Rapid cloning of high-affinity human monoclonal antibodies against influenza virus

Pre-existing neutralizing antibody provides the first line of defence against pathogens in general. For influenza virus, annual vaccinations are given to maintain protective levels of antibody against the currently circulating strains. Here we report that after booster vaccination there was a rapid and robust influenza-specific IgG+ antibody-secreting plasma cell (ASC) response that peaked at approximately day 7 and accounted for up to 6% of peripheral blood B cells. These ASCs could be distinguished from influenza-specific IgG+ memory B cells that peaked 14-21 days after vaccination and averaged 1% of all B cells. Importantly, as much as 80% of ASCs purified at the peak of the response were influenza specific. This ASC response was characterized by a highly restricted B-cell receptor (BCR) repertoire that in some donors was dominated by only a few B-cell clones. This pauci-clonal response, however, showed extensive intraclonal diversification from accumulated somatic mutations. We used the immunoglobulin variable regions isolated from sorted single ASCs to produce over 50 human monoclonal antibodies (mAbs) that bound to the three influenza vaccine strains with high affinity. This strategy demonstrates that we can generate multiple high-affinity mAbs from humans within a month after vaccination. The panel of influenza-virus-specific human mAbs allowed us to address the issue of original antigenic sin (OAS): the phenomenon where the induced antibody shows higher affinity to a previously encountered influenza virus strain compared with the virus strain present in the vaccine. However, we found that most of the influenza-virus-specific mAbs showed the highest affinity for the current vaccine strain. Thus, OAS does not seem to be a common occurrence in normal, healthy adults receiving influenza vaccination.     

Reduction and selective oxo group silylation of the uranyl dication

Uranium occurs in the environment predominantly as the uranyl dication [UO2]2+. Its solubility renders this species a problematic contaminant which is, moreover, chemically extraordinarily robust owing to strongly covalent U-O bonds. This feature manifests itself in the uranyl dication showing little propensity to partake in the many oxo group functionalizations and redox reactions typically seen with [CrO2]2+, [MoO2]2+ and other transition metal analogues. As a result, only a few examples of [UO2]2+ with functionalized oxo groups are known. Similarly, it is only very recently that the isolation and characterization of the singly reduced, pentavalent uranyl cation [UO2]+ has been reported. Here we show that placing the uranyl dication within a rigid and well-defined molecular framework while keeping the environment anaerobic allows simultaneous single-electron reduction and selective covalent bond formation at one of the two uranyl oxo groups. The product of this reaction is a pentavalent and monofunctionalized [O = U...OR]+ cation that can be isolated in the presence of transition metal cations. This finding demonstrates that under appropriate reaction conditions, the uranyl oxo group will readily undergo radical reactions commonly associated only with transition metal oxo groups. We expect that this work might also prove useful in probing the chemistry of the related but highly radioactive plutonyl and neptunyl analogues found in nuclear waste.     

Amorphous silica-like carbon dioxide

Among the group IV elements, only carbon forms stable double bonds with oxygen at ambient conditions. At variance with silica and germania, the non-molecular single-bonded crystalline form of carbon dioxide, phase V, only exists at high pressure. The amorphous forms of silica (a-SiO2) and germania (a-GeO2) are well known at ambient conditions; however, the amorphous, non-molecular form of CO2 has so far been described only as a result of first-principles simulations. Here we report the synthesis of an amorphous, silica-like form of carbon dioxide, a-CO2, which we call 'a-carbonia'. The compression of the molecular phase III of CO2 between 40 and 48 GPa at room temperature initiated the transformation to the non-molecular amorphous phase. Infrared spectra measured at temperatures up to 680 K show the progressive formation of C-O single bonds and the simultaneous disappearance of all molecular signatures. Furthermore, state-of-the-art Raman and synchrotron X-ray diffraction measurements on temperature-quenched samples confirm the amorphous character of the material. Comparison with vibrational and diffraction data for a-SiO2 and a-GeO2, as well as with the structure factor calculated for the a-CO2 sample obtained by first-principles molecular dynamics, shows that a-CO2 is structurally homologous to the other group IV dioxide glasses. We therefore conclude that the class of archetypal network-forming disordered systems, including a-SiO2, a-GeO2 and water, must be extended to include a-CO2.     

Reconstructing the early evolution of Fungi using a six-gene phylogeny

The ancestors of fungi are believed to be simple aquatic forms with flagellated spores, similar to members of the extant phylum Chytridiomycota (chytrids). Current classifications assume that chytrids form an early-diverging clade within the kingdom Fungi and imply a single loss of the spore flagellum, leading to the diversification of terrestrial fungi. Here we develop phylogenetic hypotheses for Fungi using data from six gene regions and nearly 200 species. Our results indicate that there may have been at least four independent losses of the flagellum in the kingdom Fungi. These losses of swimming spores coincided with the evolution of new mechanisms of spore dispersal, such as aerial dispersal in mycelial groups and polar tube eversion in the microsporidia (unicellular forms that lack mitochondria). The enigmatic microsporidia seem to be derived from an endoparasitic chytrid ancestor similar to Rozella allomycis, on the earliest diverging branch of the fungal phylogenetic tree.     

ANLL中转录因子GATA—2基因点突变

了解转录因子ＧＡＴＡ－２基因在ＡＮＬＬ中的表达和突变情况,方法：采用ＲＴ－ＰＣＲ检测８５例ＡＮＬＬ病人外周血单个核细胞中ＧＡＴＡ－２基因的表达情况,ＰＣＲ产物进一步经单链构象多态性（ＳＳＣＰ）分析以了解基因突变情况。结果：绝大多数ＡＮＬＬ都表达了ＧＡＴＡ－２基因（８９．４％）,ＳＳＣＰ分析发现一例Ｍ２型的ＰＣＲ产物出现异常迁移带,核苷序列分析显示在ＧＡＴＡ－２基因第８９２位的核苷酸出现点突变,即第     

Topological surface states and flat bands in the kagome superconductor CsV3Sb5

Exotic quantum phenomena may appear in material systems with multiple orders or phases,where the mutual interactions can give rise to new physics beyond that of...     

鱼类营养学研究的普遍问题及建议

在确定鱼类对潜在营养源的利用或营养物需求的研究中,常存在一些问题.本文将讨论这些问题,并提出这类实验的一般步骤. 1、饲料 商用饲料需注明饲料及厂家的全称.实验饲料需列出所有原料的标准名称及国际饲料号.精饲料及添加剂需列出原料供应商及地址.基础饲料应能提供实验鱼所需的全部营养.除了所测成分外,饲料的组成应是恒定的.最好能根据消化率数据确定饲料配方.饲料制作的步骤应描述清楚.饲料使用前应冷冻保存.饲料应采用国际上接受的方法进行化学分析. 2、实验条件 应描述饲养单元的大小、位置、水源及水处理方法、水温、流速、溶氧、水质参数、光照条件、实验鱼的详细情况及其他实验条件.养鱼系统包括流水、循环过滤水及静水系统.静水系统不能用于营养需求实验,循环水系统亦不宜用于维生素及无机盐需求量实验. 3、实验设计 每一饲养单元是一个重复,单元内的个体不能看作重复样.每一处理至少需要3个重复.实验鱼需经过一段时间暂养才能用于实验.实验周期至少8周,微营养物需求实验需要12-20周.实验期增重率应在200%-500%以上.饲养单元应随机分配于各个处理.投喂方式可以是饱食或限食.若采用限食投喂,投喂水平应接近饱食. 4、数据评价 如果对照鱼生长率较预期的低,或死亡率较高,或实验结果与预期的不同,则有必要重复实验.需求量实验应得到剂量反应曲线,否则须重新设计实验.建议采用拐点法或回归法确定最低需求量.     

龙门山造山带-川西前陆盆地系统构造事件研究

通过对构造事件形成的产物 (岩浆岩、各种矿物等 )进行氩 -氩年龄测定、裂变径迹年龄测定、ESR年龄测定和计算机模拟 ,得出龙门山造山带 -川西前陆盆地系统自印支期以来 ,共发生了 7次构造事件 :(1)卡尼期末的构造事件 (D1 ) ,形成 S1 和 F1 ,并有区域低温动热变质作用 ;(2 )诺利期末的构造事件 (安县运动 ,D2 ) ,产生川西前陆盆地上三叠统须四段与下伏地层不整合接触和须四段砾岩的分布 ,并有较强的岩浆作用 ;(3)燕山期 (12 0～ 130 Ma,D3) ,有较强的岩浆作用和变质作用 ;(4 )喜马拉雅期 6 0 Ma左右的构造事件 (D4) ;(5 )喜马拉雅期 30～ 40Ma的构造事件 (D5 ) ;(6 )喜马拉雅期 2 0～ 2 5 Ma的构造事件 (D6 ) ;(7)喜马拉雅期 10 Ma的构造事件 (D7)。后 4次构造事件使龙门山造山带 -川西前陆盆地系统发生强烈的冲断作用和隆升作用     

The Toll-like receptor 2 is recruited to macrophage phagosomes and discriminates between pathogens.

Macrophages orchestrate innate immunity by phagocytosing pathogens and coordinating inflammatory responses. Effective defence requires the host to discriminate between different pathogens. The specificity of innate immune recognition in Drosophila is mediated by the Toll family of receptors; Toll mediates anti-fungal responses, whereas 18-wheeler mediates anti-bacterial defence. A large number of Toll homologues have been identified in mammals, and Toll-like receptor 4 is critical in responses to Gram-negative bacteria. Here we show that Toll-like receptor 2 is recruited specifically to macrophage phagosomes containing yeast, and that a point mutation in the receptor abrogates inflammatory responses to yeast and Gram-positive bacteria, but not to Gram-negative bacteria. Thus, during the phagocytosis of pathogens, two classes of innate immune receptors cooperate to mediate host defence: phagocytic receptors, such as the mannose receptor, signal particle internalization, and the Toll-like receptors sample the contents of the vacuole and trigger an inflammatory response appropriate to defence against the specific organism.