Quorum Sensing

仲裁感应
  • 文章类型: Journal Article
    在人们越来越担心抗生素耐药性的情况下,创新策略在解决水产养殖中的细菌感染方面势在必行。仲裁淬火(QQ),群体感应(QS)的酶抑制,已经成为一个有希望的解决方案。本研究深入研究了益生菌菌株VelezensisD-18及其产品的QQ功能,特别是在神经弧菌507的通讯和生物膜形成中。在这项研究中,紫罗兰色杆菌MK被用作生物标志物,结果证实维氏芽孢杆菌D-18有效抑制QS。对QQ机制的进一步探索表明,维氏芽孢杆菌D-18存在内酰胺酶活性,可降解长链和短链酰基高丝氨酸内酯(AHLs)。PCR分析表明存在同源的内酰胺酶产生基因,ytnP,在维氏芽孢杆菌D-18的基因组中。该研究评估了维氏芽孢杆菌D-18对V.anguillarum507生长和生物膜形成的影响。该益生菌不仅能控制其生物膜的形成,还能显著抑制病原菌的生长。因此,VelezensisD-18通过其QQ能力证明了在水产养殖中预防V.anguillarum疾病的巨大潜力。破坏细菌通讯和控制生物膜形成的能力将B.velezensisD-18定位为在水产养殖中管理细菌性疾病的常规抗生素的有希望的生态友好的替代品。
    Amid growing concerns about antibiotic resistance, innovative strategies are imperative in addressing bacterial infections in aquaculture. Quorum quenching (QQ), the enzymatic inhibition of quorum sensing (QS), has emerged as a promising solution. This study delves into the QQ capabilities of the probiotic strain Bacillus velezensis D-18 and its products, particularly in Vibrio anguillarum 507 communication and biofilm formation. Chromobacterium violaceum MK was used as a biomarker in this study, and the results confirmed that B. velezensis D-18 effectively inhibits QS. Further exploration into the QQ mechanism revealed the presence of lactonase activity by B. velezensis D-18 that degraded both long- and short-chain acyl homoserine lactones (AHLs). PCR analysis demonstrated the presence of a homologous lactonase-producing gene, ytnP, in the genome of B. velezensis D-18. The study evaluated the impact of B. velezensis D-18 on V. anguillarum 507 growth and biofilm formation. The probiotic not only controls the biofilm formation of V. anguillarum but also significantly restrains pathogen growth. Therefore, B. velezensis D-18 demonstrates substantial potential for preventing V. anguillarum diseases in aquaculture through its QQ capacity. The ability to disrupt bacterial communication and control biofilm formation positions B. velezensis D-18 as a promising eco-friendly alternative to conventional antibiotics in managing bacterial diseases in aquaculture.
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  • 文章类型: Journal Article
    评估痰热清(TRQ)对肺炎克雷伯菌的体外和体内抗菌作用及其对细菌生物膜形成的抑制活性,探讨TRQ抑制肺炎克雷伯菌生物膜形成的作用机制。
    建立肺炎克雷伯菌体外生物膜模型,使用结晶紫染色和扫描电子显微镜(SEM)评估TRQ对生物膜形成的影响。此外,采用活板计数法评价TRQ对肺炎克雷伯菌生物膜的清除作用;采用q-RTPCR评价不同浓度TRQ对肺炎克雷伯菌生物膜相关基因表达的抑制作用;采用哈氏弧菌生物发光法检测群体感应信号分子AI-2的活性;建立肺炎克雷伯菌豚鼠肺部感染模型,在用药物治疗后,进行肺组织病理分析和肺组织细菌负荷测定。治疗组包括TRQ组,亚胺培南(IPM)组,TRQ+IPM组,无菌生理盐水组为对照组。
    通过TRQ体外实验显着抑制了肺炎克雷伯菌生物膜的形成。此外,当与IPM结合时,与TRQ组和单纯IPM组相比,生物膜中肺炎克雷伯菌的清除率显著增加.q-RTPCR分析显示,TRQ下调了肺炎克雷伯菌生物膜形成相关基因的表达,特别是luxS,wbbm,wzm,还有lsrK,并抑制细菌中AI-2分子的活性。体内实验证明TRQ能有效治疗豚鼠肺部感染,导致肺部炎症减少。此外,当与IPM结合时,肺组织中的细菌负荷显著减少.
    TRQ作为一种潜在的治疗药物,在肺炎克雷伯菌感染的治疗中起着巨大的作用,特别是与常规抗生素联合使用。TRQ可以通过抑制肺炎克雷伯菌生物膜的形成增强对细菌的清除作用,这为TRQ抗肺炎克雷伯菌感染的临床治疗提供了实验证据。
    UNASSIGNED: To evaluate the antibacterial effect of Tanreqing (TRQ) against K. pneumoniae and its inhibition activity on bacterial biofilm formation in vitro and in vivo, and to explore the mechanism of the inhibitory effects of TRQ on K. pneumoniae biofilm formation.
    UNASSIGNED: An in vitro biofilm model of K. pneumoniae was established, and the impact of TRQ on biofilm formation was evaluated using crystal violet staining and scanning electron microscopy (SEM). Furthermore, the clearance effect of TRQ against K. pneumoniae in the biofilm was assessed using the viable plate counting method; q-RT PCR was used to evaluate the inhibitory effect of different concentrations of TRQ on the expression of biofilm-related genes in Klebsiella pneumoniae; The activity of quorum sensing signal molecule AI-2 was detected by Vibrio harveyi bioluminescence assay; Meanwhile, a guinea pig lung infection model of Klebsiella pneumoniae was constructed, and after treated with drugs, pathological analysis of lung tissue and determination of bacterial load in lung tissue were performed. The treatment groups included TRQ group, imipenem(IPM) group, TRQ+IPM group, and sterile saline group as the control.
    UNASSIGNED: The formation of K. pneumoniae biofilm was significantly inhibited by TRQ in vitro experiments. Furthermore, when combined with IPM, the clearance of K. pneumoniae in the biofilm was notably increased compared to the TRQ group and IPM group alone. q-RT PCR analysis revealed that TRQ down-regulated the expression of genes related to biofilm formation in K. pneumoniae, specifically luxS, wbbm, wzm, and lsrK, and also inhibited the activity of AI-2 molecules in the bacterium. In vivo experiments demonstrated that TRQ effectively treated guinea pig lung infections, resulting in reduced lung inflammation. Additionally, when combined with IPM, there was a significant reduction in the bacterial load in lung tissue.
    UNASSIGNED: TRQ as a potential therapeutic agent plays a great role in the treatment of K. pneumoniae infections, particularly in combination with conventional antibiotics. And TRQ can enhanced the clearance effect on the bacterium by inhibiting the K. pneumoniae biofilm formation, which provided experimental evidence in support of clinical treatment of TRQ against K. pneumoniae infections.
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  • 文章类型: Journal Article
    这项研究调查了生物膜测序分批反应器(BSBR)过程中生物膜培养过程中微生物生长和代谢的机制。结果表明,污泥排放是生物膜生长的关键,因为它终止了新生生物膜和活性污泥之间对碳(C)源的竞争。对于被测试的反应堆,18d污泥排放后,P代谢和C源利用率显著提高,生物膜迅速生长。回收液的P浓度达到157.08mg/L,这足以通过矿化进一步回收磷。采用Meta-组学方法分析生物膜培养过程中微生物生长的代谢途径和功能基因。结果表明,污泥排放激活了P代谢的关键基因,抑制了C代谢的关键基因,从而加强了聚磷酸盐积累代谢(PAM)。污泥的排放不仅改变了聚磷酸盐积累生物(PAOs)的类型,而且还促进了优势PAOs的生长。污泥排放前,在不同微生物之间传播的必要代谢能力逐渐集中到少量的PAOs中,污泥排放后,它们进一步浓缩为念珠菌_Contendobacter(P3)和念珠菌_累积杆菌(P17)。信使分子C-di-GMP,主要由P3和P17产生,通过调节细胞P和C代谢促进P的富集。糖原积累生物(GAO)念珠菌_竞争性细菌分泌N-酰基高丝氨酸内酯(AHLs),刺激细胞外聚合物(EPS)中蛋白质的分泌,从而促进微生物对生物膜的粘附,并通过基于EPS的P吸附改善P代谢。在占主导地位的GAO和PAO的共同作用下,AHLs和C-di-GMP介导QS增进生物膜发展和P富集。该研究为生物膜的培养及其更广泛的应用提供了理论支持。
    This study investigated the mechanisms of microbial growth and metabolism during biofilm cultivation in the biofilm sequencing batch reactor (BSBR) process for phosphate (P) enrichment. The results showed that the sludge discharge was key to biofilm growth, as it terminated the competition for carbon (C) source between the nascent biofilm and the activated sludge. For the tested reactor, after the sludge discharge on 18 d, P metabolism and C source utilization improved significantly, and the biofilm grew rapidly. The P concentration of the recovery liquid reached up to 157.08 mg/L, which was sufficient for further P recovery via mineralization. Meta-omics methods were used to analyze metabolic pathways and functional genes in microbial growth during biofilm cultivation. It appeared that the sludge discharge activated the key genes of P metabolism and inhibited the key genes of C metabolism, which strengthened the polyphosphate-accumulating metabolism (PAM) as a result. The sludge discharge not only changed the types of polyphosphate-accumulating organisms (PAOs) but also promoted the growth of dominant PAOs. Before the sludge discharge, the necessary metabolic abilities that were spread among different microorganisms gradually concentrated into a small number of PAOs, and after the sludge discharge, they further concentrated into Candidatus_Contendobacter (P3) and Candidatus_Accumulibacter (P17). The messenger molecule C-di-GMP, produced mostly by P3 and P17, facilitated P enrichment by regulating cellular P and C metabolism. The glycogen-accumulating organism (GAO) Candidatus_Competibacter secreted N-Acyl homoserine lactones (AHLs), which stimulated the secretion of protein in extracellular polymeric substances (EPS), thus promoting the adhesion of microorganisms to biofilm and improving P metabolism via EPS-based P adsorption. Under the combined action of the dominant GAOs and PAOs, AHLs and C-di-GMP mediated QS to promote biofilm development and P enrichment. The research provides theoretical support for the cultivation of biofilm and its wider application.
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  • 文章类型: Journal Article
    微生物通过称为群体感应(QS)的集体决策机制来调节能量昂贵的表型的表达。这项研究调查了生物膜生长和QS控制的生物膜在异质多孔介质中分散的复杂动力学,采用孔隙尺度反应输运建模方法。在各种流体流动条件和生物膜生长情况下进行的模型模拟表明,QS过程不仅受到生物膜菌落的生物量密度的影响,而且受到孔结构之间复杂相互作用的影响。流速,以及生物膜生长和扩散的速率。这项研究表明,孔结构控制QS过程的启动,平流引起生物膜的振荡生长。如果生物膜动力学有利于维持足够高的信号浓度,则可以抑制这种振荡。如快速生长或缓慢扩散。通过建立一个数学框架,这项研究有助于对复杂环境中QS控制的生物膜动力学的基本理解。
    Microorganisms regulate the expression of energetically expensive phenotypes via a collective decision-making mechanism known as quorum sensing (QS). This study investigates the intricate dynamics of biofilm growth and QS-controlled biofilm dispersal in heterogeneous porous media, employing a pore-scale reactive transport modeling approach. Model simulations carried out under various fluid flow conditions and biofilm growth scenarios reveal that QS processes are influenced not only by the biomass density of biofilm colonies but also by a complex interplay between pore architecture, flow velocity, and the rates of biofilm growth and dispersal. This study demonstrates that pore architecture controls the initiation of QS processes and advection gives rise to oscillatory growth of biofilms. Such oscillation is suppressed if biofilm dynamics are in favor of sustaining a sufficiently high signal concentration, such as fast growth or slow dispersal rates. By establishing a mathematical framework, this study contributes to the fundamental understanding of QS-controlled biofilm dynamics in complex environments.
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  • 文章类型: Journal Article
    八角,或者八角,有许多用途,从烹饪到宗教。它自古以来就被用于食品工业。这项研究的主要目的是确定化学成分,抗菌,抗生物膜,以及通过加氢蒸馏八角地上部分获得的精油(EO)的抗群体感应活性。鉴定出24种组分,代表所分析的精油的92.55%。(E)-茴香脑(83.68%),柠檬烯(3.19%),α-pine烯(0.71%)是I.verumEO的主要成分。结果表明,所获得的EO对8种细菌均有不同程度的效果。关于抗生物膜活性,使用亚抑制浓度进行测试时,反式茴香脑对生物膜形成比精油更有效。针对铜绿假单胞菌PAO1测试的抗蜂群活性的结果表明,与反式-茴香脑相比,VerumEO对PAO1的蜂群行为具有更有效的抑制作用,在100µg/mL的浓度下,百分比达到38%。已鉴定的植物化合物的ADME分析证实了它们的重要药代动力学和药物相似特性。使用分子对接方法的计算机模拟研究显示,已鉴定的化合物与抗菌和抗群体感应(QS)活性中涉及的已知目标酶之间的结合得分很高。总的来说,获得的结果表明I.verumEO是一种潜在的良好抗菌剂,可以防止食品被食源性致病菌污染。
    Illicium verum, or star anise, has many uses ranging from culinary to religious. It has been used in the food industry since ancient times. The main purpose of this study was to determine the chemical composition, antibacterial, antibiofilm, and anti-quorum sensing activities of the essential oil (EO) obtained via hydro-distillation of the aerial parts of Illicium verum. Twenty-four components were identified representing 92.55% of the analyzed essential oil. (E)-anethole (83.68%), limonene (3.19%), and α-pinene (0.71%) were the main constituents of I. verum EO. The results show that the obtained EO was effective against eight bacterial strains to different degrees. Concerning the antibiofilm activity, trans-anethole was more effective against biofilm formation than the essential oil when tested using sub-inhibitory concentrations. The results of anti-swarming activity tested against P. aeruginosa PAO1 revealed that I. verum EO possesses more potent inhibitory effects on the swarming behavior of PAO1 when compared to trans-anethole, with the percentage reaching 38% at a concentration of 100 µg/mL. The ADME profiling of the identified phytocompounds confirmed their important pharmacokinetic and drug-likeness properties. The in silico study using a molecular docking approach revealed a high binding score between the identified compounds with known target enzymes involved in antibacterial and anti-quorum sensing (QS) activities. Overall, the obtained results suggest I. verum EO to be a potentially good antimicrobial agent to prevent food contamination with foodborne pathogenic bacteria.
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  • 文章类型: Journal Article
    喹啉是煤化工废水中广泛存在的难降解有机物,严重影响生态环境和人类健康。许多生化方法已经可以用于喹啉的降解,但利用微生物群落传感技术对喹啉降解的研究尚未深入。因此,本文重点研究了功能菌株在喹啉降解过程中,群体感应信号分子在生物膜形成过程中的增强机制。在本文中,信号分子C4-HSL和C6-HSL对粘附能力的影响,菌落直径,喹啉降解菌功能菌株的生物膜形成能力和生物膜形态(Ochrobactrumsp.,LC-1)进行了调查,结果表明,两种信号分子均通过表现出有效的生物强化作用,促进了喹啉降解过程中生物膜的形成。两种信号分子均能增强菌株LC-1的菌落直径,其中C4-HSL能增强菌株LC-1的生物量并刺激胞外多糖的分泌;C6-HSL能诱导菌株LC-1的粘附性能增强和胞外蛋白分泌;两种分子共同增强菌株LC-1的生物膜形成过程。本研讨对煤化工废水中喹啉的降解具有实际运用价值。
    Quinoline is a hard-to-degrade organic compound widely found in coal chemical wastewater, that seriously affects the ecological environment and human health. A number of biochemical methods are already available for quinoline degradation, but the use of microbial community sensing for quinoline degradation has not been studied in depth. Therefore, this paper focuses on the enhanced mechanism of quorum-sensing signaling molecules in the biofilm formation process during quinoline degradation by functional strains of bacteria. In this paper, the effects of the signal molecules C4-HSL and C6-HSL on the adhesion ability, colony diameter, biofilm formation ability and biofilm morphology of functional strains of quinoline degrading bacteria (Ochrobactrum sp., LC-1) were investigated, and the results showed that both signal molecules promoted the biofilm formation process during the degradation of quinoline by exhibiting an efficient biofortification effect. Both signal molecules could enhance the colony diameter of strain LC-1, where C4-HSL could enhance the biomass of strain LC-1 and stimulate the secretion of extracellular polysaccharides; and C6-HSL could induce the enhancement of adhesion performance and the secretion of extracellular proteins from strain LC-1; both molecules together enhanced the biofilm formation process of strain LC-1. This study has practical application in the degradation of quinoline in coal chemical wastewater.
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  • 文章类型: Journal Article
    群体猝灭(QQ)是细菌通讯的抑制,即,群体感应(QS)。QS是调节复杂细菌群落中生物膜形成和表型的关键机制,例如在致龋生物膜中发现的那些。尽管QQ方法被证明可以有效地减少生物量,关于它们对口腔微生物生物膜分类组成的影响的知识仍然很少。这里,我们研究了QQ内酯酶Aii20J对生物膜生物量产生和分类学组成的影响。我们从10个无龋齿和10个龋齿活跃的儿童中收集了牙龈上牙菌斑样品,并对其进行培养以产生体外生物膜。我们描述了Aii20J暴露后生物量的显著减少,通过结晶紫测定法评估。使用16SrRNA基因扩增子测序的分类学分析显示,在属水平上细菌组成没有显着变化。有趣的是,在物种水平上,Aii20J处理增加了溶血链球菌和唾液链球菌的丰度。S.cristatus和S.salivarius都表达pH缓冲酶(精氨酸脱亚胺酶和脲酶,分别)催化氨生产,从而潜在地提高局部pH并抵消生物膜的致龋潜力。在研究的局限性内,我们的发现为QQ的生物膜调节能力提供了证据,并为恢复生态失调生态系统内稳态的替代策略提供了新的见解。
    Quorum quenching (QQ) is the inhibition of bacterial communication, i.e., quorum sensing (QS). QS is a key mechanism in regulating biofilm formation and phenotype in complex bacterial communities, such as those found within cariogenic biofilms. Whereas QQ approaches were shown to effectively reduce biomass, knowledge of their impact on the taxonomic composition of oral polymicrobial biofilms remains scarce. Here, we investigate the effect of the QQ lactonase Aii20J on biomass production and taxonomical composition of biofilms. We collected supragingival plaque samples from 10 caries-free and 10 caries-active children and cultured them to generate in vitro biofilms. We describe significant biomass reductions upon Aii20J exposure, as assessed by crystal violet assays. Taxonomical profiling using 16S rRNA gene amplicon sequencing revealed no significant changes in bacterial composition at the genus level. Interestingly, at the species level Aii20J-treatment increased the abundance of Streptococcus cristatus and Streptococcus salivarius. Both S. cristatus and S. salivarius express pH-buffering enzymes (arginine deiminase and urease, respectively) that catalyze ammonia production, thereby potentially raising local pH and counteracting the biofilm\'s cariogenic potential. Within the limitations of the study, our findings provide evidence of the biofilm-modulating ability of QQ and offer novel insights into alternative strategies to restore homeostasis within dysbiotic ecosystems.
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  • 文章类型: Journal Article
    细菌对化学治疗的抗性的发展对治疗细菌感染的功效构成严重危险。抗微生物药物耐药性的关键因素之一是生物膜中细菌的生长。群体感应(QS)抑制是通过开发新的抗生物膜药物作为替代治疗产生的。细胞间通讯受到QS抑制的阻碍,其目标是QS信号通路。这项工作的目标是开发通过降低QS并充当抗生物膜剂而有效对抗铜绿假单胞菌的更新药物。在这次调查中,设计并合成了N-(苯并[d]噻唑-2-基)苯甲酰胺/N-(噻唑-2-基)苯甲酰胺衍生物3a-h,收率良好。Further,分子对接分析显示,结合亲和力值为-11.2至-7.6kcal/mol,中等至良好。通过硅方法研究了这些制备化合物的理化性质。还使用分子动力学模拟来更好地了解蛋白质和配体复合物的稳定性。比较N-(苯并[d]噻唑-2-基)苯甲酰胺3a与水杨酸(4.40±0.10),后者用作群体感应抑制剂的标准,发现N-(苯并[d]噻唑-2-基)苯甲酰胺3a(4.67±0.45)的抗QS作用大于水杨酸(4.40±0.10)。总的来说,研究结果表明,N-(苯并[d]噻唑-2-基)苯甲酰胺/N-(噻唑-2-基)苯甲酰胺衍生物3a-h可能有助于开发新的群体感应抑制剂。
    The development of bacterial resistance to chemical therapy poses a severe danger to efficacy of treating bacterial infections. One of the key factors for resistance to antimicrobial medications is growth of bacteria in biofilm. Quorum sensing (QS) inhibition was created as an alternative treatment by developing novel anti-biofilm medicines. Cell-cell communication is impeded by QS inhibition, which targets QS signaling pathway. The goal of this work is to develop newer drugs that are effective against Pseudomonas aeruginosa by decreasing QS and acting as anti-biofilm agents. In this investigation, N-(benzo[d]thiazol-2-yl)benzamide/N-(thiazol-2-yl)benzamide derivatives 3a-h were designed and synthesized in good yields. Further, molecular docking analyses revealed that binding affinity values were founded -11.2 to -7.6 kcal/mol that were moderate to good. The physicochemical properties of these prepared compounds were investigated through in-silico method. Molecular dynamic simulation was also used to know better understanding of stability of the protein and ligand complex. Comparing N-(benzo[d]thiazol-2-yl)benzamide 3a to salicylic acid (4.40±0.10) that was utilised as standard for quorum sensing inhibitor, the anti-QS action was found greater for N-(benzo[d]thiazol-2-yl)benzamide 3a (4.67±0.45) than salicylic acid (4.40±0.10). Overall, research results suggested that N-(benzo[d]thiazol-2-yl)benzamide/N-(thiazol-2-yl)benzamide derivatives 3a-h may hold to develop new quorum sensing inhibitors.
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  • 文章类型: Journal Article
    罗伊氏LimosilactobacillusZJ625和唾液杆菌ZJ614是潜在的益生菌,当作为多菌株制剂施用于宿主时,具有改善的益处。为了阐明这两种菌株之间细胞间串扰的机制,我们使用DionexNano-RSLC和融合质谱仪通过液相色谱质谱(LC-MS)研究了它们在共培养中的细胞内和细胞外蛋白质组。实验由五个生物重复组成,在指数生长中期收集样品。定量蛋白质组学图谱揭示了几种差异表达的蛋白质(DEP),它们在细胞内和细胞外蛋白质组之间和内部下调或上调。这些DEP包括合成自诱导物2的蛋白质,自诱导物2是混合培养物中群体感应过程中细胞间细菌串扰的传感器化合物。确定的其他重要DEP包括烯醇化酶,磷酸甘油酸激酶,和L-乳酸脱氢酶,在碳水化合物代谢中起作用。与转录相关的蛋白质,ATP的生产和跨膜运输,DNA修复,和那些与宿主上皮结合的潜力也被确定。与蛋白质相关的翻译后修饰包括氧化,脱酰胺,和氨的损失。重要的是,这项研究揭示了S-核糖基同型半胱氨酸裂解酶(luxS)的显着表达,参与合成在群体感应中起重要作用的自诱导因子2,在共培养中帮助细菌细胞间串扰。与罗伊氏乳杆菌ZJ625共培养时,唾液乳杆菌ZJ614的蛋白质组受影响最大。相比之下,与唾液乳杆菌ZJ614相比,从确定的培养基中省略某些培养基成分对罗伊乳杆菌ZJ625产生的影响更大。
    Limosilactobacillus reuteri ZJ625 and Ligilactobacillus salivarius ZJ614 are potential probiotic bacteria with improved benefits when administered to the host as a multi-strain preparation. To elucidate the mechanisms of cell-to-cell crosstalk between these two strains, we studied their intracellular and extracellular proteomes in co-culture by liquid-chromatography mass-spectrometry (LC-MS) using Dionex Nano-RSLC and fusion mass spectrometer. The experiment consisted of five biological replicates, and samples were collected during the mid-exponential growth phase. The quantitative proteomic profiles revealed several differentially expressed proteins (DEPs), which are down- or up-regulated between and within groups for both the intracellular and extracellular proteomes. These DEPs include proteins synthesising autoinducer-2, a sensor compound for cell-to-cell bacterial crosstalk during quorum sensing in mixed culture. Other important DEPs identified include enolase, phosphoglycerate kinase, and l-lactate dehydrogenase, which play roles in carbohydrate metabolism. Proteins associated with transcription, ATP production and transport across the membrane, DNA repair, and those with the potential to bind to the host epithelium were also identified. The post-translational modifications associated with the proteins include oxidation, deamidation, and ammonia loss. Importantly, this study revealed a significant expression of S-ribosylhomocysteine lyase (luxS) involved in synthesising autoinducer-2 that plays important roles in quorum sensing, aiding bacterial cell-to-cell crosstalk in co-cultures. The proteome of L. salivarius ZJ614 was most affected when co-cultured with L. reuteri ZJ625. In contrast, omitting some medium components from the defined medium exerted more effects on L. reuteri ZJ625 than L. salivarius ZJ614.
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  • 文章类型: Journal Article
    背景:大多数关于圆盘中细菌菌群的文献在利用传统培养方法和靶向单一细菌方面处于不利地位,粉刺杆菌。
    目的:我们的目的是使用下一代基因组工具记录正常和退化椎间盘之间细菌菌群的多样性,以入围潜在病原体。
    方法:实验病例对照研究方法:采用16S宏基因组测序来分析来自脑死亡器官自愿供体(n=20)和40个退化椎间盘的MRI正常健康椎间盘中的细菌多样性。手术(Modic(MC)=20和Non-Modic(NMC)=20)。使用通用细菌引物341F和806R扩增V3-V4区,并且使用ILUMINANOVOSEQ6000平台对文库进行测序。在具有最少100OTU(操作分类单位)的细菌处设置统计显著性,并且存在于至少70%的样品中。使用QIIME-2流水线处理QC过滤的读段。使用Greengenes/SILVA参考数据库对合并的读段进行OTU聚类和分类学分类。通过使用LC-MS/MS方法鉴定样品中的细菌代谢物来进行验证。
    结果:丰富的细菌在多样性上差异很大,正如Alpha和Beta多样性分析所证明的那样,存在于所有对照和变性样品中。对照组细菌属数量为27个(14-革兰氏阳性:13-革兰氏阴性),Modic组中的23(10-革兰氏阳性:11-革兰氏阴性),和16(11-革兰氏阳性:5-革兰氏阴性)在非Modic组中。在Modic组中,发现革兰氏阴性菌OTU占优势(占鉴定的总细菌的50%以上),而在对照组和非Modic组中,革兰氏阳性菌的OTU占优势。物种水平分析显示了大量的机会性革兰阴性病原体,如铜绿假单胞菌,Sphingomonospaucibacillus,光盘中的五重杆菌和Modic变化,超过非Modic光盘。细菌代谢物和群体感应分子如N-癸基-L-高丝氨酸内酯的存在,6-羟基烟酸,2-氨基苯乙酮,4-羟基-3-聚异戊烯基苯甲酸酯,PE(16:1(9Z)/18:0)和邻苯二甲酸验证了椎间盘中细菌的定植和细胞间通讯,排除了污染理论。痤疮杆菌并不是三组圆盘中的任何一组中的主要细菌,实际上在对照圆盘中的丰度顺序位于第16位(0.72%),在Modic光盘中排名第七(1.41%),在非Modic光盘中排名第12位(0.53%)。
    结论:这项研究发现,在退化的椎间盘中,革兰氏阴性菌占优势,并强调痤疮梭菌可能不是唯一的引起退化的细菌。这可能归因于环境,饮食,和样本人群的生活习惯。虽然这项研究没有揭示确切的病原体,这可能为未来的研究铺平道路。
    结论:这些发现要求进一步研究细菌谱与椎间盘退变表型的因果关系以及表型驱动的临床治疗方案。
    The majority of literature on bacterial flora in the disc stands disadvantaged in utilizing traditional culture methods and targeting a single bacterium, Cutibacterium acnes.
    Our objective was to document the diversity in the bacterial flora between normal and degenerated discs for shortlisting potential pathogens using next-generation genomic tools.
    Experimental case-control study.
    Researchers employed 16S metagenome sequencing to profile bacterial diversity in magnetic resonance imaging normal healthy discs from brain-dead organ voluntary donors (n=20) and 40 degenerated disc samples harvested during surgery (Modic [MC]=20 and non-Modic [NMC]=20). The V3-V4 region was amplified using universal bacterial primers 341F and 806R, and the libraries were sequenced using Illumina NovoSeq 6000 platform. Statistical significance was set at bacteria with a minimum of 100 operational taxonomic unit (OTU) and present in at least 70% of the samples. The quality check-filtered reads were processed using the QIIME-2 pipeline. The OTU clustering and taxonomic classification were carried out for the merged reads using the Greengenes/SILVA reference database. Validation was done by identification of bacterial metabolites in samples using the liquid chromatography-mass spectrometry approach.
    Abundant bacteria differing widely in diversity, as evidenced by Alfa and Beta diversity analysis, were present in all control and degenerative samples. The number of bacterial genera was 27 (14-gram-positive: 13-gram-negative) in the control group, 23 (10-gram-positive: 11-gram-negative) in the Modic group, and 16 (11-gram-positive: 5-gram-negative) in the non-Modic group. In the Modic group, gram-negative bacteria OTUs were found to be predominant (more than 50% of the total bacteria identified), whereas in control and non-Modic groups the OTUs of gram-positive bacteria were predominant. Species-level analysis revealed an abundance of opportunistic gram-negative pathogens like Pseudomonas aeruginosa, Sphingomonos paucibacillus, and Ochrobactrum quorumnocens in the discs with Modic changes, more than in non-Modic discs. The presence of bacterial metabolites and quorum-sensing molecules like N-decanoyl-L-homoserine lactone, 6-hydroxynicotinic acid, 2-aminoacetophenone, 4-hydroxy-3-polyprenylbenzoate, PE (16:1(9Z)/18:0) and phthalic acid validated the colonization and cell-cell communication of bacteria in disc ruling out contamination theory. Cutibacterium acnes was not the predominant bacteria in any of the three groups of discs and in fact was in the 16th position in the order of abundance in the control discs (0.72%), seventh position in the Modic discs (1.41%), and 12th position (0.53%) in the non-Modic discs.
    This study identified a predominance of gram-negative bacteria in degenerated discs and highlights that Cutibacterium acnes may not be the only degeneration-causing bacteria. This may be attributed to the environment, diet, and lifestyle habits of the sample population. Though the study does not reveal the exact pathogen, it may pave the way for future studies on the subject.
    These findings invite further investigation into causal relationships of bacterial profile with disc degeneration phenotypes as well as phenotype-driven clinical treatment protocols.
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