Hedgehog regulates smoothened activity by inducing a conformational switch

Hedgehog (HH) morphogen is essential for metazoan development. The seven-transmembrane protein smoothened (SMO) transduces the HH signal across the plasma membrane, but how SMO is activated remains poorly understood. In Drosophila melanogaster, HH induces phosphorylation at multiple Ser/Thr residues in the SMO carboxy-terminal cytoplasmic tail, leading to its cell surface accumulation and activation. Here we provide evidence that phosphorylation activates SMO by inducing a conformational switch. This occurs by antagonizing multiple Arg clusters in the SMO cytoplasmic tail. The Arg clusters inhibit SMO by blocking its cell surface expression and keeping it in an inactive conformation that is maintained by intramolecular electrostatic interactions. HH-induced phosphorylation disrupts the interaction, and induces a conformational switch and dimerization of SMO cytoplasmic tails, which is essential for pathway activation. Increasing the number of mutations in the Arg clusters progressively activates SMO. Hence, by employing multiple Arg clusters as inhibitory elements counteracted by differential phosphorylation, SMO acts as a rheostat to translate graded HH signals into distinct responses.     

A second human interleukin-2 binding protein that may be a component of high-affinity interleukin-2 receptors

Although activated human T and B lymphocytes express both high-affinity and low-affinity membrane receptors for interleukin-2 (IL-2), the structural features that distinguish these receptors have remained unresolved. The high-affinity receptors appear to mediate IL-2 induced T cell growth and internalization of IL-2, whereas no function has yet been ascribed to the low-affinity receptors. The Tac antigen is an IL-2 binding protein of relative molecular mass 55,000 (Mr 55K) that participates in the formation of both high- and low-affinity receptors. But Tac complementary DNA transfection and membrane fusion studies have suggested that additional T-cell components are required to produce high-affinity IL-2 receptors. In this study, we report the identification of a second human IL-2 binding protein that (1) has an Mr of approximately 70K, (2) lacks reactivity with the anti-Tac antibody, (3) binds IL-2 with intermediate affinity and (4) is present on the surface of resting T cells, large granular lymphocytes (natural killer cells), and certain T and B cell lines in the absence of the Tac antigen. Chemical crosslinking of 125I-labelled IL-2 bound to high-affinity IL-2 receptors produces labelling of both the p70 protein and the Tac antigen and the anti-Tac antibody blocks the crosslink detection of both of these proteins. Expression of Tac cDNA in a T cell line expressing the p70 protein, but lacking both Tac and high-affinity receptors, results in the reconstitution of high-affinity IL-2 receptors in these cells. Together, these findings suggest that the high-affinity human IL-2 receptor may be a membrane complex composed of at least the p70 protein and Tac antigen.     

关于滑行艇整艇参数化有限元分析通用程序的研究

采用通用有限元软件进行船舶结构分析,已经取得了很好的效果。但关于滑行艇的结构有限元分析仍存在着很多问题。例如：建模的参数化程度不高、生成的模型不便于修改、重复的工作量大等。鉴于以上情况,作者提出滑行艇有限元分析通用程序的解决方案。该方案的主要原理是先运用ANSYS二次开发语言APDL实现滑行艇结构的参数化有限元建模计算,然后利用可视化编程语言VC＋＋提供的界面接口对各参数和ANSYS进行集成、封装,形成通用的有限元计算分析程序。该程序可以实现对普通尖舭无断级滑行艇完成自动建立模型、自动加载边界条件,自动进行后处理的功能。这样就可以使复杂的问题简单化,节约大量的工作时间,也能进一步进行优化设计。     

Autoimmune diabetes as a consequence of locally produced interleukin-2.

During cell differentiation in the thymus, self-reactive T cells can be generated. The majority of these seem to be deleted after intrathymic encounter with the relevant autoantigen. As all self antigens are unlikely to be present in the thymus, some autoreactive T cells may escape censorship. Here we study the fate of these cells using transgenic mice expressing the class I molecule H-2Kb (Kb) in the insulin-producing beta-cells of the pancreas. These mice were crossed with mice transgenic for genes encoding a Kb-specific T-cell antigen receptor (TCR) which could be detected using a clonotype-specific monoclonal antibody. Although T cells expressing the highest level of transgenic TCR were deleted intrathymically in double-transgenic mice, Kb-specific T cells were detected in the periphery. These cells caused the rejection of Kb-expressing skin grafts, but ignored islet Kb antigens even after priming. But when double-transgenic mice were crossed with transgenic mice expressing the lymphokine interleukin-2 in the pancreatic beta-cells, there was a rapid onset of diabetes. These results indicate that autoreactive T cells that ignore self antigens may cause autoimmune diabetes when provided with exogenous 'help' in the form of interleukin-2.     

Basis for a ubiquitin-like protein thioester switch toggling E1-E2 affinity

Ubiquitin-like proteins (UBLs) are conjugated by dynamic E1-E2-E3 enzyme cascades. E1 enzymes activate UBLs by catalysing UBL carboxy-terminal adenylation, forming a covalent E1 throught UBL thioester intermediate, and generating a thioester-linked E2 throught UBL product, which must be released for subsequent reactions. Here we report the structural analysis of a trapped UBL activation complex for the human NEDD8 pathway, containing NEDD8's heterodimeric E1 (APPBP1-UBA3), two NEDD8s (one thioester-linked to E1, one noncovalently associated for adenylation), a catalytically inactive E2 (Ubc12), and MgATP. The results suggest that a thioester switch toggles E1-E2 affinities. Two E2 binding sites depend on NEDD8 being thioester-linked to E1. One is unmasked by a striking E1 conformational change. The other comes directly from the thioester-bound NEDD8. After NEDD8 transfer to E2, reversion to an alternate E1 conformation would facilitate release of the E2 throught NEDD8 thioester product. Thus, transferring the UBL's thioester linkage between successive conjugation enzymes can induce conformational changes and alter interaction networks to drive consecutive steps in UBL cascades.     

Structure and expression of a cloned cDNA for human interleukin-2

A cDNA coding for human interleukin-2 (IL-2) has been cloned from a cDNA library prepared from partially purified IL-2 mRNA. The DNA sequence codes for a polypeptide which consists of 153 amino acids including a putative signal sequence. A biologically active polypeptide, characteristic of human IL-2, was produced when the cDNA was fused to a simian virus 40 promoter sequence and used to transfect cultured monkey COS cells.     

A frequency-dependent switch from inhibition to excitation in a hippocampal unitary circuit

The hippocampus, a brain structure essential for memory and cognition, is classically represented as a trisynaptic excitatory circuit. Recent findings challenge this view, particularly with regard to the mossy fibre input to CA3, the second synapse in the trisynaptic pathway. Thus, the powerful mossy fibre input to CA3 pyramidal cells might mediate both synaptic excitation and inhibition. Here we show, by recording from connected cell pairs in rat entorhinal-hippocampal slice cultures, that single action potentials in a dentate granule cell evoke a net inhibitory signal in a pyramidal cell. The hyperpolarization is due to disynaptic feedforward inhibition, which overwhelms monosynaptic excitation. Interestingly, this net inhibitory synaptic response changes to an excitatory signal when the frequency of presynaptic action potentials increases. The process responsible for this switch involves the facilitation of monosynaptic excitatory transmission coupled with rapid depression of inhibitory circuits. This ability to immediately switch the polarity of synaptic responses constitutes a novel synaptic mechanism, which might be crucial to the state-dependent processing of information in associative hippocampal networks.     

Compressibility of a natural kyanite to 17.5 GPa

The compressional behaviour of a natural kyanite, (Al_1.99Fe_0.01)SiO₅, has been investigated to about 17.5 GPa at 300 K using a diamond-anvil cell and synchrotron X-ray diffraction. The pressure–volume data fitted to the third-order Birch–Murnaghan equation of state (EoS) yield an isothermal bulk modulus (K_0T) of 192 ± 6 GPa and pressure derivative () of 6 ± 1. When is fixed as 4, the derived K_0T is 201 ± 2 GPa. These values are in excellent agreement with most experimental determinations in the literature. Consequently, it can be concluded that the compressibility of kyanite under high pressures has been accurately constrained.     

Holocene dust storm variations over northern China:transition from a natural forcing to an anthropogenic forcing

Asian dust storms have long been a major environmental concern in China,affecting the lives of about one billion people.However,it is unclear whether the mechan...     

Expression of interleukin-2 receptors as a differentiation marker on intrathymic stem cells

The thymus is regarded as the primary site for T-cell lymphopoiesis, but very little is known about the lineage inter-relationships of cells within that organ. At least four subpopulations of mouse thymocytes can be defined on the basis of staining with monoclonal antibodies directed against the T-cell differentiation antigens Lyt-2 and L3T4 (ref. 2). Thus immunocompetent (medullary) thymocytes, like peripheral T cells, express either Lyt-2 (cytotoxic phenotype) or L3T4 (helper phenotype) but not both, whereas non-functional (cortical) thymocytes express both markers. In addition, a small subpopulation comprising 2-3% of cells in the thymus and expressing neither Lyt-2 nor L3T4 has recently been described. The latter cells have the properties of intrathymic 'stem cells' in that they are the first to appear in the embryonic thymus and at least some can be shown to give rise, both in vivo (ref. 4. and our unpublished data) and in vitro, to other thymocyte subpopulations. We show here that 50% of Lyt-2-/L3T4- cells in the adult thymus express receptors for the polypeptide growth hormone interleukin-2 (IL-2) whereas other cells in the thymus do not. Furthermore, immunohistochemical localization studies on frozen sections indicate a disperse distribution of IL-2 receptor-positive cells in both the cortex and medulla. These novel findings have potential implications in the context of current models of differentiation pathways within the thymus.     

T3-Ti receptor triggering of T8+ suppressor T cells leads to unresponsiveness to interleukin-2

T3-associated disulphide linked heterodimers (Tin) comprised of clonally unique alpha-chains of molecular weight (MW) 49,000-54,000 and beta chains of MW 43,000 have been identified as the antigen receptors on human cytotoxic effector and inducer T-lymphocytes. Crosslinking of Ti molecules by either the appropriate nominal antigen/MHC specificity or anti-clonotypic monoclonal antibody results in clonal expansion of such cells via induction of IL-2 receptor expression, endogenous IL-2 release and IL-2-IL-2 receptor interaction. To determine whether analogous antigen receptor molecules and autocrine growth mechanisms are utilized by suppressor T-cells, we produced an anti-clonotypic monoclonal antibody against a non-cytotoxic T8+ suppressor T-cell, T8AC6, which defines a T3-associated disulphide-linked heterodimer of similar molecular weight to the above clonotypes. We find that Te-Ti triggering of suppressor clones (T8AC6, T8AC7 or T8RW) does not result in IL-2 production or T-cell proliferation and in contrast to inducer clones, also leads to a transient IL-2 unresponsive state. We suggest that such T3-Ti receptor mediated autoregulation of suppressor T-cell growth is necessary in the facilitation of initial inducer T-cell activation following antigenic perturbation.     

Recovery from autoimmunity of MRL/lpr mice after infection with an interleukin-2/vaccinia recombinant virus

Interleukin-2 (IL-2) is a T-cell derived molecule implicated in the clonal expansion of antigen-activated T cells and in T-cell development. IL-2 is also implicated in autoimmune disease, although its role is still controversial. Murine systemic lupus erythematosus (SLE) is a good model for human SLE as most of the immunological abnormalities in the human disease also seem to be operative in the mouse. Among SLE mice, the MRL/lpr strain develops early in life autoimmune diseases such as immune complex-mediated glomerulonephritis, arthritis and arteritis. Lymphoid abnormalities associated with those diseases in this strain are thymic atrophy and abnormal proliferation of CD3+ CD4- CD8- 'double-negative' T cells, resulting in massive generalized lymph node enlargement. We have therefore now examined the effects of IL-2 on the disease progression in MRL/lpr mice using live vaccinia recombinant viruses expressing the human IL-2 gene. Vaccinated mice showed prolonged survival, decreased autoantibody and rheumatoid factor titres, marked attenuation of kidney interstitial infiltration and intraglomerular proliferation, as well as clearance of synovial mononuclear infiltrates. Inoculation with the IL-2/vaccinia recombinant virus led, in addition, to drastic reduction of the double-negative T-cell population, improved thymic differentiation and restoration of normal values of mature cells in peripheral lymphoid organs.     

含蜡天然气孔板式节流阀内蜡沉积特性数值模拟

蜡沉积是含蜡天然气在集输和处理过程中常见的一种问题,会危害到天然气管输系统的高效、安全运行。为分析含蜡天然气在孔板式节流阀内的蜡沉积问题,基于离散相模型,通过自定义函数,建立蜡沉积数值模拟模型,分析传统孔板式节流阀的蜡沉积规律。研究结果表明：节流阀内蜡沉积主要受湍动能与涡流的影响,沉积位置主要出现在喉道出口处以及距喉道出口0.17 m左右的管壁处;管道内颗粒最大沉积量随压力的增加而增加,距喉道出口0.17 m左右的沉积宽度约为0.01 m,当压力增加1 MPa,该处沉积位置向管道出口处移动约0.005 m。其中,管道内最大沉积量与节流孔径呈负相关,与颗粒直径、喉道长度呈正相关。     

藏猪白细胞介素-12基因表达对猪圆环病毒2 疫苗的免疫增强效应

为研究IL-12基因在猪体内表达对PCV2疫苗免疫应答的调控作用,本研究将藏猪IL-12基因克隆至VR1020载体,壳聚糖包裹重组质粒制备成壳聚糖纳米颗粒(VRIL-12-CNP)并与PCV2疫苗共同接种21日龄断奶仔猪.接种后第0d、7d、14d和28d采集猪前腔静脉血进行血细胞分析、PCV2抗体检测和相关免疫基因表达量检测,并记录体重.结果显示:接种VRIL-12-CNP和PCV2疫苗的实验组猪的CD3~+、CD4~+、CD8~+T细胞和PCV2抗体显著增长(P0.05),TLR2/7、IL-12/4/6/15、STAT1/3和Bcl-2基因的表达量显著高于对照组(P0.05);试验期间实验组体重净增长也明显高于对照组(P0.05).结果表明:VRIL12-CNP能增强PCV2疫苗的先天性和获得性体液、细胞免疫应答,是安全、有效的PCV2疫苗佐剂.     

封闭空间内小尺度等温竖板自然对流的三维效应

研究了封闭空间内小尺度等温竖板自然对流的三维效应、表明三维效应随着平板尺度的减小而增强，从而使换热增强．对于相同尺度的平板，采用不同的放置方式其换热状况也不相同．尺度较大时，竖放(高度方向长)比横放时换热强，但随着尺度的减小，两种放置方式的换热越来越接近．     

自然水体沉积物对Pb~(2+)和Cu~(2+)吸附研究

采用巢湖自然水体沉积物对Cu⁽²⁺⁾和Pb(2+)和Pb⁽²⁺⁾吸附研究,初步研究了投加量、温度、时间、pH值等方面对其吸附结果的影响,在吸附实验中,拟二级动力模型优于拟一级动力学模型,Pb(2+)吸附研究,初步研究了投加量、温度、时间、pH值等方面对其吸附结果的影响,在吸附实验中,拟二级动力模型优于拟一级动力学模型,Pb⁽²⁺⁾和Cu(2+)和Cu⁽²⁺⁾在缓流水体沉积物上的吸附行为更适合用Langmuir吸附等温模型来描述。红外光谱（FTIR）显示缓流水体沉积物中存在羟基、氨基、烷基、含氧基团等,这些基团对Pb(2+)在缓流水体沉积物上的吸附行为更适合用Langmuir吸附等温模型来描述。红外光谱（FTIR）显示缓流水体沉积物中存在羟基、氨基、烷基、含氧基团等,这些基团对Pb⁽²⁺⁾和Cu(2+)和Cu⁽²⁺⁾的去除起了关键性的作用;XRD显示缓流水体沉积物表面的物质为SiO_2、CaCO_3。     

废弃混凝土代替天然石灰石煅烧水泥熟料实验

从4个不同地点取得4个以破碎石灰石为粗集料的废弃混凝土样品,破碎、分离,获得以石灰石为主的颗粒,取代天然石灰石作为钙质原料煅烧出4个波特兰水泥熟料样品,并与重庆拉法基水泥厂生产的熟料和市售水泥样品进行对比.研究结果表明:废弃混凝土熟料的4种主要矿物的XRD特征峰明显;游离CaO低于1%;可以用其制备出52.5级硅酸盐水泥.     

Human interleukin-2 promotes proliferation of activated B cells via surface receptors similar to those of activated T cells

Human interleukin-2 (IL-2) is a glycoprotein of relative molecular mass (Mr) 15,000, which is released by T lymphocytes on stimulation with antigen or mitogen and functions as a T-cell growth factor (TCGF) by inducing proliferation of activated T cells. It is generally accepted that resting or activated B cells do not respond directly to IL-2 but require for their proliferation other T-cell-derived lymphokines usually referred to as B-cell growth factors (BCGFs). Recently, however, a monoclonal antibody reacting with the IL-2 receptor molecules expressed by activated T cells (anti-Tac) was shown to react also with certain B tumour cells; in addition, murine B cells proliferate in response to pure human IL-2. We now show that recombinant IL-2, derived from Escherichia coli expressing the human gene, is able to promote strong proliferation of human B cells activated with protein-A-rich Staphylococcus aureus Cowans strain I. Moreover, we demonstrate that the anti-Tac antibody also reacts with S. aureus-activated normal B cells and inhibits sharply the proliferative response of such cells to IL-2. Finally, immunoprecipitation experiments reveal that anti-Tac defines similar molecules on activated T and B cells.