131 resultados para Centrifugation, Density Gradient

em Chinese Academy of Sciences Institutional Repositories Grid Portal


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The perturbed-chain statistical associating fluid theory and density-gradient theory are used to construct an equation of state (EOS) applicable for the phase behaviors of carbon dioxide aqueous solutions. With the molecular parameters and influence parameters respectively regressed from bulk properties and surface tensions of pure fluids as input, both the bulk and interfacial properties of carbon dioxide aqueous solutions are satisfactorily correlated by adjusting the binary interaction parameter (k(ij)). Our results show that the constructed EOS is able to describe the interfacial properties of carbon dioxide aqueous solutions in a wide temperature range, and illustrate the influences of temperature, pressure, and densities in each phase on the interfacial properties.

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Density gradient ultracentrifugation (DGU) has emerged as a promising tool to prepare chirality enriched nanotube samples. Here, we assess the performance of different surfactants for DGU. Bile salts (e.g., sodium cholate (SC), sodium deoxycholate (SDC), and sodium taurodeoxycholate (TDC)) are more effective in individualizing Single Wall Carbon Nanotubes (SWNTs) compared to linear chain surfactants (e.g., sodium dodecylbenzene sulfonate (SDBS) and sodium dodecylsulfate (SDS)) and better suited for DGU. Using SC, a narrower diameter distribution (0.69-0.81 nm) is achieved through a single DGU step on CoMoCAT tubes, when compared to SDC and TDC (0.69-0.89 nm). No selectivity is obtained using SDBS. due to its ineffectiveness in debundling. We assign the reduce selectivity of dihydroxy bile salts (S DC and TDC) in comparison with trihydroxy SC to the formation of secondary micelles. This is determined by the number and position of hydroxyl ( OH) groups on the a-side of the steroid backbone. We also enrich CoMoCAT SWNT in the 0.84-0.92 nm range using the Pluronic F98 triblock copolymer. Mixtures of bile salts (SC) and linear chain surfactants (SOS) are used to enrich metallic and semiconducting laser-ablation grown SWNTs. We demonstrate enrichment of a single chirality, (6,5), combining diameter and metallic versus semiconductillg separation on CoMoCAT samples.

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Thylakoid membranes were isolated from Gymnodinium sp. and spinach, whereas the phycobilisomes were isolated and purified from red alga Porphyridium cruentum. The absorption spectra of the purified phycobilisomes (PBS) showed three peaks at 548, 564, and 624 nm, respectively, and the ratio of the fluorescence intensity at the lambda(680)(em) to lambda(80)(em5) that at was about 7.3. All these results demonstrated that the purified PBS remained intact. The thylakoid membranes were incubated with the purified phycobilisomes, and the thylakoid membranes, which harbored the phycobilisomes, were purified by sucrose density gradient centrifugation. Meantime, the conjugates of phycobilisome-thylakoid membranes were constructed using glutaraldehyde and further purified. Their characteristics were studied by measuring the absorption spectra and fluorescence emission spectra. The results showed that the phycobilisomes from Porphyridium cruentum can attach to the thylakoid membranes from Gymnodinium sp. and spinach without covalent cross-linking, but the excited energy transfer did not occur. The conjugate of phycobilisome-thylakoid. membranes with covalent cross-linking exhibits the excited energy transfer between the phycobilisomes and the thylakoid membranes.

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SNARE蛋白家族是所有真核细胞胞吐及分泌作用中的关键因子,由其介导的运输囊泡膜与靶膜的锚靠、融合为胞内蛋白的运出提供了一条重要途径。体外试验表明,Syntaxin6-Syntaxin7-Vti1b,SNAP-23-Syntaxin4等SNARE核心蛋白之间精确的相互作用是哺乳动物巨噬细胞肿瘤坏死因子α (TNF-α)运输和分泌的必备条件,在机体非特异性免疫应答反应过程中起重要作用。 本研究受上述启示,旨在揭示SNARE蛋白在海洋鱼类免疫细胞内重要细胞因子白细胞介素1β (IL-1β)的分泌过程中的作用。参照Percoll密度梯度离心技术,从鲈鱼头肾组织分离纯化巨噬细胞进行稳定培养;利用RT-PCR方法克隆出鲈鱼t-SNARE蛋白SNAP-23和Syntaxin3的部分cDNA序列,再结合先前克隆的VAMP2和已知的鲈鱼IL-1β,TNF-α和IL-8的基因序列,设计特异性引物。利用Real-time PCR技术在mRNA水平上精确测定鲈鱼巨噬细胞中上述6种基因在革兰氏阴性菌脂多糖(LPS)分子刺激下的表达变化,发现SNAP-23基因与三种细胞因子基因的表达正相关;通过免疫印迹检测SNAP-23蛋白表达变化,利用酶联免疫吸附试验(ELISA)检测IL-1β的分泌水平,在蛋白水平上验证了SNAP-23表达与IL-1β分泌的正相关性;利用5`RACE和3`RACE技术克隆出鲈鱼SNAP-23全长基因,结合定点突变策略和靶向PCR克隆手段,构建鲈鱼SNAP-23野生型融合质粒pEGFP-SNAP-23wt,Cys缺失突变融合质粒pEGFP-SNAP-23ΔCys和模拟E型肉毒神经毒素(BoNT/E)切割突变融合质粒pEGFP-SNAP-23ΔBoNT/E,以及鲈鱼IL-1β野生型融合表达质粒IL-1β-pEGFP和IL-1β-pEYFP。所有融合蛋白均在鲈鱼巨噬细胞内成功表达,结合ELISA实验结果发现,SNAP-23野生型的表达对IL-1β的分泌有促进作用,而Cys缺失突变体的表达则抑制IL-1β向胞外分泌。首次证实了鱼类巨噬细胞内SNAP-23蛋白在IL-1β分泌过程中的重要作用。此外通过与GFP共表达,定位了IL-1β分子在巨噬细胞内的分布,发现新合成的IL-1β分子很可能像TNFα一样经“内质网-胞质-伪足-胞外” 的分泌路径运出胞外。

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用平板画线法从患病栉孔扇贝(Chlamys farreri)体内分离到了一种原核生物(简称QDP)。QDP可以在改进的液体培养基MEM(含2.2%NaCl,5%小牛血清)和脑心浸液(含2.2% NaCl)中生长;菌落在显微镜下(150×)为无色、透明的小点状;革兰氏染色阴性;菌体为圆形或近似圆形。QDP在发育过程中有两种状态,一种为未成熟阶段,直径小于100nm;另一种为成熟阶段,直径变化很大,最小约60nm,最大可达4µm以上。较小的个体有拟核、核糖体和新月状的空泡,未见细胞壁;较大的个体有细胞壁,胞内大部分被空泡充满,未见拟核和核糖体。栉孔扇贝组织超簿切片电镜观查证实QDP的存在。QDP的密度随着生长发育时间的不同而有所变化,繁殖高峰期密度较大。 建立了密度梯度离心结合滤膜过滤分离技术,优化人工培养条件。最适生长温度为23℃,最适生长pH值为7.4,最适生长盐度相当于细胞培养液所需的盐浓度(0.85%NaCl)。 提取的QDP核酸能被RNase A 降解,且没有检测到DNA。以PCR、RT-PCR扩增其16SrRNA基因序列片段,PCR反应没有扩增出扩增子,而RT-PCR则扩增出了16S rRNA基因序列片段,经测定其序列全长度为1430bp,经与GENEBANK中的16S rRNA片段比较分析,与6种不同科的微生物的同源率最高的为95%-95.47%。 采用温度梯度和病原浓度梯度回归感染实验方法,较为系统地研究了QDP的致病性。研究结果表明:QDP对栉孔扇贝有强烈的致病作用,高温(23℃以上)是其致病的必要条件,证实DQP是栉孔扇贝大规模死亡的病原体之一。

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A quadtree-based adaptive Cartesian grid generator and flow solver were developed. The grid adaptation based on pressure or density gradient was performed and a gridless method based on the least-square fashion was used to treat the wall surface boundary condition, which is generally difficult to be handled for the common Cartesian grid. First, to validate the technique of grid adaptation, the benchmarks over a forward-facing step and double Mach reflection were computed. Second, the flows over the NACA 0012 airfoil and a two-element airfoil were calculated to validate the developed gridless method. The computational results indicate the developed method is reasonable for complex flows.

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A fully 3-D atomistic quantum mechanical simulation is presented to study the random dopant-induced effects in nanometer metal-oxide-semiconductor field-effect transistors. The empirical pseudopotential is used to represent the single particle Hamiltonian, and the linear combination of bulk band method is used to solve the million atom Schrodinger equation. The gate threshold fluctuation and lowering due to the discrete dopant configurations are studied. It is found that quantum mechanical effects increase the threshold fluctuation while decreasing the threshold lowering. The increase of threshold fluctuation is in agreement with the researchers' early study based on an approximated density gradient approach. However, the decrease in threshold lowering is in contrast with the density gradient calculations.

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The compatibility and crystallization behaviour of the mixtures of poly (tetrahydrofuran-methyl methacrylate) diblock copolymer (PTHF-b-PMMA) with polyvinyl chloride has been studied. We found that the compatibility of these blends, in which there is special interaction between the homopolymer and the PMMA block of the copolymer, is much better than that of the AB/A type blends; and that the crystallization rate and crystallinity of PTHF microdomain changed greatly due to the swollen by PVC homopolymer. In this paper, these changes in cryatallization are well explained according to the theories of block copolymer blends and the density gradient model presented by JIANG Ming.

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The main chlorophyll a/b light-harvesting complex (LHC 11) has been isolated directly from thylakoid membranes of marine green alga (Bryopsis corticulans Setch.) by two consecutive runs of anion exchange and gel-filtration chromatography. LHC 11 proteins in the membrane extracts treated with 3% n-Octyl-b-D-glucopyranoside (OG) obtained specific binding ability on Q Sepharose column, and thus were isolated from the thylakoid membranes in a highly selective fraction. The monomeric, trimeric and oligomeric subcomplexes of LHC 11 have been obtained by fractionation of the LHC 11 mixes with sucrose density gradient ultracentrifugation. The SDS-PAGE analysis of peptide composition and absorption spectrum showed that LHC 11 monomers, trimers and oligomers prepared through this work were intact and in high purity. Our report is the first to show that it is possible to purify LHC If directly from thylakoid membranes without extensively biochemical purification.

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The thylakoid membranes were isolated and purified from gametophyte of Porphyrayezoensis Ueda (P yezoensis) by sucrose density gradient ultracentrifugation. After R yezoensis gametophyte thylakoid membranes were solubilized with SDS, the photosystem 11 (PSII) particles were isolated and purified. The activity of PSII particles was determined with DCIP (2,6-dichloroindophenol) photoreduction reaction. The composition of purified PSII particles was detected by SDS-PAGE. As a result, seven proteins including 55 kD protein, 47 kD protein, 43 kD protein, 33 kD protein, 31 kD protein, 29 kD protein, and 18 kD protein were found. Compared with PSII particles of higher plants and other algae, they were identified as D1/D2 complex, CP47, CP43, 33 kD protein, D1, D2 and cyt c-550 respectively. Besides, other three new proteins of 20 kD, 16 kD and 14 kD respectively were found. Among these extrinsic proteins, the 16 kD and 14 kD proteins had not been reported previously, and the 20 kD protein was found for the first time in multicellular red algae.

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Thylakoid membranes were isolated and purified from diploid filamentous sporophytes of Porphyra yezoensis Ueda using sucrose density gradient ultracentrifugation (SDGUC). After thylakoid membranes were solubilized with SDS, the photosystem II (PSII) particles with high 2, 6-dichloroindophenol (DCIP) photoreduction activity were isolated by SDGUC. The absorption and fluorescence spectra, DCIP photoreduction activity and oxygen evolution activity of the thylakoid membranes and PSII particles were determined. The polypeptide composition of purified PSII particles was distinguished by SDS-PAGE. Results showed that PSII particles of sporophytes differed from the gametophytes in spectral properties and polypeptide composition. Apart from 55 kDa D1-D2 heterodimer, CP47, CP43, 33 kDa protein was also detected. However, cyt c-550, 20 kDa, 14 kDa and 16 kDa proteins found in PSII particles from gametophytes were not detected in the sporophytes.

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The highly pure and active photosystem II (PSII) complex was isolated from Bangia fusco-purpurea (Dillw) Lyngb., an important economic red alga in China, through two steps of sucrose density gradient ultracentrifugation and characterized by the room absorption and fluorescence emission spectra, DCIP (2,6-dichloroindophenol) reduction, and oxygen evolution rates. The PSII complex from B. fusco-purpurea had the characteristic absorption peaks of chlorophyll (Chl) a (436 and 676 nm) and typical fluorescence emission peak at 685 nm (Ex = 436 nm). Moreover, the acquired PSII complex displayed high oxygen evolution (139 mu mol O-2/(mg Chl h) in the presence of 2.5 mM 2,6-dimethybenzoqinone as an artificial acceptor and was active in photoreduction of DCIP (2,6-dichloroindophenol) by DPC (1,5-diphenylcarbazide) at 163 U/(mg Chl a h). SDS-PAGE also suggested that the purified PSII complex contained four intrinsic proteins (D1, D2, CP43, and CP47) and four extrinsic proteins (33-kD protein, 20-kD protein, cyt c-550, and 14-kD protein).

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MASNUM wave-tide-circulation coupled numerical model (MASNUM coupled model, hereinafter) is developed based on the Princeton Ocean Model (POM). Both POM and MASNUM coupled model are applied in the numerical simulation of the upwelling off Yangtze River estuary and in Zhejiang coastal waters in summer. The upwelling mechanisms are analyzed from the viewpoint of tide, and a new mechanism is proposed. The study suggests that the tidally inducing mechanism of the upwelling includes two dynamic aspects: the barotropic and the baroclinic process. On the one hand, the residual currents induced by barotropic tides converge near the seabed, and upwelling is generated to maintain mass conservation. The climbing of the residual currents along the sea bottom slope also contributes to the upwelling. On the other hand, tidal mixing plays a very important role in inducing the upwelling in the baroclinic sea circumstances. Strong tidal mixing leads to conspicuous front in the coastal waters. The considerable horizontal density gradient across the front elicits a secondary circulation clinging to the tidal front, and the upwelling branch appears near the frontal zone. Numerical experiments are designed to determine the importance of tide in inducing the upwelling. The results indicate that tide is a key and dominant inducement of the upwelling. Experiments also show that coupling calculation of the four main tidal constituents(M-2, S-2, K-1, and O-1), rather than dealing with the single M-2 constituent, improves the modeling precision of the barotropic tide-induced upwelling.

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本研究分为三个部分:1.以坛紫菜(Porphyra haitanesis Chang et Zheng)的叶状体和丝状体为研究对象,比较坛紫菜叶状体和丝状体的光合色素、色素蛋白的组成,并提取纯化藻红蛋白、藻蓝蛋白、藻胆体及类囊体膜和光系统。研究结果表明坛紫菜叶状体和丝状体色素及色素蛋白的含量不同,藻红蛋白是主要的色素蛋白,坛紫菜叶状体和丝状体的藻红蛋白的含量分别为2.9mg藻红蛋白/g鲜重、4.2mg藻红蛋白/g鲜重,这表明坛紫菜叶状体和丝状体藻红蛋白含量丰富,是提取藻红蛋白很好的材料。藻胆体的性质差异不大,但类囊体膜差异显著,从坛紫菜叶状体中分离到了两种不同的类囊体膜带,光系统Ⅰ(PSⅠ)和PSⅡ分别结合在两条类囊体膜带上,但从坛紫菜丝状体中也分离到两条类囊体膜带,它们的光谱性质和蛋白组成相似,仅放氧速率和DCIP活性有差异,从坛紫菜丝状体中我们仅分离到PSⅡ。坛紫菜叶状体PSⅡ有5种外在蛋白(33、20、Cytc 550、15、12kDa蛋白),而坛紫菜丝状体外在蛋白仅有4条,缺少12kDa蛋白。2. 以在中国江苏部分地区进行了大规模的商业化栽培的突变体条斑紫菜(Porphyra yezoensis Ueda)和野生型条斑紫菜为研究对象,比较其色素及色素蛋白组成、对不能光质的利用率及藻胆体的组成。条斑紫菜和突变型条斑紫菜对不同的光质利用效果有差异,在白光的照射下,野生型紫菜的放氧速率最大,而突变型紫菜在黄光照射下的放氧速率最大。条斑紫菜野生型与突变型色素含量上有明显的差异,突变型紫菜的藻红蛋白含量明显减少而藻蓝蛋白的含量增加。通过杂交的方法证实诱变所获得条斑紫菜突变体为细胞质突变,但是突变型紫菜却发生了由细胞核编码的γ亚基的缺失,这表明突变型紫菜藻红蛋白含量和性质发生了明显的变化。3. 为了找出淡水红藻-深紫美芒藻(Compsopogon coeruleus (Balbis) Montagne)分布狭窄及生物产量低的原因,本文对深紫美芒藻在不同的盐离子浓度下的放氧速率及藻胆体色素组成和结构上进行研究。结果显示:微量的NaCl(0.1mM)促进深紫美芒藻放氧,而深紫美芒藻在较高的NaCl(1、10mM), NaH2PO4 (0.1、1、10mM)和 NH4NO3(0.1、1、10mM)溶液中却没有检测到氧气的产生。这与深紫美芒藻生长的环境一致即深紫美芒藻生活在低盐浓度、低营养的泉水中。深紫美芒藻的藻胆体是由藻红蛋白、藻蓝蛋白及别藻蓝蛋白组成,上面结合α、β和γ亚基,含有藻红胆素、藻篮胆素,但缺乏缺少藻尿胆素。

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InAs was deposited by molecular beam epitaxy (MBE) on a GaAs substrate with an intentional temperature gradient from centre to edge. Two-dimensional (2D) to three-dimensional (3D) morphology evolution was found along the direction in which the substrate temperature was decreasing. Quantum dots (QDs) with density as low as similar to 8 x 10(6) cm(-2) were formed in some regions. We attribute the morphological evolution to the temperature-dependent desorption of deposited indium and the intermixing between deposited indium and gallium from the buffer.