Effects of glucose,glutamine, and malate on the metabolism of spodoptera frugiperda clone 9 (sf9) cells

Optimal design and operation of bioreactors for insect cell culture is facilitated by functional relations providing quantitative information on cellular metabolite consumption kinetics, as well as on the specific cell growth rates (μ_G). Initial specific consumption rates of glucose, malate, and oxygen, and associated changes in μ_G, were measured forSpodoptera frugiperda clone 9 (Sf9) cells grown in batch suspension culture in medium containing 7–35 mM glucose, 0–16 mM malate, and 4–16 mM glutamine. The initial specific glucose consumption rate (q _G ) could be described by a modified Michaelis-Menten equation treating malate as a “competitive” inhibitorK ₁ = 6.5 mM) and glutamine as a “noncompetitive” inhibitorK _I = 14 mM) ofq _G , with aK _m of 7.1 mM for glucose. All three carbon sources were found to increase μ_G in a saturable manner, and a modified Monod equation was employed to describe this relationship (μ_Gmax = 0.047 h^-1). The initial specific oxygen consumption rate (q_O2) in Sf9 cells could be related to μ_G by the maintenance energy model, and it was calculated that, under typical culture conditions, about 15–20% of the cellular energy demand comes from functions not related to growth. Fitted parameters in mathematical expression for μg: K₄, Monod constant for glucose (mM); K₅, modified Monod constant for malate (mM); K₆, Monod constant for glutamine (mM); m_o2, specific consumption rate of oxygen by the cells under zero-growth conditions (nmol/cell/h); q_F, initial specific fumarate production rate (nmol/cell/ h);q _G , initial specific glucose consumption rate (nmol/cell/h); q_Gmax, maximum initial specific glucose consumption rate (nmol/cell/h);q _M , initial specific malate consumption rate (nmol/cell/h); q_o2, initial specific oxygen consumption rate (nmol/cell/h); Y_o2, cell yield on oxygen (cells/nmol); μ, initial specific cell growth rate (h^-1); μ_g, initial specific cell growth rate (h^-1); μG_max, maximum initial specific cell growth rate (h^-1).     

阿尔芬波加热研究的进展

本文概述了阿尔芬波加热的理论和实验情况。讨论了阿尔芬波加热的基本物理问题,并介绍了某些重要的实验结果。对在大托卡马克上采用阿尔芬波加热的可能性进行了全面论述。     

A competent extraction method of plant proteins for 2-D gel electrophoresis

The efficient extraction of high‐quality proteins is a key factor for a successful proteomic analysis approach. In the method suggested here, absolute ethanol containing 10 mM DTT was used to precipitate the proteins in plant tissue homogenates followed by their resuspension in a urea‐/thiourea‐ and NP‐40‐containing solution. Protein profiles were examined on pH 3–11 non‐linear IEF strips and SDS‐PAGE and compared with extracts using the established method of acetone‐10% TCA/0.07% 2‐mercaptoethanol precipitation (V. Méchin et al., Methods Mol. Biol. 2006, 355, 1–8). In addition to protein profile similarity for the two extracts, the acidic part of the acetone containing 10% TCA/0.07% 2‐mercaptoethanol extraction showed protein spots with high molecular weight in the range of 250–150 kDa, while the ethanol containing 10 mM DTT extracts indicated extra proteins spots at the basic part of the gels with molecular weights in the range of 25–15 kDa. The MALDI‐TOF‐MS of differential spots from acetone containing 10% TCA/0.07% 2‐mercaptoethanol precipitation method and absolute ethanol containing 10mM DTT indicated no similarity, ruling out the possibility that the two clusters shown represent identical proteins. The described method is easy in implementation, chemicals used are less toxic and proteins are easier to resuspend therefore presents an additional choice to implement towards finding the optimum method for extraction.     

Liquid phase microextraction applications in food analysis

Over the last years, liquid-phase microextraction (LPME) in its different application modes (single drop microextraction, dispersive liquid-liquid microextraction and hollow fiber-LPME) has been increasingly applied for the extraction of both inorganic and organic analytes from different matrices. Its advantages over conventional extraction procedures (simplicity, effectiveness, rapidity and low consumption of organic solvents) has also attracted its application in the complex food analysis field, in which it has clearly provided good and challenging results. A comprehensive review dealing with those articles published since its introduction till the end of March 2011 is presented, offering also a critical vision of the analytical potential of LPME for the analysis of foods.     

快放电直线型变压器驱动源磁芯脉冲损耗特性

建立了快脉冲磁芯损耗特性测试平台,对比研究了50 m的DG6硅钢和25 m的2605TCA非晶两种材料磁芯损耗特性;采用一种新的特征参量(磁芯单位面积上激磁电压陡度)来规范磁芯的激磁电压条件,使得实验结果与快放电直线型变压器驱动源实际工作条件下磁芯性能具有可比性;通过测量初级漏电流及次级开路电压,获得了相同激磁条件下两种磁芯等效损耗电阻的大小,50 m 的DG6硅钢磁芯损耗约为25 m的2605TCA非晶磁芯损耗的4倍;计算了两种材料磁芯总损耗中涡流损耗所占的比例,50 m的DG6硅钢磁芯涡流损耗占总损耗的75%,25 m的2605TCA非晶磁芯涡流损耗占总损耗的28%。     

The effect of 2-hydroxypropyl-β-cyclodextrin on the excited triplet state of promazine and chlorpromazine

Chlorpromazine hydrochloride (CPZ) is a widely used anti-psychotic drug that induces skin photosensitization and photoallergy response after systematic use or topical applications. The photoallergic mechanism is still unknown. However, it has been proposed that the triplet excited state (³CPZ*) could participate in the photodamaging effects. In this work, we report the photophysical properties of the triplet excited state of CPZ and its parent derivative promazine hydrochloride (PZ) in the presence of 2-hydroxypropyl-β-cyclodextrin (HPC). Absorption measurements indicate that PZ and CPZ form an inclusion complex with HPC through a 1:1 stoichiometry. The equilibrium constant at 25 °C is (2.55 ± 0.09) × 10³ M⁻¹ and (3.27 ± 0.07) × 10³ M⁻¹ for PZ and CPZ, respectively. The CPZ and PZ triplet excited state properties changed in the presence of HPC. The triplet lifetime increases with HPC concentration that is related to the amount of drug bound. In addition, the triplet intersystem crossing quantum yield was determined to be 0.45 and 0.17 for PZ and CPZ, respectively, when more than 95% of the drug molecules are bound to HPC. Altogether, these results suggest that the microenvironment plays a crucial role in the ³CPZ* and ³PZ* properties and thus it can modulate their photosensitizing effects.     

An improved temporal clustering analysis method applied to whole-brain data in fMRI study

Temporal clustering analysis (TCA) has been proposed as a method to detect the brain responses of an fMRI time series when the time and location of the activation are completely unknown. But TCA is still incompetent in dealing with the time series of the whole brain due to the existence of many inactive pixels. If only active pixels are considered, the sensitivity of TCA will be improved greatly and it could be applied to the whole brain. In this study, some modifications were made to TCA to remove inactive pixels, and the applicability of the modified TCA to the whole brain was validated with a set of visual fMRI data. Based on the time series of the modified TCA, activations of the whole brain corresponding to the visual stimulation were detected. Compared with the previous TCA, the modified TCA method shows a significant improvement in the sensitivity to detect activation peaks of the whole brain.     

Trichloroacetic acid in the foliage of forest trees

Trichloroacetic acid (TCA) is an atmospheric oxidation product of airborne C₂-chlorocarbons and a widespread air pollutant. It was widely used as a herbicide in the fifties and sixties, in some countries, still today. It occurs in the foliage of conifers and deciduous trees of mountain forests in Germany, e. g. the Black Forest, the Northern Alps or the Erzgebirge. Quantitative determination in the ppb-range is achieved by gas chromatography using thick film capillary columns and electron-capture detection. The levels in spruce needles at two forest sites correlate with the extent of needle loss. The data are supportive of the hypothesis that TCA is causally involved in the induction of mountain forest decline.