N-acetylmuramic acid 6-phosphate lyases (MurNAc etherases): role in cell wall metabolism, distribution, structure, and mechanism

MurNAc etherases cleave the uniqued-lactyl ether bond of the bacterial cell wall sugar N-acetylmuramic acid (MurNAc). Members of this newly discovered family of enzymes are widely distributed among bacteria and are required to utilize peptidoglycan fragments obtained either from the environment or from the endogenous cell wall (i.e., recycling). MurNAc etherases are strictly dependent on the substrate MurNAc possessing a free reducing end and a phosphoryl group at C6. They carry a single conserved sugar phosphate isomerase/sugar phosphate- binding (SIS) domain to which MurNAc 6-phosphate is bound. Two subunits form an enzymatically active homodimer that structurally resembles the isomerase module of the double-SIS domain protein GlmS, the glucosamine 6-phosphate synthase. Structural comparison provides insights into the two-step lyase-type reaction mechanism of MurNAc etherases: β-elimination of the D-lactic acid substituent proceeds through a 2,3-unsaturated sugar intermediate to which water is subsequently added. Received 31 August 2007; received after revision 12 October 2007; accepted 1 November 2007     

The cytotoxicity of eosinophil cationic protein/ribonuclease 3 on eukaryotic cell lines takes place through its aggregation on the cell membrane

Human eosinophil cationic protein (ECP)/ ribonuclease 3 (RNase 3) is a protein secreted from the secondary granules of activated eosinophils. Specific properties of ECP contribute to its cytotoxic activities associated with defense mechanisms. In this work the ECP cytotoxic activity on eukaryotic cell lines is analyzed. The ECP effects begin with its binding and aggregation to the cell surface, altering the cell membrane permeability and modifying the cell ionic equilibrium. No internalization of the protein is observed. These signals induce cell-specific morphological and biochemical changes such as chromatin condensation, reversion of membrane asymmetry, reactive oxygen species production and activation of caspase-3-like activity and, eventually, cell death. However, the ribonuclease activity component of ECP is not involved in this process as no RNA degradation is observed. In summary, the cytotoxic effect of ECP is attained through a mechanism different from that of other cytotoxic RNases and may be related with the ECP accumulation associated with the inflammatory processes, in which eosinophils are present. Received 26 October 2007; accepted 23 November 2007     

Regulation of phagocyte migration and recruitment by Src-family kinases

Src-family kinases (SFKs) regulate different granulocyte and monocyte/macrophage responses. Accumulating evidence suggests that members of this family are implicated in signal transduction pathways regulating phagocytic cell migration and recruitment into inflammatory sites. Macrophages with a genetic deficiency of SFKs display marked alterations in cytoskeleton dynamics, polarization and migration. This same phenotype is found in cells with either a lack of SFK substrates and/or interacting proteins such as Pyk2/FAK, c-Cbl and p190RhoGAP. Notably, SFKs and their downstream targets also regulate monocyte recruitment into inflammatory sites. Depending on the type of assay used, neutrophil migration in vitro may be either dependent on or independent of SFKs. Also neutrophil recruitment in in vivo models of inflammation may be regulated differently by SFKs depending on the tissue involved. In this review we will discuss possible mechanisms by which SFKs may regulate phagocytic cell migratory abilities.     

On the mechanism of inhibition of p27 degradation by 15-deoxy-Δ12,14-prostaglandin J 2 in lymphoblasts of Alzheimer’s disease patients

It has been proposed that neuroinflammation, among other factors, may trigger an aberrant neuronal cell cycle re-entry leading to neuronal death. Cell cycle disturbances are also detectable in peripheral cells from Alzheimer’s disease (AD) patients. We previously reported that the anti-inflammatory 15- deoxy-Δ^12,14-prostaglandin J ₂ (15d-PGJ ₂) increased the cellular content of the cyclin-dependent kinase inhibitor p27, in lymphoblasts from AD patients. This work aimed at elucidating the mechanisms of 15d-PGJ ₂-induced p27 accumulation. Phosphorylation, half-life, and the nucleo-cytoplasmic traffic of p27 protein were altered by 15d-PGJ2 by mechanisms dependent on PI3K/Akt activity. 15d-PGJ ₂ prevents the calmodulin-dependent Akt overactivation in AD lymphoblasts by blocking its binding to the 85-kDa regulatory subunit of PI3K. These effects of 15d-PGJ ₂ were not mimicked by 9,10-dihydro-15-deoxy-Δ^12,14- prostaglandin J ₂, suggesting that 15d-PGJ ₂ acts independently of peroxisome proliferator-activated receptor γ activation and that the α,β-unsaturated carbonyl group in the cyclopentenone ring of 15d-PGJ ₂ is a requisite for the observed effects. Received 14 July 2008; received after revision 2 September 2008; accepted 12 September 2008     

柴油机燃用生物柴油燃烧与排放

研究了柴油机燃用0号柴油和生物柴油的燃烧放热规律．通过对燃烧特征参数的计算分析,发现生物柴油的燃烧始点有所提前,滞燃期缩短;燃烧初期放热尖峰出现时刻对应的曲轴转角有所提前,瞬时放热率峰值下降;燃烧持续期延长．同时还比较了柴油机燃用生物柴油和0号柴油的经济性和排放特性,发现燃油消耗率增加12％,而各种排放污染物除NOx略有上升外,CO、HC和颗粒物PM均显著下降．     

基于PEMS的混合动力客车排放和油耗性能评价

在中国城市客车行驶工况条件下,运用车栽排放测试技术（portable emission measurement system,简称PEMS）研究了某重型混合动力客车在怠速运转时采取停机和不停机两种不同控制策略下的实际排放因子和燃油消耗率．试验研究结果表明,停机模式比不停机模式可提高该车燃油经济性4．6％,但CO排放因子却增加了53．6％;怠速不停机时,HC＋NOx排放增加20．6％．同时说明对于混合动力车辆的油耗和排放,特别是排放的评价,应当采取基于整车的评价方法．     

无环境信息下多尺度网格细胞群空间表征模型

针对无环境信息条件下网格细胞空间表征问题,提出一种多尺度网格细胞群的空间表征模型.通过减少吸引子网络模型网格细胞的偏好朝向,使网格细胞具有特定的感应方向,仅对特定方向上的速度产生响应.基于改进的吸引子网络模型,构建多尺度网格细胞群,并对空间进行表征.仿真结果表明,改进后的网格细胞模型只响应相应方向的速度,准确整合速度信...     

在Hep G2细胞中利用siRNA沉默GFP基因的实验探讨

目的:通过探讨小分子双链RNA(siRNA)通过RNA干扰(RNAi)机制沉默肝癌细胞株Hep G2荧光素报告基因GFP的各项生物学指标,为进一步实验和临床研究提供依据.方法:通过体外实验,对siRNA和siRNA脂质体的稳定性和细胞毒性进行评估;通过荧光标记技术,研究不同时间siRNA脂质体的转染效率;通过荧光显微镜下观察转染细胞和RT-PCR检测靶基因mRNA表达,确定不同种类、形式、剂量的siRNA对靶基因的沉默效能.结果:在空白培养液中,siRNA和siRNA脂质体可稳定至4h,在5%血清中2～4h后明显降解;100nM的siRNA和siRNA脂质体无明显细胞毒性;siRNA脂质体转染细胞株的效率随时间不同而不同,4h可达90%;非特异性siRNA/脂质体无沉默靶基因表达作用,特异性siRNA/脂质体对靶基因的沉默作用在一定范围内随剂量升高(20nM、50nM、100nM)而升高.结论:siRNA的稳定性可满足实验需要,且无明显细胞毒性,特异性siRNA转染肝癌细胞株能有效抑制靶基因表达,用脂质体转染可提高转染效率.siRNA通过RNAi机制沉默靶基因表达,为肿瘤的治疗和基础研究提供了新的工具和思路.     

磁固载雷尼镍在固定床反应器中的催化性能

采用强-弱双磁性载体固载雷尼镍催化剂,在自制的鼓泡固定床反应器中对对羟基亚苄基海因和邻硝基乙苯体系进行氢化实验.结果表明,雷尼镍催化剂的活性和稳定性均有所提高.对羟基苄基海因的平均收率达到90%以上,邻硝基乙苯的平均转化率超过99%,催化剂寿命可提高到300 h以上.     

120kW级燃料电池可变喉口引射器的设计及特性

引射器是质子交换膜燃料电池氢气循环系统中的关键部件，依靠超音速流动的射流工质实现阳极排气的再循环。传统的固定结构引射器通常针对电堆额定工况设计，工作在非设计工况时性能较差；而可变喉口引射器可以动态地改变其喉口大小，有效扩大其工作范围。基于Sokolov设计理论，搭建了引射器的一维模型并探究了其工作特性，在此基础上提出了一种可变喉口引射器的设计方法。研究结果表明，引射器的工作特性主要受其操作压力、喉口直径和混合室直径影响；通过缩小喉口、提高工作流体压力可以使工作喷嘴保持临界状态，这对大负载下的引射性能影响较小，但能够有效提高电堆小负载下的引射比，并使引射器的工作范围向小负载扩展。