Structure of oxidized alpha-haemoglobin bound to AHSP reveals a protective mechanism for haem

The synthesis of haemoglobin A (HbA) is exquisitely coordinated during erythrocyte development to prevent damaging effects from individual alpha- and beta-subunits. The alpha-haemoglobin-stabilizing protein (AHSP) binds alpha-haemoglobin (alphaHb), inhibits the ability of alphaHb to generate reactive oxygen species and prevents its precipitation on exposure to oxidant stress. The structure of AHSP bound to ferrous alphaHb is thought to represent a transitional complex through which alphaHb is converted to a non-reactive, hexacoordinate ferric form. Here we report the crystal structure of this ferric alphaHb-AHSP complex at 2.4 A resolution. Our findings reveal a striking bis-histidyl configuration in which both the proximal and the distal histidines coordinate the haem iron atom. To attain this unusual conformation, segments of alphaHb undergo drastic structural rearrangements, including the repositioning of several alpha-helices. Moreover, conversion to the ferric bis-histidine configuration strongly and specifically inhibits redox chemistry catalysis and haem loss from alphaHb. The observed structural changes, which impair the chemical reactivity of haem iron, explain how AHSP stabilizes alphaHb and prevents its damaging effects in cells.     

Positive selection of CD4+ T cells mediated by MHC class II-bearing stromal cell in the thymic cortex

T lymphocytes differentiate in the thymus, where functionally immature, CD4+CD8+ (double positive) thymocytes develop into functionally mature CD4+ helper cells and CD8+ cytotoxic (single positive) T cells. The thymus is the site where self-reactive T cells are negatively selected (clonally deleted) and where T cells with the capacity to recognize foreign antigens in association with self-proteins encoded by the major histocompatibility complex (MHC) are positively selected. The net result of these developmental pathways is a T-cell repertoire that is both self-tolerant and self-restricted. One unresolved issue is the identity of the thymic stromal cells that mediate the negative and positive selection of the T-cell repertoire. Previous work has pointed to a bone-marrow-derived macrophage or dendritic cell as the inducer of tolerance, whereas a radiation-resistant, deoxyguanosine-resistant thymic cell seems to mediate the positive selection of self-MHC restricted T cells. Thymic stromal cells in the cortex interact with the T-cell antigen receptor on thymocytes. Using several strains of transgenic mice that express the class II MHC molecule I-E in specific regions of the thymus, we show directly that the positive selection of T cells is mediated by an I-E-bearing cell in the thymic cortex.     

材料内部裂纹自修复中组织生长机制

为探索高温塑性变形过程中裂纹修复规律 ,采用高温物理模拟方法研究了 2 0 Mn Mo材料内部孔隙性裂纹的自修复过程。实验中发现在裂纹修复过程中普遍存在着缺陷自由面上组织生长现象 ,该现象解释了自修复消除孔隙的原因 ,证实了裂纹修复的再结晶机制。研究结果对揭示裂纹演化和修复机理具有重要意义 ,在实际生产中能够作为修复裂纹的指导原则。该成果对于控制塑性加工中缺陷的变化过程具有重要的理论意义和实用价值     

Wnt proteins are lipid-modified and can act as stem cell growth factors

Wnt signalling is involved in numerous events in animal development, including the proliferation of stem cells and the specification of the neural crest. Wnt proteins are potentially important reagents in expanding specific cell types, but in contrast to other developmental signalling molecules such as hedgehog proteins and the bone morphogenetic proteins, Wnt proteins have never been isolated in an active form. Although Wnt proteins are secreted from cells, secretion is usually inefficient and previous attempts to characterize Wnt proteins have been hampered by their high degree of insolubility. Here we have isolated active Wnt molecules, including the product of the mouse Wnt3a gene. By mass spectrometry, we found the proteins to be palmitoylated on a conserved cysteine. Enzymatic removal of the palmitate or site-directed and natural mutations of the modified cysteine result in loss of activity, and indicate that the lipid is important for signalling. The purified Wnt3a protein induces self-renewal of haematopoietic stem cells, signifying its potential use in tissue engineering.     

pMOSFET的NBTI退化机理及内在影响因素

研究了P型MOSFET的NBTI效应退化机理,以及栅氧化层电场和沟道载流子浓度对NBTI效应的影响.首先,通过电荷泵实验对NBTI应力带来的p M OSFET的界面损伤进行了测试,并利用TCAD仿真软件对测试结果进行分析,结果表明该器件的NBTI退化主要由其沟道区的界面态产生引起,而电荷注入的影响相对可以忽略.然后,通过施加衬底偏置电压的方法实现了增加器件栅氧化层电场但保持沟道载流子浓度不变的效果,进而研究了栅氧化层电场和沟道载流子浓度2个内在因素分别对NBTI退化的影响.最后,通过对比不同栅极电压和不同衬底偏置电压条件下器件的2个内在影响因素变化与NBTI退化的关系,证明了p MOSFET的NBTI效应主要由器件的栅氧化层电场决定,沟道载流子浓度对器件NBTI效应的影响可以忽略.     

Evidence for low sulphate and anoxia in a mid-Proterozoic marine basin

Many independent lines of evidence document a large increase in the Earth's surface oxidation state 2,400 to 2,200 million years ago, and a second biospheric oxygenation 800 to 580 million years ago, just before large animals appear in the fossil record. Such a two-staged oxidation implies a unique ocean chemistry for much of the Proterozoic eon, which would have been neither completely anoxic and iron-rich as hypothesized for Archaean seas, nor fully oxic as supposed for most of the Phanerozoic eon. The redox chemistry of Proterozoic oceans has important implications for evolution, but empirical constraints on competing environmental models are scarce. Here we present an analysis of the iron chemistry of shales deposited in the marine Roper Basin, Australia, between about 1,500 and 1,400 million years ago, which record deep-water anoxia beneath oxidized surface water. The sulphur isotopic compositions of pyrites in the shales show strong variations along a palaeodepth gradient, indicating low sulphate concentrations in mid-Proterozoic oceans. Our data help to integrate a growing body of evidence favouring a long-lived intermediate state of the oceans, generated by the early Proterozoic oxygen revolution and terminated by the environmental transformation late in the Proterozoic eon.     

国企财务运行机制设计的因素分析

摘要：现行国有企业的财务运行机制已不适应新形势下企业发展的需要。因此，必须结合企业生存的内外环境构建新型的财务运行机制。本文从体制、法制、市场等因素入手分析了构建新型财务运行机制的外部环境，并提出应从构建有效的资金管理机制、设置新型财务管理机构、提高财务人员专业及综合素质等方面完善财务运行机制的内部环境。     

Ca2+ release from endoplasmic reticulum is mediated by a guanine nucleotide regulatory mechanism

Ca2+ accumulation and release from intracellular organelles is important for Ca2+-signalling events within cells. In a variety of cell types, the active Ca2+-pumping properties of endoplasmic reticulum (ER) have been directly studied using chemically permeabilized cells. The same preparations have been extensively used to study Ca2+ release from ER, in particular, release mediated by the intracellular messenger inositol 1,4,5-trisphosphate (InsP3). So far, these studies and others using microsomal membrane fractions have revealed few mechanistic details of Ca2+ release from ER, although a recent report indicated that InsP3-mediated Ca2+ release from liver microsomes may be dependent on GTP. In contrast to the latter report, we describe here the direct activation of a specific and sensitive guanine nucleotide regulatory mechanism mediating a substantial release of Ca2+ from the ER of cells of the neuronal cell line N1E-115. These data indicate the operation of a major new Ca2+ gating mechanism in ER which is specifically activated by GTP, deactivated by GDP, and which appears to involve a GTP hydrolytic cycle.     

Basal body movements as a mechanism for mitochondrial genome segregation in the trypanosome cell cycle

The mitochondrial genome of Trypanosoma brucei is organized in the form of a complex catenated network of circular DNA molecules. This mass of DNA, known as the kinetoplast, is present at a unique site in the single mitochondrion, and is replicated in a discrete, periodic S phase of the cell cycle. The single-copy nature of the kinetoplast suggests that there is a mechanism ensuring segregation fidelity of replicated copies to each daughter cell. Historically, speculation regarding the nature of this mechanism has often attributed significance to the close association between the kinetoplast and the flagellum basal body. We provide here direct evidence that this mitochondrial DNA complex is indeed linked to the basal body, and segregation of the kinetoplast DNA is dependent on a microtubule-mediated separation of the new and old flagellar basal bodies during the cell cycle. This unique system may represent the remnants of an evolutionarily archaic mechanism for genome segregation.     

Non-viral gene carrier mediated short hairpin RNA interference for inhibition of tumor cells growth

A tumor-targeting gene vector G250mAb-PEI-PEG has been prepared by modification of polyethylenimine （PEI） with polyethyleneglycol （PEG） and G250, a monoclonal antibody against the G250 antigen on tumor cell surface. The transfection efficiency was as high as 70% in G250 positive HeLa cells, whereas the transfection efficiency was relatively low （30%） in normal NIH3T3 cells. A plasmid encoding the short hairpin RNA （shRNA） specific for nucleostemin gene （NS） was efficiently transfected into the HeLa cells with this nonviral gene vector. RNA interference down-regulated the expression of NS gene in HeLa cells, inhibited cells proliferation and induced apoptosis. However, the growth and activity of the NIH3T3 cells were not affected under the same treatment. These results indicate that the reported nonviral gene vector, G250mAb-PEI-PEG, can target and efficiently deliver genes into HeLa cells, and has the potential for the cervical cancer treatment.     

Requirement for mast cell growth factor for primordial germ cell survival in culture.

Mast-cell growth factor (MGF) is encoded by the murine steel (Sl) locus and is a ligand for the tyrosine kinase receptor protein encoded by the proto-oncogene c-kit at the murine dominant white spotting (W) locus. Mutations at both these loci affect mast cells, primordial germ cells (PGCs), haemopoietic stem cells and melanocytes. In many Sl and W mutants, the rapid proliferation of PGC that normally occurs between day 7 and 13.5 of embryonic development fails to occur. As c-kit is expressed in PGCs while MGF is expressed in the surrounding mesenchyme, MGF might promote the proliferation of PGCs. Here we report that MGF is essential for PGC survival in culture, but does not stimulate PGC proliferation. Moreover, whereas both the transmembrane and soluble proteolytic cleavage forms of MGF stimulate mast-cell proliferation, soluble MGF has a relatively limited ability to support survival of PGCs in culture, thus explaining the sterility in mice carrying the steel-dickie (Sld) mutation, which encodes only a soluble form of MGF, and providing a functional role for a transmembrane growth factor.     

Crystal structure of a lectin-like natural killer cell receptor bound to its MHC class I ligand

Natural killer (NK) cell function is regulated by NK receptors that interact with MHC class I (MHC-I) molecules on target cells. The murine NK receptor Ly49A inhibits NK cell activity by interacting with H-2D(d) through its C-type-lectin-like NK receptor domain. Here we report the crystal structure of the complex between the Ly49A NK receptor domain and unglycosylated H-2D(d). The Ly49A dimer interacts extensively with two H-2D(d) molecules at distinct sites. At one interface, a single Ly49A subunit contacts one side of the MHC-I peptide-binding platform, presenting an open cavity towards the conserved glycosylation site on the H-2D(d) alpha2 domain. At a second, larger interface, the Ly49A dimer binds in a region overlapping the CD8-binding site. The smaller interface probably represents the interaction between Ly49A on the NK cell and MHC-I on the target cell, whereas the larger one suggests an interaction between Ly49A and MHC-I on the NK cell itself. Both Ly49A binding sites on MHC-I are spatially distinct from that of the T-cell receptor.     

Structure of a Ran-binding domain complexed with Ran bound to a GTP analogue: implications for nuclear transport

The protein Ran is a small GTP-binding protein that binds to two types of effector inside the cell: Ran-binding proteins, which have a role in terminating export processes from the nucleus to the cytoplasm, and importin-beta-like molecules that bind cargo proteins during nuclear transport. The Ran-binding domain is a conserved sequence motif found in several proteins that participate in these transport processes. The Ran-binding protein RanBP2 contains four of these domains and constitutes a large part of the cytoplasmic fibrils that extend from the nuclear-pore complex. The structure of Ran bound to a non-hydrolysable GTP analogue (Ran x GppNHp) in complex with the first Ran-binding domain (RanBD1) of human RanBP2 reveals not only that RanBD1 has a pleckstrin-homology domain fold, but also that the switch-I region of Ran x GppNHp resembles the canonical Ras GppNHp structure and that the carboxy terminus of Ran is wrapped around RanBD1, contacting a basic patch on RanBD1 through its acidic end. This molecular 'embrace' enables RanBDs to sequester the Ran carboxy terminus, triggering the dissociation of Ran x GTP from importin-beta-related transport factors and facilitating GTP hydrolysis by the GTPase-activating protein ranGAP. Such a mechanism represents a new type of switch mechanism and regulatory protein-protein interaction for a Ras-related protein.     

Botany: a new self-pollination mechanism

Pollen grains from most flowering plants are transported by wind or animals and deposited on the receptive surface of the stigma of a different individual, but self-pollination is also common. We have discovered a new process for self-pollination in the laterally orientated flowers of a Chinese herb, in which a film of pollen is transported from the anther (pollen sacs) by an oily emulsion that slides sideways along the flower's style and into the individual's own stigma. This mode of self-pollination is a new addition to the broad range of genetic and morphological mechanisms that have evolved in flowering plants (angiosperms), and may be common in species growing in shady, windless and insect-poor habitats.     

一类Helmholtz方程Cauchy问题的最优误差界

考虑一类Helmholtz方程Cauchy问题,给出这个问题的最优误差界.用谱正则化方法和修正的Tikhonov正则化方法来求解这个问题,得到Holder型误差估计.根据正则化的最优理论,误差估计是阶数最优的.