Bimetallic ferrocenyl derivatives: Molecular second order hyperpolarizabilities and second harmonic generation

The ground and excited states dipole moments and the second order hyperpolarizabilities of a series of oxazolinyl-ferrocenyl derivatives have been measured. After complexation of the oxazoline group with a second metal ion, the new bimetallic complexes show an increased hyperpolarizability, as determined by the EFISHG technique. A copolymer of the same bimetallic complex with methyl methacrylate has been synthesized, and the second order susceptibility has been measured by second harmonic generation at 1.06µm.     

洪水脉冲理论及其应用

洪水脉冲理论是继河流连续统理论之后的第4个河流生态理论,主要阐述洪水脉冲驱动下,河流与其洪泛区之间的横向水力联系对河流洪泛区系统进程的重要性,强调洪水脉冲的重要性及河流洪泛区系统的整体性。本文论述洪水脉冲理论的概念模型、主要观点及其在河流洪泛区系统的应用进展。洪水脉冲理论的应用主要体现在三方面在洪水塑造河流洪泛区系统地貌进程方面,侧重于河流形态的“主导径流”研究,成果不断提升;在生物地球化学循环上,以对氮的研究为主,提出洪水脉冲是河流洪泛区新陈代谢及生物地球化学循环的重要驱动力;在生物进程上,集中在对大型植物和以水鸟和鱼类为代表的动物研究上。文中提出要提高河滨生物量,并重建全局的生物多样性,应提倡人为的洪水脉冲。对于干旱半干旱区湿地水鸟的保护核心是洪水发生频率和淹没范围对水鸟繁殖及避难的作用,以及水在湿地景观中的分配,这一点对于中国松嫩平原西部干旱半干旱区湿地水鸟的保护具有重要意义。洪水脉冲理论的提出是基于对热带亚马逊河的观察实验,随着其在其它类型区域的推广应用,除洪水脉冲以外的其它因素的作用突现出来,人类干扰洪水脉冲的生态效应亦愈来愈显现。因此,有必要加强洪水脉冲与其它环境因子的耦合作用,及这种作用在人类干扰活动下,导致的河流洪泛区系统进程变化的研究,提出更具有普适性的洪水脉冲理论。     

Molecular engineering of epoxide hydrolase and its application to asymmetric and enantioconvergent hydrolysis

Safety and regulatory issues favor increasing use of enantiopure compounds in pharmaceuticals. Enantiopure epoxides and diols are valuable intermediates in organic synthesis for the production of optically active pharmaceuticals. Enantiopure epoxide can be prepared using epoxide hydrolase (EH)-catalyzed asymmetric hydrolysis of its racemate. Enantioconvergent hydrolysis of racemic epoxides by EHs possessing complementary enantioselectivity and regioselectivity can lead to the formation of enantiopure vicinal diols with high yield. EHs are cofactor-independent and easy-to-use catalysts. EHs will attract much attention as commercial biocatalysts for the preparation of enantiopure epoxides and diols. In this paper, recent progress in molecular engineering of EHs is reviewed. Some examples and prospects of asymmetric and enantioconvergent hydrolysis reactions are discussed as supplements to molecular engineering to improve EH performance.     

Glycolytic pathway as an ATP generation system and its application to the production of glutathione and NADP

Efficient ATP generation is required to produce glutathione and NADP. Hence, the generation of ATP was investigated using the glycolytic pathway of yeast. Saccharomyces cerevisiae cells immobilized using polyacrylamide gel generated ATP from adenosine, consuming glucose and converting it to ethanol and carbon dioxide. Under optimal conditions, the ATP-generating activity of immobilized yeast cells was 7.0 μmol h^?1 ml^?1 gel. A column packed with these immobilized yeast cells was used for continuous ATP generation. The half-life of the column was 19 days at a space velocity of (SV) 0.3 h^?1 at 30°C. The properties of glutathione- and NADP-producing reactions coupled with the ATP-generating reaction were investigated. Escherichia coli cells with glutathione synthesizing activity and Brevibacterium ammoniagenes cells with NAD kinase activity were immobilized in a polyacrylamide gel lattice. Under optimal conditions, the immobilized E. coli cells and immobilized B. ammoniagenes cells produced glutathione and NADP at the rates of 2.1 and 0.65 μmol h^?1 ml^?1 gel, respectively, adding ATP to the reaction mixture. In order to produce glutathione and NADP economically and efficiently, the glutathione- and NADP-producing reactions were finally coupled with the ATP-generating reaction catalysed by immobilized S. cerevisiae cells. To compare the productivities of glutathione and NADP, and to compare the efficiency of ATP utilization for the production of these two compounds, the two reactor systems, co-immobilized cell system and mixed immobilized cell system, were designed. As a result, these two compounds were also found to be produced by these two kinds of reactor systems. Using the data obtained, the feasibility and properties of ATP generation by immobilized yeast cells are discussed in terms of the production of glutathione and NADP.     

Sensitivity of second harmonic generation from styryl dyes to transmembrane potential

In this article we present results from the simultaneous nonlinear (second harmonic generation and two-photon excitation fluorescence) imaging and voltage clamping of living cells. Specifically, we determine the sensitivity to transmembrane potential of second harmonic generation by ANEP-chromophore styryl dyes as a function of excitation wavelength and dye structure. We have measured second harmonic sensitivities of up to 43% per 100 mV, more than a factor of four better than the nominal voltage sensitivity of the dyes under "one-photon" fluorescence. We find a dependence of voltage sensitivity on excitation wavelength that is consistent with a two-photon resonance, and there is a significant dependence of voltage sensitivity on the structure of the nonchromophore portion of the dyes.     

A probabilist model of the relations between pollen and climate and its application to 80 European annual spectra

The relation between pollen and climate is commonly computed by regressing the climatical factor. The disadvantage of this method is that it does not respect the ordinal and intermittent nature of field data. This paper overcomes the artefacts created by this problem by using a probabilist calibration, that quantifies the ecological linkage between a taxon T and a factor with a general parameter, the probability PROX for an abundance A being confined near the rank F of the factor. Confining simulates the effect of the factor upon the concentration of presences and ordering of abundances, and calibrates the climatical behaviour of a taxon with the set of PROX for all possible pairs (A,F). It summarizes a behaviour with the probable position of each abundance A in the range of the factor. Calibration was applied to 130 pollen taxa observed in a network of 80 standardized annual aeropollinic spectra. Spectra were mostly from France, the rest being from a transect stretching from Sweden to Algeria. Spectra were characterized by the values of 10 climatic factors, as well as the presence and abundance of 130 pollen taxa. The influence of geographical climate differences upon pollen content in the atmosphere was quantified by comparing the spectra. Pairs from different localities but the same year were compared. The reliability of indicator taxa was tested by estimating the climate in the 80 spectra using calibration. For all the taxa observed in a spectrum, the envelope of confinings generally followed an unimodal gradient, whose mode was the probable position of the spectrum. Reliability of the estimate was measured by its accuracy, being the agreement between estimates and measures; and by its stability, being the agreement between two estimates from the same climate according to two different flora (the two halves of a spectrum for instance). Average accuracy was 72%, and average stability 87%.     

Characterization of a novel laccase from the isolated Coltricia perennis and its application to detoxification of biomass

A highly efficient laccase-producing fungus was isolated from soil and identified as Coltricia perennis SKU0322 by its morphology and by comparison of its internal transcribed spacer (ITS) rDNA gene sequence. Extracellular laccase (Cplac) from C. perennis was purified to homogeneity by anion-exchange and gel filtration chromatography. Cplac is a monomeric glycoprotein with 12% carbohydrate content and a molecular mass of 66 kDa determined by polyacrylamide-gel electrophoresis. Ultraviolet-visible absorption spectroscopy observed type 1 and type 3 copper signals from Cplac. The enzyme acted optimally at pH 3–4 and 75 °C. Its optimal activity was with 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonate) (ABTS), it also oxidized various lignin-related phenols. The enzyme was characterized as a multi-copper blue laccase by its substrate specificity and internal amino acid sequence. It showed a higher catalytic efficiency towards ABTS (k_cat/K_m = 18.5 s^?1 μM^?1) and 2,6-dimethoxyphenol (k_cat/K_m = 13.9 s^?1 μM^?1) than any other reported laccase. Its high stability and catalytic efficiency suggest its suitability for industrial applications: it detoxified phenolic compounds in acid-pretreated rice straw and enhanced saccharification yield.     

分子模拟方法及其在分子生物学中的应用

常用的分子模拟方法有 :量子力学法、分子力学方法、蒙特卡洛法和分子动力学法。四种方法各有优势 ,共同成为分子模拟的组成部分。综述了分子模拟法在分子生物学中的应用 ,最后介绍了分子模拟的发展方向 ,并预测了其未来的发展趋势。常用的分子模拟方法有 :量子力学法、分子力学方法、蒙特卡洛法和分子动力学法。四种方法各有优势 ,共同成为分子模拟的组成部分。综述了分子模拟法在分子生物学中的应用 ,最后介绍了分子模拟的发展方向 ,并预测了其未来的发展趋势。     

A modified Zeeman model for producing HRV signals and its application to ECG signal generation

Developing a mathematical model for the artificial generation of electrocardiogram (ECG) signals is a subject that has been widely investigated. One of the challenges is to generate ECG signals with a wide range of waveforms, power spectra and variations in heart rate variability (HRV)--all of which are important indexes of human heart functions. In this paper we present a comprehensive model for generating such artificial ECG signals. We incorporate into our model the effects of respiratory sinus arrhythmia, Mayer waves and the important very low-frequency component in the power spectrum of HRV. We use a new modified Zeeman model for generating the time series for HRV, and a single cycle of ECG is produced by using a simple neural network. The importance of the work is the model's ability to produce artificial ECG signals that resemble experimental recordings under various physiological conditions. As such the model provides a useful tool to simulate and analyse the main characteristics of ECG, such as its power spectrum and HRV under different conditions. Potential applications of this model include using the generated ECG as a flexible signal source to assess the effectiveness of a diagnostic ECG signal-processing device.     

Top-down elasticity analysis and its application to energy metabolism in isolated mitochondria and intact cells

This paper reviews top-down elasticity analysis, which is a subset of metabolic control analysis. Top-down elasticity analysis provides a systematic yet simple experimental method to identify all the primary sites of action of an effector in complex systems and to distinguish them from all the secondary, indirect, sites of action. In the top-down approach, the complex system (for example, a mitochondrion, cell, organ or organism) is first conceptually divided into a small number of blocks of reactions interconnected by one or more metabolic intermediates. By changing the concentration of one intermediate when all others are held constant and measuring the fluxes through each block of reactions, the overall kinetic response of each block to each intermediate can be established. The concentrations of intermediates can be changed by adding new branches to the system or by manipulating the activities of blocks of reactions whose kinetics are not under investigation. To determine how much an effector alters the overall kinetics of a block of reactions, the overall kinetic response of the block to the intermediate is remeasured in the presence of the effector. Blocks that contain significant primary sites of action will display altered kinetics; blocks that change rate only because of secondary alterations in the concentrations of other metabolites will not. If desired, this elasticity analysis can be repeated with the primary target blocks subdivided into simpler blocks so that the primary sites of action can be defined with more and more precision until, with sufficient subdivision, they are mapped onto individual kinetic steps. Top-down elasticity analysis has been used to identify the targets of effectors of oxygen consumption in mitochondria, hepatocytes and thymocytes. Effectors include poisons such as cadmium and hormones such as tri-iodothyronine. However, the method is more general than this; in principle it can be applied to any metabolic or other steady-state system.     

An in-library ligation strategy and its application in CRISPR/Cas9 screening of high-order gRNA combinations

Simultaneous targeting multiple genes is a big advantage of CRISPR (clustered regularly interspaced short palindromic repeats) genome editing but challenging to achieve in CRISPR screening. The crosstalk among genes or gene products is a common and fundamental mechanism to ensure cellular stability and functional diversity. However, the screening approach to map high-order gene combinations to the interesting phenotype is still lacking. Here, we developed a universal in-library ligation strategy and applied it to generate multiplexed CRISPR library, which could perturb four pre-designed targets in a cell. We conducted in vivo CRISPR screening for potential guide RNA (gRNA) combinations inducing anti-tumor immune responses. Simultaneously disturbing a combination of three checkpoints in CD8+ T cells was demonstrated to be more effective than disturbing Pdcd1 only for T cell activation in the tumor environment. This study developed a novel in-library ligation strategy to facilitate the multiplexed CRISPR screening, which could extend our ability to explore the combinatorial outcomes from coordinated gene behaviors.     

High-efficiency system for the construction of adenovirus vectors and its application to the generation of representative adenovirus-based cDNA expression libraries

We here describe a convenient system for the production of recombinant adenovirus vectors and its use for the construction of a representative adenovirus-based cDNA expression library. The system is based on direct site-specific insertion of transgene cassettes into a replicating donor virus. The transgene is inserted into a donor plasmid containing the viral 5' inverted terminal repeat, the complete viral packaging signal, and a single loxP site. The plasmid is then transfected into a Cre recombinase-expressing packaging cell line that has been infected with a donor virus containing a partially deleted packaging signal flanked by loxP sites. Cre recombinase, by two steps of action, sequentially catalyzes the generation of a nonpackageable donor virus acceptor substrate and the generation of the desired recombinant adenovirus vector. Due to its growth impairment, residual donor virus can efficiently be counterselected during amplification of the recombinant adenovirus vector. By using this adenovirus construction system, a plasmid-based human liver cDNA library was converted by a single step into an adenovirus-based cDNA expression library with about 10(6) independent adenovirus clones. The high-titer purified library was shown to contain about 44% of full-length cDNAs with an average insert size of 1.3 kb. cDNAs of a gene expressed at a high level (human alpha(1)-antitrypsin) and a gene expressed at a relatively low level (human coagulation factor IX) in human liver were isolated from the adenovirus-based library using an enzyme-linked immunosorbent assay-based screening procedure.     

Spectrum of the nonstationary electromyographic signal modelled with integral pulse frequency modulation and its application to estimating neural drive information

The spectrum of nonstationary electromyographic signal (EMG) is investigated, from which the error for neural drive information estimation from nonstationary EMG is studied in terms of signal-to-noise ratio (SNR), in analytical, numerical simulation, and experimental work. The signal refers to the neural drive information embedded within the nonstationary EMG, and noise refers to other portions of EMG that induce error in the estimation. The analytical expressions for the SNRs of force-modulated EMG with both single and multiple motor units (MU) are derived based on a sinusoidal integral pulse frequency modulation (IPFM) model. It is shown that the previously developed SNR expressions for stationary (unmodulated) EMG are special cases of the formulas presented here. The SNR results obtained from numerical simulated EMG agree very well with the analytical result. Results from nonstationary (modulated) surface EMG obtained from seven subjects also match the analytical and simulation results reasonably well. The results obtained from this work establish an analytical framework in studying and estimating the neural drive information contained in the EMG in the context of anisotonic and isometric contractions. Through the analytical study, the effects of different physiological parameters are identified, thus providing theoretical guidelines for developing advanced signal processing methods for nonstationary EMG in applications such as prosthesis control.     

水稻高产形成的分子生态学基础及其应用研究

介绍了10多年来在水稻产量形成的发育遗传及其分子生态学基础的研究成果,分析了传统上应用比较生理学研究作物高产形成的优点与不足,强调应积极引进其它学科的研究内容及其方法,深化对作物栽培科学问题的认识,提出了根据作物不同发育阶段的基因作用规律及其性质,研究制定相应措施。及时调控那些受显性遗传效应机制和加性效应基因与环境互作同向表达的性状是实现高产稳产的技术关键,认为这是作物遗传生态栽培学的主要特色与技术创新,并就作物栽培向分子栽培学的发展及其前景作了初步探讨。     

一株新分离的云芝栓孔菌Z-1应用于木质素降解及染料脱色

木质素是一种非结晶性的复杂三维网状酚类高分子聚合物,被认为是木质纤维素生物质抗降解的天然屏障.探索并开发高效降解木质素的微生物资源已成为近些年来的研究焦点.本研究对新分离的一株具有潜在木质素降解能力的菌株Z-1开展了系列研究.首先通过形态学和系统发育分析将菌株Z-1鉴定为云芝栓孔菌Trametes versicolor...