅度高ＶＰＴ，但ｐｎ结击穿电压低于２０Ｖ，不能应用于高压ＭＯＳ管；采用适量的防穿通注入和适当增大沟道长度为最理想的工艺途径。     

Sex pheromone specificity as a reproductive isolating mechanism among the sibling speciesArchips argyrospilus andA. mortuanus and other sympatric tortricine moths (Lepidoptera: Tortricidae)

MaleArchips argyrospilus (Walker) were maximally attracted to a 60404200 admixture of (Z)-11-tetradecenyl, (E)-11-tetradecenyl, (Z)-9-tetradecenyl and dodecyl acetates. The identification of (Z)-9-tetradecenyl acetate, in addition to the other 3 components reported previously, is the first report of a 4-component pheromone system in moths.Archips mortuanus Kearfoot, a sibling species, was maximally attracted to a blend of the same components, but in a 90101 mix of the tetradecenyl acetates. Among a number of tortricine moths that co-occur on apples and other hosts in New York, British Columbia, and elsewhere, differences in diel rhythms of attraction and seasonal distribution are insufficient to maintain reproductive isolation. Although the attractant systems of these species often possess components in common, males are maximally attracted to a speciesspecific blend. In tortricine moths attraction specificity appears to be a paramount mechanism for species partitioning.     

高校体育教学中学生创新能力的培养

从创新能力和体育教学的关系、体育教学的特性及创造教育实施原则等方面入手,对高校体育教学中学生创新能力的培养进行分析,并阐述其基本方法和途径。     

ZrO2对CaO-SiO2系玻璃析晶的影响

通过DSC曲线研究了不同含量的ZrO2对CaO-SiO2系玻璃析晶的影响,根据JMA方程计算动力学参数(如析晶活化能E、频率因子ν、晶化指数n)。从计算结果可以看出,随着ZrO2含量的增加,析晶活化能E减少,而频率因子ν和晶化指数n增加。当ZrO2含量为7%时,E值最小:235.58kJ/mol;n值最大:4.70。说明ZrO2在CaO-SiO2玻璃体系是有效的晶核剂。     

民族传统体育项目引入高校体育课的研究

在高校开展少数民族传统优秀体育项目教学,不仅能使少数民族传统体育走向规范化、科学化,较好地展现学生的运动能力,培养民族文化情感,激发民族热情,更重要的是还能提高学生的身心健康和综合素质,有利于对民族传统体育的继承、发展和广泛地传播。     

基于FPGA的TDM over IP的设计

设计一种TDM over IP的技术系统。该系统用FPGA将E1数据包通过串并转换到千兆以太网数据包,经IP网传输到终端设备,再通过并串转换将以太网数据包转换成E1数据包完成传输。经过软件和硬件测试验证了此系统在千兆以太网上传输的可行性,系统没有出现误码,固有输出抖动是0.55UI,最大峰值抖动指标均满足ITU-G.823标准要求。