Direct gene transfer into Actinidia deliciosa protoplasts: analysis of transient expression of the CAT gene using TLC autoradiography and a GC-MS-based method

Chloramphenicol acetyl transferase (CAT) gene was used as a reporter gene to assess the conditions for polyethylene glycol (PEG)-mediated transfection of kiwifruit protoplasts. The effect of plasmid concentration and the presence of carrier DNA were each assessed by analysing CAT activity in transfected protoplasts using thin-layer chromatography (TLC) autoradiographic detection of acetylated chloramphenicol. A gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) non-radioactive method was developed for monitoring CAT gene activity. This method provides a high speed of analysis (30 min) and precise means of detecting acetylated products at the nanomolar level, enabling quantification at very low transfection rates. Using this method we optimized plasmid and PEG concentration and also assessed the effect of heat shock on transfection. The best CAT activity was obtained using 30% polyethylene glycol 4000 and by submitting protoplasts to heat shock (45 °C, 5 min) prior to transfection.     

A short history of nuclear activation analysis

The major developments in the field of nuclear activation analysis, from 1936 to 1989, are discussed. The developments are grouped into five consecutive time periods. The impact of various scientists on the development of the field in the first 35 years is also discussed.     

Enhanced ethylene production by primary roots of Zea mays L. in response to sub-ambient partial pressures of oxygen

Ethylene production by primary roots of 72–h-old intact seedlings of Zea mays L. cv. LG11 was studied under ambient and sub-ambient oxygen partial pressures (pO₂) using a gas flow-through system linked to a photoacoustic laser detector. Despite precautions to minimize physical perturbation to seedlings while setting-up, ethylene production in air was faster during the first 6h than later, in association with a small temporary swelling of the roots. When roots were switched from air (20–8kPa O₂) to 3 or 5kPa O₂ after 6h, ethylene production increased within 2—3 h. When, the roots were returned to air 16 h later, ethylene production decreased within 2—3 h. The presence of 10kPa CO₂ did not interfere with the effect of 3kPa O₂. Transferring roots from air to 12–5kPa did not change ethylene production, while a reduction to 1 kPa O₂ induced a small increase. The extra ethylene formed in 3 and 5 kPa O₂ was associated with plagiotropism, swelling, root hair production, and after 72 h, increased amounts of intercellular space (aerenchyma) in the root cortex. Root extension was also slowed down, but the pattern of response to oxygen shortage did not always match that of ethylene production. On return to air, subsequent growth patterns became normal within a few hours. In the complete absence of oxygen, no ethylene production was detected, even when anaerobic roots were returned to air after 16 h.     

采用Reichardt运动检测器和Boltzmann Machine神经网络的运动计算

建立了一个探讨灵长类视皮层从Ｖ１区到ＭＴ区的运动信息加工原理的计算模型,这个过程的突出特征是视觉运动信息经过了从局部检测进步到整体感知。模型的第一层由用于抽提运动模式的局部速度以及结构性质的Ｒｅｉｃｈａｒｄｔ运动检测器组成,进一步的加工是通过Ｂｏｌｔｚｍａｎｎ Ｍａｃｈｉｎｅ神经网络来实现的。这种网络的学习算法具有局部更新的显著性质,在学习阶段,网络不断地修改联结权重以形成对于记录在网络的显单元上     

Molecular Characteristics of Bovine Serum Albumin-Dextran Conjugates

Bovine serum albumin (BSA)-dextran conjugates were prepared by using the Maillard reaction; depending on the ratio of dextran to BSA used, about 0.5–1 mol of dextran could be bound to 1 mol of native BSA. SDS–PAGE patterns revealed that BSA and dextran had been covalently bonded. Structural analyses by fluorescence spectroscopy and circular dichroism indicated that the BSA surface in each conjugate was covered with dextran without any great disruption of the native conformation. The conjugates could be grouped into two fractions on the basis of the weight-average molecular mass measured: the main fraction at 1.95–2.35×10⁵ g/mol and a less-abundant fraction with aggregates greater than 1.50×10⁶ g/mol. High-performance size-exclusion chromatography in conjunction with multi-angle laser light scattering detection revealed that the BSA-dextran conjugates prepared by using the Maillard reaction had various molar masses and radii.     

城市群新冠疫情时空分布格局与分异机制的地理探测

自从2019年12月湖北武汉爆发新型冠状病毒（COVID-19）以来,疫情在全国范围内迅速传播并蔓延。城市群既是我国新型城镇化主体特定的空间组织形态,也是相互连接的城市网,集合了相当数量不同性质、类型和等级规模的城市,城市群经济发展程度较高、人口密度大,交通发达,是疫情传染扩散-关联的重点区域,因而跨区域感染风险高,联防联控难度大。以疫情集中地区城市群为研究对象,通过核密度分析聚焦重点研究的城市群,从多层空间尺度对疫情在数量、人口迁移和空间分布上进行测度,利用空间自相关分析疫情分布特征,运用地理探测器方法客观地测度城市群疫情发展的差异性主导因素,从景观格局中挖掘相关主导因子,为生态、安全的空间规划和治理提供科学依据和思路。结果显示：①全国范围感染人数核密度高值区的空间分布与长江中游城市群和三大沿海城市群范围耦合,首位核心城市确诊感染数在城市群中占比均较高,城市群疫情扩散分布呈现典型的由核心城市向外辐射的特征,湖北地区迁出的人数对迁往地区的疫情形势构成一定程度的影响;②全国范围的COVID-19感染人数和感染增长率分布存在显著空间集聚特征,城市群范围感染人数的空间聚集性从长江中游城市群向其他城市群依次减弱;③地理探测器识别出人口密度、城乡建设、交通、卫生、科技、生态绿地为疫情的主导因素,并且交互后所产生的共同作用增加了对疫情的解释力;④生态与建设用地景观格局对疫情的解释能力较强,其中建设用地的聚集性和生态用地的蔓延性为主要因子。     

基于地理探测器的中国农业生态效率时空分异及其影响因素

研究农业生态效率的时空分异及其影响因素对实现中国农业生态高质量发展具有重要意义。基于2000—2018年中国30个省/区/市的面板数据,采用超效率SBM模型测算省际农业生态效率,在时间序列、空间可视化及趋势面分析揭示农业生态效率时空演变规律的基础上,进一步利用地理探测器模型识别影响农业生态效率空间分异的主导因子及其交互作用。结果表明: 2000—2018年,中国农业生态效率整体呈现稳定上升的趋势,但仍然处于较低水平,存在较大提升空间;中国农业生态效率具有显著的空间分异特征,总体上呈现出东西部地区较高、而中部地区较低的空间分布格局;中国农业生态效率的空间分异受到农业资源禀赋、社会经济、自然生态环境等多种因素的影响,不同因子对农业生态效率空间分异的影响存在明显差异且因子间交互作用会增强其空间分异。综上,要关注农业生态效率时空分异的主导因子,并注重区域间的协同合作,以实现农业的高质量发展。     

Characterization of the mouse ClC-K1/Barttin chloride channel

Several Cl⁻ channels have been described in the native renal tubule, but their correspondence with ClC-K1 and ClC-K2 channels (orthologs of human ClC-Ka and ClC-Kb), which play a major role in transcellular Cl⁻ absorption in the kidney, has yet to be established. This is partly because investigation of heterologous expression has involved rat or human ClC-K models, whereas characterization of the native renal tubule has been done in mice. Here, we investigate the electrophysiological properties of mouse ClC-K1 channels heterologously expressed in Xenopus laevis oocytes and in HEK293 cells with or without their accessory Barttin subunit. Current amplitudes and plasma membrane insertion of mouse ClC-K1 were enhanced by Barttin. External basic pH or elevated calcium stimulated currents followed the anion permeability sequence Cl⁻ > Br⁻ > NO₃⁻ > I⁻. Single-channel recordings revealed a unit conductance of ~ 40 pS. Channel activity in cell-attached patches increased with membrane depolarization (voltage for half-maximal activation: ~ − 65 mV). Insertion of the V166E mutation, which introduces a glutamate in mouse ClC-K1, which is crucial for channel gating, reduced the unit conductance to ~ 20 pS. This mutation shifted the depolarizing voltage for half-maximal channel activation to ~ + 25 mV. The unit conductance and voltage dependence of wild-type and V166E ClC-K1 were not affected by Barttin. Owing to their strikingly similar properties, we propose that the ClC-K1/Barttin complex is the molecular substrate of a chloride channel previously detected in the mouse thick ascending limb (Paulais et al., J Membr. Biol, 1990, 113:253–260).     

福建省主要入侵植物空间分异及其影响因素

在实地调查数据的基础上,本研究结合GIS空间分析技术和地理探测器模型,分析福建省入侵植物空间分布情况,以及地理与社会环境因子及其交互作用对入侵植物分布的影响.结果表明:福建省共记录入侵植物82种,其中,优势科为菊科,小蓬草、藿香蓟和空心莲子草出现频次最高.沿海区域的入侵植物物种数量多于内陆区域,福州和厦门为福建省外来入侵植物的两大热点地区.入侵植物在不同海拔均有分布,但入侵植物的种类随着海拔的升高总体呈下降趋势.地理探测器分析显示,自然环境因子中降水和社会经济因子中路网密度、人口密度是入侵植物空间分布的主要影响因子.各因子的空间交互作用会正向影响入侵植物的空间分布,这反映出入侵植物空间分布影响要素的复杂性.综上,将地理探测器应用到入侵植物研究领域是可行的,筛选出的环境指示因子可用于监测福建省入侵植物的适生区,从而为采取有效的防控措施提供科学依据.