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1. Cephalosporins Interfere With Quorum Sensing and Improve the Ability of Caenorhabditis elegans to Survive Pseudomonas aeruginosa Infection

   https://doi.org/10.3389/fmicb.2021.598498  

   Kumar, Lokender; Brenner, Nathanael; Brice, John; Klein-Seetharaman, Judith; Sarkar, Susanta K. (January 2021, Frontiers in Microbiology)

   null (Ed.)

   Pseudomonas aeruginosa utilizes the quorum sensing (QS) system to strategically coordinate virulence and biofilm formation. Targeting QS pathways may be a potential anti-infective approach to treat P. aeruginosa infections. In the present study, we define cephalosporins’ anti-QS activity using Chromobacterium violaceum CV026 for screening and QS-regulated mutants of P. aeruginosa for validation. We quantified the effects of three cephalosporins, cefepime, ceftazidime, and ceftriaxone, on (1) pyocyanin production using spectrophotometric assay, (2) bacterial motility using agar plate assay, and (3) biofilm formation using scanning electron microscopy. We also studied isogenic QS mutant strains of PAO1 (Δ lasR ,Δ rhlR ,Δ pqsA , and Δ pqsR) to compare and distinguish QS-mediated effects on the motility phenotypes and bacterial growth with and without sub-MIC concentrations of antibiotics. Results showed that cephalosporins have anti-QS activity and reduce bacterial motility, pyocyanin production, and biofilm formation for CV026 and PAO1. Also, sub-MICs of cefepime increased aminoglycosides’ antimicrobial activity against P. aeruginosa PAO1, suggesting the advantage of combined anti-QS and antibacterial treatment. To correlate experimentally observed anti-QS effects with the interactions between cephalosporins and QS receptors, we performed molecular docking with ligand binding sites of quorum sensing receptors using Autodock Vina. Molecular docking predicted cephalosporins’ binding affinities to the ligand-binding pocket of QS receptors (CviR, LasR, and PqsR). To validate our results using an infection model, we quantified the survival rate of C aenorhabditis elegans following P. aeruginosa PAO1 challenge at concentrations less than the minimum inhibitory concentration (MIC) of antibiotics. C. elegans infected with PAO1 without antibiotics showed 0% survivability after 72 h. In contrast, PAO1-infected C. elegans showed 65 ± 5%, 58 ± 4%, and 49 ± 8% survivability after treatment with cefepime, ceftazidime, and ceftriaxone, respectively. We determined the survival rates of C. elegans infected by QS mutant strains Δ lasR (32 ± 11%), Δ rhlR (27 ± 8%), Δ pqsA (27 ± 10%), and Δ pqsR (37 ± 6%), which suggest essential role of QS system in virulence. In summary, cephalosporins at sub-MIC concentrations show anti-QS activity and enhance the antibacterial efficacy of aminoglycosides, a different class of antibiotics. Thus, cephalosporins at sub-MIC concentrations in combination with other antibiotics are potential candidates for developing therapies to combat infections caused by P. aeruginosa. 

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2. Effects of fluid shear stress on oral biofilm formation and composition and the transcriptional response of Streptococcus gordonii

   https://doi.org/10.1111/omi.12481  

   Nairn, Brittany_L; Lima, Bruno_P; Chen, Ruoqiong; Yang, Judy_Q; Wei, Guanju; Chumber, Ashwani_K; Herzberg, Mark_C (August 2024, Molecular Oral Microbiology)

   Abstract Biofilms are subjected to many environmental pressures that can influence community structure and physiology. In the oral cavity, and many other environments, biofilms are exposed to forces generated by fluid flow; however, our understanding of how oral biofilms respond to these forces remains limited. In this study, we developed a linear rocker model of fluid flow to study the impact of shear forces onStreptococcus gordoniiand dental plaque‐derived multispecies biofilms. We observed that as shear forces increased,S. gordoniibiofilm biomass decreased. Reduced biomass was largely independent of overall bacterial growth. Transcriptome analysis ofS. gordoniibiofilms exposed to moderate levels of shear stress uncovered numerous genes with differential expression under shear. We also evaluated an ex vivo plaque biofilm exposed to fluid shear forces. LikeS. gordonii, the plaque biofilm displayed decreased biomass as shear forces increased. Examination of plaque community composition revealed decreased diversity and compositional changes in the plaque biofilm exposed to shear. These studies help to elucidate the impact of fluid shear on oral bacteria and may be extended to other bacterial biofilm systems. 

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3. Superparamagnetic nanoparticles for the treatment of periodontal disease

   https://doi.org/10.1117/12.2654656  

   Fritz, Shayden; Armijo, Leisha M.; Simpson, Kayla; Lopez, Kimberly; Osinski, Marek; Bandara, Nihal; Smyth, Hugh D.; Sheng, Jian; Xu, Wei (March 2023, Colloidal Nanoparticles for Biomedical Applications XVIII)

   Osiński, Marek; Kanaras, Antonios G. (Ed.)

   Periodontal diseases are prevalent worldwide and are linked to numerous other health conditions due to dysbiosis and chronic inflammatory state. Most periodontal diseases are caused by pathogenic bacteria that colonize dental tissues in the form of biofilm. Eradication of bacterial biofilms can be difficult to achieve due to the complex architecture of the teeth and gums which complicates the removal. Orthodontic wires and dental devices introduce additional hurdles to the adequate removal of biofilms by traditional methods since mechanical disruption via direct contact with toothbrush bristles, floss, and abrasive toothpaste is limited. Magnetically activated nanoparticles (NPs), specifically iron oxide nanoparticles (IONPs) that can be functionalized as antimicrobial particles and remotely controlled by magnetic fields, are of interest for oral biofilm eradication. We present data in multi-species bacterial cultures, established biofilms, human gingival keratinocytes, and human gingival fibroblast cells alone and in the presence of multispecies biofilm co-cultures to determine the safest, most efficacious IONP size ranges and treatment concentrations of active magnetic NPs for removal of dental biofilms. We report enhanced efficacy for IONPs coated with alginate vs. dextran, and small sizes (~8 nm vs. >20 nm in size) appear to exhibit enhanced antimicrobial efficacy. Human gingival keratinocyte (TIGK) cells in co-culture with treated and untreated multispecies biofilms in an in-vitro periodontitis model also exhibited a trend of reduced inflammatory markers in wells with IONP-treated biofilms. 

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4. Biofabrication and characterization of multispecies electroactive biofilms in stratified paper-based scaffolds

   https://doi.org/10.1039/d2an01059c  

   Elhadad, Anwar; Choi, Seokheun (September 2022, The Analyst)

   Bioelectrochemical technologies have attracted significant scientific interest because the effective bacterial electron exchange with external electrodes can provide a sustainable solution that joins environmental remediation and energy recovery. Multispecies electroactive bacterial biofilms are catalysts that will drive the operation of bioelectrochemical devices. Unfortunately, there is a lack of understanding of key mechanisms determining their electron-generating capabilities and syntrophic relations within microbial communities in biofilms. This is because there are no universally standardized models for simple, rapid, reliable, and cost-effective fabrication and characterization of electroactive multispecies biofilms. The heterogeneous and long-term nature of biofilm formation has hampered the development of those models. This work develops novel biofabrication and analysis platforms by creating innovative, paper-based 3-D systems that accurately recapitulate the structure, function, and physiology of living multispecies biofilms. Multiple layers of paper containing bacterial cells were stacked to simulate different layered 3-D biofilm models with defined cellular compositions and microenvironments. Overall bacterial electrogenic capabilities through the biofilm structures were characterized by thoroughly monitoring collective electron flows through different external resistors. Changes in the type of species and order of stacking created biofilm modeling which allowed for the study of their electrogenic performance via variation in electron flow rate output. Furthermore, multi-laminate structures allowed for straightforward de-stacking and layer-by-layer separation for analyses of pH distribution and cellular viability. Our multi-laminate structures provide a new strategy for (i) controlling the biofilm geometry of 3-D bacterial cultures, (ii) monitoring the microbial electoral properties, and (iii) constructing an artificial biofilm layer by layer. 

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5. Synergy between c-di-GMP and Quorum-Sensing Signaling in Vibrio cholerae Biofilm Morphogenesis

   https://doi.org/10.1128/jb.00249-22  

   Prentice, Jojo A.; Bridges, Andrew A.; Bassler, Bonnie L. (January 2022, Journal of Bacteriology)

   O’Toole, George (Ed.)

   ABSTRACT Transitions between individual and communal lifestyles allow bacteria to adapt to changing environments. Bacteria must integrate information encoded in multiple sensory cues to appropriately undertake these transitions. Here, we investigate how two prevalent sensory inputs converge on biofilm morphogenesis: quorum sensing, which endows bacteria with the ability to communicate and coordinate group behaviors, and second messenger c-di-GMP signaling, which allows bacteria to detect and respond to environmental stimuli. We use Vibrio cholerae as our model system, the autoinducer AI-2 to modulate quorum sensing, and the polyamine norspermidine to modulate NspS-MbaA-mediated c-di-GMP production. Individually, AI-2 and norspermidine drive opposing biofilm phenotypes, with AI-2 repressing and norspermidine inducing biofilm formation. Surprisingly, however, when AI-2 and norspermidine are simultaneously detected, they act synergistically to increase biofilm biomass and biofilm cell density. We show that this effect is caused by quorum-sensing-mediated activation of nspS - mbaA expression, which increases the levels of NspS and MbaA, and in turn, c-di-GMP biosynthesis, in response to norspermidine. Increased MbaA-synthesized c-di-GMP activates the VpsR transcription factor, driving elevated expression of genes encoding key biofilm matrix components. Thus, in the context of biofilm morphogenesis in V. cholerae, quorum-sensing regulation of c-di-GMP-metabolizing receptor levels connects changes in cell population density to detection of environmental stimuli. IMPORTANCE The development of multicellular communities, known as biofilms, facilitates beneficial functions of gut microbiome bacteria and makes bacterial pathogens recalcitrant to treatment. Understanding how bacteria regulate the biofilm life cycle is fundamental to biofilm control in industrial processes and in medicine. Here, we demonstrate how two major sensory inputs—quorum-sensing communication and second messenger c-di-GMP signaling—jointly regulate biofilm morphogenesis in the global pathogen Vibrio cholerae. We characterize the mechanism underlying a surprising synergy between quorum-sensing and c-di-GMP signaling in controlling biofilm development. Thus, the work connects changes in cell population density to detection of environmental stimuli in a pathogen of clinical significance. 

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