Actin polymerization-driven molecular movement of mDia1 in living cells

mDia1, a Rho effector, belongs to the Formin family of proteins, which shares the conserved tandem FH1-FH2 unit structure. Formins including mDia1 accelerate actin nucleation while interacting with actin filament fast-growing ends. Here our single-molecule imaging revealed fast directional movement of mDia1 FH1-FH2 for tens of microns in living cells. The movement of mDia1 FH1-FH2 was blocked by actin-perturbing drugs, and the speed of mDia1 FH1-FH2 movement appeared to correlate with actin elongation rates. In vitro, mDia1 FH1-FH2 associated persistently with the growing actin barbed end. mDia1 probably moves processively along the growing end of actin filaments in cells, and Formins may be a molecular motility machinery that is independent from motor proteins.     

Rocket launcher mechanism of collaborative actin assembly defined by single-molecule imaging

Interacting sets of actin assembly factors work together in cells, but the underlying mechanisms have remained obscure. We used triple-color single-molecule fluorescence microscopy to image the tumor suppressor adenomatous polyposis coli (APC) and the formin mDia1 during filament assembly. Complexes consisting of APC, mDia1, and actin monomers initiated actin filament formation, overcoming inhibition by capping protein and profilin. Upon filament polymerization, the complexes separated, with mDia1 moving processively on growing barbed ends while APC remained at the site of nucleation. Thus, the two assembly factors directly interact to initiate filament assembly and then separate but retain independent associations with either end of the growing filament.     

Role of formins in actin assembly: nucleation and barbed-end association

Nucleation of branched actin filaments by the Arp2/3 complex is a conserved process in eukaryotic cells, yet the source of unbranched actin filaments has remained obscure. In yeast, formins stimulate assembly of actin cables independently of Arp2/3. Here, the conserved core of formin homology domains 1 and 2 of Bni1p (Bni1pFH1FH2) was found to nucleate unbranched actin filaments in vitro. Bni1pFH2 provided the minimal region sufficient for nucleation. Unique among actin nucleators, Bni1pFH1FH2 remained associated with the growing barbed ends of filaments. This combination of properties suggests a direct role for formins in regulating nucleation and polarization of unbranched filamentous actin structures.     

仿生法评定饲料干物质消化率的影响因素

【目的】为确立饲料养分生物学效价仿生法的评定操作规程,探讨仿生法测定饲料干物质消化率的影响因素。【方法】试验采用单因素完全随机设计,共设6个处理,每个处理5个重复,每个重复使用1根仿生消化管,其中处理1—2、3—4、5—6分别考察透析袋的前处理方式、消化残渣脱脂和蠕动泵泵入方式对饲料体外干物质消化率的影响。【结果】①透析袋使用两种前处理方式,玉米、豆粕和麦麸干物质消化率无显著影响（P＞0.05）;②脱脂显著提高玉米、豆粕和麦麸的干物质和粗脂肪的消化率,干物质消化率提高约3%,粗脂肪消化率达到90%以上（P＜0.05）,但脱脂未对饲料其它养分的消化率产生影响（P＞0.05）;③蠕动泵泵入缓冲液方式对饲料干物质消化率有显著影响,注入式所测得的玉米、豆粕和麦麸的饲料干物质消化率显著低于吸入式（P＜0.05）,但是其测试的平行性较好。【结论】仿生法测试饲料体外干物质消化率时,消化残渣应进行脱脂,透析袋采用两种前处理方式均可,蠕动泵以注入方式更符合测试精度的要求。     

Requirement for coronin 1 in T lymphocyte trafficking and cellular homeostasis

The evolutionarily conserved actin-related protein (Arp2/3) complex is a key component of actin filament networks that is dynamically regulated by nucleation-promoting and inhibitory factors. Although much is known about actin assembly, the physiologic functions of inhibitory proteins are unclear. We generated coronin 1-/- mice and found that coronin 1 exerted an inhibitory effect on cellular steady-state F-actin formation via an Arp2/3-dependent mechanism. Whereas coronin 1 was required for chemokine-mediated migration, it was dispensable for T cell antigen receptor functions in T cells. Moreover, actin dynamics, through a mitochondrial pathway, was linked to lymphocyte homeostasis.     

Hierarchical self-assembly of F-actin and cationic lipid complexes: stacked three-layer tubule networks

We describe a distinct type of spontaneous hierarchical self-assembly of cytoskeletal filamentous actin (F-actin), a highly charged polyelectrolyte, and cationic lipid membranes. On the mesoscopic length scale, confocal microscopy reveals ribbonlike tubule structures that connect to form a network of tubules on the macroscopic scale (more than 100 micrometers). Within the tubules, on the 0.5- to 50-nanometer length scale, x-ray diffraction reveals an unusual structure consisting of osmotically swollen stacks of composite membranes with no direct analog in simple amphiphilic systems. The composite membrane is composed of three layers, a lipid bilayer sandwiched between two layers of actin, and is reminiscent of multilayered bacterial cell walls that exist far from equilibrium. Electron microscopy reveals that the actin layer consists of laterally locked F-actin filaments forming an anisotropic two-dimensional tethered crystal that appears to be the origin of the tubule formation.     

所罗门群岛海域长鳍金枪鱼繁殖生物学初步研究

根据2012年9-12月所罗门群岛海域采集的307尾长鳍金枪鱼样本,分析其性别比、性腺成熟度和性腺指数等繁殖生物学参数。结果表明：繁殖群体总性别比和性成熟个体性别比均不符合1：1（P〈0．05）,雄性个体比例大于雌性,性别比在产卵期间出现一定波动,但普遍小于1：1。性别比与个体大小密切相关,雄性比例随叉长增长而降低。性成熟度分析表明,产卵期间性成熟个体比例达到70％以上,其中存在大量的产卵后群体,未成熟比例较低。雌性不同性成熟度大小差异明显（P〈0．05）,但雄性并不明显（P〉0．05）。相同性成熟度雄性个体平均叉长均大于雌性,但并没有体现出一致的变化。雌、雄个体初次性成熟叉长分别为84．88cm和85．61cm。性腺指数（Gsi）在各性成熟度和叉长组间存在显著差异（P〈0．05）,雌性GSI大于1．2几乎全部为成熟个体,尤其GSI大于2普遍为即将产卵（V期）和产卵后（Ⅵ期）群体;雄性个体GSI普遍小于2,各叉长组雌性GSI普遍高于雄性,这表明雌性性腺发育较快。而调查期间9月GSI均值明显高于其他3月（P〈0．05）,综合分析认为该海域长鳍金枪鱼9—10月达到产卵高峰期。     

Actin polymerization and ATP hydrolysis

F-actin is the major component of muscle thin filaments and, more generally, of the microfilaments of the dynamic, multifunctional cytoskeletal systems of nonmuscle eukaryotic cells. Polymeric F-actin is formed by reversible noncovalent self-association of monomeric G-actin. To understand the dynamics of microfilament systems in cells, the dynamics of polymerization of pure actin must be understood. The following model has emerged from recent work. During the polymerization process, adenosine 5'-triphosphate (ATP) that is bound to G-actin is hydrolyzed to adenosine 5'-diphosphate (ADP) that is bound to F-actin. The hydrolysis reaction occurs on the F-actin subsequent to the polymerization reaction in two steps: cleavage of ATP followed by the slower release of inorganic phosphate (Pi). As a result, at high rates of filament growth a transient cap of ATP-actin subunits exists at the ends of elongating filaments, and at steady state a stabilizing cap of ADP.Pi-actin subunits exists at the barbed ends of filaments. Cleavage of ATP results in a highly stable filament with bound ADP.Pi, and release of Pi destabilizes the filament. Thus these two steps of the hydrolytic reaction provide potential mechanisms for regulating the monomer-polymer transition.     

濆江流域不同土地利用方式下土壤磷积累特征及流失风险

采用实地采样调查、室内分析与数理统计法研究了濆江小流域耕地、果园、茶园、人工林地、自然林地、饲草地和荒草地7种土地利用方式下表层土壤磷素的积累及吸附特征。结果表明,人工林地、饲草地与耕地土壤全磷>1.0 g·kg-1,呈现积累趋势;饲草地和耕地的Olsen-P≥40.0 mg·kg-1,超过磷素流失临界值;人工林地、饲草地和耕地土壤Ca Cl2-P>4.5 mg·kg-1,也超过磷素流失临界值。茶园土壤吸附固定磷能力极强,耕地、果园地、林地(尤其是人工林地)与草地土壤的吸附固定磷能力较弱。人工林地、耕地与饲草地土壤磷素流失风险较高,宜采用补偿性施磷或维持性磷肥法;果园、荒草地、自然林地的风险较低,茶园土壤磷素几乎无流失风险。     

两品系萼花臂尾轮虫的生活史特征和受精率研究

应用单个体培养和群体培养方法,在25℃条件下,对萼花臂尾轮虫Brachionus calyciflorus南京品系(NJ)和美国佛罗里达品系(FL)的幼体发育时间、胚胎发育时间、繁殖量、种群增长参数及品系间、品系内受精率进行了研究.结果表明:萼花臂尾轮虫FL品系与NJ品系的幼体发育时间、第一个幼体孵出时间和繁殖量之间均无显著性差异(P＞0.05),而NJ品系的胚胎发育时间比FL品系显著缩短6.9％(P＜0.05); NJ品系的净生殖率(R0)、内禀增长率(rm)、世代时间(T)和生命期望(e0)均显著大于FL品系(P＜0.05);萼花臂尾轮虫FL和NJ品系雌雄交配试验结果表明,两品系间的受精率低于各自品系内交配的受精率.本研究表明,南京品系和佛罗里达品系萼花臂尾轮虫的发育和种群增长参数存在显著差异,但两品系之间不存在完全的生殖隔离,未形成姐妹种.     

作物细胞质雄性不育与肌动蛋白

本文比较了小麦、玉米及白莱的细胞质雄性不育系与保持系花药或花粉中的肌动蛋白。SDS 聚丙烯酰胺凝胶电泳结果表明,保持系含有明显的肌动蛋白区带;而雄性不育系的区带不明显。将凝胶进行光密度扫描亦表明,保持系比不育系所含肌动蛋白数量多。并比较了花药或花粉中的肌动蛋白与兔骨骼肌的肌动蛋白,其电泳迁移率完全相同。     

Observations of the liquid-crystal analog of the abrikosov phase

Freeze-fracture transmission electron micrographs of the smectic A(*) phase confirm the twist grain boundary model of Renn and Lubensky. The fracture surface has an undulating structure with a 0.5-micrometer helical pitch parallel to 4.1-nanometer smectic layers. The layers are disrupted by a lattice of screw dislocations oriented normal to the helical axis. Optical diffraction shows that rotation of smectic blocks occurs in discrete steps of about 17 degrees ; hence, the screw dislocations are 14 to 15 nanometers apart and the grain boundaries are 24 nanometers apart. These observations show that the SmA(*) phase is the liquid-crystal analog of the Abrikosov phase in superconductors.     

全履带式再生稻收割机行走底盘碾压率的模拟与分析

为减少全履带式再生稻收割机收获再生稻头季时行走底盘对留桩的碾压率(履带碾压面积与收割面积的比值),以利于提高再生季水稻产量、并改善再生季稻米品质,基于履带式车辆设计理论,以割幅Z、轨距B、履带接地长度L、履带宽度b、转向半径R0、底盘中心轴线与割台割刀纵向距离X为影响因素,建立了全履带式再生稻收割机行走底盘结构模型及其田间直行转弯碾压模型,以种植行距i,株距c,穴径br的水稻为对象,对上述各参数对碾压率的影响规律进行了分析,结果表明,其他参数相同条件下,直行时,碾压率δ1随割幅Z与履带宽度b的比值增加而减小;转弯时,全履带式再生稻收割机碾压率δ2随转向角度θ增大而减小,随转向半径R0的增大而减小;碾压率不受底盘中心轴线与割台割刀纵向距离X的影响;轨距B以及割幅Z与轨距B之差为行距和株距的公倍数时有利于减少碾压率;在相同接地比压条件下,割幅Z增加有利于减少碾压率。为减少碾压率,全履带式再生稻收割机结构设计时,在满足接地比压前提下,应减少履带宽度b和接地长度L,增大割幅Z,轨距B取行距和株距的公倍数,割幅Z与轨距B之差为行距和株距的最小公倍数,采用回转式行走路径;结合田块形状与面积,优先选用较大转向半径R0;在农艺上,建议水稻种植行距与株距有整数倍关系。     

Mechanical rotation of the c subunit oligomer in ATP synthase (F0F1): direct observation

F0F1, found in mitochondria or bacterial membranes, synthesizes adenosine 5'-triphosphate (ATP) coupling with an electrochemical proton gradient and also reversibly hydrolyzes ATP to form the gradient. An actin filament connected to a c subunit oligomer of F0 was able to rotate by using the energy of ATP hydrolysis. The rotary torque produced by the c subunit oligomer reached about 40 piconewton-nanometers, which is similar to that generated by the gamma subunit in the F1 motor. These results suggest that the gamma and c subunits rotate together during ATP hydrolysis and synthesis. Thus, coupled rotation may be essential for energy coupling between proton transport through F0 and ATP hydrolysis or synthesis in F1.     

长期施肥紫色土有效磷变化及其对稻麦轮作产量的影响

【目的】通过总结分析长期施肥处理下紫色土稻麦轮作土壤有效磷的变化特征,以及土壤磷素变化对作物产量的影响,为紫色土稻麦轮作磷素管理提供理论依据。【方法】依托国家肥力监测网紫色土肥力监测试验站27年的稻麦轮作定位试验,选取10种不同施肥处理：CK处理（只种作物不施肥）;N、NP、NK、PK、NPK为不同氮（N）、磷（P）、钾（K）化肥配施处理;M、NPKS、NPKM、1.5NPK+M为有机肥（M）、秸秆还田（S）及其与化肥配施处理。试验数据涵盖1991—2018年,测定不同施肥处理下土壤有效磷含量和作物产量,计算100 kg籽粒磷素吸收量和磷肥利用率,分析土壤磷素变化对累积磷盈亏的响应,采用不同模型计算土壤磷素农学阈值。【结果】长期施用磷肥能够显著提高土壤有效磷含量,各施磷处理有效磷年均增量为0.80—2.32 mg·kg^-1;而不施磷处理CK、N、NK和单施有机肥处理M的土壤有效磷含量则逐年下降至平稳状态。不施磷处理土壤磷素一直处于亏缺状态,施磷各处理27年后土壤累积磷盈余量为244.8—698.2 kg P·hm^-2,其中1.5NPK+M处理累积磷盈余量最高;施磷处理土壤累积盈余量与土壤Olsen-P增量呈显著线性相关,土壤每盈余磷100 kg·hm^-2,土壤有效磷含量提高4.27—6.5 mg·kg^-1。磷肥施用能显著提升稻麦轮作系统作物产量和吸磷量,100 kg水稻籽粒需磷量为0.17—0.41 kg,100 kg小麦籽粒需磷量为0.25—0.57 kg;试验各处理的磷肥利用率为10.3%—39.7%;4种模型（线性-平台模型、双直线模型、BoxLucas模型和米切里西模型）均能较好地拟合作物产量与紫色土有效磷含量的响应关系,其中双直线模型的拟合度最好,其计算的水稻和小麦的土壤有效磷农学阈值分别为13.28和9.93 mg·kg^-1。 【结论】在紫色土水稻-小麦轮作体系中,合理施用磷肥能显著提高作物吸磷量、产量以及土壤有效磷含量。推荐双直线模型用于计算紫色土稻麦轮作体系下土壤有效磷的农学阈值,生产上应根据土壤有效磷含量及其农学阈值调整磷肥施用量。     

Localized stabilization of microtubules by integrin- and FAK-facilitated Rho signaling

Microtubule (MT) stabilization is regulated by the small guanosine triphosphate (GTP)-binding protein Rho and its effector, mammalian homolog of Diaphanous (mDia), in migrating cells, but factors responsible for localized stabilization at the leading edge are unknown. We report that integrin-mediated activation of focal adhesion kinase (FAK) at the leading edge is required for MT stabilization by the Rho-mDia signaling pathway in mouse fibroblasts. MT stabilization also involved FAK-regulated localization of a lipid raft marker, ganglioside GM1, to the leading edge. The integrin-FAK signaling pathway may facilitate Rho-mDia signaling through GM1, or through a specialized membrane domain containing GM1, to stabilize MTs in the leading edge of migrating cells.     

Countercurrent chromatography with flow-through coil planet centrifuge

We have developed a new method of countercurrent chromatography which employs a vertical helical tube in the centrifugal field. The helical tube is arranged so that it does not rotate as it revolves, thus eliminating the need for rotating seals. When the gyrating tube is filled with either phase and the other phase is introduced into the tube in the proper direction, an equilibrium state results in which the two phases are split into multiple alternating segments within the coil. Each phase oscillates to and fro with the rotation as the moving phase is steadily eluted out through the other end of the tube. Consequently, solutes introduced into the tube are subjected to a rapid partition process, resulting in an efficient chromatographic separation without the complications arising from solid supports. The method is illustrated by the microanalytical separation of dinitrophenyl amino acids and can be used on a preparative scale.     

Cytochalasin B: does it affect actin-like filaments?

An in vitro system was used to test the purported action of cytochalasin B. At concentrations 100 times those used for experiments in vivo, cytochalasin B did not cause the breakdown of F-actin, did not inhibit the transformation of G-actin to F-actin, did not inhibit the binding of heavy meromyosin to F-actin, and did not inhibit the adenosine triphosphate-induced release of heavy meromyosin from F-actin.     

Mechanics of stimulated neutrophils: cell stiffening induces retention in capillaries

The effect of peptide chemoattractants on neutrophil mechanical properties was studied to test the hypothesis that stimulated neutrophils (diameter, 8 micrometers) are retained in pulmonary capillaries (5.5 micrometers) as a result of a decreased ability of the cell to deform within the capillary in response to the hydrodynamic forces of the bloodstream. Increased neutrophil stiffness, actin assembly, and retention in both 5-micrometer pores and the pulmonary vasculature were seen in response to N-formyl-methionyl-leucyl-phenylalanine. These changes were abolished in cells that had been incubated with 2 micromolar cytochalasin D, an agent that disrupts cellular actin organization. A monoclonal antibody directed at the CD11-CD18 adhesive glycoprotein complex did not inhibit the increase in stiffness or retention in pores. These data suggest that neutrophil stiffening may be both necessary and sufficient for the retention that is observed. Hence, neutrophil sequestration in lung and other capillaries in the acute inflammatory process may be the result of increased stiffness stimulated by chemoattractants.     

中华沙鳅个体生殖力研究

为了研究中华沙鳅(Botia superciliaris)个体生殖力及其与多项生物学指标的关系,2009年5～9月从沱江资中段和长江宜宾一南溪段分别收集性腺Ⅳ期中华沙鳅雌鱼32尾和22尾进行研究.结果如下:①沱江资中段中华沙鳅产卵群体由2、3龄鱼组成;绝对生殖力(F)为 560～4 401粒,平均1 915粒,体长相对生殖力(FL)为63～458粒/cm,平均217粒/cm,体重相对生殖力(FW)为39～424粒/g,平均230粒/g;绝对生殖力、体长相对生殖力与体长、体重、净体重和成熟系数相关性极显著,体重相对生殖力与体重和成熟系数相关性显著,个体生殖力与肥满度相关性不显著.②长江宜宾一南溪段产卵群体由2、3、4龄鱼组成;绝对生殖力(F)为1 593～14 892粒,平均4 991粒,体长相对生殖力(FL)为168～1 568粒/cm,平均509粒/cm,体重相对生殖力(FW)为49～294粒/g,平均123粒/g;绝对生殖力、体长相对生殖力和体重相对生殖力与体重、净体重、成熟系数和肥满度相关性极显著,与体长和年龄相关性不显著.研究结果表明沱江资中段中华沙鳅繁殖群体低龄化、小型化,建议加强其资源保护.