Energy metabolism of <Emphasis Type="Italic">Heliobacterium modesticaldum</Emphasis> during phototrophic and chemotrophic growth

Background  

Heliobacterium modesticaldum is a gram-positive nitrogen-fixing phototrophic bacterium that can grow either photoheterotrophically or chemotrophically but not photoautotrophically. Surprisingly, this organism is lacking only one gene for the complete reverse tricarboxylic acid (rTCA) cycle required for autotrophic carbon fixation. Along with the genomic information reported recently, we use multiple experimental approaches in this report to address questions regarding energy metabolic pathways in darkness, CO₂ fixation, sugar assimilation and acetate metabolism.     

Genome-wide investigation reveals high evolutionary rates in annual model plants

Background  

Rates of molecular evolution vary widely among species. While significant deviations from molecular clock have been found in many taxa, effects of life histories on molecular evolution are not fully understood. In plants, annual/perennial life history traits have long been suspected to influence the evolutionary rates at the molecular level. To date, however, the number of genes investigated on this subject is limited and the conclusions are mixed. To evaluate the possible heterogeneity in evolutionary rates between annual and perennial plants at the genomic level, we investigated 85 nuclear housekeeping genes, 10 non-housekeeping families, and 34 chloroplast genes using the genomic data from model plants including Arabidopsis thaliana and Medicago truncatula for annuals and grape (Vitis vinifera) and popular (Populus trichocarpa) for perennials.     

Antibiotic susceptibility of coagulase-negative staphylococci isolated from very low birth weight babies: comprehensive comparisons of bacteria at different stages of biofilm formation

Background

Coagulase-negative staphylococci are major causes of bloodstream infections in very low birth weight babies cared for in Neonatal Intensive Care Units. The virulence of these bacteria is mainly due to their ability to form biofilms on indwelling medical devices. Biofilm-related infections often fail to respond to antibiotic chemotherapy guided by conventional antibiotic susceptibility tests.

Methods

Coagulase-negative staphylococcal blood culture isolates were grown in different phases relevant to biofilm formation: planktonic cells at mid-log phase, planktonic cells at stationary phase, adherent monolayers and mature biofilms and their susceptibilities to conventional antibiotics were assessed. The effects of oxacillin, gentamicin, and vancomycin on preformed biofilms, at the highest achievable serum concentrations were examined. Epifluorescence microscopy and confocal laser scanning microscopy in combination with bacterial viability staining and polysaccharide staining were used to confirm the stimulatory effects of antibiotics on biofilms.

Results

Most coagulase-negative staphylococcal clinical isolates were resistant to penicillin G (100%), gentamicin (83.3%) and oxacillin (91.7%) and susceptible to vancomycin (100%), ciprofloxacin (100%), and rifampicin (79.2%). Bacteria grown as adherent monolayers showed similar susceptibilities to their planktonic counterparts at mid-log phase. Isolates in a biofilm growth mode were more resistant to antibiotics than both planktonic cultures at mid-log phase and adherent monolayers; however they were equally resistant or less resistant than planktonic cells at stationary phase. Moreover, for some cell-wall active antibiotics, concentrations higher than conventional MICs were required to prevent the establishment of planktonic cultures from biofilms. Finally, the biofilm-growth of two S. capitis isolates could be enhanced by oxacillin at the highest achievable serum concentration.

Conclusion

We conclude that the resistance of coagulase-negative staphylococci to multiple antibiotics initially remain similar when the bacteria shift from a planktonic growth mode into an early attached mode, then increase significantly as the adherent mode further develops. Furthermore, preformed biofilms of some CoNS are enhanced by oxacillin in a dose-dependent manner.     

抑制肿瘤生长活性的内皮生长抑制剂EDI-8t

对内源性的内皮生长抑制剂的序列来源分析后,用RT-PCR方法从人脐带中获得了417bp的cDNA,并克隆到毕赤酵母表达载体pPIC9,然后转化毕赤酵母GS115获得重组表达菌株。表达菌株通过甲醇诱导表达后,将发酵液通过分离纯化获得重组蛋白纯品,命名为EDI-8t。体外实验结果表明,这个胶原蛋白VIII来源的rhEDI-8t蛋白能特异性地抑制牛主动脉内皮细胞的增殖和迁移,并能引起内皮细胞的凋亡。动物体内实验结果表明,rhEDI-8t蛋白样品可有效抑制裸鼠皮下肝癌肿瘤的生长,抑制效果和参照品endostatin相同。但在小鼠黑色素瘤肺转移模型中,rhEDI-8t显示了高于参照品的抑癌效果。     

秋水仙碱对微管蛋白的作用机制及其细胞效应研究进展

秋水仙碱诱导染色体同源加倍的生物学机理与构成纺锤体微管蛋白密切相关. 秋水仙碱作用于细胞的根本效应是改变细胞微管的状态,使微管解聚或停止组装;秋水仙碱作用于微管的方式是其分子结构中的 A 环与β微管蛋白354半胱氨酸结合、C环结合在239半胱氨酸和N末端氨基酸;不同植物种类微管蛋白的处理效应有明显的差异.秋水仙碱的处理效应影响到细胞一切与微管活动有关的功能,具体表现为改变细胞的发育进程、阻断染色体的分裂及细胞器不能正常运动,除此之外,秋水仙碱还可以诱导染色体结构变异.本文主要综述了秋水仙碱作用于微管蛋白的机制及秋水仙碱处理的细胞效应等研究进展,为该领域的研究提供信息资料.     

人类巨细胞病毒编码US28受体拮抗性多肽的筛选鉴定及体外活性研究

立足US28的广谱趋化因子结合活性,寻找拮抗剂多肽的先导物．通过预测US28的结合模拟位设计多肽序列,合成相关多肽,再利用自建的25种趋化因子随机构成的噬菌体12肽库筛选结合模拟位,竞争性ELISA方法验证,从而获得30个阳性克隆,测序并推导氨基酸序列,得到7种序列：GSESLNAHCALW、EIDGFNAHCALL、VIARLNAHCALR、ATECLNAHCALW、VIESLNAHCALW、DNGSINAHCALL及VKKTLNAHCALR．所有序列富含疏水氨基酸,其中8个克隆有LNAHCAL保守序列．采用趋化抑制实验检测多肽对生理性趋化因子引起的细胞迁移活性的影响,流式细胞仪检测细胞内钙离子浓度变化．结果表明,利用人源趋化因子序列构建肽库并筛选有效序列的方法取得成功,筛选出的阳性克隆保守序列LNAHCAL可以模拟Human MIP-1β与US28 N端的结合位,从而与合成多肽H22结合,多肽H22可以阻断受体结合生理性趋化因子形成的趋化作用,本身不引起趋化运动,不影响胞内信号转导和细胞自然活性,具有广谱趋化因子拮抗剂的可能性．     

井冈山杜鹃叶片抗氧化系统对高温胁迫的响应

以井冈山杜鹃(Rhododendron jinggangshanicum Tam)4年生实生苗为材料,通过人工气候箱盆栽实验,研究了不同高温处理下其叶片形态及生理生化指标的变化,以阐明高温胁迫下井冈山杜鹃抗氧化系统的防御机理.结果显示:(1)30℃和38℃处理6 d后,叶片过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)及过氧化氢酶(CAT)活性分别比对照极显著(P<0.01)升高44.44%和127.16%、33.12%和129.35%、106.52%和188.64%;超氧化物歧化酶(SOD)活性增幅较小,分别比对照增加3.77%和5.11%.(2)随着胁迫温度的升高,可溶性蛋白质、脯氨酸(Pro)含量均明显上升,而抗坏血酸(AsA)含量却显著降低;除30℃处理与对照之间的可溶性蛋白质含量差异显著(P<0.05)外,其它处理间差异均达到极显著(P<0.01).(3)随着胁迫温度的升高,叶片过氧化氢(H2O2)、丙二醛(MDA)含量大量积累,各处理间差异达到极显著或显著;而形态上则表现为受害加重,植株出现轻度萎蔫、老叶褐化脱落、新叶坏死等现象.研究表明,井冈山杜鹃受到高温胁迫的氧化伤害可能与AsA含量显著下降和H2O2、MDA含量大幅增加有关,其植物体能诱导增加抗氧化保护酶活性及Pro含量以提高其抗氧化能力,从而减轻高温引发的活性氧毒害.     

串联质谱图谱从头测序算法研究进展

近年来,基于质谱技术的高通量蛋白质组学研究发展迅速,利用串联质谱图谱鉴定蛋白质是其数据处理中一个基础而又重要的环节．由于不需要利用蛋白质序列数据库,从头测序方法能够分析新物种或者基因组未测序物种的串联质谱数据,具有数据库搜索方法不可替代的优势．简要介绍高通量串联质谱图谱从头测序问题及其研究现状．归纳出几种典型的计算策略并分析了各种策略的优缺点．总结常用的从头测序算法和软件,介绍算法评估的各种指标和常用评估数据集,概括各种算法的特点,展望未来研究可能的发展方向．     

线粒体内膜解偶联蛋白UCP1在大肠杆菌中的活性表达

线粒体的呼吸耗氧偶联着ATP的合成,而位于线粒体内膜上的跨膜蛋白解偶联蛋白(uncoupling protein,UCP)能够破坏这种偶联关系.在大肠杆菌中表达了有生物活性的鼠源解偶联蛋白1(rUCP1).重组rUCP1的表达导致大肠杆菌宿主细胞生长变慢;在电子显微镜下观察免疫标记的结果显示,重组rUCP1主要表达在细菌膜上;同时将rUCP1重构到脂质体中也能够测到质子转运活性.这些结果说明,真核生物UCP1能够在原核生物中表达出有生物活性的形式,且能纯化得到足量的rUCP1蛋白用于进一步的结构生物学研究.     

寒冷诱导产热时大鼠棕色脂肪能量代谢相关蛋白谱的变化

棕色脂肪组织(BAT)的生理作用与白色脂肪显著不同,它以产热的形式释放能量而不是将能量以ATP的形式储存．线粒体是在能量代谢和维持细胞稳态中具有重要功能的细胞器．为了更好地了解棕色脂肪中的能量代谢过程,运用双向电泳及质谱相结合的技术,分离了大鼠白色和棕色脂肪线粒体,对其差异蛋白质谱进行了系统分析和鉴定．参与脂肪和氨基酸代谢、三羧酸循环及线粒体呼吸链的蛋白质在棕色脂肪线粒体中的表达明显高于白色脂肪线粒体,在寒冷诱导下这些蛋白质的表达进一步上调．此外,参与辅酶Q合成的一系列COQ 基因在棕色脂肪中经寒冷适应后表达明显上调．该研究表明,辅 酶Q合成的增高在非颤栗性产热中具有重要作用,为进一步了解棕色脂肪特异性的能量代谢提供了新的思路．