华北低丘山区栓皮栎生态系统氧同位素日变化及蒸散定量区分

利用稳定同位素技术对华北低丘山区栓皮栎生态系统氧同位素日变化及蒸散定量区分进行研究,为华北低丘山区森林生态系统水汽交换研究提供基础。试验采用离轴积分腔输出光谱技术(OA-ICOS)连续测定生态系统不同高度水汽浓度和δ18O值,同时采用真空提取和液态水同位素分析仪测定枝条和土壤的δ18O值。结果显示,4个晴天中大气水汽浓度日变化复杂,变化趋势差异大,而δ18O日变化均成高-低-高的"V"型变化,最小值出现在12:00—18:00。Keeling方程在10:00—12:00的相关系数R2均大于0.71,方程达到极显著水平,表明此时段蒸腾速率较高,满足植物蒸腾的同位素稳定态假设。利用Keeling方程估算的栓皮栎生态系统δET值有相似的低-高-低日变化,与大气的δv值变化趋势相反。同位素分割结果显示栓皮栎生态系统中蒸腾占蒸散比例日变化呈现低-高-低的趋势,10:00—14:00蒸腾占蒸散比例达到90%以上,尽管6:00—10:00和14:00—18:00的蒸腾占蒸散比例下降,但平均值仍高达69.38%,表明华北低丘山区栓皮栎生态系统的蒸散主要来源于植物蒸腾。     

Two-stage carbon distribution and cofactor generation for improving l-threonine production of Escherichia coli

L -Threonine, a kind of essential amino acid, has numerous applications in food, pharmaceutical, and aquaculture industries. Fermentative l -threonine production from glucose has been achieved in Escherichia coli. However, there are still several limiting factors hindering further improvement of l -threonine productivity, such as the conflict between cell growth and production, byproduct accumulation, and insufficient availability of cofactors (adenosine triphosphate, NADH, and NADPH). Here, a metabolic modification strategy of two-stage carbon distribution and cofactor generation was proposed to address the above challenges in E. coli THRD, an l -threonine producing strain. The glycolytic fluxes towards tricarboxylic acid cycle were increased in growth stage through heterologous expression of pyruvate carboxylase, phosphoenolpyruvate carboxykinase, and citrate synthase, leading to improved glucose utilization and growth performance. In the production stage, the carbon flux was redirected into l -threonine synthetic pathway via a synthetic genetic circuit. Meanwhile, to sustain the transaminase reaction for l -threonine production, we developed an l -glutamate and NADPH generation system through overexpression of glutamate dehydrogenase, formate dehydrogenase, and pyridine nucleotide transhydrogenase. This strategy not only exhibited 2.02- and 1.21-fold increase in l -threonine production in shake flask and bioreactor fermentation, respectively, but had potential to be applied in the production of many other desired oxaloacetate derivatives, especially those involving cofactor reactions.     

Proteomic signatures of serum albumin-bound proteins from stroke patients with and without endovascular closure of PFO are significantly different and suggest a novel mechanism for cholesterol efflux

Background

The anatomy of PFO suggests that it can allow thrombi and potentially harmful circulatory factors to travel directly from the venous to the arterial circulation – altering circulatory phenotype. Our previous publication using high-resolution LC-MS/MS to profile protein and peptide expression patterns in plasma showed that albumin was relatively increased in donor samples from PFO-related than other types of ischemic strokes. Since albumin binds a host of molecules and acts as a carrier for lipoproteins, small molecules and drugs, we decided to investigate the albumin-bound proteins (in a similar sample cohort) in an effort to unravel biological changes and potentially discover biomarkers related to PFO-related stroke and PFO endovascular closure.

Methods

The method used in this study combined albumin immuno-enrichment with high resolution LC-MS in order to specifically capture and quantify the albumin-bound proteins. Subsequently, we measured cholesterol and HDL in a larger, separate cohort of PFO stroke patients, pre and post closure.

Results

The results demonstrated that a number of proteins were specifically associated with albumin in samples with and without endovascular closure of the PFO, and that the protein profiles were very different. Eight proteins, typically associated with HDL were common to both sample sets and quantitatively differently abundant. Pathway analysis of the MS results suggested that enhanced cholesterol efflux and reduced lipid oxidation were associated with PFO closure. Measurement of total cholesterol and HDL in a larger cohort of PFO closure samples using a colorimetric assay was consistent with the proteomic predictions.

Conclusions

The collective data presented in this study demonstrate that analysis of albumin-bound proteins could provide a valuable tool for biomarker discovery on the effects of PFO endovascular closure. In addition, the results suggest that PFO endovascular closure can potentially have effects on HDL, cholesterol and albumin-bound ApoA-I abundance, therefore possibly providing benefits in cardioprotective functions.

Electronic supplementary material

The online version of this article (doi:10.1186/1559-0275-12-2) contains supplementary material, which is available to authorized users.     

Tailor-made approach for selective isolation and elution of low-density lipoproteins by immunoaffinity sorbent on silica

Immunoaffinity procedure was developed for isolation of low density lipoprotein (LDL) from biological samples by using silica-derived immunoaffinity sorbent. Sorbent was prepared by immobilization of monoclonal anti-apoB-100 antibody onto macroporous silica particles, using carefully optimized binding chemistry. Binding capacity of the sorbent towards LDL was determined by batch extraction experiments with solutions of isolated LDL in phosphate-buffered saline, and found to be 8 mg LDL/g. The bound LDL fraction was readily released from the sorbent by elution with ammonia at pH 11.2. The total time needed for isolation procedure was less than 1 h, with LDL recoveries being essentially quantitative for samples containing less than 0.3 mg LDL/mL. With higher concentrations, recoveries were less favorable, most probably due to irreversible adsorption caused by LDL aggreggation. However, reusability studies with isolated LDL at concentration 0.2 mg/mL indicate that the developed immunoaffinity material may be used for multiple binding-release cycles, with minor losses in binding capacity. Finally, the sorbent was successfully applied to isolation of LDL from diluted plasma. Apart from its practical implications for LDL isolation, this study provides crucial insights into issues associated with LDL-sorbent interactions, and may be useful in future efforts directed to development of lipoprotein isolation approaches.     

季节性雪被对青藏高原东缘高寒草甸2种优势植物碳、氮积累和分配的影响

依据2006~2008冬季的自然雪被分布状况,在青藏高原东缘的高寒草甸中设置3条样带（即深雪、中雪和浅雪）。在2009年的生长季,在3个雪梯度样带中,分别测定了2种优势植物圆穗蓼（Polygonum macrophyllum）和黑褐穗苔草（Carex atrofusca subsp. Minor）生物量和碳氮营养积累及分配的动态特征。结果表明,深雪能够促进圆穗蓼和黑褐穗苔草生物量和碳氮养分的积累,可能使它们产生的凋落物数量更多且质量更好。深雪更有利于圆穗蓼（非禾本科草本植物）根系生物量、碳氮养分的积累;深雪不仅同时促进黑褐穗苔草（禾本科植物）地上部分和根系生物量、碳氮积累,而且还使其种子产量增加和质量提高,潜在地增强了黑褐穗苔草种子的繁殖能力,可能使得黑褐穗苔草的种间竞争能力增强。可以预见,未来季节性雪被的变化,必定会引起青藏高原东缘高寒草甸的初级生物量及其结构、植物群落物种组成等均发生相应变化。     

可溶性淀粉合成酶SSⅢ基因对马铃薯的遗传转化

马铃薯是淀粉生产中重要的农作物之一,而可溶性淀粉合成酶SSⅢ是可溶性淀粉合成酶的主要活性成分,通过基因工程的手段来研究SSⅢ基因在淀粉合成中的功能可以用于改良马铃薯淀粉的品质.本研究采用根癌农杆菌介导法将强组成型表达启动子CaMV 35S驱动的可溶性淀粉合成酶SSⅢ基因的RNA干扰表达载体导入马铃薯栽培品种克新1号和克新4号中,获得了65株卡那霉素抗性植株.对抗性植株PCR检测结果表明,SSⅢ基因的干扰片段已整合到马铃薯基因组中,RT-PCR检测表明SSⅢ基因在转录水平上受到了明显抑制.该研究为马铃薯淀粉品质的改良奠定了基础.     

盐地碱蓬液泡膜Ca2+/H+逆转运蛋白SsCAX1 N末端原核表达和多克隆抗体的制备

采用PCR技术,从盐地碱蓬液泡膜Ca2+/H+逆转运蛋白SsCAX1的cDNA中扩增了其N末端亲水区段(1～210 bp),并将其插入到原核表达载体pGEX-6p-3中进行诱导表达.获得分子量约33.5 kDa的可溶性融合蛋白,经纯化后从免疫家兔中得到了SsCAX1的特异性抗体.     

A transversely isotropic viscoelastic constitutive equation for brainstem undergoing finite deformation

The objective of this study was to define the constitutive response of brainstem undergoing finite shear deformation. Brainstem was characterized as a transversely isotropic viscoelastic material and the material model was formulated for numerical implementation. Model parameters were fit to shear data obtained in porcine brainstem specimens undergoing finite shear deformation in three directions: parallel, perpendicular, and cross sectional to axonal fiber orientation and determined using a combined approach of finite element analysis (FEA) and a genetic algorithm (GA) optimizing method. The average initial shear modulus of brainstem matrix of 4-week old pigs was 12.7 Pa, and therefore the brainstem offers little resistance to large shear deformations in the parallel or perpendicular directions, due to the dominant contribution of the matrix in these directions. The fiber reinforcement stiffness was 121.2 Pa, indicating that brainstem is anisotropic and that axonal fibers have an important role in the cross-sectional direction. The first two leading relative shear relaxation moduli were 0.8973 and 0.0741, respectively, with corresponding characteristic times of 0.0047 and 1.4538 s, respectively, implying rapid relaxation of shear stresses. The developed material model and parameter estimation technique are likely to find broad applications in neural and orthopaedic tissues.