High-performance size-exclusion chromatography studies on the formation and distribution of polar compounds in camellia seed oil during heating

Camellia seed oil （CSO） is rich in oleic acid and has a high number of active components, which give the oil high nutritional value and a variety of biological activity. The aim of the present study was to determine the changes in the content and distribution of total polar compounds （TPC） in CSO during heating. TPC were isolated by means of preparative flash chromatography and further analyzed by high-performance size-exclusion chromatography （HPSEC） The TPC content of CSO increased from 4.74% to 25.29%, showing a significantly lower formation rate as compared to that of extra virgin olive oil （EVOO） and soybean oil （SBO） during heating. Furthermore, heating also resulted in significant differences （P〈0.05） in the distribution of TPC among these oils. Though the content of oxidized triacyl- glycerol dimers, oxidized triacylglycerol oligomers, and oxidized triacylglycerol monomers significantly increased in all these oils, their increased percentages were much less in CSO than those in EVOO, indicating that CSO has a greater ability to resist oxidation. This work may be useful for the food oil industry and consumers in helping to choose the correct oil and to decide on the useful lifetime of the oil.

Efficient purification of cell culture-derived classical swine fever virus by ultrafiltration and size-exclusion chromatography

Large-scale production of cell culture-based classical swine fever virus(CSFV)vaccine is hampered by the adverse reactions caused by contaminants from host cell and culture medium.Hence,we have developed an efficient method for purifying CSFV from cell-culture medium.Pure viral particles were obtained with two steps of tangential-flow filtration(TFF)and size-exclusion chromatography(SEC),and were compared with particles from ultracentrifugation by transmission electron microscopy(TEM),infectivity and recovery test,and real time fluorescent quantitative PCR(FQ-PCR).TFF concentrated the virus particles effectively with a retention rate of 98.5%,and 86.2%of viral particles were obtained from the ultrafiltration retentate through a Sepharose 4 F F column on a biological liquid chromatography system.CSFV purified by TFF-SEC or ultracentrifugation were both biologically active from 1.0×10–4.25 TCID50·mL^(–1) to 3.0×10^(–6.25) TCID50·mL^(–1),but the combination of TFF and SEC produced more pure virus particles than by ultracentrifugation alone.In addition,pure CSFV particles with the expected diameter of 40–60 nm were roughly spherical without any visible contamination.Mice immunized with CSFV purified by TFF-SEC produced higher antibody levels compared with immunization with ultracentrifugation-purified CSFV(P<0.05).The purification procedures in this study are reliable technically and feasible for purification of large volumes of viruses.

一种新颖的体积排阻色谱加宽效应的去除方法Ⅰ理论测试

分子量及其分布是表征聚合物特征的重要参数,体积排阻色谱(Size-exclusion chromatography)广泛用于测量聚合物分子量和分子量分布。然而,由于带加宽效应的存在,导致实际得到的分子量分布曲线形状受到了一定程度的扭曲,从而影响了分析的准确性。目前国际上基于实验或理论提出的解决方案的缺点不是实验代价昂贵就是理论方法的近似度太高而不能够被普遍采用。本文结合了退卷积方法和蒙特卡洛Metropolis拟合技术,开发了一种有效且简便的带加宽效应去除方法,本文对这种方法进行了严密的理论验证,证明了该方法可以准确获得聚合物分子量分布信息。

Effects of Rice Variety and Growth Location in Cambodia on Grain Composition and Starch Structure

The effects of variety and growth location on grain composition and starch structures were investigated using three rice （Oryza sativa L.） cultivars （Phka Romduol, Sen Pidao and IR66） with different amylose contents. All the three cultivars were planted in three different agro-climatic zones （Phnom Penh, Coastal and Plateau） of Cambodia. The protein content of polished grains increased when rice was planted at a location with higher average temperature, but their lipid content decreased. The amylose content and degree of branching were not greatly affected by the minor temperature differences among the growing locations. Starch fine structures characterized by the chain-length distribution were significantly different among the cultivars, but not significantly among different locations. The results suggested that protein and lipid biosyntheses were more sensitive to the environmental temperature than that of starch in rice grains.

Interaction of Coal Humic Acids with Fungal Laccase

Humic acids (HA) are one of the main environmental factors controlling the fate and behavior of the compounds released into the environment. In particular, they are universally considered of great importance in determining soil extracellular enzyme activity and stability via association with essential soil enzymes. The objective of this study was to investigate the interaction of coal HA with an extracellular multicopper oxidase laccase (EC 1.10.3.2) that catalyze the oxidation of a wide range of reducing substances in the environment. Using size-exclusion chromatography analysis and monitoring laccase activity, the formation of a stable and an enzymatically active complex between HA and laccase was shown. Basing the data obtained by isoelectric focusing of HA-laccase complex, non-covalent character of laccase association with HA was considered and binding of laccase to HA by weak dispersive forces such as van der Waals, hydrophobic, π-π, CH-π and others was hypothesized.

CHROMATOGRAPHIC REFOLDING OF PROTEINS:MOLECULAR ACTION AND COLUMN CONTROL

Protein expression in E coil often results in the formation of a kind of protein aggregate called inclusion body Conversion of the inactive protein aggregate into biologically active protein is a key step in production of recombinant products Convenlional dilution refolding technique suffers from disadvantages of low recovery and low concentration Various chromatographic refolding techniques have been developed over the last few years These include size-exclusion chromatography, ion exchange chromatography, hydrophobic interaction chromatography and different affinity chromatography. A successful strategy is the use of gradient elution in column control which provides a gentle and gradual change of the solution environment for the macromolecule to rsfold at nano-scale, The gradient refolding at column scale could minimize misfolding and aggregation which are induced by sudden change of the solution in conventional refolding operation.

The conditional mitochondrial protein complexome in the Arabidopsis thaliana root and shoot

Protein complexes are important for almost all biological processes.Hence,to fully understand how cells work,it is also necessary to characterize protein complexes and their dynamics in response to various cellular cues.Moreover,the dynamics of protein interaction play crucial roles in regulating the(dis)association of protein complexes and,in turn,regulating biological processes such as metabolism.Here,mitochondrial protein complexes were investigated by blue native PAGE and size-exclusion chromatography under conditions of oxidative stress in order to monitor their dynamic(dis)associations.Rearrangements of enzyme interactions and changes in protein complex abundance were observed in response to oxidative stress induced by menadione treatment.These included changes in enzymatic protein complexes involving g-amino butyric acid transaminase(GABA-T),D-ornithine aminotransferase(D-OAT),or proline dehydrogenase 1(POX1)that are expected to affect proline metabolism.Menadione treatment also affected interactions between several enzymes of the tricarboxylic acid(TCA)cycle and the abundance of complexes of the oxidative phosphorylation pathway.In addition,we compared the mitochondrial complexes of roots and shoots.Considerable differences between the two tissues were observed in the mitochondrial import/export apparatus,the formation of super-complexes in the oxidative phosphorylation pathway,and specific interactions between enzymes of the TCA cycle that we postulate may be related to the metabolic/energetic requirements of roots and shoots.

Dextran-modified iron oxide nanoparticles

Dextran-modified iron oxide nanoparticles were prepared by precipitation of Fe（Ⅱ） and Fe（Ⅲ） salts with ammonium hydroxide by two methods. Iron oxide was precipitated either in the presence of dextran solution, or the dextran solution was added after precipitation. In the second method, the iron oxide particle size and size distribution could be controlled depending on the concentration of dextran in the solution. The nanoparticles were characterized by size-exclusion chromatography, transmission electron microscopy and dynamic light scattering. Optimal conditions for preparation of stable iron oxide colloid particles were determined, The dextran/iron oxide ratio 0-0,16 used in precipitation of iron salts can be recommended for synthesis of nanoparticles suitable for biomedical applications, as the colloid does not contain excess dextran and does not coagulate.