Recombinant dengue virus type 3 envelope domain III protein from Escherichia coli

Dengue is a public health problem of global significance for which there is neither an effective antiviral therapy nor a preventive vaccine. The envelope protein of dengue virus is the major antigen to elicit neutralizing antibody response and protective immunity in hosts. Optimization of culture media was carried out for enhanced production of recombinant dengue virus type 3 envelope domain III (rDen 3 EDIII) protein in E. coli. Further, batch and fed-batch cultivation process were also developed in optimized medium. After fed-batch cultivation, the dry cell weight was about 22.80 g/L of culture. The rDen 3 EDIII protein was purified using immobilized metal affinity chromatography. This process produced ～649 mg of purified rDen 3 EDIII protein per liter of culture. The purity of the protein was determined by SDS-PAGE analysis and the reactivity was checked by Western blotting as well as ELISA. These results show that the purified protein may be used for the dengue diagnosis or further prophylactic studies for dengue infection.     

Modeling the impact of periodic bottlenecks, unidirectional mutation, and observational error in experimental evolution

Antibiotic resistant bacteria are a constant threat in the battle against infectious diseases. One strategy for reducing their effect is to temporarily discontinue the use of certain antibiotics in the hope that in the absence of the antibiotic the resistant strains will be replaced by the sensitive strains. An experiment where this strategy is employed in vitro [5] produces data which showed a slow accumulation of sensitive mutants. Here we propose a mathematical model and statistical analysis to explain this data.The stochastic model elucidates the trend and error structure of the data. It provides a guide for developing future sampling strategies, and provides a framework for long term predictions of the effects of discontinuing specific antibiotics on the dynamics of resistant bacterial populations.This Research is part of the Initiative in Bioinformatics and Evolutionary Studies (IBEST) at the University of Idaho. Funding was provided by NSF EPSCoR EPS-0080935, NSF EPSCoR, EPS-0132626, and NIH NCRR grant NIH NCRR- 20RR016448. Paul Joyce is also funded by NSF DEB-0089756, and NSF DMS-0072198.     

非线性动力学在脑电信号分析中的应用

EEG是由大脑产生的非线性时间序列,体现出混沌行为。近年来迅速发展的非线性动力学理论为脑电信号分析开创了一个新的领域。本文综述了近年来非线性动力学在脑电信号研究中（睡眠阶段,麻醉深度,认知过程,精神分裂,痴呆及癫痫）的进展,以期对脑神经动力学有更好的理解。     

Optimal Production of Poly-γ-glutamic Acid by Metabolically Engineered Escherichia coli

Metabolically-engineered Escherichia coli strains were developed by cloning poly-γ-glutamic acid (γ-PGA) biosynthesis genes, consisting of pgsB, pgsC and pgsA, from Bacillus subtilis The metabolic and regulatory pathways of γ-PGA biosynthesis in E. coli were analyzed by DNA microarray. The inducible trc promoter and a constitutive promoter (P_HCE) derived from the d-amino acid aminotransferase (D-AAT) gene of Geobacillus toebii were employed. The constitutive HCE promoter was more efficient than inducible trc promoter for the expression of γ-PGA biosynthesis genes. DNA microarray analysis showed that the expression levels of several NtrC family genes, glnA, glnK, glnG, yhdX, yhdY, yhdZ, amtB, nac, argT and cbl were up-regulated and sucA, B, C, D genes were down-regulated. When (NH₄)₂SO₄ was added at 40 g/l into the feeding solution, the final γ-PGA concentration reached 3.7 g/l in the fed-batch culture of recombinant E. coli/pCOpgs.     

Migration, trapping and local dynamics of whiteflies (Homoptera: Aleyrodidae)

1 The ability to quantify whitefly migration provides a tool that can contribute to an improved understanding of the epidemic development of whitefly‐transmitted viruses. 2 In an attempt to develop a protocol for estimating whitefly immigration and emigration rates in an annual crop, new traps and sampling devices were tested in the field and models for population dynamics were developed. 3 An estimate of immigration rate was derived from the growth of a natural population of Trialeurodes vaporariorum (Westwood) in the beginning of a crop cycle before offspring of immigrants contributed to population growth. 4 A model for changes in whitefly density during an entire bean (Phaseolus vulgaris L.) crop cycle, including an immigration parameter, was also developed. 5 Non‐attractant window traps surrounding an annual field crop were assumed to intercept whiteflies immigrating into and emigrating away from the crop. Captures on these traps could not categorically be identified as immigrants or emigrants, but the cumulated captures nevertheless explained 66% of the variation in population density found within the field. Hence, window traps may be used as an efficient and reliable alternative to yellow sticky traps, aspirator methods and leaf‐turn methods, etc., for estimating whitefly densities in field crops.     

A Polished and Reinforced Thinned-skull Window for Long-term Imaging of the Mouse Brain

In vivo imaging of cortical function requires optical access to the brain without disruption of the intracranial environment. We present a method to form a polished and reinforced thinned skull (PoRTS) window in the mouse skull that spans several millimeters in diameter and is stable for months. The skull is thinned to 10 to 15 μm in thickness with a hand held drill to achieve optical clarity, and is then overlaid with cyanoacrylate glue and a cover glass to: 1) provide rigidity, 2) inhibit bone regrowth and 3) reduce light scattering from irregularities on the bone surface. Since the skull is not breached, any inflammation that could affect the process being studied is greatly reduced. Imaging depths of up to 250 μm below the cortical surface can be achieved using two-photon laser scanning microscopy. This window is well suited to study cerebral blood flow and cellular function in both anesthetized and awake preparations. It further offers the opportunity to manipulate cell activity using optogenetics or to disrupt blood flow in targeted vessels by irradiation of circulating photosensitizers.     

不同强度及时间窗骨骼肌缺血后处理对兔心肌的保护作用

目的:通过比较不同强度及时间窗骨骼肌缺血后处理对兔缺血/再灌注心肌的保护效能,试图寻找最佳强度和时间窗的骨骼肌缺血后处理方案。方法:健康新西兰大白兔42只(雄性)随机分为7组(n=6):①假手术组(Sham)、②缺血对照组(CON)、③标准骨骼肌后处理组(SP)、④延迟6min骨骼肌后处理组(6M-DSP)、⑤延迟1 min骨骼肌后处理组(1M-DSP)、⑥骨骼肌后处理加强组(SSP)、⑦骨骼肌后处理减弱组(WSP)。以开胸结扎冠状动脉左室支固定部位方法制作缺血/再灌注模型,以游离并夹闭双侧腹股沟髂外动脉固定位置方法造成骨骼肌缺血,再灌注末以TTC法确定心肌梗死范围,并分别于心肌缺血前、后及再灌注1 h、2 h,以生化法测定血清肌酸激酶(CK)及乳酸脱氢酶(LDH)水平。结果:和CON组相比,1M-DSP组心肌梗死重量比及面积比分别下降了42.32%及42.68%、SP组分别下降了49.97%及43.78%、SSP组分别下降了48.36%及48.86%,(P均<0.05),但三组之间相比,心梗范围未见明显差异;而6M-DSP、WSP组与CON组相比未见心肌保护作用;肌酸激酶(CK)的水平和梗死范围变化趋势一致。结论:兔在心肌缺血/再灌注之前完成骨骼肌5 min缺血/1 min再灌注1次循环的缺血后处理,可以起到明显的心肌保护作用。     

Removal of surfactant dodecylbenzenesulfonate by consecutive use of ozonation and biodegradation

Successful surfactant removal from wastewater is often limited by the high concentration of the surfactant. The use of advanced oxidation processes can be the key to aid biological treatment of water containing high amounts of surfactants. The present study analyzes the biodegradation of the anionic surfactant sodium dodecylbenzenesulfonate (SDBS) and the effects of its combination with ozonation. SDBS pre‐ozonation favors the metabolism by microorganisms. Experimental results indicate that the application of a concentration of up to 60 μM of ozone for 60 min, prior to contact with microorganisms, increases the percentage of SDBS removed by biodegradation alone. These results demonstrate that the removal of SDBS and of the total organic carbon is increased by the consecutive use of ozonation and biodegradation.     

政府投资审批制度下园林建设程序中的问题探讨

政府使用公共财政投资建设公益性的工程,需遵守一定的审批与管理流程。由于缺少园林工程投资估算指标体系,以及园林工程特有的艺术性,使审批流程与设计流程产生了一些矛盾。审批过程中,评审者关注投资的合理性,设计者关注园林美学观感,两者的不同立场导致对部分园林绿地建设内容必要性的分歧。为了在保证建设资金投入合理性前提下最大限度地实现园林绿地的完整功能与艺术水平,对有关的问题进行梳理,提出平衡设计与评审的原则,并强调在审批与建设过程中各方面调整思维方式,进行换位思考的必要性。     

不同手术方式对腰椎间盘突出症治疗的效果比较分析

目的:探讨应用微创技术治疗腰椎间盘突出症的疗效.方法:对我院脊柱骨科自2005年l1月～2010年10月收治的128例腰椎间盘突出症患者应用不同手术方式进行治疗,其中应用APLD( automated percutaneous lumbar discectomy)治疗单纯腰椎间盘突出症48例;应用MED(microendoscopic discectomy)治疗复杂型腰椎间盘突出症42例;应用传统后路椎板开窗技术治疗单纯腰椎间盘突出症38例;比较各组手术时间、术中出血、术后住院时间、疗效及并发症.术后均随访8个月～3年,观察复发情况.结果:三组不同手术方式手术时间及优良率比较差异无统计学意义(P＞0.05),APLD及MED组术中出血量及术后住院时间与传统手术组比较,差异有显著性(P＜0.05);术后随访疼痛全部缓解,无复发.结论:在严格掌握适应症的基础上,采用微创技术和采用传统手术治疗腰椎间盘突出症疗效相当,但微创技术创伤小、术中出血量少,术后住院日短,恢复快,优于传统手术.