浙江西门岛海洋特别保护区大型底栖动物功能群特征及其与环境的关系

为研究浙江西门岛海洋特别保护区大型底栖动物功能群的变化规律及其与环境因子的关系,作者分别于2010年4月(春季)、11月(秋季),2011年8月(夏季)和2012年2月(冬季)进行了4个航次的大型底栖动物调查,共鉴定出大型底栖动物78种,根据其食性类型划分为浮游生物食者、植食者、肉食者、杂食者、碎屑食者5种功能群.各功能群平均密度从高到低依次为浮游生物食者＞肉食者＞植食者＞碎屑食者＞杂食者,平均生物量从高到低依次为浮游生物食者＞碎屑食者＞肉食者＞杂食者＞植食者.单因素方差分析结果表明,大型底栖动物各功能群的密度和生物量季节间均无显著性差异.典范对应分析结果表明,影响大型底栖动物功能群的主要环境因子包括温度、溶解氧、溶解态无机磷和表层沉积物的中值粒径,排序轴对功能群-环境关系的贡献率计算结果表明环境变量可以较好地解释功能群的变化情况.     

甲酸乙酯对松材线虫的熏蒸效果

【背景】在溴甲烷面临禁用的情形下,探讨新熏蒸剂甲酸乙酯对松材线虫的处理效果,可以科学地评估甲酸乙酯的使用前景。【方法】采用松材线虫分离液和带疫松木段,设置甲酸乙酯5个剂量梯度、5个处理温度和5个处理时间,测定其对松材线虫的毒力及CT值。【结果】在25℃下处理3、6、12、24、48h,甲酸乙酯对松材线虫的Lc50分别为2．63、1．60、0．99、0．41、0．20mg·L-1。温度对毒力有显著影响,在10～29℃,随温度升高,甲酸乙酯对松材线虫的毒力降低,19和29℃下,1．85mg·L-1甲酸乙酯处理松材线虫的死亡率分别为63％和100％。甲酸乙酯熏蒸12h内,能完全杀灭木段中的松材线虫。在23℃下处理6和12h,松材线虫死亡率达到99％时,甲酸乙酯的cr值分别为453．94和424．14mg·h-1·L-1。【结论与意义】甲酸乙酯可用于松材线虫的检疫处理。     

大鼠骨骼肌损伤后中性粒细胞、巨噬细胞和肌成纤维细胞数量分析研究

目的 研究大鼠骨骼肌损伤后中性粒细胞、巨噬细胞和肌成纤维细胞数量的变化情况,为今后骨骼肌损伤修复的病理学机制研究打下坚实的基础.方法 建立大鼠骨骼肌机械性损伤动物模型,随机分为伤后6h、12h、1d、3d、7d、10d、14d及正常对照组.应用免疫组织荧光染色和免疫组织化学染色,检测大鼠骨骼肌损伤后不同时间点中性粒细胞、巨噬细胞和肌成纤维细胞的数量.结果 伤后6h-12h,损伤区可见中性粒细胞和巨噬细胞浸润,中性粒细胞数量达到高峰.伤后1d,损伤区巨噬细胞数量急剧增加,迅速达到高峰,而中性粒细胞数量开始下降.伤后3d,中性粒细胞和巨噬细胞数量都显著下降.伤后7d,肌成纤维细胞开始出现.到伤后10d-14d,损伤区主要以肌成纤维细胞为主,偶见巨噬细胞.结论 大鼠骨骼肌损伤区中性粒细胞、巨噬细胞和肌成纤维细胞数量呈时间规律性变化,以期为骨骼肌损伤修复的病理学机制研究提供参考资料.     

A highly sensitive and selective optical sensor for Pb(2+) by using conjugated polymers and label-free oligonucleotides

The detection of Pb(2+) with DNA-based biosensor is usually susceptible to severe interference from Hg(2+) because of the T-Hg(2+)-T interaction between Hg(2+) and T residues. In this study, we developed a rapid, sensitive, selective and label-free sensor for the detection of Pb(2+) in the presence of Hg(2+) based on the Pb(2+)-induced G-quadruplex formation with cationic water-soluble conjugated polymer (PMNT) as a "polymeric stain" to transduce optical signal. We selected a specific sequence oligonucleotide, TBAA (5'-GGAAGGTGTGGAAGG-3'), which can form a G-quadruplex structure upon the addition of Pb(2+). This strategy provided a promising alternative to Pb(2+) determination in the presence of Hg(2+) instead of the universal masking agents of Hg(2+) (such as CN(-), SCN(-)). Based on this observation, a simple "mix-and-detect" optical sensor for the detection of Pb(2+) was proposed due to the distinguishable optical properties of PMNT-ssDNA and PMNT-(G-quadruplex) complexes. By this method, we could identify micromolar Pb(2+) concentrations within 5min even with the naked eye. Furthermore, the detection limit was improved to the nanomolar range by the fluorometric method. We also successfully utilized this biosensor for the determination of Pb(2+) in tap water samples.     

Immunosensor based on magnetic relaxation switch and biotin-streptavidin system for the detection of Kanamycin in milk

A rapid, sensitive, and simple immunosensor was developed for the detection of Kanamycin (KM) in milk. This immunosensor is based on magnetic relaxation switch (MRS) assay and biotin-streptavidin system (B-SA system). The target analyte (KM) competed with those on the surface of the superparamagnetic iron oxide (SPIO) nanoparticles and hence affected the formation of SPIO aggregates. The dispersed and aggregated states of SPIO can modulate the spin-spin relaxation time (T(2)) of the neighboring water molecule. T(2) was then changed as an effect of the target analyte. The B-SA system was used to amplify the SPIO binding, thus enhance the sensitivity. The detection working was 1.5 to 25.2ngmL(-1) and limit of detection (LOD) was determined to be 0.1ngmL(-1). The LOD of the immunosensor decreased tenfold, and its analysis time (45min) was much shorter than that of enzyme-linked immunosorbent assay (6h to 8h). The average recoveries of the KM at various spiking levels ranged from 80.2% to 85.6% with a relative standard deviation (RSD) below 4.0%. The results showed that the MRS immunosensor was a promising platform for the determination of small molecular residues because of its high sensitivity, specificity, homogeneity, and speed.     

One-step electrochemically co-assembled redox-active [Ru(bpy)(2)(tatp)](2+)-BSA-SWCNTs hybrid film for non-redox protein biosensors

A redox-active [Ru(bpy)(2)(tatp)](2+)-BSA-SWCNTs (bpy=2,2'-bipyridine, tatp=1,4,8,9-tetra-aza-triphenylene, BSA=bovine serum albumin, SWCNTs=single-walled carbon nanotubes) hybrid film is fabricated on an indium-tin oxide (ITO) electrode via one-step electrochemical co-assembly approach. BSA is inherently dispersive and therefore served as the linking mediator of SWCNTs, which facilitate the redox reactions of [Ru(bpy)(2)(tatp)](2+) employed as a reporter of BSA. The evidences from differential pulse voltammetry, cyclic voltammetry, scanning electron microscope, emission spectroscopy and fluorescence microscope reveal that the [Ru(bpy)(2)(tatp)](2+)-BSA-SWCNTs hybrid can be electrochemically co-assembled on the ITO electrode, showing two pairs of well-defined Ru(II)-based redox waves. Furthermore, the electrochemical co-assembly of the [Ru(bpy)(2)(tatp)](2+)-BSA-SWCNTs hybrid is found to be strongly dependent on the simultaneous presence of BSA and SWCNTs, indicating a good linear response to BSA in the range from 6 to 50mgL(-1). The results from this study provide an electrochemical co-assembly method for the development of non-redox protein biosensors.     

Needle δ13C and mobile carbohydrates in Pinus koraiensis in relation to decreased temperature and increased moisture along an elevational gradient in NE China

A tree’s crown interacts with atmospheric variables such as CO₂, temperature, and humidity. Physioecology of leaves/needles (e.g. δ¹³C, mobile carbohydrates, and nitrogen) is, therefore, strongly affected by microclimate in and surrounding a tree crown. To understand the physiological responses of leaves to changes in air temperature and moisture, we measured δ¹³C, soluble sugars, starch, and total nitrogen (N) concentrations in current year and 1-yr-old needles of Pinus koraiensis trees, and compared the growing season air temperature and relative humidity within and outside P. koraiensis crowns along an elevational gradient from 760 to 1,420 m a.s.l. on Changbai Mountain, NE China. Our results indicated that needle N and mobile carbohydrates concentrations, as well as needle δ¹³C values changed continuously with increasing elevation, corresponding to a continuous decrease in air temperature and an increase in relative humidity. Needle carbon and nitrogen status is highly significantly negatively correlated with temperature, but positively correlated with relative humidity. These results indicate that increases in air temperature in combination with decreases in relative humidity may result in lower levels of N and mobile carbohydrates in P. koraiensis trees, suggesting that future climate changes such as global warming and changes in precipitation patterns will directly influence the N and carbon physiology at P. koraiensis individual level, and indirectly affect the competitive ability, species composition, productivity and functioning at the stand and ecosystem level in NE China. Due to the relatively limited range of the transect (760–1,420 m) studied, further research is needed to explain whether the present results are applicable to scales across large elevational gradients.     

Triple mutated antibody scFv2F3 with high GPx activity: insights from MD, docking, MDFE, and MM-PBSA simulation

By combining computational design and site-directed mutagenesis, we have engineered a new catalytic ability into the antibody scFv2F3 by installing a catalytic triad (Trp²⁹–Sec⁵²–Gln⁷²). The resulting abzyme, Se-scFv2F3, exhibits a high glutathione peroxidase (GPx) activity, approaching the native enzyme activity. Activity assays and a systematic computational study were performed to investigate the effect of successive replacement of residues at positions 29, 52, and 72. The results revealed that an active site Ser⁵²/Sec substitution is critical for the GPx activity of Se-scFv2F3. In addition, Phe²⁹/Trp–Val⁷²/Gln mutations enhance the reaction rate via functional cooperation with Sec⁵². Molecular dynamics simulations showed that the designed catalytic triad is very stable and the conformational flexibility caused by Tyr¹⁰¹ occurs mainly in the loop of complementarity determining region 3. The docking studies illustrated the importance of this loop that favors the conformational shift of Tyr⁵⁴, Asn⁵⁵, and Gly⁵⁶ to stabilize substrate binding. Molecular dynamics free energy and molecular mechanics-Poisson Boltzmann surface area calculations estimated the pK _a shifts of the catalytic residue and the binding free energies of docked complexes, suggesting that dipole–dipole interactions among Trp²⁹–Sec⁵²–Gln⁷² lead to the change of free energy that promotes the residual catalytic activity and the substrate-binding capacity. The calculated results agree well with the experimental data, which should help to clarify why Se-scFv2F3 exhibits high catalytic efficiency.