细菌形成“生物被膜”还是“生物膜”?

“Biofilm”是指黏附于非生物或生物表面后,细菌通过分泌的胞外聚合物(extracellular polymeric substances, EPS)将自身包裹其中而形成的细菌聚集体膜状物。“Biofilm”的形成主要包括黏附、定殖、发育、成熟与主动消散5个阶段。“Biofilm”是具有复杂结构与多重功能的“动态细菌组织”,因此在相对稳定微环境中的细菌具有更强的抗逆性,从而导致病原菌更难去除,而益生菌则更易在宿主体内存活与定殖。目前,对于“biofilm”的中文翻译存在“生物被膜”和“生物膜”两种名词混用的现象,少有文献对此深入讨论,并明确指出哪种翻译更为准确。基于此,本文对“biofilm”的形成过程、生理功能和使用现状进行了分析与讨论,并认为“biofilm”翻译为“生物被膜”更为严谨、准确。     

细菌“定植”还是“定殖”？

细菌定植/殖(Bacterial Colonization),是指来自不同环境的细菌接触到机体,并在特定部位黏附、生长和繁殖的现象。无论是共生菌对肠道的保护作用,还是致病菌引起的感染,往往都要依赖细菌定植/殖。目前,教材与期刊等对于"细菌定植"和"细菌定殖"这2个科技名词的使用不一,并存在两者混用的现象,尚未有明确的定论指出哪种使用正确。为了避免以上问题,并对"Bacterial Colonization"的正确翻译提供参考,本文对细菌定植/殖的条件和过程、意义、使用现状进行了分析与讨论,最终认为"细菌定殖"更适用于"Bacterial Colonization"的翻译。     

“严谨反应”还是“严紧反应”?

“Stringent response”是指细菌在遭受营养饥饿与环境胁迫时,由代谢酶RelA/SpoT催化合成信号分子鸟苷四/五磷酸[(p)ppGpp],从而诱导细菌细胞关闭rRNA、tRNA及核糖体蛋白基因转录,停止多种蛋白质的翻译,严控大部分代谢活动的一系列适应性基因表达过程。“Stringent response”几乎是所有细菌应对逆境的重要调节机制。目前,国内文献对“stringent response”的中文翻译存在“严谨反应”和“严紧反应”混用的现象。基于此,本文对“stringent response”的调控机制、生理功能及字面含义进行了分析,认为“stringent response”翻译为“严紧反应”更为合理、准确。     

细菌“特殊结构”之浅见

细菌的结构是病原生物学课程教学中最为基础的教学内容.随着科学技术和研究方法的发展,人们对细菌结构的认识也愈加深入.对国内教材普遍采用的细菌“特殊结构”概念提出不同的观点,试与同行切磋探讨.     

此“壁酸”非彼“壁酸”—关于“细胞生物学”教材中细菌细胞壁特征描述的商榷

目前,相当数量的"细胞生物学"教材中关于革兰氏阳性菌(G+)与革兰氏阴性菌(G–)细胞壁特征差异的描述不够准确,尤其是有关"壁酸"的论述,存在一些不妥与混乱。这给学生的学习带来了较大困惑。该文根据最新文献查证,讨论了细菌细胞壁壁酸的确切含义,并对革兰氏阳性菌与革兰氏阴性菌之间细胞壁差异提出了更准确的表述。     

四君子汤对免疫抑制小鼠肝脏细菌易位的影响

目的观察四君子汤对免疫抑制小鼠免疫功能及肝脏细菌易位的影响,探讨神经-内分泌-代谢-微生态-免疫网络之间的关系。方法应用氢化可的松制备小鼠免疫功能抑制模型,造成肝脏细菌易位;观察四君子汤对小鼠吞噬细胞功能的影响和细菌易位的控制,同时设自然恢复组和正常对照组进行比较。结果氢化可的松灌喂3d后,小鼠吞噬细胞的吞噬功能明显下降,肝脏出现大量细菌易位;经四君子汤治疗6h后,小鼠吞噬细胞吞噬功能显著提高。肝脏细菌易位明显减少。结论四君子汤能有效地控制免疫抑制小鼠肝脏细菌易位。     

溶解固氮蓝藻的细菌

固氮蓝藻作为水稻新肥源已获得良好增产效果。但在藻种培养池中有时发现藻体溶解,溶解区呈现红圈,因而称之为“红圈病”。此种病状在文献中尚未见有记载。我们对此病病原及分离出的溶解固氮蓝藻的细菌——M7820、C798菌株进行了研究。本文描述了病征及溶解细菌的某些生物学特性和影响溶藻能力的因素。     

甲醛固定对Live/Dead BacLight Bacterial Viability Kit死活细菌染液荧光显微计数海洋细菌的影响

利用Live/Dead BacLight Bacterial Viability Kit 死活细菌染液对采自湛江东海大堤海水、沉积物细菌和大型海藻拟刚毛藻（Cladophoropsis zollingeri）内生细菌数量进行了甲醛固定处理前后的荧光显微计数对比分析。结果表明,新鲜样品（不加甲醛固定）、甲醛刚固定样品、甲醛固定1 周样品和甲醛固定2 周样品中海洋细菌数量差异不显著（p>0.05）。甲醛固定对Live/DeadBacLight Bacterial Viability Kit 死活细菌染液荧光显微计数海洋细菌数量无显著影响,固定后的样品可在2 周内完成计数。     

微重力环境下的细菌生物学效应

本文主要根据国内外微重力环境下细菌生物学效应的研究文献,从细菌生长繁殖、抗生素敏感性、毒力变化、新陈代谢等方面进行总结,并概述了微重力环境改变细菌生物学性状的作用机制,展望了微重力环境细菌生物学技术的未来发展。     

多器官功能不全综合征时肠道细菌易位及通里攻下法影响的实验研究

肠道是全身感染的起源,细菌易位在ＭＯＤＳ的发生发展中具有重要作用。本实验以放射性同位素３５Ｓ标记致病Ｅｃｏｌｉ作示踪剂,研究不同剂量酵母多糖腹腔注射所致ＭＯＤＳ时肠道细菌易位的途径和程度,并观察通里攻下中药和抗生素对细菌易位的影响。结果表明,酵母多糖腹腔注射能造成肠道屏障损伤引起细菌易位,细菌易位的程度具有剂量依赖性。细菌易位途径主要有两条,低剂量时细菌易位以肠系膜淋巴结途径为主,高剂量时以门静脉途径为主。予先应用新霉素和灭滴灵给肠道脱污染反而加重细菌易位,抗生素治疗对细菌移位效果不明显。以大承气汤为代表的通里攻下中药对控制细菌易位有显著效果     

A central interdomain protein joint in elongation factor G regulates antibiotic sensitivity, GTP hydrolysis, and ribosome translocation

The antibiotic fusidic acid potently inhibits bacterial translation (and cellular growth) by lodging between domains I and III of elongation factor G (EF-G) and preventing release of EF-G from the ribosome. We examined the functions of key amino acid residues near the active site of EF-G that interact with fusidic acid and regulate hydrolysis of GTP. Alanine mutants of these residues spontaneously hydrolyzed GTP in solution, bypassing the normal activating role of the ribosome. A conserved phenylalanine in the switch II element of EF-G was important for suppressing GTP hydrolysis in solution and critical for catalyzing translocation of the ribosome along mRNA. These experimental results reveal the multipurpose roles of an interdomain joint in the heart of an essential translation factor that can both promote and inhibit bacterial translation.     

Translocation of DNA across bacterial membranes.

DNA translocation across bacterial membranes occurs during the biological processes of infection by bacteriophages, conjugative DNA transfer of plasmids, T-DNA transfer, and genetic transformation. The mechanism of DNA translocation in these systems is not fully understood, but during the last few years extensive data about genes and gene products involved in the translocation processes have accumulated. One reason for the increasing interest in this topic is the discussion about horizontal gene transfer and transkingdom sex. Analyses of genes and gene products involved in DNA transfer suggest that DNA is transferred through a protein channel spanning the bacterial envelope. No common model exists for DNA translocation during phage infection. Perhaps various mechanisms are necessary as a result of the different morphologies of bacteriophages. The DNA translocation processes during conjugation, T-DNA transfer, and transformation are more consistent and may even be compared to the excretion of some proteins. On the basis of analogies and homologies between the proteins involved in DNA translocation and protein secretion, a common basic model for these processes is presented.     

Mechanistic insights into translation inhibition by aminoglycoside antibiotic arbekacin

How aminoglycoside antibiotics limit bacterial growth and viability is not clearly understood. Here we employ fast kinetics to reveal the molecular mechanism of action of a clinically used, new-generation, semisynthetic aminoglycoside Arbekacin (ABK), which is designed to avoid enzyme-mediated deactivation common to other aminoglycosides. Our results portray complete picture of ABK inhibition of bacterial translation with precise quantitative characterizations. We find that ABK inhibits different steps of translation in nanomolar to micromolar concentrations by imparting pleotropic effects. ABK binding stalls elongating ribosomes to a state, which is unfavorable for EF-G binding. This prolongs individual translocation step from ∼50 ms to at least 2 s; the mean time of translocation increases inversely with EF-G concentration. ABK also inhibits translation termination by obstructing RF1/RF2 binding to the ribosome. Furthermore, ABK decreases accuracy of mRNA decoding (UUC vs. CUC) by ∼80 000 fold, causing aberrant protein production. Importantly, translocation and termination events cannot be completely stopped even with high ABK concentration. Extrapolating our kinetic model of ABK action, we postulate that aminoglycosides impose bacteriostatic effect mainly by inhibiting translocation, while they become bactericidal in combination with decoding errors.     

A SecY homologue is involved in chloroplast-encoded D1 protein biogenesis

We have used the photosystem II reaction center D1 protein as a model to study the mechanisms of targeting and insertion of chloroplast-encoded thylakoid membrane proteins. The unusually high turnover rate and distinct pausing intermediates during translation make the D1 protein biogenesis particularly suitable for these purposes. Here we show that cpSecY, a chloroplast homologue of bacterial essential translocon component SecY, interacts tightly with thylakoid membrane-bound ribosomes, suggesting its involvement in protein translocation and insertion. Co-immunoprecipitation and cross-linking experiments indicated that cpSecY resides in the vicinity of D1 elongation intermediates and provided evidence for a transient interaction of cpSecY with D1 elongation intermediates during the biogenesis of D1. After termination of translation, such interactions no longer existed. Our results indicate that, in addition to a well characterized role of cpSecY in posttranslational translocation of nuclear-encoded proteins, it seems to be also involved in cotranslational membrane protein translocation and insertion in chloroplasts.     

Cleavage of the sarcin–ricin loop of 23S rRNA differentially affects EF-G and EF-Tu binding

Ribotoxins are potent inhibitors of protein biosynthesis and inactivate ribosomes from a variety of organisms. The ribotoxin α-sarcin cleaves the large 23S ribosomal RNA (rRNA) at the universally conserved sarcin–ricin loop (SRL) leading to complete inactivation of the ribosome and cellular death. The SRL interacts with translation factors that hydrolyze GTP, and it is important for their binding to the ribosome, but its precise role is not yet understood. We studied the effect of α-sarcin on defined steps of translation by the bacterial ribosome. α-Sarcin-treated ribosomes showed no defects in mRNA and tRNA binding, peptide-bond formation and sparsomycin-dependent translocation. Cleavage of SRL slightly affected binding of elongation factor Tu ternary complex (EF-Tu•GTP•tRNA) to the ribosome. In contrast, the activity of elongation factor G (EF-G) was strongly impaired in α-sarcin-treated ribosomes. Importantly, cleavage of SRL inhibited EF-G binding, and consequently GTP hydrolysis and mRNA–tRNA translocation. These results suggest that the SRL is more critical in EF-G than ternary complex binding to the ribosome implicating different requirements in this region of the ribosome during protein elongation.     

Co-translational targeting and translocation of the amino terminus of opsin across the endoplasmic membrane requires GTP but not ATP

The tight coupling between ongoing translation and translocation across the mammalian endoplasmic reticulum has made it difficult to determine the requirements that are specific for translocation. We have developed an in vitro assay that faithfully mimics the co-translational targeting and translocation of the amino terminus of opsin without ongoing translation. Using this system we demonstrate that this post-translational targeting and translocation requires nucleotide triphosphates but not cytosolic proteins. The addition of GTP alone was sufficient to fully restore targeting. The addition of ATP was not specifically required, and non-hydrolyzable analogs of ATP that blocked 90% of the ATPase activity also had no inhibitory effect on translocation.     

线粒体核糖体——细胞器中的翻译机器

线粒体核糖体作为细胞器中的翻译机器,与细菌核糖体以及真核细胞质核糖体在rRNA和蛋白质组分、拓扑结构、来源等方面差异显著。本文综述线粒体核糖体研究进展,对比分析其理化性质和实验结构的相似性与特殊性。基于线粒体核糖体的结构和生物学功能进一步推测:经过与tRNA的相互识别和空间取向,mRNA链构象能否影响其编码产物——新生肽链的构象,期望揭示mRNA在翻译过程中可能的作用机理。     

Recognition of the Agrobacterium tumefaciens VirE2 translocation signal by the VirB/D4 transport system does not require VirE1

Agrobacterium tumefaciens uses a type IV secretion system to deliver a nucleoprotein complex and effector proteins directly into plant cells. The single-stranded DNA-binding protein VirE2, the F-box protein VirF and VirE3 are delivered into host cells via this VirB/D4 encoded translocation system. VirE1 functions as a chaperone of VirE2 by regulating its efficient translation and preventing VirE2-VirE2 aggregation in the bacterial cell. We analyzed whether the VirE1 chaperone is also essential for transport recognition of VirE2 by the VirB/D4 encoded type IV secretion system. In addition, we assayed whether translocation of VirF and VirE3, which also forms part of the virE operon, is affected by the absence of VirE1. We employed the earlier developed CRAFT (Cre recombinase Reporter Assay For Translocation) assay to detect transfer of Cre::Vir fusion proteins from A. tumefaciens into plants, monitored by stable reconstitution of a kanamycin resistance marker, and into yeast, screened by loss of the URA3 gene. We show that the C-terminal 50 amino acids of VirE2 and VirE3 are sufficient to mediate Cre translocation into host cells, confirming earlier indications of a C-terminal transport signal. This transfer was independent of the presence or absence of VirE1. Besides, the translocation efficiency of VirF is not altered in a virE1 mutant. The results unambiguously show that the VirE1 chaperone is not essential for the recognition of the VirE2 transport signal by the transport system and the subsequent translocation across the bacterial envelope into host cells.     

基于翻译调控的细菌耐药研究进展

由于抗生素的大量使用,细菌耐药问题凸显,直接威胁人类生命健康和世界经济发展。过去对于细菌耐药的遗传和分子机制研究较为透彻,而对应的调控机制研究相对较少。翻译调控作为生命体最重要的调控方式之一,在细菌耐药研究领域的重要性尚未被学术界充分重视。本文介绍了影响翻译过程的抗生素的主要作用机制,重点从核糖体的修饰和突变、tRNA总量的动态调控、tRNA氨酰化、tRNA甲基化、核糖体保护蛋白和翻译因子这几个方面概述了基于翻译调控的细菌耐药研究进展,为研究者们提供了一个基于翻译调控角度研究细菌耐药的新视角,同时也为开发靶向细菌翻译调控的新型抗生素提供一些新思路。