Effect of Micro-Patterned Mucin on Quinolone and Rhamnolipid Profiles of Mucoid Pseudomonas aeruginosa under Antibiotic Stress.
Effect of Micro-Patterned Mucin on Quinolone and Rhamnolipid Profiles of Mucoid Pseudomonas aeruginosa under Antibiotic Stress.
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微图案化黏蛋白在抗生素胁迫下对黏液型铜绿假单胞菌喹诺酮及鼠李糖脂特征的影响
DOI：
10.1021/acsinfecdis.2c00519
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发表时间：
2023-01-13
影响因子：
5.3
通讯作者：
Bohn, Paul W.
中科院分区：
文献类型：
--
作者：
Jia, Jin;Parmar, Dharmeshkumar;Ellis, Joanna F.;Cao, Tianyuan;Cutri, Allison R.;Shrout, Joshua D.;V. Sweedler, Jonathan V.;Bohn, Paul W.
关键词：
铜绿假单胞菌通常与医院获得性感染有关，它在各种表面形成生物膜的能力和导致的抗生素耐药性增强严重限制了治疗选择。由于表面附着使铜绿假单胞菌对群体感应(QS)敏感，并通过化学和机械线索诱导毒力，我们利用结合共聚焦拉曼显微镜(CRM)和基质辅助激光解吸/电离-质谱仪(MALDI-MS)的多模式化学成像，通过空间模式粘蛋白，结合亚抑制浓度的妥布霉素，研究表面特性对粘液型和非粘液型铜绿假单胞菌群体感应(QS)和毒力因子的影响。在不同的时间点，样品由固-水界面的表面附着静态生物膜、上清液和气-水界面的膜状生物膜组成。尽管在上清液中存在亚抑制浓度的妥布霉素会抑制静态生物膜的生长和发展，而不依赖于菌株和表面粘蛋白模式，但我们观察到粘液型和非粘液型菌株的行为明显不同。在非粘液型菌株中，喹诺酮类药物信号被诱导得更早，并且受到粘蛋白表面图案的影响，其程度在所测试的粘液型菌株中是不存在的。此外，在粘液型和非粘液型菌株的膜生物膜中都观察到吩嗪毒力因子，如绿青素(PYO)，但在这两种菌株的静态生物膜中都没有检测到，这突显了膜生物膜和静态生物膜在应激反应方面的差异。粘液型和非粘液型菌株之间的差异与它们特定的物候一致，在这种物候中，粘液型菌株形成了高度保护的生物膜。TOC图像显示了样品准备和分析工作流程的关键步骤。图像从左到右描绘了生物膜样品采集、拉曼和质谱学成像的介质和时间点，并提取了喹诺酮类药物信号的拉曼和离子图像。利用非破坏性共聚焦拉曼成像技术对各种细菌产物进行了表征。生物膜表面的MSI表征允许评估细菌信号分子的分布模式和空间定位。从样品表面连续获取MS数据有助于重建与每个像素或激光消融位置所观察到的峰值强度相对应的图像。在我们的研究中，我们分析了铜绿假单胞菌产生的生物分子，以了解在存在亚抑制浓度的抗生素的情况下，表面微环境如何影响喹诺酮类药物和鼠李糖脂的分泌。
Pseudomonas aeruginosa is commonly implicated in hospital acquired infections where its capacity to form biofilms on a variety of surfaces and the enhanced antibiotic resistance that results seriously limit treatment choices. Because surface attachment sensitizes P. aeruginosa to quorum sensing (QS) and induces virulence through both chemical and mechanical cues, we investigate the effect of surface properties through spatially-patterned mucin, combined with sub-inhibitory concentrations of tobramycin on QS and virulence factors in both mucoid and non-mucoid P. aeruginosa strains using multi-modal chemical imaging combining confocal Raman microscopy (CRM) and matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS). Samples are comprised of surface-adherent static biofilms at a solid-water interface, supernatant liquid, and pellicle biofilms at an air-water interface at various time points. Although the presence of a sub-inhibitory concentration of tobramycin in the supernatant retards growth and development of static biofilms independent of strain and surface mucin patterning, we observe clear differences in the behavior of mucoid and non-mucoid strains. Quinolone signals in a non-mucoid strain are induced earlier and are influenced by mucin surface patterning to a degree not exhibited in the mucoid strain tested. Additionally, phenazine virulence factors, such as pyocyanin (PYO), are observed in the pellicle biofilms of both mucoid and non-mucoid strains, but are not detected in the static biofilms from either strain, highlighting the differences in stress response between pellicle and static biofilms. Differences between mucoid and nonmucoid strains are consistent with their strain-specific phenology, in which the mucoid strain develops highly protected biofilms. TOC image shows key steps of the sample preparation and analysis workflow. Images from left to right depict media and time points of biofilm sample collection, Raman and mass spectrometry imaging, and extracted Raman and ion images of quinolone signal. Non-destructive confocal Raman imaging was used to characterize various bacterial products. MSI characterization of biofilm surfaces allows assessment of distribution patterns and spatial localization of bacterial signaling molecules. Sequential acquisition of MS data from the sample surface facilitates reconstruction of an image that corresponds to the peak intensities observed per each pixel or location of laser ablation. In our study, we analyzed biomolecules produced by P. aeruginosa to understand how the secretion of quinolones and rhamnolipids are impacted by the surface microenvironment in the presence of a sub-inhibitory concentration of antibiotic.
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影响因子：
3.7
作者：
Babić F;Venturi V;Maravić-Vlahovicek G
通讯作者：
Maravić-Vlahovicek G
DOI：
10.1099/00221287-13-3-572
发表时间：
1955-01-01
期刊：
JOURNAL OF GENERAL MICROBIOLOGY
影响因子：
--
作者：
HOLLOWAY, BW
通讯作者：
HOLLOWAY, BW
影响因子：
5.3
作者：
Jia J;Ellis JF;Cao T;Fu K;Morales-Soto N;Shrout JD;Sweedler JV;Bohn PW
通讯作者：
Bohn PW
影响因子：
4
作者：
Bhattacharjee, Arunima;Nusca, Tyler D.;Hochbaum, Allon I.
通讯作者：
Hochbaum, Allon I.
影响因子：
4.3
作者：
Brewer, Luke K.;Jones, Jace W.;Kane, Maureen A.
通讯作者：
Kane, Maureen A.