982 resultados para Aaa-atpase


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在回转器模拟微重力刺激导致盐生杜氏藻细胞代谢特性变化的基础上,通过几种代谢抑制剂和激活剂的实验及其结果分析,发现细胞质膜可能是盐生杜氏藻细胞最直接的微重力感受体;质膜磷脂-蛋白、PMH+-ATPase、膜电位、Ca2+及钙调蛋白在其信号传导与响应过程中起到重要的作用.构建了盐生杜氏藻对微重力刺激感受、传导与响应的初步模型.

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Ure2p is the protein determinant of the Saccharomyces cerevisiae prion state [URE3]. Constitutive overexpression of the HSP70 family member SSA1 cures cells of [URE3]. Here, we show that Ssa1p increases the lag time of Ure2p fibril formation in vitro in the presence or absence of nucleotide. The presence of the HSP40 co-chaperone Ydj1p has an additive effect on the inhibition of Ure2p fibril formation, whereas the Ydj1p H34Q mutant shows reduced inhibition alone and in combination with Ssa1p. In order to investigate the structural basis of these effects, we constructed and tested an Ssa1p mutant lacking the ATPase domain, as well as a series of C-terminal truncation mutants. The results indicate that Ssa1p can bind to Ure2p and delay fibril formation even in the absence of the ATPase domain, but interaction of Ure2p with the substrate-binding domain is strongly influenced by the C-terminal lid region. Dynamic light scattering, quartz crystal microbalance assays, pull-down assays and kinetic analysis indicate that Ssa1p interacts with both native Ure2p and fibril seeds, and reduces the rate of Ure2p fibril elongation in a concentration-dependent manner. These results provide new insights into the structural and mechanistic basis for inhibition of Ure2p fibril formation by Ssa1p and Ydj1p.

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Iron is an essential trace element for biological requirements of phytoplankton. Effects of iron on physiological and biochemical characteristics of Microcystis wesenbergii were conducted in this study. Results showed that 0.01 mu M [Fe3+] seriously inhibited growth and chlorophyll synthesis of M. wesenbergii, and induced temporary increase of ATPase activities, however, NR. ACP and ALP activities were restrained by iron limitation. Interestingly, iron addition on day 8 resulted in the gradual restoration of structures and functions of above enzymes and resisted a variety of stresses from iron limitation. M. wesenbergii in 10 mu M [Fe3+] treatment group grew normally. enzymes maintained normal levels, and residual phosphate contents in cultures first sharply decreased, then smoothly as M. wesenbergii has a characteristic of luxury consumption of phosphorus. Above parameters in 100 mu M [Fe3+] treatment group were almost same with those in 10 mu M [Fe3+] treatment group except for NR, ACP and ALP activities. In 100 mu M [Fe3+] treatment group, activities of ACP and ALP had temporary increase because phosphate and ferric iron could form insoluble compound - ferric phosphate (Fe3PO4) through adsorption effect. resulting in lack of bioavailable phosphate in culture media. The experiment suggested that too low or too high iron can affect obviously physiological and biochemical characteristics of M. wesenbergii.

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This study examined the toxic effects of microcystins on mitochondria of liver and heart of rabbit in vivo. Rabbits were injected i.p. with extracted microcystins (mainly MC-RR and -LR) at two doses, 12.5 and 50 MCLReq. mu g/kg bw, and the changes in mitochondria of liver and heart were studied at 1, 3,12, 24 and 48 h after injection. MCs induced damage of mitochondrial morphology and lipid peroxidation in both liver and heart. MCs influenced respiratory activity through inhibiting NADH dehydrogenase and enhancing succinate dehydrogenase (SDH). MCs altered Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities of mitochondria and consequently disrupted ionic homeostasis, which might be partly responsible for the loss of mitochondrial membrane potential (MMP). MCs were highly toxic to mitochondria with more serious damage in liver than in heart. Damage of mitochondria showed reduction at 48 h in the low dose group, suggesting that the low dose of MCs might have stimulated a compensatory response in the rabbits. (C) 2008 Elsevier Inc. All rights reserved.

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Tributyltin (TBT) is widely used as antifouling paints, agriculture biocides, and plastic stabilizers around the world, resulting in great pollution problem in aquatic environments. However, it has been short of the biomonitor to detect TBT in freshwater. We constructed the suppression subtractive hybridization library of Tetrahymena thermophila exposed to TBT, and screened out 101 Expressed Sequence Tags whose expressions were significantly up- or down-regulated with TBT treatment. From this, a series of genes related to the TBT toxicity were discovered, such as glutathione-S-transferase gene (down-regulated), plasma membrane Ca2+ ATPase isoforms 3 gene (up-regulated) and NgoA (up-regulated). Furthermore, their expressions under different concentrations of TBT treatment (0.5-40 ppb) were detected by real time fluorescent quantitative PCR. The differentially expressed genes of T thermophila in response to TBT were identified, which provide the basic to make Tetrahymena as a sensitive, rapid and convenient TBT biomonitor in freshwater based on rDNA inducible expression system. (c) 2006 Elsevier B.V. All rights reserved.

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The effects of salt stress on carbohydrate metabolism in Microcoleus vaginatus Gom., a cyanobacterium isolated from desert algal crusts, were investigated in the present study. Extracellular total carbohydrates and exopolysaccharides (EPS) in the culture medium produced by M. vaginatus increased significantly during the growth phase and reached a maximum during the stationary phase. The production of extracellular carbohydrates also significantly increased under higher salt concentrations, which was attributed to an increase in low molecular weight carbohydrates. In the presence of NaCl, the production of cellular total carbohydrates decreased and photosynthetic activity was impaired, whereas cellular reducing sugars, water-soluble sugars and sucrose content and sucrose phosphate synthase activity increased, reaching a maximum in the presence of 200 mmol/L NaCl. These parameters were restored to original levels when the algae were transferred to a non-saline medium. Sodium and K+ concentrations of stressed cells decreased significantly and H+-ATPase activity increased after the addition of exogenous sucrose or EPS. The results suggest that EPS and sucrose are synthesized to maintain the cellular osmotic equilibrium between the intra- and extracellular environment, thus protecting algal cells from osmotic damage, which was attributed to the selective exclusion of cellular Na+ and K+ by H+-ATPase.

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Clinorotation experiments were established to simulate microgravity on ground. It was found that there were obvious changes of Dunaliella salina FACHB435 cells and their metabolic characteristics during clinorotation. The changes included the increases of glycerol content, the rate of H+ secretion and PM H+-ATPase activity, and the decrease of ratio of the plasma membrane (PM) phospholipid to PM protein. These results indicated that microgravity was a stress environment to Dunaliella salina. It is deduced that it would be possible to attribute the effect of microgravity on algal cells to the secondary activation of water stress.

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随着网络带宽、计算机处理能力和存储容量的迅速提高,以及各种视频信息处理技术的出现,全程数字化、网络化的视频监控系统优势愈发明显。其高度的开放性、集成性和灵活性为视频监控系统和设备的整体性能提升创造了必要的条件;同时也为整个安防产业的发展提供了更加广阔的发展空间,崭新的应用模式和市场机遇不断涌现。视频监控系统过程向着大型、连续、综合化发展,形成了复杂监控过程,监测控制的要求越来越高,需要更高性能的系统和采用更优秀的控制手段,面临着不能用传统方法解决的新问题。本文概述了目前视频监控中面临的挑战,简要介绍了与视频监控相关的研究领域和研究现状,研究了视频监控中若干亟待解决的问题,主要取得了以下几个方面的研究成果: 第一,提出了基于AdaBoost的改进的人脸检测算法,针对AdaBoost算法的训练速度慢的问题,提出了基于阈值控制的训练方法;同时研究了AdaBoost算法人脸检测方法,利用肤色模型检测人脸区域,并对颜色模型进行了光照补偿。实验结果表明本文的算法具有较好的检测结果。 第二,提出了基于Canny算法的一般目标检测算法,提出了改进的Canny边缘检测算法,研究了Canny算法中噪声抑制的方法,采用改进中心加权的MTM算法有效的抑制噪声。针对Canny检测算法中阈值设置的问题,提出改进的Canny阈值补偿的方法。实验结果表明,改进的Canny算法相比原算法具有更好的目标检测性能。 第三,提出了一种基于均值漂移(Mean Shift)的改进的目标跟踪算法,通过搜索窗口带宽的计算,加权背景信息以及卡尔曼滤波器建模改进了跟踪算法,避免了均值漂移算法中的一些关键问题。对比实验结果表明,本文的改进方法相比原算法具有较好的性能。 第四,研究了视频监控中基于可扩展视频编码(SVC)的技术。首先讨论了视频监控中采用可扩展视频编码(SVC)的优势,探讨了视频监控中采用可扩展视频编码(SVC)的框架。然后针对于视频质量评估问题,设计并实现了基于可扩展视频编码(SVC)的视频质量评估系统Evalvid-SVC,研究了基于可扩展视频编码(SVC)的视频质量评估。 第五,研究并实现了视频数据安全传输技术。提出了基于Diameter的统一认证方案。任何用户想要获取视频资源,都必须通过AAA子系统的认证和授权,授予合法用户以特定的方式使用资源。另外,为了保证监控数据从监控前端安全地传送到视频监控客户端,本文提出了一种有效的保证视频数据安全传输的方案。设计的数据加密算法应用DES算法对前端设备采集到的音视频数据进行加密,并通过定时更新和RSA加密的方式保护和传输DES密钥。

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针对传统分布式入侵检测系统组件之间依赖程度大、系统不够健壮且入侵检测系统自身结构固定不能适应入侵的变化的问题,提出了一种基于Agent的自适应的分布式入侵检测系统(简称AAA-DIDS)·AAADIDS采用Agent概念重新构造系统的组件,改进了分布式入侵检测系统由于高层节点单一无冗余而产生的可靠性差的缺陷,从构造上克服了分布式入侵检测系统的脆弱性·同时,AAADIDS系统采用智能技术构建了自适应的入侵检测系统模型,增加了系统应对入侵行为变化的智能性·AAA-DIDS系统相对于传统的分布式入侵检测系统有效地提高了系统自身的可靠性和针对外界变化的适应能力·

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超支化聚合物是一类高度支化的具有三维椭球状立体构造的大分子。由于具有传统线形聚合物所没有的低粘度、高流变、良好的溶解性及大量末端官能团等物理化学特性,近年来已成为高分子科学界研究的一个热点。十多年来,人们在合成方法、表征手段、应用及理论研究等方面取得了可喜的成就。但是,目前超支化聚合物的发展还存在着合成方法和所合成的聚合物种类有限,成本较高及结构可控性差等问题。设计合成了五个系列的新型ABx单体,二经基苯氯代苯酞亚胺、二乙酞氧基苯甲酸苯酞亚胺、三乙酞氧基苯酞胺酸,二经基苯基联苯酰胺酸和多轻基烷基联苯酞胺酸,再分别通过缩聚反应一步成功制备了新型芳香超支化聚醚酞亚胺、芳香超支化聚酷酰亚胺,可降解的超支化聚酷酞胺、芳香和半芳香超支化聚酯酞胺。通过傅立叶红外光谱(FT)、核磁共振波谱(NMR)凝胶渗透色谱(GPC)、热差(DSC)和热重(TGA)等分析手段,详细研究了它们的结构和性质,这些聚合物都具有较低的粘度、良好的溶解性和热稳定性。末端基团的种类和性质在很大程度上影响聚合物的性质。通过小角X一射线散射仪和紫外一可见光谱研究了由天然原料制备的超支化聚酷酞胺的降解行为。设计了由商品化原料,二梭酸酐(AAA,型)与二乙醇胺归32型)、脂肪二梭酸配与多经基伯胺(CBx型)和二酸(Az型)与多轻基伯胺一步合成超支化聚合物的新方法,成功地合成了二十四种不同结构的新型超支化聚酷酰胺。通过FTIR、NMR和DEPT NMR、GPC、基质辅助激光解析时间飞行质谱(MALDI-TOF-MS)、DSC和TGA等分析手段,详细研究了它们的结构和性质。这些聚合物都具有较低的粘度、良好的溶解性。

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以贾第虫、毛滴虫、内变形虫和微抱子虫等为代表的几类原生生物,不仅因为他们的寄生致病性而在医学上长期备受关注,它们的进化地位也是一个十分令人注目的问题。因为曾认为它们不具线粒体等细胞器,再加上一些分子系统学研究表明它们处在真核生物的最基部,因此不少人认为它们是在线粒体产生之前即已分化的极原始真核生物,其进化地位是处在原核生物向真核生物的过渡阶段,并有人称之为achezoa。这一发现一度被认为对探讨真核细胞(生物)的起源进化极为重要,是进化生物学上的重要突破。然而,近年来不断有新的证据对此提出质疑,其进化地位也就存在较大争议。本文首先利用PCR扩增、测序和基因组数据库搜索等技术方法鉴定了蓝氏贾第虫(Giardialamblia)、阴道毛滴虫(Trichomonasvaginalis)和痢疾内变形虫(entamoebahistolytica)的II型DNA拓扑异构酶基因序列。RT-PCR和序列分析表明它们均不具内含子。蛋白质序列搜索的结果表明它们与其它真核生物的DNA拓扑异构酶H是高度同源的。用生物信息学的方法,我们还对这些酶的性质进行了初步分析。分析还表明蓝氏贾第虫的DNA拓扑异构酶H具有一些不同于其.宿主的特征,如在ATPase区和中间区有六个插入,中间区要长大约100个氨基酸,而C端区又短大约200个氨基酸且富含带电荷的氨基酸残基。这些结果对研制以该酶为靶分子的专一性抗贾第虫药物具有指导意义。其次,将上述获得的序列数据结合GenBank数据库中已有的脑炎微抱子虫(Encephalitozooncuniculi)和其它一系列处在不同进化地位的真核生物的相应序列数据,用多种方法构建出分子系统树,对这些"无线粒体"原生生物的进化地位进行了探讨,并对"长枝吸引"对系统树的影响进行了分析。结果表明,由于DNA拓扑异构酶H的特点和可以克服"长枝吸引"等以往分子系统分析中的不足,所构建的系统树不仅能有效地反映出已普遍接受的真核生物各主要类群的系统关系,而且显示出这些"无线粒体"原生动物不同于以前系统树所反映的进化地位:它们并非是最早分支出来的真核生物,而是在具有线粒体的生物如动基体类或菌虫类等之后才分化的、分别属于不同进化地位的类群。结合近来它们中发现了类似线粒体细胞器等证据,我们认为这些所谓"无线粒体"的原生生物虽然其中有些种类(如以贾第虫为代表的双滴虫类)进化地位很低等,对探讨真核细胞的早期进化具有一定意义,但总体上它们并非过去所认为的那么极端原始,它们应该是线粒体产生之后才分别分化出来的不同生物类群

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近年来,随着温室气体体积分数不断上升,研究CO2和O3体积分数升高对植物的影响已取得一定进展,但二者对植物的复合作用及生理研究不够深入。文章利用开顶式气室研究了大气CO2和O3体积分数升高对银杏(Ginkgo biloba L.)光合特性的影响。结果表明,在整个生长季内,与对照相比,在大气CO2体积分数为700×10-6条件下,银杏叶片净光合速率显著增加(P<0.05),希尔反应活力增大,Ca2+/Mg2+-ATPase活性增强,光合产物可溶性糖和淀粉含量增多;而在O3体积分数为80×10-9的情况下,银杏叶片净光合速率下降,希尔反应活力减小,Ca2+/Mg2+-ATPase活性减弱,光合产物可溶性糖和淀粉含量减少;在CO2和O3复合作用(700×10-6+80×10-9)条件下,银杏叶片净光合速率、希尔反应活力、可溶性糖和淀粉均有所增加,且淀粉含量增加极显著(P<0.01),而Ca2+-ATPase活性先增强后减弱,Mg2+-ATPase活性先减弱后增强。说明CO2可缓解O3对银杏的负效应,而O3亦对CO2的正效应有削弱作用。

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毫秒延迟发光测定结果表明低温弱光处理黄瓜叶片导致类囊体原位 (in situ)耦联度显著降低。DCCD可以恢复低温弱光处理的黄瓜叶片的毫秒延迟发光的慢相强度和反映类囊体膜质子吸收的 9- AA(9- Aminoacridine)荧光猝灭能力 ,说明类囊体耦联度降低的原因是质子由 CF0 大量快速渗漏。进一步研究结果表明 ,活性氧和 CF1的脱落不是低温弱光引起黄瓜类囊体耦联度降低的根本原因。

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拓扑异构酶(topoisomerase)是一类控制和修改双螺旋DNA复制和转录过程中的拓扑结构的酶,是生命活动中最重要的酶。以IA类和II类拓扑异构酶中共存的toprim以及CAP-like结构域为研究对象,对拓扑异构酶中的三大类酶的分子进化情况进行了分析。结果显示在IA类和II类酶之间序列保守性很低,但是具有两个保守的结构域,在IIB类拓扑异构酶中toprim结构域中存在着和其他toprim结构域相同的四个保守位点,而在CAP-like结构域中IA类和II类中存在较大差异,没有明显的序列保守性,IIB类和IIA类的CAP-like结构域在二级结构上非常相似。从toprim结构域系统进化研究中我们发现IIA类和皿类中toprim结构域的进化关系很近,两类酶的toprim结构域在亲缘关系上和primase较远,而以上三者和IA类的进化关系最远。CAP-like结构域的系统进化研究发现IIA类以及IA类的domain4的CAP-like结构域进化关系比较近,IIB类和他们之间关系稍微远一些,IA类的domain3和以上几个结构域的关系较远,这也与他们的二级结构上的一致性是相同的。通过分析,IIA、IIB类起源于类似IA类的古老的拓扑异构酶,'在IA类进化中经过基因复制产生了两个不同的CAP-like结构域。然后祖先拓扑异构酶发生了变化,N'端加入了ATPase结构域和DNAgyrase/Mutlsecond结构域,形成了严格依赖ATP供能的真核生物IIA类,在细菌中断开成为两个亚基的细菌中IIA类。IIB类是祖先细胞的IIA类的一个或者是两个亚单位在古细菌以及真核生物中通过复制、重组和缺失造成的,IIB类中的toprim结构域很接近IIA类,可以认为,llB类中的toPrim结构域直接由IIA类转移而来,而IIB类中的cAP一1汰e结构域较IIA类中产生更早一些,应该是由拓扑异构酶祖先中产生的二级结构为aβaaββ的CAP-like结构域直接进化而来。然而,两个结构域的基因在连接到一起时候发生了不同于一般顺序的拼接,于是nB类中两个结构域形成了不同于现在的IA类和IIA类的顺序。

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The assembly and disassembly of RecA-DNA nucleoprotein filaments on double-stranded DNA (dsDNA) or single-stranded DNA (ssDNA) are important steps for homologous recombination and DNA repair. The assembly and disassembly of the nucleoprotein filaments are sensitive to the reaction conditions. In this work, we investigated different morphologies of the formed nucleoprotein filaments at low temperature under different solution conditions by atomic force microscopy (AFM). We found that low temperature and long keeping time could induce the incomplete disassembly of the formed nucleoprotein filaments.