Assembly and Maturation of the Bacteriophage Lambda Procapsid: gpC Is the Viral Protease

Viral capsids are robust structures designed to protect the genome from environmental insults and deliver it to the host cell. The developmental pathway for complex double-stranded DNA viruses is generally conserved in the prokaryotic and eukaryotic groups and includes a genome packaging step where viral DNA is inserted into a pre-formed procapsid shell. The procapsids self-assemble from monomeric precursors to afford a mature icosahedron that contains a single “portal” structure at a unique vertex; the portal serves as the hole through which DNA enters the procapsid during particle assembly and exits during infection. Bacteriophage λ has served as an ideal model system to study the development of the large double-stranded DNA viruses. Within this context, the λ procapsid assembly pathway has been reported to be uniquely complex involving protein cross-linking and proteolytic maturation events. In this work, we identify and characterize the protease responsible for λ procapsid maturation and present a structural model for a procapsid-bound protease dimer. The procapsid protease possesses autoproteolytic activity, it is required for degradation of the internal “scaffold” protein required for procapsid self-assembly, and it is responsible for proteolysis of the portal complex. Our data demonstrate that these proteolytic maturation events are not required for procapsid assembly or for DNA packaging into the structure, but that proteolysis is essential to late steps in particle assembly and/or in subsequent infection of a host cell. The data suggest that the λ-like proteases and the herpesvirus-like proteases define two distinct viral protease folds that exhibit little sequence or structural homology but that provide identical functions in virus development. The data further indicate that procapsid assembly and maturation are strongly conserved in the prokaryotic and eukaryotic virus groups.     

Human RegIV Protein Adopts a Typical C-Type Lectin Fold but Binds Mannan with Two Calcium-Independent Sites

Human RegIV protein, which contains a sequence motif homologous to calcium-dependent (C-type) lectin-like domain, is highly expressed in mucosa cells of the gastrointestinal tract during pathogen infection and carcinogenesis and may be applied in both diagnosis and treatment of gastric and colon cancers. Here, we provide evidence that, unlike other C-type lectins, human RegIV binds to polysaccharides, mannan, and heparin in the absence of calcium. To elucidate the structural basis for carbohydrate recognition by NMR, we generated the mutant with Pro91 replaced by Ser (hRegIV-P91S) and showed that the structural property and carbohydrate binding ability of hRegIV-P91S are almost identical with those of wild-type protein. The solution structure of hRegIV-P91S was determined, showing that it adopts a typical fold of C-type lectin. Based on the chemical shift perturbations of amide resonances, two calcium-independent mannan-binding sites were proposed. One site is similar to the calcium-independent sugar-binding site on human RegIII and Langerin. Interestingly, the other site is adjacent to the conserved calcium-dependent site at position Ca-2 of typical C-type lectins. Moreover, model-free analysis of ¹⁵N relaxation parameters and simplified Carr-Purcell-Meiboom-Gill relaxation dispersion experiments showed that a slow microsecond-to-millisecond time-scale backbone motion is involved in mannan binding by this site, suggesting a potential role for specific carbohydrate recognition. Our findings shed light on the sugar-binding mode of Reg family proteins, and we postulate that Reg family proteins evolved to bind sugar without calcium to keep the carbohydrate recognition activity under low-pH environments in the gastrointestinal tract.     

Structural Determination of Monosialyl Trisaccharides Obtained From Caprine Colostrum

Acidic oligosaccharides were separated by dialysis, ion-exchange, preparative paper and gel chromatography from caprine colostrum. Four sialyl trisaccharides were characterized by ¹H-NMR spectrometry as follows: α-N-acetylneuraminyl-(2,6)-β-d-galactopyranosyl-(1,4)-2-N-acetamido-2-deoxy-d-glucopyranose (Neu5Ac α 2-6Gal β 1-4GlcNAc), α-N-acetylneuraminyl-(2,3)-β-d-galactopyranosyl-(1,4)-d-glucopyranose (Neu5Ac α 2-3Gal β-1-4Glc), α-N-acetylneuraminyl-(2,6)-β-d-galactopyranosyl-(1,4)-d-glucopyranose (Neu5Ac α 2-6Gal β 1-4Glc) and α-N-glycolylneuraminyl-(2,6)-β-d-galactopyranosyl-(1,4)-d-glucopyranose (Neu5Gc α 2-6Gal β 1-4Glc).     

ICU中心发生胸腔感染的影响因素分析及预防对策

目的:探讨重症监护病房(ICU)中心发生胸腔感染的情况及其影响因素,并提出相应的预防对策。方法:选择2015年2月至2017年2月我院ICU中心收治的98例患者进行研究,均为全麻下行开胸术后住ICU者。收集所有患者临床资料,分析胸腔感染的发生情况,通过比较发生/未发生胸腔感染患者的临床资料,探讨ICU中心发生胸腔感染的危险因素,并提出相应的预防对策。结果:在98例患者中,有15例发生胸腔感染,发生率为15.31%,以铜绿假单胞菌所占比例最高,为40.00%。单因素分析结果显示:性别、术前抗菌药物的使用、胸管类型和ICU中心胸腔感染无相关性(P0.05),而年龄、手术时间、术前肺功能、引流管留置时间、手术创口污染、原发病灶蔓延均和ICU中心胸腔感染密切相关(P0.05);多因素logistic回归分析结果显示:年龄≥60岁、手术时间≥2h、术前肺功能、引流管留置时间≥3d、手术创口污染、原发病灶蔓延均是造成ICU中心胸腔感染的独立危险因素(OR=3.485、3.714、3.571、5.731、6.172、6.081,P0.05)。结论:ICU中心发生胸腔感染会对患者病情恢复造成较大影响,在今后临床工作中,需重视围术期管理,积极采取合理的预防措施,降低胸腔感染的发生率。     

基于景观地形的小流域单元减灾调控评价研究

近年来,中国诸多大中城市内涝灾害频发。武汉市作为城市内涝频发的典型城市,其建成区地表高度（20.0~24.0 m）大多低于外江常年所处的洪水位高度（23.9 m）,在区域性暴雨条件下,极易引发内涝灾害。武汉由于其内涝典型性,2013年被列为全国31个重点防洪城市之一,2015年被列入首批海绵城市试点城市之一。从城市内涝发生机理出发,以水文角度的小流域单元作为内涝风险区划的基本单元,利用小流域单元分析武汉市景观地形要素（地表高程、地表起伏度、地表粗糙度）与城市内涝积水密度的相关性及其影响程度,量算统计出小流域单元的内涝风险等级。基于此,提出武汉市小流域单元减灾地形调控评价。以武汉市作为城市内涝问题的研究案例,以期为武汉市内涝缓解提供新的思考路径,具有典型性和必要性。     

Population genetic structure and conservation of marbled murrelets (<Emphasis Type="Italic">Brachyramphus marmoratus</Emphasis>)

Marbled murrelets (Brachyramphus marmoratus) are coastal seabirds that nest from California to the Aleutian Islands. They are declining and considered threatened in several regions. We compared variation in the mitochondrial control region, four nuclear introns and three microsatellite loci among194 murrelets from throughout their range except Washington and Oregon. Significant population genetic structure was found: nine private control region haplotypes and three private intron alleles occurred at high frequency in the Aleutians and California; global estimates of F _ST or Φ_ST and most pairwise estimates involving the Aleutians and/or California were significant; and marked isolation-by-distance was found. Given the available samples, murrelets appear to comprise five genetic management units: (1) western Aleutian Islands, (2) central Aleutian Islands, (3) mainland Alaska and British Columbia, (4) northern California, and (5) central California.     

Effect of Exposure Period and Algal Concentration on the Frequency of Infection of Aposymbiotic Ciliates by Symbiotic Algae from Paramecium bursaria

The effect of exposure period and concentration of algae on the frequency of infection of aposymbiotic ciliates by algae obtained from the same clone of Paramecium bursaria syngen 2, was studied. The frequency of infection was roughly proportional to the algal concentration and to the exposure time of ciliates to algae. The relationship of algal concentration to infection frequency closely fitted the Poisson distribution curve for N = 1, suggesting that the minimum number of algae required to infect a single ciliate is 1. However, the data also strongly suggested that the average number of algae required to initiate infection of an average ciliate was ? 1,000. Three possible resolutions of this situation are: (a) the selection by the ciliate of a rare infective variant from a heterogeneous population: (b) the rare escape of an alga from digestion by the ciliate; and (c) the requirement for a large number of algae-ciliate contacts to induce susceptibility in the ciliate. Splitting the exposure of ciliates to algae into 2 periods of 0.5 h, separated by 5 h in the absence of algae, produced a much higher frequency of infection than a single l-h exposure, supporting the suggestion that the large number of algae is required to induce susceptibility in the ciliate which can then be infected by as few as a single algal cell.     

TWEAK and the Central Nervous System

Tumor necrosis factor-like weak inducer of apoptosis (TWEAK) is a member of the tumor necrosis factor superfamily that acts on responsive cells via binding to a cell surface receptor named fibroblast growth factor-inducible 14 (Fn14). TWEAK can regulate numerous cellular responses in vitro and in vivo. Recent studies have indicated that TWEAK and Fn14 are expressed in the central nervous system (CNS), and that in response to a variety of stimuli, including cerebral ischemia, there is an increase in TWEAK and Fn14 expression in perivascular astrocytes, microglia, endothelial cells, and neurons with subsequent increase in the permeability of the blood–brain barrier (BBB) and cell death. Furthermore, there is a growing body of evidence indicating that TWEAK induces the activation of the NF-κB in the CNS with release of proinflammatory cytokines and matrix metalloproteinases. In addition, inhibition of TWEAK activity by either treatment with a Fn14-Fc fusion protein or neutralizing anti-TWEAK antibodies has shown therapeutic efficacy in animal models of ischemic stroke, cerebral edema, and multiple sclerosis.     

Atomistic and coarse-grained analysis of double spectrin repeat units: the molecular origins of flexibility

Spectrin is an ubiquitous protein in metazoan cells, and its flexibility is one of the keys to maintaining cellular structure and organization. Both alpha-spectrin and beta-spectrin polypeptides consist primarily of triple coiled-coil modular repeat units, and two important factors that determine spectrin flexibility are the bending flexibility between two consecutive repeat units and the conformational flexibility of individual repeat units. Atomistic molecular dynamics (MD) simulations are used here to study double spectrin repeat units (DSRUs) from the human erythrocyte beta-spectrin (HEbeta89) and the chicken brain alpha-spectrin (CBalpha1617). From the results of MD simulations, a highly conserved Trp residue in the A-helix of most repeat units that has been suggested to be important in conferring stability to the coiled-coil structures is found not to have a significant effect on the conformational flexibility of individual repeat units. Characterization of the bending flexibility for two consecutive repeats of spectrin via atomistic simulations and coarse-grained (CG) modeling indicate that the bending flexibility is governed by the interactions between the AB-loop of the first repeat unit, the BC-loop of the second repeat unit and the linker region. Specifically, interactions between residues in these regions can lead to a strong directionality in the bending behavior of two repeat units. The biological implications of these finding are discussed.     

60Coγ射线与EMS复合处理对山黧豆抗氧化酶活力及ODAP含量的影响

1　引　　言山黧豆抗寒、抗旱、耐贫瘠 ,尤其适于干旱、半干旱地区种植 .但山黧豆中含有的毒素 β ＯＤＡＰ会导致下肢瘫痪 .由于这个原因限制了山黧豆的大面积栽培 .我们采用 60 Ｃｏγ射线和ＥＭＳ单因子、复因子诱变的方法选育丰产、低毒、无毒山黧豆新品系 .近年来对生物辐射敏感性的研究表明 ,其抗性除与本身ＤＮＡ损伤修复能力有关外 ,还与体内所含防护物质有关 ,ＳＯＤ、ＰＯＤ、ＣＡＴ即是重要的保护物质 .关于抗氧化酶活性与辐射剂量、诱变剂浓度关系的研究很少 ,因此本实验对山黧豆成苗率、抗氧化酶活性、和 β ＯＤＡＰ含量进行…