Intragenic recombination leads to pilus antigenic variation in Neisseria gonorrhoeae

The pilus of the bacterium Neisseria gonorrhoeae is a fimbriate surface structure which promotes attachment of the bacterium to host epithelial cells. Gonococcal pilus phase variation is characterized by a rapid on/off switch in which piliated (P+) cells throw off non-piliated (P-) variants and vice versa. Two regions of the gonococcal chromosome (pilE1 and pilE2) act as pilin expression loci, reminiscent of the MAT locus in the yeast Saccharomyces cerevisiae, while several other chromosomal regions contain silent (non-expressing) pilin sequences. Biochemical and antigenic diversity is seen in pili from a wide variety of clinical isolates. Pilins (pilus subunits) are composed of conserved N-terminal and variable C-terminal regions; the conserved region of gonococcal pilin is also found in pilins produced by widely disparate bacteria. We show here that the gonococcal pilin undergoes antigenic variation in vitro and in vivo. The protein consists of constant, semi-variable and hypervariable regions. This antigenic variation probably involves gene conversion of mini-cassettes of pilin information.     

DNA transformation leads to pilin antigenic variation in Neisseria gonorrhoeae

Many pathogenic bacteria express pili (fimbriae) on their cell surfaces. These structures mediate binding of bacteria to host tissues, and may also be involved in other aspects of pathogenesis. Neisseria gonorrhoeae pili are mainly composed of a single protein, pilin, whose expression is controlled at chromosomal expression loci (pilE). An intact pilin gene and promoter sequences are only found at pilE. Strain MS11 contains two expression sites (pilE1 and pilE2), whereas several of its derivatives and other clinical isolates contain only one. Silent pilin loci (pilS1-pilS7) contain truncated variant pilin genes lacking the promoter and conserved pilin gene sequences. Pilin antigenic variation in N. gonorrhoeae occurs by DNA recombination between one of he silent partial variant gene segments in pilS and an expressed pilin gene in pilE. The recombination reactions are nonreciprocal, and therefore the mechanism has been classified as gene conversion. We report that much of the recombination between pilin loci actually occurs after transformation of living piliated cells by DNA liberated from lysed cells within a population. This constitutes a new molecular mechanism for an antigenic variation system, as well as the first specific function for a DNA transformation system.     

Binding of a non-beta-lactam antibiotic to penicillin-binding proteins

In the search for new beta-lactam antibiotics of natural origin, the discoveries of cephamycins and sulfazecins (monobactams) were important turning points in that they accelerated many screening efforts aimed at other new compounds. In our target-directed screening for beta-lactam antibiotics using beta-lactam hypersensitive mutants, we have examined Gram-negative bacteria isolated from natural habitats and have recently reported several types of beta-lactam antibiotics such as cephabacins and formadicins. Here we report a novel antibiotic, lactivicin, found using this system. Although lactivicin has various biological activities commonly observed in beta-lactam antibiotics, it does not possess a beta-lactam ring in its molecule, but has the unique structure of a dicyclic dipeptide.     

Gene structure and extracellular secretion of Neisseria gonorrhoeae IgA protease

Several human bacterial pathogens, including the Gram-negative diplococcus Neisseria gonorrhoeae, produce extracellular proteases that are specific for human immunoglobulin IgA1. Immunoglobulin A (IgA) proteases have been studied extensively and the genes of some species cloned in Escherichia coli, but their role in pathogenesis remains unclear. Recently we derived a DNA fragment of 5 kilobases (kb) from N. gonorrhoeae MS11 directing extracellular active enzyme in E. coli. Although the mature enzyme of strain MS11 was shown to have a relative molecular mass of 106,000 (Mr 106K) in gels, the DNA sequence of this cloned fragment reveals a single gene coding for a 169K precursor of IgA protease. The precursor contains three functional domains, the amino-terminal leader which is assumed to initiate the inner membrane transport of the precursor, the protease, and a carboxyl-terminal 'helper' domain apparently required for extracellular secretion (excretion). Based on the structural features of the precursor, we propose a model in which the helper serves as a pore for excretion of the protease domain through the outer membrane. IgA protease acquires an active conformation as its extracellular transport proceeds and is released as a proform from the membrane-bound helper by autoproteolysis. The soluble proform further matures into the 106 K IgA protease and a small stable alpha-protein.     

Reassortment of pilin genes in Neisseria gonorrhoeae occurs by two distinct mechanisms

Phase and antigenic variation of pilin expression in Neisseria gonorrhoeae result from recombination events in which variant sequences from one of the silent loci (pilS) are transferred to the expression locus (pilE). Such rearrangements were originally thought to be gene conversions, but findings showing that phase variation is partially inhibited by DNase I, that piliated (P+) cells are highly competent for DNA uptake and that gonococci readily undergo autolysis in culture, led to the suggestion that pilin variation occurs through transformation by exogenous DNA. We have developed a simple method for the selection of non-piliated (P-) cells and have evaluated naturally occurring P+ to P- transitions. Two primary pathways of pilin variation can be distinguished--transformation-mediated recombination, which is influenced by culture conditions and inhibited by DNase I, and intragenomic reciprocal recombination, which is unaffected by DNase I. Furthermore, we demonstrate that both piliated and revertible P- cells are competent for DNA uptake, an essential prerequisite of the first pathway.     

Determination of the genus-specific antigens in outer membrane proteins from the strains of Leptospira interrogans and Leptospira biflexa with different virulence

Objective:To determine the existence of genus-specific antigens in outer membrane proteins (OMPs) of leptospira with different virulence. Methods: Microscope agglutination test (MAT) was applied to detect the agglutination between commercial rabbit antiserum against leptospiral genus-specific TR/Patoc I antigen and 17 strains of Leptospira interrongans belonging to 15 serogroups and 2 strains of Leptospira biflexa belonging to 2 serogroups.The outer envelopes (OEs) of L.interrogans serogroup Icterohaemorrhagiae serovar lai strain lai (56601) with strong virulence and serogroup Pomona serovar pomona strain Luo (56608) with low virulence,and L.biflexa serogroup Semaranga serovar patoc strain Patoc I without virulence were prepared by using the method reported in Auran et al.(1972).OMPs in the OEs were obtained by treatment with sodium deoxycholate. SDS-PAGE and western blot were used for analyzing the features of the OMPs on electrophoretic pattern and the immunoreactivity to the antiserum against TR/Patoc I antigen, respectively. Results:All the tested strains belonging to different leptospiral serogroups agglutinated to the antiserum against leptospiral genus-specific TR/Patoc I antigen with agglutination titers ranging from 1:256-1:512. A similar SDS-PAGE pattern of the OMPs from the three strains of leptospira with different virulence was shown and the molecular weight of a major protein fragment in the OMPs was found to be approximately 60 KDa.A positive protein fragment with approximately 32 KDa confirmed by Western blot,was able to react with the antiserum against leptospiral genus-specific TR/Patoc I antigen, and was found in each the OMPs of the three stains of leptospira.Conclusion: There are genus-specific antigens on the surface of L.interrogans and L.biflexa. The OMP with molecular weight of 32 KDa may be one of the genus-specific protein antigens of leptospira.     

双抗转CP基因线辣椒对非克隆病毒的抗病性

本试验以转化黄瓜花叶病毒外壳蛋白基因 (CMV- CP)和烟草花叶病毒外壳蛋白基因(TMV- CP)的线辣椒纯合系作试材 ,比较了转化植株对接种 CMV非克隆株系后抗病性的变化以及转化植株对接种 PVX,PVY和 TEV等非克隆病毒后的抗病性变化 .结果表明 :转化线辣椒能抵抗 CMV非克隆株系的侵染 ,抗病表达特点为系统症状延迟出现 ,显症株率和病害严重度级别大幅度降低 ,接种叶片与新生叶片中的病毒含量明显减少 .对 PVX,PVY和 TEV等非克隆病毒也表达了相同的抗病特点     

混合增压柴油机增压系统参数模拟计算研究

该文优化调整模拟计算时确定的混合增压系统的参数,扩大了增压系统涡轮的流量,减小了高速电机功率,电机功率下降直接带来发动机的轻量化和增压器转动惯量的大幅减小。同时,涡轮流量的增加还提高了车用发动机在常用工况下的经济性和低速高负荷工况下的潜在动力性。与原废气旁通增压柴油机相比,参数调整后的混合增压柴油机减小了排气提前角调整不当对发动机燃油经济性的扰动。     

Resistance to beta-lactam antibiotics by re-modelling the active site of an E. coli penicillin-binding protein

The beta-lactam antibiotics kill bacteria by inhibiting a set of penicillin-binding proteins (PBPs) that catalyse the final stages of peptidoglycan synthesis. In some bacteria the development of intrinsic resistance to beta-lactam antibiotics by the reduction in the affinity of PBPs causes serious clinical problems. The introduction of beta-lactam antibiotics that are resistant to hydrolysis by beta-lactamases may also result in the emergence of intrinsic resistance among the Enterobacteriaceae. The clinical problems that would arise from the emergence of resistant PBPs in enterobacteria have led us to examine the ease with which Escherichia coli can gain resistance to beta-lactams by the production of altered PBPs. The development of resistant PBPs also provides an interesting example of enzyme evolution, since it requires a subtle re-modeling of the enzyme active centre so that it retains affinity for its peptide substrate but excludes the structurally analogous beta-lactam antibiotics. We show here that only four amino-acid substitutions need to be introduced into PBP 3 of E. coli to produce a strain possessing substantial levels of resistance to a wide variety of cephalosporins. We also show that transfer of the gene encoding the resistant PBP 3 from the chromosome to a plasmid could result in the spread of intrinsic resistance not only to other strains of E. coli but also to other enterobacterial species.     

A群脑膜炎球菌发酵工艺的优化

目的:优化A群脑膜炎球菌的发酵工艺,提高荚膜多糖产量。方法:在30L发酵罐中,通过调整溶氧(DO)、pH值、培养温度、接种终浓度、葡萄糖补料方式等变化,考察其对A群脑膜炎球菌菌体生长及荚膜多糖产量的影响,优化发酵工艺。结果:在发酵过程中,DO在10%～20%范围内对多糖合成无明显影响;在pH7.0,培养温度36℃,有利于多糖合成;发酵中间补加葡萄糖可增加多糖合成,固定速率补糖更有利于多糖合成。结论:通过参数优化,确定了A群脑膜炎球菌的最佳发酵工艺。     

混合状态Petri网及其在过程控制混杂系统中的应用

为了处理混杂系统的建模问题，提出了一种称为混合状态Petri网(HSPN)的混合Petri网，给出了HSPN的定义及其变迁规则。采用一个实际化工过程为例建立了HSPN模型，该模型可以用于混合控制器的设计，仿真结果表明了其有效性。本文还简要地讨论了HSPN的一些性质。     

Nuclear transplantation in different strains of zebrafish

Single later blastula nuclei from AB strain of zebrafish (Danio rerio) were transplanted into enucleated unfertilized eggs of Long fin strain. Of 1119 cloning embryos, 14 reconstructed embryos developed into fry. DNA fingerprinting systems of the cloned fish were similar to those of the nuclear donor fish, but were distinctly different from those of the nuclear recipient fish. It confirmed that the genetic material originated from nuclear donor cell other than from nuclear recipient egg. The research suggested that the basic technique for nuclear transplantation performed with different strains of zebrafish has made a breakthrough. It should be helpful for the study of some important developmental problems such as gene function, the regulation ogene expression during animal development, the developmental potential of a nucleus and the interactions between the donor nucleus and the recipient cytoplasm, etc.     

Hybrid Forwarding in Cooperative Communications

A cooperative communication scheme was developed in which the relay node performs hybrid forwarding, where the relay node will not forward data for a transmission pair unless the corresponding direct transmission fails, and the relay node will not forward data for a transmission pair unless the quality of the channel between the corresponding source and the relay is good. A performance analysis of the outage probability and the diversity-multiplexing tradeoff for the scheme shows that the forwarding mechanism efficiently reduces fading in wireless channels by achieving full diversity order of magnitude improvement. A considerable signal-to-noise ratio gain is provided by this scheme compared with existing cooperation schemes with fixed forwarding. This flexible forwarding mechanism enables the scheme to provide the optimal diversity-multiplexing tradeoff performance.     

混合系统基于模型诊断建模问题研究

基于模型的诊断是为了克服传统故障诊断方法的缺点而兴起的一项新型的智能诊断推理技术,其应用越来越广泛.而混合系统是当前应用较多的动态系统之一,其故障诊断问题是当前研究领域的热门.在此介绍两种对混合系统进行诊断的建模方法,并对这两种建模方法进行比较找出各自适用方向,特别是应用MBD技术对混合系统进行诊断的建模方法,根据对实际航天生态系统建模说明混合键合图基本原理,为进一步进行基于模型的故障检测和故障追踪直至故障隔离和鉴别提供基础模型.     

功率分流式混合动力汽车复合电源系统设计

为解决功率分流式混合动力汽车单一蓄电池功率密度小、循环寿命短等问题,引入超级电容-蓄电池复合电源系统,利用AVL-Cruise/Simulink联合仿真平台搭建了功率分流式混合动力汽车的动力系统模型,在基于发动机最优工作曲线的能量管理控制策略中加入了复合电源功率分配策略,该功率分配策略能够缓冲起停发动机、制动工况下的电机工作时的大电流对电池的冲击,使电池尽可能工作在高效率区间来提高车辆的燃油经济性.在此基础上,对蓄电池组和超级电容进行了参数匹配,仿真结果表明蓄电池的放电过程得到了优化,所设计的复合电源系统能够提高车辆的燃油经济性.