Enriched accumulation and biotransformation of selenium in the edible seaweed Laminaria japonica

Accumulations of selenium in kelp Laminaria japonica cultured in seawater was achieved by adding selenite (Na2SeO3) with or without N-P (NaNO3 + NaH2PO4) nutrients at different concentrations. Biotransformation of selenium in the kelp was investigated through measuring the selenium of biological samples and different biochemical fractionations. The results showed that the optimal selenite-enrichment concentration is 200 mg L-1, which can allow the kelp to accumulate a total selenium content from 0.51 +/- 0.15 to 26.23 +/- 3.12 mug g(-1) of fresh weight (fw). Selenium composition analysis of kelp (control group) showed that selenium is present as organic selenium, which is up to 86.22% of the total selenium, whereas inorganic selenium is barely 4.85%. When L. japonica was exposed for 56 h in seawater containing 200 mg L-1 Na2SeO3, the organic selenium was 16.70 mug g(-1) of fw (68.23%) and inorganic selenium was 4.71 mug g(-1) of fw (19.26%). The capability of accumulation of selenium was further enhanced by adding N-P nutrients to the selenite-enriched medium. Total selenium is increased to be 33.65 mug g(-1) of fw at optimal concentration of N-P nutrient (150 mg L-1 NaNO3 and 25 mg L-1 NaH2PO4), whereas the inorganic selenium was not increased and remained at 4.597 mug g(-1) of fw (13.36%), and the increased part of selenium was organic selenium. This implied that kelp L. japonica could effectively transform inorganic selenium into organic selenium through metabolism.

Effect of carbon source on growth and lipid accumulation in Chlorella sorokiniana GXNN01

Heterotrophic culture of microalgae to develop methods of increasing biomass productivity and storage lipids has brought new insight to commercial biodiesel production. To understand the relationship between heterotrophy and lipid production, the effects of carbon sources on the growth and lipid accumulation of Chlorella sorokiniana GXNN01 was studied. The alga exhibited an increased growth rate in response to the addition of carbon sources, which reached the stationary phase after 48 h at 30A degrees C. In addition, glucose and NaAc had a significant effect on the lipid accumulation during the early-stationary phase. Specifically, the lipid content was 0.237 +/- 0.026 g g(-1) cell dry weight and 0.272 +/- 0.041 g L-1 when glucose was used as the carbon source, whereas the lipid content reached 0.287 +/- 0.018 g g(-1) cell dry weight and 0.288 +/- 0.008 g L-1 when NaAc was used as the carbon source. The neutral lipid content was found to first decrease and then increase over time during the growth phase. A glucose concentration of 20 mmol L-1 gave the maximal lipid yield and the optimum harvest time was the early-stationary phase.

Effect of iron on growth and lipid accumulation in Chlorella vulgaris

The economic feasibility of algal mass culture for biodiesel production is enhanced by the increase in biomass productivity and storage lipids. Effect of iron on growth and lipid accumulation in marine microalgae Chlorella vulgaris were investigated. In experiment I, supplementing the growth media with chelated FeCl3 in the late growth phase increased the final cell density but did not induce lipid accumulation in cells. In experiment II, cells in the late-exponential growth phase were collected by centrifugation and re-inoculated into new media supplemented with five levels of Fe3+ concentration. Total lipid content in cultures supplemented with 1.2 x 10(-5) mol L-1 FeCl3 was up to 56.6% biomass by dry weight and was 3-7-fold that in other media supplemented with lower iron concentration. Moreover, a simple and rapid method determining the lipid accumulation in C. vulgaris with spectrofluorimetry was developed. (c) 2007 Elsevier Ltd. All rights reserved.

The Shandong mud wedge and post-glacial sediment accumulation in the Yellow Sea

Two well-defined deltaic sequences in the Bohai Sea and in the South Yellow Sea represent post-glacial accumulation of Yellow River-derived sediments. Another prominent depocenter on this epicontinental shelf, a pronounced clinoform in the North Yellow Sea, wraps around the northeastern and southeastern end of the Shandong Peninsula, extending into the South Yellow Sea. This Shandong mud wedge is 20 to 40 m thick and contains an estimated 300 km(3) of sediment. Radiocarbon dating, shallow seismic profiles, and regional sea-level history suggest that the mud wedge formed when the rate of post-glacial sea-level rise slackened and the summer monsoon intensified, at about 11 ka. Geomorphic configuration and mineralogical data indicate that present-day sediment deposited on the Shandong mud wedge comes not only from the Yellow River but also from coastal erosion and local rivers. Basin-wide circulation in the North Yellow Sea may transport and redistribute fine sediments into and out of the mud wedge.

靶向斑马鱼VEGF的shRNA表达载体的构建及其对斑马鱼胚胎血管发育的影响

血管内皮生长因子(vascular endothelial growth factor, VEGF)是一种多功能的细胞因子，其主要作用是促进血管内皮细胞增殖和增加血管通透性，是肿瘤及正常组织血管生成的中心调控因素，以VEGF为靶点的肿瘤血管靶向性治疗成为近几年肿瘤治疗的新途径。RNAi是近年来新发展的一项反向遗传学技术，是一种研究基因功能的有力工具。斑马鱼作为一种重要的模式生物，被广泛用于胚胎的分子发育机制、疾病模型的构建以及药物筛选等研究中。然而在斑马鱼中运用RNAi技术进行基因功能研究是一个相对较新的领域，研究资料较少，并且目前进行的斑马鱼RNAi实验中，siRNA大都是通过化学方法或体外转录合成的。体外合成的siRNA在进入体内后会被降解而无法达到持久阻抑基因表达的目的。因此本研究旨在探讨VEGF特异性siRNA表达载体对斑马鱼VEGF基因的沉默作用，通过分析表型及相关细胞因子的变化，阐明VEGF对斑马鱼胚胎血管生成的影响及作用机制。 研究通过计算机辅助设计软件，针对斑马鱼VEGF mRNA不同位点设计合成了4段含siRNA特异序列的DNA单链，经退火，克隆入pSilencer 4.1-CMV neo载体CMV启动子下游，构建了重组质粒pS1-VEGF、pS2-VEGF、pS3-VEGF及pS4-VEGF。 通过显微注射的方法将载体导入1-2细胞期斑马鱼体内，于胚胎发育的48 h采用RT-PCR的方法检测VEGF基因的表达量，研究不同干扰序列对VEGF基因表达的干涉作用。结果显示，针对不同位点的表达载体对VEGF基因表达的抑制效率有显著差异。它们对VEGF mRNA的抑制率分别为80.5%，42.8%，12.5%，40.7%。通过筛选我们得到了一条具有高效抑制作用的载体pS1-VEGF，该载体的相应序列靶向斑马鱼两个主要异构体VEGF165和VEGF121的共有外显子序列。 形态学检测结果显示，注射了pS1-VEGF的胚胎出现了心包膜水肿、血流速度减慢、循环红细胞堆积等症状。定量碱性磷酸酶染色显示，注射pS1-VEGF能够抑制斑马鱼胚胎新生血管的形成，当注射剂量为0.4 ng时，血管生成的抑制率为31.8%。NBT/BCIP血管染色显示，注射该载体后72 h，50%的斑马鱼肠下静脉、节间血管以及其它血管的发育受到不同程度的抑制。随着注射剂量的加大，血管发育受抑制的情况也随之加重，当注射剂量为1 ng时，只有心脏、头部及卵黄有血液循环。对干扰效果的特异性进行了研究，结果表明pS1-VEGF对斑马鱼内源基因胸苷酸合成酶（thymidylate synthase, TS）基因的表达没有明显的抑制作用。针对TS基因的shRNA表达载体及与斑马鱼没有同源性的对照载体对VEGF基因表达也没有明显的抑制作用。浓度梯度实验表明在0-1.2 ng的范围内干扰效果具有剂量依赖性。 以胚胎整体原位杂交的方法检测质粒对VEGF基因受体NRP1基因表达的影响，发现VEGF特异性shRNA表达载体能够引起NRP1基因表达的降低，说明斑马鱼中VEGF所介导的血管生成作用至少在部分上是依赖于NRP通路所调节的。 本研究工作为进一步研究斑马鱼基因功能、VEGF调控网络提供了一个快速、有效的手段，为阐明斑马鱼的血管生成机制提供了新的资料，为采用RNAi技术，以VEGF为靶点，以斑马鱼为模型对肿瘤进行基因治疗研究奠定了基础。

Accumulation of petroleum hydrocarbons and heavy metals in clams (Ruditapes philippinarum) in Jiaozhou Bay, China

Accumulation and distributions of aliphatic and polyaromatic hydrocarbons (PAHs) and heavy metals were measured in tissues of the clam Ruditapes philippinarum collected from 5 sites in Jiaozhou Bay, Qingdao, China. The concentrations of total aliphatic hydrocarbon and PAHs ranged from 570 to 2 574 ng/gdw (gram dry weight) and from 276 to 939 ng/gdw, in the most and least polluted sites, respectively. The bio-accumulation of hydrocarbons and PAHs in the clams appeared to be selective. Aliphatic hydrocarbons were predominantly represented by short chain (< nC(23)) n-alkanes, suggesting that petroleum hydrocarbons were likely the major contamination source. The selective uptake of 3 and 4 ring PAHs, such as naphthalene, fluorene, phenanthrene, fluoranthene and pyrene, by the clams was probably related to the physiological and bio-kinetic processes that were energetically favorable for uptake of compounds with fewer rings. Accumulation of the metals Cd, Cu, Zn, Pb, Cr, Hg, and As in the clam tissues also showed high variability, ranging from 0.043 to 87 A mu g/gdw. Among the 7 detected metals, Zn, Cd, Cu, and As had a particularly high potential of accumulation in R. philippinarum. In general, a positive correlation was found between the tissue concentrations and sediment concentrations of hydrocarbons and of some metals. Our study suggests that moderate contamination with polyaromatic hydrocarbons, and low to moderate contamination with metals, currently exists for clam R. philippinarum in Jiaozhou Bay, in comparison with other regional studies. A long-term monitoring program is certainly needed for assessment of the potential ecological influence and toxicity of these contaminants of R. philippinarum in Jiaozhou Bay.

Influence of lanthanum on the accumulation of trace elements in chloroplasts of cucumber seedling leaves

Influence of La3+ on the accumulation of trace elements (Se-75, Co-56, Rb-83, V-48, (95)mTc, and Ga-67) in chloroplasts of cucumber seedling leaves was studied by a radioactive multitracer technique. At the same time, chloroplast contents of different concentrations of La3+ treatment were calculated. It was observed that chloroplast contents peaked at 0.02 mM La3+ treatment and that the uptake and distribution of these trace elements in chloroplasts of cucumber seedling leaves are different under different La3+, treatments. With the increase of lanthanum concentrations from 0.002 to 2 mM, the uptake percentages of Se-75, Co-56, and Rb-83 presented an obvious increase and then sharply decreased in contrast to the nonlanthanum treatment, whereas there appeared a sharp decrease and then restored control level in the uptake of V-48. The other two trace elements, namely Tc-95m and Ga-67, were accumulated only in the presence of 0.02 mM La3+. The results indicate that lanthanum treatments to growing the cucumber lead to the change of trace element uptake in the chloroplasts of leaves, which suggest that lanthanum might influence the accumulation of trace elements in chloroplasts of cucumber seedling leaves by regulation of various ion transport mechanisms, thus affecting the photosystem of leaves.