Probabilistic Modeling of Rosette Formation

英文摘要: Rosetting, or forming a cell aggregate between a single target nucleated cell and a number of red blood cells (RBCs), is a simple assay for cell adhesion-mediated by specific receptor-ligand interaction. For example, rosette formation between sheep RBC and human lymphocytes has been used to differentiate T cells from B cells. Rosetting assay is commonly used to determine the interaction of Fc gamma-receptors (Fc gamma R) expressed on inflammatory cells and IgG-coated on RBCs. Despite its wide use in measuring cell adhesion, the biophysical parameters of rosette formation have not been well characterized. Here we developed a probabilistic model to describe the distribution of rosette sizes, which is Poissonian. The average rosette size is predicted to be proportional to the apparent two-dimensional binding affinity of the interacting receptor-ligand pair and their site densities. The model has been supported by experiments of rosettes mediated by four molecular interactions: Fc gamma RIII interacting with IgG, T cell receptor and coreceptor CD8 interacting with antigen peptide presented by major histocompatibility molecule, P-selectin interacting with P-selectin glycoprotein ligand 1 (PSGL-1), and L-selectin interacting with PSGL-1. The latter two are structurally similar and are different from the former two. Fitting the model to data enabled us to evaluate the apparent effective two-dimensional binding affinity of the interacting molecular pairs: 7.19x10(-5) mu m(4) for Fc gamma RIII-IgG interaction, 4.66x10(-3) mu m(4) for P-selectin-PSGL-1 interaction, and 0.94x10(-3) mu m(4) for L-selectin-PSGL-1 interaction. These results elucidate the biophysical mechanism of rosette formation and enable it to become a semiquantitative assay that relates the rosette size to the effective affinity for receptor-ligand binding.

黄海沉积物多细胞趋磁原核生物的特性研究

多细胞趋磁原核生物（Multicellular magnetotactic prokaryotes，MMPs） 是一类由7～45 个含有磁小体的革兰氏阴性细胞聚集而成的球形或者椭圆形 的细胞聚集体，是研究生命起源与进化、细胞分化和生物矿化的模式生物， 目前仅在大西洋沿岸具有一定盐度的层化水体或沉积物中发现。 本文通过光学显微镜和电子显微镜研究了黄海沉积物MMPs 的超微结 构、运动特点和分裂方式等生物学特征，调查了MMPs 的生态分布特征，并 对其尝试培养。 根据形态差别，黄海沉积物的MMPs 可分为花瓣型MMPs（rosette-like MMPs）、菠萝型MMPs（pineapple-like MMPs）和松球型MMPs（pinecone-like MMPs）。花瓣型MMPs 是由23±4 个卵圆形的细胞螺旋形排列而成的球形聚 集体，直径为5.4±0.8 μm，鞭毛周生。细胞内外膜附近有子弹头形/和方形的 铁氧化物型磁小体。菠萝型MMPs 是由39±9 个方形细胞组成的大小为9.6±1.2 μm ×7.8±0.9 μm 的椭圆形聚集体，鞭毛周生。这类MMPs 由多环细胞组成的， 从椭圆体的赤道面向两极，细胞环的直径变小；在每一环内，细胞像书本似 并列相连；相邻两环的细胞为交错式相连。这种结构比花瓣型MMPs 的更为 紧密。菠萝型MMPs 的磁小体均为子弹头形铁氧化物，磁小体的排列与MMPs 的长轴近似平行。松球型MMPs 是由多个长条形的细胞围绕中心的一个凹陷 辐射排列而成的球形聚集体，直径在9.0～14.2 μm 之间。尼罗红和DAPI 染 色发现三种MMPs 均具有脂类颗粒，花瓣型MMPs 和菠萝型MMPs 在聚集体 的表面具有一层外膜，这说明MMPs 的细胞排列具有高度组织性，在一定程 度证明它属于多细胞生物。 花瓣型MMPs 和菠萝型MMPs 分裂时均保持多细胞形式，但花瓣型 MMPs 沿着聚集体的短轴分开，而菠萝型MMPs 沿着长轴分开。两种MMPs 具有MMPs 典型的逃逸运动，花瓣型MMPs 和菠萝型MMPs 的运动速度分别 为55±26 μm/s 和99±50 μm/s。 黄海花瓣型MMPs 的超微结构、运动方式和分裂特点与大西洋沿岸多个 地区发现的MMPs 相似，花瓣型MMPs 可能是MMPs 的优势类群。菠萝型 MMPs 从整体形态、细胞排列和分裂方式上与花瓣型MMPs 显著不同，是一 类新的MMPs。松球型MMPs 是一类尚未报道的MMPs。 对MMPs 的生态分布调查发现，花瓣型MMPs 广泛分布于砂质沉积物中， 最大丰度出现在氧化还原跃层（redoxcline）。菠萝型MMPs 多分布在砾石沉 积物的表层。两种MMPs 占据不同的生态位，暗示着两者可能具有不同的生 理代谢途径。 对MMPs 的培养发现，在实验室内MMPs 可存活8 个月，MMPs 丰度随 着时间变化出现周期性的变化，推测其繁殖周期可能是10～15 天。 本文为太平洋沿岸MMPs 的首次研究，支持MMPs 在全球广泛分布的观 点，并展示了MMPs 的形态多样性。

Seed germination and seedling morphology of the seagrass, Halophila engelmannii (Hydrocharitaceae)

Seeds of Halophila engelmannii Aschers., that were collected in Redfish Bay, Texas, at weekly intervals from mid-May to mid-June 1986, began to germinate 3–4 weeks after collection. Most of the collections subsequently showed an increase in the rate of germination under increased light intensity and all had a stoppage of germination after transfer to darkness, indicating a light requirement to break endogenous seed dormancy. During the 5 weeks after seeds germinated, seedlings in soil culture produced a rosette of six leaves before the appearance of a rhizome bud in the axil of the third leaf. The first node of the rhizome produced a root and an upright shoot with a pseudowhorl of three to five leaves.

Aging behavior and mechanical properties of Mg-Gd-Ho alloys

The Mg-8Gd-0.6Zr-xHo (x = 1, 3 and 5, mass%) alloys were prepared by casting technology, and structures, aging strengthening mechanism and mechanical properties of the alloys were investigated. The age behaviors and the mechanical properties are improved by adding Ho addition. The structures of the alloys are characterized by the present of rosette-shaped equiaxed grains. The peak hardness value of the Mg-8Gd-0.6Zr-3Ho alloy is 100 Hv, which is about 30% higher than that of Mg-8Gd-0.6Zr alloy.

Age hardening and mechanical properties of Mg-Gd-Er alloy

The Mg-8Gd-0.6Zr-xEr (x = 1, 3 and 5 mass%) alloys were prepared by casting technology, and the microstructures, age hardening behaviors and mechanical properties of alloys have been investigated. Microstructures of the alloys are characterized by the presence of rosette-shaped equiaxed grains. The age hardening behaviors and the tensile properties are enhanced by adding Er element. The maximum aged hardness of Mg-8Gd-0.6Zr-5Er alloy is 97, it is nearly 1.24 times higher than that of Er-free alloy.

黄海纤毛虫的丰度和生物量

海洋浮游纤毛虫是在海洋中浮游生活的一类单细胞原生动物，主要是指寡毛类纤毛虫(Oligotrich ciliates)，隶属原生动物界(Protozoa)、纤毛门(Ciliophora)、寡毛纲(Oligotrichea)，分属于Oligotrichina 和Tintinnina两个亚纲。它们个体微小，粒径在5-200 µm之间，是微型浮游动物和海洋微食物环(Marine Microbial Food Web)的重要组成部分。 2006年4月至2007年12月，在黄海(包括胶州湾)采样分析海洋浮游纤毛虫的种类组成(砂壳纤毛虫)、丰度和生物量，分析纤毛虫在这一海区的季节变化和空间变化。 纤毛虫丰度和生物量的研究方法为：Rosette采水器(胶州湾用Niskin采水器)采集水样，取1 L水样，加Lugol’s试剂固定(终浓度1%)，Utermöhl方法100倍镜检。测量虫体的体长、体宽，按最接近的几何形状(圆柱体、球体和圆锥体)计算体积。生物量由体积乘转换系数(0.19 pgC/µm3)得到，砂壳纤毛虫的肉体体积按照壳体积的1/3近似。 本文的结果表明，胶州湾各站纤毛虫平均丰度于6月达到全年最高值6065 ind./L，12月为全年丰度最低值843 ind./L。平均生物量8月达全年最高值(18.5 µg C/L)，6月为全年最低值(0.6 µg C/L)。砂壳纤毛虫种丰富度于8月达到最高值，共发现25种砂壳纤毛虫，1月种类最少(6种)。湾内站位的纤毛虫平均丰度比湾外的高(6月和8月除外)。砂壳纤毛虫在纤毛虫总丰度中的比例较小，平均为25%，范围为8-57%，分别于1月和8月达到最低和最高值。 两次冷水团大面调查结果表明，4月表层纤毛虫平均丰度(1490 ind./L)要高于10月(972 ind./L)。10月表层纤毛虫生物量0.14-5.33 µg C/L，14194站、15694站和15894站生物量较高，为4.08-5.33 µg C/L。无壳纤毛虫优势种Laboea strobila在两个航次中均呈现斑块分布，4月航次丰度0-10000 ind./L，10月航次丰度11-350 ind./L；砂壳纤毛虫优势种Ptychocylis obtusa仅在4月航次发现，最大丰度2895 ind./L，10月航次未发现。4月航次砂壳纤毛虫有百乐拟铃虫(Tintinnopsis beroidea)，丰度为0-1920 ind./L；卡拉直克拟铃虫(Tintinnopsis karajacensis)，丰度很小(10-93 ind./L)。10月航次砂壳纤毛虫优势种Tintinnidium primitivum，丰度为35-700 ind./L；也出现了尖底类瓮虫(Amphorellopsis acuta)和网纹虫(Favella spp.)，但丰度不大(0-210 ind./L)；运动类铃虫(Codonellopsis mobilis)、筒状拟铃虫(Tintinnopsis tubulosoides)和Eutintinnus sp.丰度也较低(35-105 ind./L)；Craterella torulata丰度为0-120 ind./L，主要分布于15694站。10月航次已经出现了温跃层，位于30 m左右水层，纤毛虫主要分布于温跃层之上。 六次黄海断面航次表明：温跃层在5月已经出现，到12月消失。在有温跃层的5月、6月、8月、9月，纤毛虫主要分布于温跃层(30 m左右)之上。其中8月份航次纤毛虫丰度最高，表层平均丰度3103 ind./L。12月份纤毛虫丰度最低，表层平均丰度406 ind./L。纤毛虫生物量春夏季为0.02-5.5 µg C/L，冬季为0.04-1.99 µg C/L。小型无壳纤毛虫占优势，砂壳纤毛虫东方拟铃虫(Tintinnopsis orientalis)、筒状拟铃虫、运动类铃虫、Craterella torulata和Tintinnidium primitivum几乎在各个航次均有分布。

一种无缆自动返回式采水器的设计与研究

海洋环境污染监测、海水化学与地质调查和生物采样，大多数情况下需要现场采集水样。有时需要在不同深度依次采样，获取梯度剖面的现场水体信息，以保证分析结果的统一性、规范性和类比性。近几十年来，水体取样和保存技术迅速发展，各式各样的采水器纷纷问世并获得成功应用。常规CTD葵花式采水器（CTD rosette sampler）（配备尼斯金采样瓶（Niskin bottle）或郭福洛采样瓶（Go-Flo bottle））和船用吸泵均为目前广泛使用的梯度水样采集装置。但其应用各有一定缺陷。因此，设计一种既适用于大型科考船又适用于小型船只、采样深度可精确控制、采水操作简便、无沾污的采水器在海洋科学研究中有较大实用意义。 本课题所设计的无缆自动返回式采水器（a New Auto-Returned Column Water Sampling Device-‘ARWAD’）是一种新型垂直剖面采水器，该采水器外挂四个1.5L卡盖式结构的HOUSKIN采水瓶。为采水器加载适当重量的配置体，使其在水中受到负浮力下潜。采水是在释放配置体后，采水器受到正浮力上升的过程中依次完成的。 本文的主要内容为：（1） 本课题详述了该无缆自动返回式采水器的机械机构与密封方案。采水器的主壳体为硬铝合金圆柱状防水耐压设计，外挂四个卡盖式结构的采水瓶，其机械机构主要包括①配置体的悬挂与释放机构；②采水瓶关闭采水机构；（2） 详述该采水器的控制系统，包括硬件设计方案和软件设计，并对控制系统进行了调试。控制系统装置于主体的防水耐压仓内，以单片机为核心，装载压力传感器获知水深。下位机程序控制在到达指定深度时释放配置体和关闭各采样瓶，上位机程序实现在甲板上的自检、设定配置体的释放深度和各采水瓶的关闭采样深度；（3） 用解析法和经验公式对该采水器承压零件尺寸进行设计，同时用数值计算方法对其强度校核；（4） 为保证采水器的采水精度和确保其工作安全，对采水器进行水动力学计算，确定阻力及阻力系数等参数，使采水器在上升采水过程中最大速度小于2.0m/s，以为外形设计提供参考；（5） 采水器采样性能验证。现场采集水样后，在实验室测定悬浮颗粒物的质量浓度和体积浓度分布。在采集水样的同时，应用激光粒度仪获取现场悬浮物粒径分布信息。通过与激光粒度仪现场测定结果对比，证明该自动返回式采水器采水深度准确，质量较好。 该采水器使用时无需钢丝绳，而是装载压力传感器获知水深，由此避免了使用绞车和由钢丝绳计量深度带来的深度误差；采样时也无须连接电缆，节省电力；自由下潜和上升，无需配备绞车深度补偿设施，避免出现深度逆变现象。主要应用于驾驶小规模船只在较浅水域（湖泊或河口）采水，亦可用于大型科考船，其结构轻便、造价低廉以及操作方便等性能使其成为CTD葵花式采水器和泵采系统的有益补充。

A new species of Saussurea (Asteraceae) from Tibet and its systematic position based on ITS sequence analysis

A new species of Saussurea, S. erecta S. W Liu, J. T Pan A J. Q. Liu sp. nov., is described from Tibet. It resembles S. kingii but may be distinguished by having distinct stems and glabrous achenes. Saussurea kingii was placed in sect. Pseudoeriocoryne of subgen. Eriocoryne; this section was circumscribed by acaulescence and an inflorescence with congested capitula surrounded by a rosette of leaves. The discovery of S. erecta with distinct stems, cauline leaves and corymbose capitula blurred the delimitation of sect. Pseudoeriocoryne and suggested that the section may be polyphyletic. Both the close relationship and the significant difference between S. erecta and S. kingii were confirmed by analyses of nrDNA ITS sequences. The resulting phylogenies based on ITS data further suggest that Saussurea sect. Pseudoeriocoryne, as traditionally defined, does not constitute a monophyletic group. The rapid radiation and speciation of Saussurea in the Qinghai-Tibetan Plateau, as inferred from ITS phylogeny, are discussed. (c) 2005 The Linnean Society of London.

Phylogenetic analyses of Saussurea sect. Pseudoeriocoryne (Asteraceae : Cardueae) based on chloroplast DNA trnL-F sequences

The genus Saussurea is distributed mainly in the temperate and subarctic regions of Eurasia and consists of about 300 species classified into six subgenera and 20 sections. Sect. Pseudoeriocoryne in the subgenus Eriocoryne comprises four species, and is delimited mainly by acaulescence and an inflorescence with congested capitula surrounded by a rosette of leaves. All of these species are endemic to the and Qinghai-Tibet Plateau. Sequences from the chloroplast DNA trnL-F region were obtained for the four species in this section and 26 other species from four subgenera of Saussurea to resolve phylogenetic relationships among these species and to determine whether the shared characters that define sect. Pseudoeriocoryne are synapomorphic or were acquired by convergent evolution. The resulting phylogenies indicated that Saussurea sect. Pseudoeriocoryne as traditionally defined does not constitute a monophyletic group and that each of its species belongs to separate clades. Furthermore, none of these species showed a close relationship with the other species of subgenus Eriocoryne. Our results further indicated that none of the investigated subgenera are monophyletic, and that species from different subgenera clustered together. All these conclusions are provisional and their confirmation would require stronger phylogenetic support. Two possible explanations are suggested for low sequence divergence, poor resolution of internal clades and clustering of species with the rather distinct morphology of Saussurea detected in the present study. The first is rapid radiation and diversification triggered by fast habitat fragmentation due to the recent lifting of the Qinghai-Tibet Plateau and the Quaternary climate oscillations. This could have led to rapid morphological divergence while sequences diverged very little, and also caused the convergent acquisition of similar characteristics in unrelated lineages due to similar selection pressures. The second possible explanation is that both introgressive hybridization and reticulate evolution might have caused the transferring of cpDNA sequences between morphologically dissimilar species, thus leading to homogenization of sequences between lineages. (C) 2004 Elsevier Ltd. All rights reserved.