The prevalence of plasmid‐mediated quinolone resistance determinants among clinical isolates of ESBL or AmpC‐producing Escherichia coli from Chinese pediatric patients

Three kinds of PMQR determinants (qnr genes, aac(6’)‐Ib‐cr, and qepA) have been discovered and shown to be widely distributed among clinical isolates. To characterize the prevalence of PMQR determinants in ESBL or AmpC‐producing E. coli clinical isolates in Chinese children, a total of 292 ESBL or AmpC‐producing E. coli clinical isolates collected from five children's hospitals in China from 2005 to 2006 were screened for PMQR determinants by PCR. Twenty (6.8%) of the 292 isolates were positive for PMQR determinants. A total of 12 (4.1%) isolates were positive for qnr genes, comprising three positive for qnrA (1.0%), three for qnrB (1.0%), and six for qnrS (2.1%). Twenty‐four (8.2%) isolates were positive for aac(6’)‐Ib, of which 10 (3.4% of 292) had the –cr variant. There was no qepA gene detected in the isolates. Conjugation revealed that qnr, aac(6’)‐Ib‐cr, and ESBL‐encoding genes were transferred together.     

Simple and rapid detection of Salmonella serovar Enteritidis under field conditions by loop‐mediated isothermal amplification

Aim: The objective of this study is to develop a serovar‐specific loop‐mediated isothermal amplification (LAMP) method for sensitive, rapid, and inexpensive detection of Salmonella serovar Enteritidis under field conditions. Methods: A set of six specific primers was designed with Salmonella Enteritidis DNA as the target. LAMP conditions were optimized by incubating the target DNA with the Bst DNA polymerase large fragment in a simple water bath. The sensitivity and specificity of LAMP was then compared with those of fluorescent quantitative real‐time polymerase chain reaction (FQ‐PCR). Results: The results were as follows. (1) Serovar‐specific Salmonella Enteritidis DNA was amplified at 65°C in as early as 20 min in a water bath. (2) A colour change visible to the naked eye indicated a positive amplification reaction. (3) The detection limit of the LAMP assay was 4 copies μl^?1; thus, the sensitivity and specificity of this assay is similar to those of the FQ‐PCR. Conclusions: LAMP is a high‐throughput detection technique with high sensitivity, specificity, and simplicity; these factors make it suitable for specifically detecting Salmonella Enteritidis under field conditions and in laboratory settings. Thus, LAMP eliminates the need for complicated equipment and technical training in the detection of this specific serovar. Significance and impact of the study: This is the first study involving the use of LAMP to detect Salmonella serovar‐specific DNA sequences. It is also the first to report an ideal method of distinguishing between Salmonella Enteritidis and other Salmonella under field conditions.     

RalA-exocyst interaction mediates GTP-dependent exocytosis

Many secretory cells utilize a GTP-dependent pathway, in addition to the well characterized Ca2+-dependent pathway, to trigger exocytotic secretion. However, little is currently known about the mechanism by which this may occur. Here we show the key signaling pathway that mediates GTP-dependent exocytosis. Incubation of permeabilized PC12 cells with soluble RalA GTPase, but not RhoA or Rab3A GTPases, strongly inhibited GTP-dependent exocytosis. A Ral-binding fragment from Sec5, a component of the exocyst complex, showed a similar inhibition. Point mutations in both RalA (RalA(E38R)) and the Sec5 (Sec5(T11A)) fragment, which abolish RalA-Sec5 interaction also abolished the inhibition of GTP-dependent exocytosis. Moreover, transfection with wild-type RalA, but not RalA(E38R), enhanced GTP-dependent exocytosis. In contrast the RalA and the Sec5 fragment showed no inhibition of Ca2+-dependent exocytosis, but cleavage of a SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) protein by Botulinum neurotoxin blocked both GTP- and Ca2+-dependent exocytosis. Our results indicate that the interaction between RalA and the exocyst complex (containing Sec5) is essential for GTP-dependent exocytosis. Furthermore, GTP- and Ca2+-dependent exocytosis use different sensors and effectors for triggering exocytosis whereas their final fusion steps are both SNARE-dependent.     

Application of impedance spectroscopy for monitoring colloid Au-enhanced antibody immobilization and antibody-antigen reactions

We used colloidal Au to enhance the amount of antibody immobilized on a gold electrode and ultimately monitored the interaction of antigen-antibody by impedance measurement. Self-assembly of 6 nm (diameter) colloidal Au onto the self-assembled monolayers (SAMs) of 4-aminothiophenol modified gold electrode resulted in an easier attachment of antibody. The redox reactions of [Fe(CN)6](4-)/[Fe(CN)6](3-) on the gold surface were blocked due to the procedures of self-assembly of 4-aminothiophenol and antibody immobilization, which were investigated by cyclic voltammetry and impedance spectroscopy. The interaction of antigen with grafted antibody recognition layers was carried out by soaking the modified electrode into a phosphate buffer at pH 7.4 with various concentrations of antigen at 37 degrees C for 30 min. The antibody recognition layers and their interactions with various concentrations of antigen could be detected by measurements of the impedance change. The results show that this method has good correlation for detection of Hepatitis B virus surface antigen in the range of 0.5-200 microg/l and a detection limit of about 50 ng/l.     

NaCl胁迫对毛竹叶片的电阻抗图谱参数及膜透性的影响

以NaCl胁迫下毛竹（Phyllostachys edulis）实生苗叶片为材料,测定其电阻抗图谱参数和膜透性的变化,通过膜透性与电阻抗图谱参数间的相关性,来证明电阻抗图谱法研究竹子受胁迫程度的有效性。结果表明：随着盐浓度的升高,胞外电阻、胞内电阻和弛豫时间呈现先减小、后增加、再减小的特征,而弛豫时间分布系数表现恰好相反;叶片细胞膜相对透性逐渐增大,脯氨酸的含量先逐渐增大,然后减小。相关分析表明：胞内电阻、胞外电阻、弛豫时间与脯氨酸含量呈极显著负相关（p〈0.01）;胞外电阻、弛豫时间与膜相对透性呈显著负相关（p〈0.05）。电阻抗图谱参数能够有效地表示毛竹受NaCl胁迫的程度,电阻抗图谱法将是竹子逆境胁迫研究的一种有效方法。     

北部湾带鱼的摄食习性

2008年8月至2009年9月,对北部湾带鱼逐月渔获物胃含物进行鉴定和统计分析,研究其摄食习性.结果表明：北部湾带鱼捕食种类包括鱼类、头足类、底栖甲壳类以及浮游动物等,属于广食性动物.优势种类有少鳞犀鳕(37.99%)、蓝圆鲹(16.42%)和中国毛虾(10.03%).蓝圆鲹和尖吻小公鱼全年在全湾带鱼的食物中均有出现,是重要的饵料指标种类.摄食强度与饱满指数的季节差异十分显著(P<0.001),食物多样性无显著季节性差异(P>0.05),秋季最高,全年平均为1.97.聚类分析表明,北部湾带鱼在达到50%性成熟肛长值(190 mm)时,饵料由幼鱼的以小型浮游动物、中上层鱼类及甲壳类为主向成鱼的以较大型鱼类和头足类为主转换.     

我国主要类型昆虫对CO_2升高响应的研究进展

大气CO2浓度增加已经受到国内外的极大关注。自2002年以来,在自行设计、组装的一系列密闭式动态CO2气室和开顶式CO2浓度控制箱基础上,研究了我国主要类型昆虫对CO2浓度升高响应的特征。结果显示,大气CO2浓度升高降低了棉铃虫Helicoverpa armigera的适合度和对棉花的危害作用,增加了棉花对棉铃虫为害的补偿作用,使以咀嚼式口器昆虫为代表的棉铃虫种群发生与危害下降;但大气CO2浓度升高改变了植物组织营养物质的组成与含量,提高了蚜虫Aphis gosypii对氨基酸营养的利用与补偿效率,降低了3种麦蚜的种间竞争,导致蚜虫种群发生与危害严重;而对烟粉虱Bemisia tabaci的种群特征影响较少。大气CO2浓度升高对天敌昆虫的影响存在种的特异性,表现出种群上升、下降和变化不大等特征。未来大气CO2浓度升高下,由于作物生长发育加快,生物量增加,蚜虫种群增多,导致吡虫啉农药防治效果下降,由此未来大气CO2浓度升高下农民将被迫使用更多的化学农药防治蚜虫类害虫,进而加重环境污染。     

内蒙古科尔沁沙地两种沙生植物冷蒿和差不嘎蒿竞争策略的生理生态学依据

研究了中国内蒙古科尔沁沙地两种优势灌木冷蒿(Artemisiafrigida Willd)和差不嘎蒿(Artemisia halodendronTurcz.ex Bess)在不同土壤水分状况条件下的气体交换、水分关系和叶片的化学特性.测定设置了5个土壤水分梯度:土壤最大含水量(体积含水量,30%)、田间持水量(对照,20%)、轻度水分胁迫(10%)、极端干旱(＜4%)和旱后复水(20%).冷蒿的净光合速率(Pn)、蒸腾速率(TR)和水势(ψw低于差不嘎蒿,相对水分亏缺(RWD)、束缚水含量(BWC)、束缚水与自由水的比值(BWC/FWC)和综合抗旱性指数(DI)高于差不嘎蒿.两种灌木对不同土壤含水量的响应不同,随着土壤含水量的逐渐下降,差不嘎蒿的ψw、BWC和BWC/FWC出现大幅度的波动,波动幅度远远的大于冷蒿,冷蒿则显示了一个较差不嘎蒿高的持水能力和抗旱能力.冷蒿和差不嘎蒿的脯氨酸和总可溶性糖含量随土壤含水量的降低均呈增加的趋势,冷蒿增加的幅度大于差不嘎蒿,说明冷蒿的渗透调节能力在干旱过程中有较大提高.在长期极端干旱条件下,两种灌木的ψw,RWD,BWC和BWC/FWC的终极值相近;有机物、脯氨酸和总可溶性糖含量大量累积;冷蒿的脯氨酸和总可溶性糖含量增加的幅度和可溶性蛋白质降解的幅度远远超过差不嘎蒿;我们认为此时累积的物质主要作为营养物质,以供植物的旱后恢复,此时冷蒿的恢复能力超过差不嘎蒿,这是极端干旱条件下差不嘎蒿死亡而冷蒿存活的主要原因之一.我们的研究结果显示,相对于差不嘎蒿而言,冷蒿在水分胁迫条件下的生理生态学特性更有利于其在固定沙地的生长,差不嘎蒿则由于对水资源微弱的竞争丧失了生存优势,是引起科尔沁沙地植被演替过程中冷蒿替代差不嘎蒿的主要原因.     

Cyclin/CDK regulates the nucleocytoplasmic localization of the human papillomavirus E1 DNA helicase

Cyclin-dependent kinases (CDKs) play key roles in eukaryotic DNA replication and cell cycle progression. Phosphorylation of components of the preinitiation complex activates replication and prevents reinitiation. One mechanism is mediated by nuclear export of critical proteins. Human papillomavirus (HPV) DNA replication requires cellular machinery in addition to the viral replicative DNA helicase E1 and origin recognition protein E2. E1 phosphorylation by cyclin/CDK is critical for efficient viral DNA replication. We now show that E1 is phosphorylated by CDKs in vivo and that phosphorylation regulates its nucleocytoplasmic localization. We identified a conserved regulatory region for localization which contains a dominant leucine-rich nuclear export sequence (NES), the previously defined cyclin binding motif, three serine residues that are CDK substrates, and a putative bipartite nuclear localization sequence. We show that E1 is exported from the nucleus by a CRM1-dependent mechanism unless the NES is inactivated by CDK phosphorylation. Replication activities of E1 phosphorylation site mutations are reduced and correlate inversely with their increased cytoplasmic localization. Nuclear localization and replication activities of most of these mutations are enhanced or restored by mutations in the NES. Collectively, our data demonstrate that CDK phosphorylation controls E1 nuclear localization to support viral DNA amplification. Thus, HPV adopts and adapts the cellular regulatory mechanism to complete its reproductive program.     

Analysis of B-Class Genes NAP3L3 and NAP3L4 in Narcissus tazetta var. chinensis

Very few flower organ identity genes have been characterized in Chinese narcissus (Narcissus tazetta var. chinensis), which has petaloid sepals. Here, we report the cloning of two full-length B-class genes, namely NAP3L3 and NAP3L4, that are orthologs of the DEFICIENS lineage. Both genes are highly expressed in the second whorl of the perianth and in the stamens. NAP3L4 is also expressed strongly in the ovule. The functions of these two genes were further analyzed using transgenic plants. Ectopic expression of either gene in Arabidopsis gave no obvious floral organ transformation phenotypes. In yeast two-hybrid assays, NAP3L3 and NAP3L4 failed to homodimerize and interacted weakly with each other. The data suggest that these two genes might not be involved in the formation of petaloid sepals. Isolation and functional analysis of other B-class paralogs should be conducted to fully understand petaloid tepal development in Chinese narcissus.