Biological and Potential Therapeutic Roles of Sirtuin Deacetylases

Sirtuins comprise a unique class of nicotinamide adenine dinucleotide (NAD⁺)-dependent deacetylases that target multiple protein substrates to execute diverse biological functions. These enzymes are key regulators of clinically important cellular and organismal processes, including metabolism, cell division and aging. The desire to understand the important determinants of human health and lifespan has resulted in a firestorm of work on the seven mammalian sirtuins in less than a decade. The implication of sirtuins in medically important areas such as diabetes, cancer, cardiovascular dysfunction and neurodegenerative disease has further catapulted them to a prominent status as potential targets for nutritional and therapeutic development. Here, we present a review of published results on sirtuin biology and its relevance to human disease. Received 25 June 2008; received after revision 20 August 2008; accepted 29 August 2008     

Zinc binding to peptide analogs of the structural zinc site in alcohol dehydrogenase: Implications for an entatic state

Zinc binding to the peptide replica and analogs to residues 93–115 of horse liver alcohol dehydrogenase (ADH) was examined by competition of the peptides and the chromophoric chelator 4-(2- pyridylazo)resorcinol for zinc and X-ray absorption fine structure analysis of the zinc ligands. In the enzyme, zinc is coordinated by four Cys residues. In the peptide replica, zinc is bound to three Cys and one His residue. A four-Cys zinc coordination is observed only when His is removed, leading to increased zinc stability. ADH crystal structures reveal that the ε-amino group of the conserved residue Lys323 is within H-bond distance of the backbone amide oxygens of residues 103, 105 and 108, likely stabilizing the zinc coordination in the enzyme. The peptide data thus indicate structural strain and increased energy in the zinc-binding site in the protein, characteristic of an entatic state, implying a functional nature for this zinc site. Received 3 July 2008; received after revision 11 August 2008; accepted 1 September 2008     

Na+ binding to meizothrombin desF1

Meizothrombin is the physiologically active intermediate generated by a single cleavage of prothrombin at R320 to separate the A and B chains. Recent evidence has suggested that meizothrombin, like thrombin, is a Na(+)-activated enzyme. In this study we present the first X-ray crystal structure of human meizothrombin desF1 solved in the presence of the active site inhibitor PPACK at 2.1 A resolution. The structure reveals a Na(+) binding site whose architecture is practically identical to that of human thrombin. Stopped-flow measurements of Na(+) binding to meizothrombin desF1 document a slow phase of fluorescence change with a k(obs) decreasing hyperbolically with increasing [Na(+)], consistent with the existence of three conformations in equilibrium, E*, E and E:Na(+), as for human thrombin. Evidence that meizothrombin exists in multiple conformations provides valuable new information for studies of the mechanism of prothrombin activation.     

Dissection of a novel molecular determinant mediating Golgi to trans-Golgi network transition

Two major functions of the Golgi apparatus (GA) are formation of complex glycans and sorting of proteins destined for various subcellular compartments or secretion. To fulfill these tasks proper localization of the accessory proteins within the different sub-compartments of the GA is crucial. Here we investigate structural determinants mediating transition of the two glycosyltransferases β-1,4- galactosyltransferase 1 (gal-T1) and the α-1,3-fucosyltransferase 6 (fuc-T6) from the trans-Golgi cisterna to the trans-Golgi network (TGN). Upon treatment with the ionophore monensin both glycosyltransferases are found in TGN-derived swollen vesicles, as determined by confocal fluorescence microscopy and density gradient fractionation. Both enzymes carry a signal consisting of the amino acids E₅P₆ in gal-T1 and D₂P₃ in fuc-T6 necessary for the transition of these glycosyltransferases from the trans-Golgi cisterna to the TGN, but not for their steady state localization in the trans-Golgi cisterna. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Received 30 July 2008; received after revision 17 September 2008; accepted 29 September 2008     

ERK activation by Ca2+ ionophores depends on Ca2+ entry in lymphocytes but not in platelets, and does not conduct membrane scrambling

Rapid Ca²⁺-dependent phospholipid (PL) reorganization (scrambling) at the plasma membrane is a mechanism common to hematopoietic cells exposing procoagulant phosphatidylserine (PS). The aim of this research was to determine whether activation of the extracellular signal-regulated kinase (ERK) pathway was required for PL scrambling, based on a single report analyzing both responses induced by Ca²⁺ ionophores in megakaryoblastic HEL cells. Ca²⁺ ionophore-stimulated ERK phosphorylation was induced in platelets without external Ca²⁺, whereas exogenous Ca²⁺ entry was crucial for ERK activation in Jurkat T cells. In both cells, membrane scrambling only occurred following Ca²⁺ entry and was not blocked by inhibiting ERK phosphorylation. Furthermore, ERK proteins are strongly phosphorylated in transformed B lymphoblastic cell lines, which do not expose PS in their resting state. Overall, the data demonstrated that ERK activation and membrane scrambling are independent mechanisms. A. Arachiche, I. Badirou: These authors contributed equally to this work. Received 18 June 2008; received after revision 24 September 2008; accepted 1 October 2008     

基于圆盘极点约束的系统不确定项容忍区间分析

在线性系统中基于Lyapunov稳定性理论,讨论了双不确定项同时波动时其容忍区间的算法,并结合闭环极点配置和线性矩阵不等完成圆盘极点配置问题中系统不确定项的冗余设计。数值仿真验证了本文控制器设计方法的可行性。     

大跨径预应力混凝土连续梁施工控制技术

大跨径预应力混凝土连续箱梁悬臂浇筑施工过程中，施工控制是个复杂的动态系统工程，是实现大桥成桥线形、内力满足设计要求的重要手段。结合南京长江第二大桥北汊桥主桥施工控制实践，阐述了悬臂浇筑施工过程中的施工控制原理、方法和分析计算理论，研究了双幅桥混凝土箱梁的温度分布特性以及高强混凝土水化热温度的控制，科学地指导了施工。使全桥10个合拢段的合拢轴线误差小于或等于4mm，标高误差小于或等于23mm，成桥线形均在设计允许误差范围内。克服了高强度混凝土水化热产生的龟裂和强度损失的现象。     

Novel matched filtering method and its application

The method of using a narrowband filter to realize matched filtering is derived. A novel method of using spectrum sampling to realize matched filtering is presented, and the method can conquer the disadvantages that the narrowband filter cannot adopt the adaptive scheduling of phased array radars and realize matched filtering for several waveforms. A novel error extraction method is proposed, which uses a time division multipath method to realize the intermediate frequency extraction. This method can save lots of space for vehicle-born radar systems, reduce the influence of amplitude and phase distortion caused by devices, and enhance the system reliability. Simulation results show that the spectrum sampling method is applicable, and the implementation of frequency spectrum sampling is elaborated.     

斜齿圆柱齿轮螺旋角的测绘方法

分析齿轮主要参数,提出用测量跨棒(球)距和公法线长度计算斜齿圆柱齿轮螺旋角的测绘方法,并推导出螺旋角的计算公式。该方法经过多次实际验证,误差较小。     

对角型拟线性双曲方程组周期解的Blowup

本文考虑具周期初始数据的对角型拟线性双曲方程组Cauchy问题 ,证明其经典周期解一定在有限时间内破裂 ,且给出了经典解生命跨度上界估计 .