使用电子断层技术对IgG1抗体逐个分子的三维重构与结构分析

抗体(antibody)又称免疫球蛋白(immunoglobulin,Ig),是人体免疫反应的重要参与者.了解抗体的结构和结构动态特征,是理解人体免疫作用机理、修复或提高免疫能力、定向设计抗体以治疗各种疾病的基础.本文以人体IgG1抗体为对象,综述了使用透射电子显微学方法研究IgG1抗体结构方向的最新进展.详细介绍了使用逐个分子的电子断层三维重构技术(individual-particle electron tomography,IPET)对抗体进行结构研究的方法,包括样品制备、图像处理和数据分析等.并描述了利用该技术,在研究抗体结合肽分子后的结构形变和通过收集不同构象来研究抗体动态结构特征方面所取得的阶段性成果.最后,对尚待解决的关键问题与该技术未来的发展方向进行了讨论与展望.     

Application of white cellulose acetate sheets on fossil wood investigation

Generally, transparent cellulose acetate sheets as a peel technique material are used in the identification of fossils, whereas white cellulose acetate sheets as a biochemistry technique material are applied in serum protein electrophoresis (SPE). Here we report the application of white cellulose acetate sheets for identifying a polished fossil wood from the Upper Mesozoic of West Liaoning, China. Based on the characters of transverse, radial, and tangential sections, the fossil wood is ascribed to a taxon of Protoglyptostroboxylon sp. Compared with transparent cellulose acetate sheets, white cellulose acetate sheets not only provide the similar information as that of the former, but also are more easily acquired in the Chinese market than the former. Because peel technique supported by white cellulose acetate sheets has the advantages of simple, time-saving, safe, reliable, and practical operation with lower material loss, it is a very good choice for the polished fossil wood investigation in labs, museums, geological parks, and handicraft shops. It is also a convenient approach to training students to learn the anatomic structure of fossil wood.     

Methods to characterize the structure of food powders – a review

Food powders can exist in amorphous, crystalline or mixed structure depending on the order of molecular arrangement in the powder particle matrices. In food production, the structure of powders has a greatly effect on their stability, functionality, and applicability. The undesirable structure of powders can be accidentally formed during production. Therefore, characterization of powder structure as well as quantification of amorphous–crystalline proportions presenting in the powders are essential to control the quality of products during storage and further processing. For these purposes, many analytical techniques with large differences in the degree of selectivity and sensitivity have been developed. In this review, differences in the structure of food powders are described with a focus being placed on applications of amorphous powders. Essentially, applicability of common analytical techniques including X-ray, microscopic, vapor adsorption, thermal, and spectroscopic approaches for quantitative and qualitative structural characterization of food powders is also discussed.     

CO2 emissions from an undrained tropical peatland: Interacting influences of temperature,shading and water table depth

Emission of CO₂ from tropical peatlands is an important component of the global carbon budget. Over days to months, these fluxes are largely controlled by water table depth. However, the diurnal cycle is less well understood, in part, because most measurements have been collected daily at midday. We used an automated chamber system to make hourly measurements of peat surface CO₂ emissions from chambers root‐cut to 30 cm. We then used these data to disentangle the relationship between temperature, water table and heterotrophic respiration (R_het). We made two central observations. First, we found strong diurnal cycles in CO₂ flux and near‐surface peat temperature (<10 cm depth), both peaking at midday. The magnitude of diurnal oscillations was strongly influenced by shading and water table depth, highlighting the limitations of relying on daytime measurements and/or a single correction factor to remove daytime bias in flux measurements. Second, we found mean daily R_het had a strong linear relationship to the depth of the water table, and under flooded conditions, R_het was small and constant. We used this relationship between R_het and water table depth to estimate carbon export from both R_het and dissolved organic carbon over the course of a year based on water table records. R_het dominates annual carbon export, demonstrating the potential for peatland drainage to increase regional CO₂ emissions. Finally, we discuss an apparent incompatibility between hourly and daily average observations of CO₂ flux, water table and temperature: water table and daily average flux data suggest that CO₂ is produced across the entire unsaturated peat profile, whereas temperature and hourly flux data appear to suggest that CO₂ fluxes are controlled by very near surface peat. We explore how temperature‐, moisture‐ and gas transport‐related mechanisms could cause mean CO₂ emissions to increase linearly with water table depth and also have a large diurnal cycle.     

Monitoring Protein Adsorption with Solid-state Nanopores

Solid-state nanopores have been used to perform measurements at the single-molecule level to examine the local structure and flexibility of nucleic acids ^1-6, the unfolding of proteins ⁷, and binding affinity of different ligands ⁸. By coupling these nanopores to the resistive-pulse technique ^9-12, such measurements can be done under a wide variety of conditions and without the need for labeling ³. In the resistive-pulse technique, an ionic salt solution is introduced on both sides of the nanopore. Therefore, ions are driven from one side of the chamber to the other by an applied transmembrane potential, resulting in a steady current. The partitioning of an analyte into the nanopore causes a well-defined deflection in this current, which can be analyzed to extract single-molecule information. Using this technique, the adsorption of single proteins to the nanopore walls can be monitored under a wide range of conditions ¹³. Protein adsorption is growing in importance, because as microfluidic devices shrink in size, the interaction of these systems with single proteins becomes a concern. This protocol describes a rapid assay for protein binding to nitride films, which can readily be extended to other thin films amenable to nanopore drilling, or to functionalized nitride surfaces. A variety of proteins may be explored under a wide range of solutions and denaturing conditions. Additionally, this protocol may be used to explore more basic problems using nanopore spectroscopy.     

Begehbare Freiflugvoliere für Störche, Ibisse und Enten im Opel-Zoo Kronberg: Walk-through aviary for storks, ibisses and ducks at the Opel-Zoo Kronberg

The new “Walk-through aviary” at the Opel-Zoo Kronberg, Germany, which was opened in September 2008 after a ten months construction period, is described. Covering around 1450m² and being up to 12.5 m high this aviary is one of the biggest in German zoos. The technical innovation is the first use of a stainless steel mash, which was permanent darkened using a chemical reaction. This method is named: INOX-Spectral ® - technique.The author explains the decision-making for the construction and gives an overview of the first year of keeping the birds in this aviary. The public reaction to this building, which is unlikely for the overall animal keeping attitude at the Opel-Zoo, is also covered by this article.     

Efficacy of Tower Medfly Eclosion Systems

A 16-repetition experiment was conducted to evaluate the performance of the “tower” system for eclosion of sterile medflies, Ceratitis capitata (Wiedemann). This system has now replaced the PARC system previously used in Florida S.I.T. programs. In addition to testing the efficacy of these eclosion systems, as compared to the PARC system, quality control was also monitored and evaluated. No significant differences were found between either system in regards to C. capitata yield, weight or flight ability (p=0.05). Based on these comparative trials, the tower eclosion system appears to be an efficient alternative to the PARC system.     

Optimizing Diversity Assessment Protocols for High Canopy Ants in Tropical Rain Forest

The canopies of tropical rain forests support highly diverse, yet poorly known, animal and plant communities. It is vital that researchers who invest the time needed to gain access to the high canopy are able efficiently to survey the animals and plants that they find there. Here, we develop diversity assessment protocols for one of the most ecologically important canopy animal groups, the ants, in lowland dipterocarp rain forest in Sabah, Malaysia. We design and test a novel trap (the purse‐string trap) that can be remotely collected, thus avoiding disturbance to ants. We compare this modified trap with two other methods for surveying canopy ants: precision insecticide fogging and baited pitfall trapping. In total, we collected 39,351 ants belonging to 173 species in 38 genera. Fogging collected the most individuals and species, followed by purse‐string trapping with baited pitfall trapping catching the fewest. Fogging also resulted in samples with a different species composition to purse‐string trapping and baited pitfall trapping, which were not different from one another. Using a ‘greedy algorithm’, which guides the selection of inventory methods in order to maximize new species discovered per researcher‐hour, we show that projects allocating fewer than 132 researcher‐hours to canopy ant collection and identification should sample exclusively using fogging. Those with more time should use a combination of methods. This prioritization technique could be used to accelerate species discovery in future rapid biodiversity assessments. Abstract in Malay is available in the online version of this article.     

Differential regulation of K+ channels in Arabidopsis epidermal and stelar root cells

The patch clamp technique was applied to protoplasts isolated from the epidermis and pericycle of Arabidopsis roots and their plasma membrane currents investigated. In the whole cell configuration, all protoplasts from the epidermis exhibited depolarization‐activated time‐dependent outwardly rectifying (OR) currents whereas OR currents were present in only 50% of cells from the pericycle. The properties of the OR currents in the epidermis and pericycle were compared with respect to their selectivity, pharmacology and gating. The time‐dependent activation kinetics, selectivity and sensitivity to extracellular tetraethyl ammonium of the OR current in each cell type were not significantly different. The reversal potential (E_rev) of the OR currents indicated that they were primarily due to the movement of K⁺. However, the gating properties of the OR currents from the epidermis differed markedly from those exhibited in the pericycle. Although both cell types displayed OR currents with voltage‐dependent gating modulated in a potassium‐dependent fashion [i.e. the activation threshold (V_0.5) was displaced to more positive voltages as extracellular K⁺ increased], the OR currents in the epidermis also displayed voltage‐independent gating by extracellular K⁺ which dramatically regulated current density. In the present study, reducing extracellular K⁺ activity from 40 to 0.87 mm reduced the OR current density in epidermal cells by approximately 80%. The chord conductance of the OR current saturated as a function of extracellular K⁺ and could be fitted with a Michaelis–Menten function to yield a binding constant (K_m) of 10.5 mm . The ability of other monovalent cations to substitute for K⁺‐gating of the OR currents was also investigated and shown to exhibit a relative sequence of K⁺ ≥ Rb⁺ > Cs⁺ > Na⁺ ≥ Li⁺ (Eisenmann sequence IV) with respect to efficacy of gating. Furthermore, single channel recordings demonstrated that channel activity rather than the single channel conductance was modulated by extracellular K⁺. In contrast, OR current density in the pericycle was largely independent of extracellular K⁺. It is suggested that the contrasting gating properties of the K⁺ channels in the epidermis and pericycle reflect their different physiological roles, particularly with respect to their role in K⁺ (nutrient) transport from the soil solution to the shoot.