Neutrophil beta2 integrin inhibition by enhanced interactions of vasodilator-stimulated phosphoprotein with S-nitrosylated actin

Production of reactive species in neutrophils exposed to hyperoxia causes S-nitrosylation of β-actin, which increases formation of short actin filaments, leading to alterations in the cytoskeletal network that inhibit β(2) integrin-dependent adherence (Thom, S. R., Bhopale, V. M., Mancini, D. J., and Milovanova, T. N. (2008) J. Biol. Chem. 283, 10822-10834). In this study, we found that vasodilator-stimulated protein (VASP) exhibits high affinity for S-nitrosylated short filamentous actin, which increases actin polymerization. VASP bundles Rac1, Rac2, cyclic AMP-dependent, and cyclic GMP-dependent protein kinases in close proximity to short actin filaments, and subsequent Rac activation increases actin free barbed end formation. Using specific chemical inhibitors or reducing cell concentrations of any of these proteins with small inhibitory RNA abrogates enhanced free barbed end formation, increased actin polymerization, and β(2) integrin inhibition by hyperoxia. Alternatively, incubating neutrophils with formylmethionylleucylphenylalanine or 8-bromo-cyclic GMP activates either cyclic AMP-dependent or cyclic GMP-dependent protein kinase, respectively, outside of the short F-actin pool and phosphorylates VASP on serine 153. Phosphorylated VASP abrogates the augmented polymerization normally observed with S-nitrosylated actin, VASP binding to actin, elevated Rac activity, and elevated formation of actin free barbed ends, thus restoring normal β(2) integrin function.     

土壤因子对府谷清水川流域砒砂岩区刺槐和沙棘AMF的影响

应用相关性分析和通径分析研究了府谷清水川流域砒砂岩区土壤因子对刺槐(Robinia pseudoacacia)和沙棘(Hippophae rhamnoides)根际丛枝菌根真菌(AMF)的影响.结果表明:AMF可以与刺槐和沙棘形成良好的共生关系;从两种植物根际土中共分离鉴定出AMF 13种,其中摩西囊霉(Glomus mosseae)、地球囊霉(Glomus geospo-rum)和缩球囊霉(Glomus constrictum)为优势种;AMF平均侵染率为90.6%,平均孢子密度为559.1个?100 g-1干土;AMF的孢子密度、侵染率和种类之间没有显著的相关性;刺槐和沙棘根际AMF侵染率均与pH呈显著正相关,在刺槐根际,pH和有效氮通过直接作用影响侵染率,有机质和有效磷通过间接作用影响侵染率;在沙棘根际,pH和有效磷通过直接作用影响侵染率,有机质和有效氮通过间接作用影响侵染率;在沙棘根际,有机质和pH主要是通过直接作用影响AMF种类,有效氮主要通过有效磷的间接作用影响AMF种类.     

EFFECTS OF LIGHT ON PHOTOSYNTHESIS,GRAZING, AND POPULATION DYNAMICS OF THE HETEROTROPHIC DINOFLAGELLATE PFIESTERIA PISCICIDA (DINOPHYCEAE)1

In studying how environmental factors control the population dynamics of Pfiesteria piscicida Steidinger et Burkholder, we examined the influence of light regime on kleptoplastidic photosynthesis, growth, and grazing. Prey (Rhodomonas sp.)‐saturated growth rate of P. piscicida increased (0.67 ± 0.03 d^?1 to 0.91 ± 0.11 d^?1) with light intensity varying from 0 to 200 μmol photons·m^?2·s^?1. No significant effect was observed on grazing, excluding the possibility that light enhanced P. piscicida growth through stimulating grazing. Light‐grown P. piscicida exhibited a higher gross growth efficiency (0.78 ± 0.10) than P. piscicida incubated in the dark (0.32 ± 0.16), and photosynthetic inhibitors significantly decreased growth of recently fed populations. These results demonstrate a role of kleptoplastidic photosynthesis in enhancing growth in P. piscicida. However, when the prey alga R. sp. was depleted, light's stimulating effect on P. piscicida growth diminished quickly, coinciding with rapid disappearance of Rhodomonas‐derived pigments and RUBISCO from P. piscicida cells. Furthermore, the effect of light on growth was reversed after extended starvation, and starved light‐grown P. piscicida declined at a rate significantly greater than dark‐incubated cultures. The observed difference in rates of decline appeared to be attributable to light‐dependent cannibalism. Using a 5‐chloromethylfluorescein diacetate staining technique, cannibalistic grazing was observed after 7 days of starvation, at a rate four times greater under illumination than in the dark. The results from this study suggest that kleptoplastidy enhances growth of P. piscicida only in the presence of algal prey. When prey is absent, P. piscicida populations may become vulnerable to light‐stimulated cannibalism.     

利用体细胞核移植技术制作人胰岛素原转基因牛 （英文）

通过体细胞核移植技术制作了人胰岛素原转基因牛。在CMV启动子指导下以内部核糖体进入位点序列(IRES)连接的新霉素抗性基因和绿色荧光蛋白基因组成了双重标记基因的筛选系统,用于转基因细胞的富集以及细胞和植入前胚胎的筛选。转基因通过电穿孔的方法(900V/cm,5ms)转入体外培养的牛胎儿成纤维细胞,基因转染细胞在添加G418 (800μg/mL)的培养基中培养10天以富集转基因细胞。选择表达绿色荧光蛋白的转基因细胞作为核供体进行体细胞核移植,重构胚经体外培养至囊胚阶段,选择表达绿色荧光蛋白的囊胚进行胚胎移植。为比较基因转染以及供体细胞所处周期对转基因细胞核移植胚胎发育的影响,用作核移植供体的转基因细胞或非转基因细胞先饥饿培养2—4天(0.5 ?S) ,然后恢复培养(10?S) 10 h使细胞同步化于G1期,以正常培养的细胞作为对照进行核移植。结果表明,转基因细胞作为核供体得到的核移植胚胎的体外囊胚发育率低于以非转基因细胞为核供体的对照组(23.2% VS 35.2 %,P<0.05) ;转基因细胞周期同步化处理与否对其克隆胚囊胚发育率无显著影响(23.2% VS 18.9 %,P>0.05)。胚胎移植后2个月直肠检查发现7头受体牛(每头移植2—4枚胚胎)中有一头妊娠,并最终发育足月产下一头小牛。聚合酶链反应(PCR)检测和DNA测序分析表明其为转人胰岛素原基因的转基因克隆牛。     

The roles of veratryl alcohol and nonionic surfactant in the oxidation of phenolic compounds by lignin peroxidase

Veratryl alcohol (VA) at higher concentration stimulated the lignin peroxidase (LiP)-catalyzed oxidation of phenolic compounds remarkably. This novel phenomenon was due to its competition with the phenols for the active site of the enzyme and to the high reactivity of the formed cation radical of VA (VA+*) which resulted in an additional oxidation of the phenols. The influence of the nonionic surfactant Tween 80 on the VA-enhanced LiP-catalyzed oxidation of phenols depended on its concentration. At lower concentration it had a small synergetic effect but at higher concentration it decreased the initial rate. Studies of the capillary electrophoretic behavior of LiP in the presence of Tween 80 showed that this effect was caused by the surfactant aggregation on LiP which, at higher surfactant concentrations, might impede the access of VA to its binding site on LiP and, consequently, the VA+* formation.