Quorum-sensing Molecules Research Articles

A new strategy for greenhouse gas emission reduction in the anaerobic/anoxic/oxic biological treatment process using exogenous N-acyl-homoserine lactones, a quorum-sensing signaling molecules

ABSTRACT In this study, the impact of exogenous N-acyl-homoserine lactones (AHLs) on greenhouse gas (GHG) emissions in anaerobic/anoxic/oxic (A/A/O) systems was analyzed by manipulating the type and dosage of AHLs. The mechanism behind AHLs’ effects on GHG emissions was explored through changes in microbial community structure. Findings revealed that N-octanoyl-homoserine lactone (C8-HSL) and high-dose N-dodecanoyl-homoserine lactone (C12-HSL) increased GHG emissions, while low-dose C12-HSL decreased them. Moreover, C8-HSL and high-dose C12-HSL promoted methane (CH4) and nitrous oxide (N2O) production by affecting sludge particle size. Bacterial community analysis highlighted Acinetobacter and Flavobacterium's roles in N2O emissions and acetate methanogens in methane synthesis. Metabolic pathway analysis showed that the acetic acid (CH3COOH) methanogenic pathway was the main methanogenic pathway; C8-HSL and C12-HSL influenced methane emission by affecting the methanogenic pathway and N2O emission by changing nitrous oxide reductase (Nos) abundance. This research underscores AHL-based quorum sensing's potential in mitigating GHG emissions during activated sludge wastewater treatment, offering insights into their application and impact on key microbial activities. Limitations include the absence of methane emission reduction by signaling molecules and the need for further investigation into their effects on sludge accumulation.

Tracing the path of Quorum sensing molecules in cystic fibrosis mucus in a biomimetic in vitro permeability platform

P. aeruginosa employs specific quorum sensing (QS) mechanisms to orchestrate biofilm formation, enhancing resistance to host defences. In physiological conditions, QS molecules permeate the lung environment and cellular membrane to reach the cytoplasmic Aryl Hydrocarbon Receptor (AhR) that is pivotal for activating the immune response against infection. In pathological conditions like cystic fibrosis (CF) this interkingdom communication is altered, favouring P. aeruginosa persistence and chronic infection. Here, we aim to investigate the molecular journey of QS molecules from CF-like environments to the cytoplasm by quantifying via HPLC-MS the permeability of selected QS molecules (quinolones, lactones, and phenazines) through in vitro models of the two main biological lung barriers: CF-mucus and cellular membrane. While QS molecules not activating AhR exhibit intermediate permeability through the cellular membrane model (PAMPA) (1.0–4.0 × 10−6 cm/s), the AhR-activating molecule (pyocyanin) shows significantly higher permeability (8.6 ± 1.4 × 10−6 cm/s). Importantly, combining the CF mucus model with PAMPA induces a 50% decrease in pyocyanin permeability, indicating a strong mucus-shielding effect with pathological implications in infection eradication. This study underscores the importance of quantitatively describing the AhR-active bacterial molecules, even in vitro, to offer new perspectives for understanding P. aeruginosa virulence mechanisms and for proposing new antibacterial therapeutic approaches.

Global transcriptional response of Pseudomonas aeruginosa to UVA radiation.

Ultraviolet A (UVA) radiation is the major fraction of UV radiation reaching the Earth's surface. Its harmful effects on microorganisms, due mainly to oxidative damage, have been exploited for development of natural solar and commercial UVA-based disinfection methods. In this work, the global transcriptional response of Pseudomonas aeruginosa exposed to ultraviolet A (UVA) radiation was analyzed. To conduct this study, we analyzed the whole transcriptome of the PAO1 strain grown to logarithmic phase under sublethal doses of UVA or in the dark. We found that a total of 298 genes responded to UVA with a change of at least two-fold (5.36% of the total P. aeruginosa genome), and showed equal amount of induced and repressed genes. An important fraction of the induced genes were involved in the response to DNA damage and included induction of SOS, prophage and pyocins genes. The results presented in this study suggest that one of the main UVA targets are proteins carrying [Fe-S] clusters since several genes involved in the processes of synthesis, trafficking and assembly of these structures were upregulated. The management of intracellular iron levels also seems to be a robust response to this stress factor. The strong induction of genes involved in denitrification suggest that this pathway and/or reactive nitrogen species such as nitric oxide could have a role in the response to this radiation. Regarding the down-regulated genes, we found many involved in the biosynthesis of PQS, a quorum-sensing signal molecule with a possible role as endogenous photosensitizer.

In Situ Enrichment of Anammox Bacteria from Pig Farm Anoxic Sludge Through Co-Cultivation with a Quorum-Sensing Functional Strain Pseudomonas aeruginosa

Anaerobic ammonium oxidation (anammox), as an efficient and low-carbon method for nitrogen removal from wastewater, faces the challenge of slow enrichment of functional bacteria. In this study, the enrichment of anammox bacteria Candidatus Brocadia was successfully accelerated by co-culturing with the quorum-sensing strain Pseudomonas aeruginosa and anoxic sludge from a pig farm. Experimental results showed that the R2, which had Pseudomonas aeruginosa added, exhibited chemical reaction ratios RS (NO2−-N consumption/NH4+-N consumption) and RP (NO3−-N production/NH4+-N consumption) closer to the theoretical values of the anammox reaction since Phase Ⅱ. Bacterial community analysis indicated that the abundance of Candidatus Brocadia in R2 reached 1.63% in cycle 20, significantly higher than the 0.45% in R1. More quorum-sensing signaling molecules, primarily C6-HSL, were detected in R2. C6-HSL was positively correlated with processes such as the secretion of anammox extracellular polymers (EPS) and the regulation of nitric oxide reductase (Nir), which may explain the reason behind the accelerated increase in the abundance of Candidatus Brocadia through co-culturing. Moreover, the metabolism of the dominant genus Paracoccus within the two groups of reactors also showed positive regulation by C6-HSL, with its abundance trend similar to that of Candidatus Brocadia, jointly completing the nitrogen removal process in the reactors. However, it is still unknown which genera secrete large amounts of C6-HSL after inoculation with Pseudomonas aeruginosa. This research provides a novel and low-cost method for the enrichment of anammox bacteria.

Chemical signaling in biofilm-mediated biofouling.

Biofouling is the undesirable accumulation of living organisms and their metabolites on submerged surfaces. Biofouling begins with adhesion of biomacromolecules and/or microorganisms and can lead to the subsequent formation of biofilms that are predominantly regulated by chemical signals, such as cyclic dinucleotides and quorum-sensing molecules. Biofilms typically release chemical cues that recruit or repel other invertebrate larvae and algal spores. As such, harnessing the biochemical mechanisms involved is a promising avenue for controlling biofouling. Here, we discuss how chemical signaling affects biofilm formation and dispersion in model species. We also examine how this translates to marine biofouling. Both inductive and inhibitory effects of chemical cues from biofilms on macrofouling are also discussed. Finally, we outline promising mitigation strategies by targeting chemical signaling to foster biofilm dispersion or inhibit biofouling.

Quorum-sensing molecules regulate biochar-assisted anaerobic digestion system for methane production: Single-stage vs. two-stage digestion

Promoting methane production performance from waste activated sludge is of great significance for energy self-sufficiency operation of wastewater treatment plants. This study evaluated the methane production enhancement strategies of biochar and quorum-sensing molecules like N-acylated homoserine lactone (AHL) in single-stage and two-stage anaerobic digestion systems. The methane yield increased to 134.9 mL/g volatile suspended solids in single-stage digestion system, which was 41.7 % higher than that of control group, while it in two-stage digestion system was not only lower than that obtained in single-stage digestion system but also less than that observed in groups adding AHL either at the beginning or on the third day of experiment period. The mechanism studies indicated that the single-stage digestion strategy promoted solubilization, hydrolysis, acetogenesis, and methanogenesis stages and improved the conversion and utilization of organic matters with the enhanced syntrophic metabolisms between microorganisms. In addition, the electron transfer activity between microorganisms was promoted, with an increase ratio of 54.7 %, meanwhile, the syntrophic relationships between methanogens such as Methanosarcina and electroactive bacteria such as norank_f__Bacteroidetes_vadinHA17 were promoted. As a result, both acetotrophic and hydrogenotrophic methanogenesis pathways were improved. This study provided some novel approaches for promoting anaerobic digestion performance while using biochar as the additive.

Metabolic activities of marine ammonia-oxidizing archaea orchestrated by quorum sensing.

Ammonia-oxidizing archaea (AOA) play crucial roles in marine carbon and nitrogen cycles by fixing inorganic carbon and performing the initial step of nitrification. Evaluation of carbon and nitrogen metabolism popularly relies on functional genes such as amoA and accA. Increasing studies suggest that quorum sensing (QS) mainly studied in biofilms for bacteria may serve as a universal communication and regulatory mechanism among prokaryotes; however, this has yet to be demonstrated in marine planktonic archaea. To bridge this knowledge gap, we employed a combination of metabolic activity markers (amoA, accA, and grs) to elucidate the regulation of AOA-mediated nitrogen, carbon processes, and their interactions with the surrounding heterotrophic population. Through co-transcription investigations linking metabolic markers to potential key QS genes, we discovered that QS molecules could regulate AOA's carbon, nitrogen, and lipid metabolisms under different conditions. Interestingly, specific AOA ecotypes showed a preference for employing distinct QS systems and a distinct QS circuit involving a typical population. Overall, our data demonstrate that QS orchestrates nitrogen and carbon metabolism, including the exchange of organic metabolites between AOA and surrounding heterotrophic bacteria, which has been previously overlooked in marine AOA research.

Synergistic effects of quorum-sensing molecules and antimicrobials against Candida albicans and Pseudomonas aeruginosa biofilms: in vitro and in vivo studies.

Candida albicans can form polymicrobial biofilms with other microorganisms, such as Pseudomonas aeruginosa, at infection sites. As biofilms are highly resistant to antibiotics there is a need for new antibiofilm agents that have unique targets and modes of action. In this study the antibiofilm effects of two quorum-sensing molecules (QSMs), farnesol and tyrosol, were investigated alone and in combination with antibiotics (aztreonam, colistin, tobramycin) and antifungals (fluconazole, amphotericin B, caspofungin), against single- and dual-species biofilms of C. albicans and P. aeruginosa in in vitro and in vivo systems. It was observed that QSMs alone, especially farnesol, showed at least a 1-log reduction against preformed single- and dual-species biofilms of C. albicans and P. aeruginosa. Combination of QSMs with colistin or fluconazole was found to be effective against both single- and dual-species biofilms in vitro. Increased survival was observed in C. elegans when treated with colistin or fluconazole in combination with QSMs, compared with no treatment. Additionally, the QSMs and colistin and farnesol combinations effectively inhibited biofilm formation by C. albicans and P. aeruginosa on bronchial epithelial cells, and reduced IL-1β expression in lung bronchial epithelial cells. There is a need for effective treatments for bacterial-fungal biofilm infections and, to our knowledge, there have been no studies of QSMs and antimicrobial combinations against dual-species biofilms involving C. albicans and P. aeruginosa. Hence these findings will make a significant contribution to the literature.

Nanotopography Influences Host-Pathogen Quorum Sensing and Facilitates Selection of Bioactive Metabolites in Mesenchymal Stromal Cells and Pseudomonas aeruginosa Co-Cultures.

Orthopedic implant-related bacterial infections and resultant antibiotic-resistant biofilms hinder implant-tissue integration and failure. Biofilm quorum sensing (QS) communication determines the pathogen colonization success. However, it remains unclear how implant modifications and host cells are influenced by, or influence, QS. High aspect ratio nanotopographies have shown to reduce biofilm formation of Pseudomonas aeruginosa, a sepsis causing pathogen with well-defined QS molecules. Producing such nanotopographies in relevant orthopedic materials (i.e., titanium) allows for probing QS using mass spectrometry-based metabolomics. However, nanotopographies can reduce host cell adhesion and regeneration. Therefore, we developed a polymer (poly(ethyl acrylate), PEA) coating that organizes extracellular matrix proteins, promoting bioactivity to host cells such as human mesenchymal stromal cells (hMSCs), maintaining biofilm reduction. This allowed us to investigate how hMSCs, after winning the race for the surface against pathogenic cells, interact with the biofilm. Our approach revealed that nanotopographies reduced major virulence pathways, such as LasR. The enhanced hMSCs support provided by the coated nanotopographies was shown to suppress virulence pathways and biofilm formation. Finally, we selected bioactive metabolites and demonstrated that these could be used as adjuncts to the nanostructured surfaces to reduce biofilm formation and enhance hMSC activity. These surfaces make excellent models to study hMSC-pathogen interactions and could be envisaged for use in novel orthopedic implants.

Design of Thermoresponsive Genetic Controls with Minimal Heat-Shock Response.

As temperature serves as a versatile input signal, thermoresponsive genetic controls have gained significant interest for recombinant protein production and metabolic engineering applications. The conventional thermoresponsive systems normally require the continuous exposure of heat stimuli to trigger the prolonged expression of targeted genes, and the accompanied heat-shock response is detrimental to the bioproduction process. In this study, we present the design of thermoresponsive quorum-sensing (ThermoQS) circuits to make Escherichia coli record transient heat stimuli. By conversion of the heat input into the accumulation of quorum-sensing molecules such as acyl-homoserine lactone derived from Pseudomonas aeruginosa, sustained gene expressions were achieved by a minimal heat stimulus. Moreover, we also demonstrated that we reprogrammed the E. coli Lac operon to make it respond to heat stimuli with an impressive signal-to-noise ratio (S/N) of 15.3. Taken together, we envision that the ThermoQS systems reported in this study are expected to remarkably diminish both design and experimental expenditures for future metabolic engineering applications.

Electrochemical Detection of Pseudomonas aeruginosa Quorum Sensing Molecule (S)-N-Butyryl Homoserine Lactone Using Molecularly Imprinted Polymers.

Pseudomonas aeruginosa is a multidrug-resistant Gram-negative bacterium that poses a significant threat to public health, necessitating rapid and on-site detection methods for rapid recognition. The goal of the project is therefore to indirectly detect the presence of P. aeruginosa in environmental water samples targeting one of its quorum-sensing molecules, namely, (S)-N-butyryl homoserine lactone (BHL). To this aim, molecularly imprinted polymers (MIPs) were synthesized via bulk free-radical polymerization using BHL as a template molecule. The obtained MIP particles were immobilized onto screen-printed electrodes (MIP-SPEs), and the BHL rebinding was analyzed via electrochemical impedance spectroscopy (EIS). To study the specificity of the synthesized MIPs, isotherm curves were built after on-point rebinding analysis performed via LC-MS measurements for both MIPs and NIPs (nonimprinted polymers, used as a negative control), obtaining an imprinting factor (IF) of 2.8 (at C f = 0.4 mM). The MIP-SPEs were integrated into an electrochemical biosensor with a linear range of 1 × 101-1 × 103 nM and a limit of detection (LoD) of 31.78 ± 4.08 nM. Selectivity measurements were also performed after choosing specific interferent molecules, such as structural analogs and potential interferents, followed by on-point analysis performed in spiked tap water to prove the sensor's potential to detect the presence of the quorum-sensing molecule in environmentally related real-life samples.

Microbead-Encapsulated Luminescent Bioreporter Screening of P. aeruginosa via Its Secreted Quorum-Sensing Molecules.

Pseudomonas aeruginosa is an opportunistic Gram-negative bacterium that remains a prevalent clinical and environmental challenge. Quorum-sensing (QS) molecules are effective biomarkers in pinpointing the presence of P. aeruginosa. This study aimed to develop a convenient-to-use, whole-cell biosensor using P. aeruginosa reporters individually encapsulated within alginate-poly-L-lysine (alginate-PLL) microbeads to specifically detect the presence of bacterial autoinducers. The PLL-reinforced microbeads were prepared using a two-step method involving ionic cross-linking and subsequent coating with thin layers of PLL. The alginate-PLL beads showed good stability in the presence of a known cation scavenger (sodium citrate), which typically limits the widespread applications of calcium alginate. In media containing synthetic autoinducers-such as N-(3-oxo dodecanoyl) homoserine lactone (3-oxo-C12-HSL) and N-butanoyl-L-homoserine lactone (C4-HSL), or the cell-free supernatants of planktonic or the flow-cell biofilm effluent of wild P. aeruginosa (PAO1)-the encapsulated bacteria enabled a dose-dependent detection of the presence of these QS molecules. The prepared bioreporter beads remained stable during prolonged storage at 4 and -80 °C and were ready for on-the-spot sensing without the need for recovery. The proof-of-concept, optical fiber-based, and whole-cell biosensor developed here demonstrates the practicality of the encapsulated bioreporter for bacterial detection based on specific QS molecules.

Absence of farnesol salvage in Candida albicans and probably in other fungi.

The absence of farnesol salvage constitutes a major difference in the metabolic capabilities of fungi. In terms of fungal physiology, the lack of farnesol salvage pathways relates to how farnesol acts as a quorum-sensing molecule in Candida albicans and why farnesol should be investigated for use in combination with other known antifungal antibiotics. Its absence is essential for a model (K. W. Nickerson et al., Microbiol Mol Biol Rev 88:e00081-22, 2024), wherein protein farnesylation, protein chaperones, and the unfolded protein response are combined under the unifying umbrella of a cell's intracellular farnesol (Fi). In terms of human health, farnesol should have at least two different modes of action depending on whether those cells have farnesol salvage. Because animals have farnesol salvage, we can now see the importance of dietary prenols as well as the potential importance of farnesol in treating neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease, and multiple sclerosis.

The Candida albicans quorum-sensing molecule farnesol alters sphingolipid metabolism in human monocyte-derived dendritic cells.

Candida albicans, an opportunistic fungal pathogen, produces the quorum-sensing molecule farnesol, which we have shown alters the transcriptional response and phenotype of human monocyte-derived dendritic cells (DCs), including their cytokine secretion and ability to prime T cells. This is partially dependent on the nuclear receptor peroxisome proliferator-activated receptor gamma (PPAR-γ), which has numerous ligands, including the sphingolipid metabolite sphingosine 1-phosphate. Sphingolipids are a vital component of membranes that affect membrane protein arrangement and phagocytosis of C. albicans by DCs. Thus, we quantified sphingolipid metabolites in monocytes differentiating into DCs by High-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Farnesol increased the activity of serine palmitoyltransferase, leading to increased levels of 3-keto-dihydrosphingosine, dihydrosphingosine, and dihydrosphingosine 1-phosphate and inhibited dihydroceramide desaturase by inducing oxidative stress, leading to increased levels of dihydroceramide and dihydrosphingomyelin species and reduced ceramide levels. Accumulation of dihydroceramides can inhibit mitochondrial function; accordingly, farnesol reduced mitochondrial respiration. Dihydroceramide desaturase inhibition increases lipid droplet formation, which we observed in farnesol-treated cells, coupled with an increase in intracellular triacylglycerol species. Furthermore, inhibition of dihydroceramide desaturase with either farnesol or specific inhibitors impaired the ability of DCs to prime interferon-γ-producing T cells. The effect of farnesol on sphingolipid metabolism, triacylglycerol synthesis, and mitochondrial respiration was not dependent on PPAR-γ. In summary, our data reveal novel effects of farnesol on sphingolipid metabolism, neutral lipid synthesis, and mitochondrial function in DCs that affect their instruction of T cell cytokine secretion, indicating that C. albicans can manipulate host cell metabolism via farnesol secretion.IMPORTANCECandida albicans is a common commensal yeast, but it is also an opportunistic pathogen which is one of the leading causes of potentially lethal hospital-acquired infections. There is growing evidence that its overgrowth in the gut can influence diseases as diverse as alcohol-associated liver disease and COVID-19. Previously, we found that its quorum-sensing molecule, farnesol, alters the phenotype of dendritic cells differentiating from monocytes, impairing their ability to drive protective T cell responses. Here, we demonstrate that farnesol alters the metabolism of sphingolipids, important structural components of the membrane that also act as signaling molecules. In monocytes differentiating to dendritic cells, farnesol inhibited dihydroceramide desaturase, resulting in the accumulation of dihydroceramides and a reduction in ceramide levels. Farnesol impaired mitochondrial respiration, known to occur with an accumulation of dihydroceramides, and induced the accumulation of triacylglycerol and oil bodies. Inhibition of dihydroceramide desaturase resulted in the impaired ability of DCs to induce interferon-γ production by T cells. Thus, farnesol production by C. albicans could manipulate the function of dendritic cells by altering the sphingolipidome.

Effects of chlorogenic acid-grafted-chitosan on biofilms, oxidative stress, quorum sensing and c-di-GMP in Pseudomonas fluorescens

This study determined the inhibitory mechanism as well as anti-biofilm activity of chlorogenic acid-grafted-chitosan (CS-g-CA) against Pseudomonas fluorescens (P. fluorescens) in terms of biofilm content, oxidative stress, quorum sensing and cyclic diguanosine monophosphate (c-di-GMP) concentration, and detected the changes in the expression levels of related genes by quantitative real-time PCR (qRT-PCR). Results indicated that treatment with sub-concentrations of CS-g-CA for P. fluorescens led to reduce the biofilm size of large colonies, decrease the content of biofilm and extracellular polymers, weaken the motility and adhesion of P. fluorescens. Moreover, CS-g-CA resulted in higher ROS levels, diminished catalase activity (CAT), and increased superoxide dismutase (SOD) in P. fluorescens. CS-g-CA reduced the production of quorum-sensing signaling molecules (AHLs) and the concentration of c-di-GMP in bacteria. Genes for flagellar synthesis (flgA), the resistance to stress (rpoS and hfq), and pde (phosphodiesterases that degrade c-di-GMP) were significantly down-regulated as determined by RT-PCR. Overall, CS-g-CA leads to the accumulation of ROS in bacteria via P. fluorescens environmental resistance genes and decreases the activity of enzymes in the bacterial antioxidant system, and interferes with the production and reception of quorum-sensing signaling molecules and the synthesis of c-di-GMP in P. fluorescens, which regulates the generation of biofilms.

Unlocking fungal quorum sensing: Oxylipins and yeast interactions enhance secondary metabolism in monascus

Exploring the symbiotic potential between fungal and yeast species, this study investigates the co-cultivation dynamics of Monascus, a prolific producer of pharmacologically relevant secondary metabolites, and Wickerhamomyce anomalous. The collaborative interaction between these microorganisms catalyzed a substantial elevation in the biosynthesis of secondary metabolites, prominently Monacolin K and natural pigments. Central to our discoveries was the identification and enhanced production of oxylipins (13S-hydroxyoctadecadienoic acid，13S-HODE), putative quorum-sensing molecules, within the co-culture environment. Augmentation with exogenous oxylipins not only boosted Monacolin K production by over half but also mirrored morphological adaptations in Monascus, affecting both spores and mycelial structures. This augmentation was paralleled by a significant upregulation in the transcriptional activity of genes integral to the Monacolin K biosynthetic pathway, as well as genes implicated in pigment and spore formation. Through elucidating the interconnected roles of quorum sensing, G-protein-coupled receptors, and the G-protein-mediate signaling pathway, this study provides a comprehensive view of the molecular underpinnings facilitating these metabolic enhancements. Collectively, our findings illuminate the profound influence of Wickerhamomyces anomalous co-culture on Monascus purpureus, advocating for oxylipins as a pivotal quorum-sensing mechanism driving the observed symbiotic benefits.

Pseudomonas aeruginosa N-3-Oxododecanoyl Homoserine Lactone Disrupts Endothelial Integrity by Activating the Angiopoietin-Tie System.

The activation of the angiopoietin (Angpt)-Tie system is linked to endothelial dysfunction during sepsis. Bacterial quorum-sensing molecules function as pathogen-associated molecular patterns. However, their impact on the endothelium and the Angpt-Tie system remains unclear. Therefore, this study investigated whether treatment with N-3-oxododecanoyl homoserine lactone (3OC12-HSL), a quorum-sensing molecule derived from Pseudomonas aeruginosa, impaired endothelial function in human umbilical vein endothelial cells. 3OC12-HSL treatment impaired tube formation even at sublethal concentrations, and immunocytochemistry analysis revealed that it seemed to reduce vascular endothelial-cadherin expression at the cell-cell interface. Upon assessing the mRNA expression patterns of genes associated with the Angpt-Tie axis, the expressions of Angpt2, Forkhead box protein O1, Tie1, and vascular endothelial growth factor 2 were found to be upregulated in the 3OC12-HSL-treated cells. Moreover, western blot analysis revealed that 3OC12-HSL treatment increased Angpt2 expression. A co-immunoprecipitation assay was conducted to assess the effect of 3OC12-HSL on the IQ motif containing GTPase activating protein 1 (IQGAP1) and Rac1 complex and the interaction between these proteins was consistently maintained regardless of 3OC12-HSL treatment. Next, recombinant human (rh)-Angpt1 was added to assess whether it modulated the effects of 3OC12-HSL treatment. rh-Angpt1 addition increased cellular viability, improved endothelial function, and reversed the overall patterns of mRNA and protein expression in endothelial cells treated with 3OC12-HSL. Additionally, it was related to the increased expression of phospho-Akt and the IQGAP1 and Rac1 complex. Collectively, our findings indicated that 3OC12-HSL from Pseudomonas aeruginosa can impair endothelial integrity via the activation of the Angpt-Tie axis, which appeared to be reversed by rh-Angpt1 treatment.

Quorum sensing in bacteria: in silico protein analysis, ecophysiology, and reconstruction of their evolutionary history

BackgroundQuorum sensing (QS) is a sophisticated cell-to-cell signalling mechanism that allows the coordination of important processes in microbial populations. The AI-1 and AI-2 autoinducer systems are among the best characterized bacterial QS systems at the genetic level.ResultsIn this study, we present data derived from in silico screening of QS proteins from bacterial genomes available in public databases. Sequence analyses allowed identifying candidate sequences of known QS systems that were used to build phylogenetic trees. Eight categories were established according to the number of genes from the two major QS systems present in each genome, revealing a correlation with specific taxa, lifestyles or metabolic traits. Many species had incomplete QS systems, encoding the receptor protein but not the biosynthesis of the quorum sensing molecule (QSMs). Reconstruction of the evolutionary history of the LuxR family and prediction of the 3D structure of the ancestral protein suggested their monomeric configuration in the absence of the signal molecule and the presence of a cavity for its binding.ConclusionsHere we correlate the taxonomic affiliation and lifestyle of bacteria from different genera with the QS systems encoded in their genomes. Moreover, we present the first ancestral reconstruction of the LuxR QS receptors, providing further insight in their evolutionary history.

Effect of Quorum Sensing Molecules on the Quality of Bacterial Nanocellulose Materials.

Bacterial nanocellulose (BNC) biofilms, produced by various bacterial species, such as Gluconacetobacter xylinus, represent a highly promising multifunctional material characterized by distinctive physiochemical properties. These biofilms have demonstrated remarkable versatility as nano biomaterials, finding extensive applications across medical, defense, electronics, optics, and food industries. In contrast to plant cellulose, BNC biofilms exhibit numerous advantages, including elevated purity and crystallinity, expansive surface area, robustness, and excellent biocompatibility, making them exceptional multifunctional materials. However, their production with consistent morphological properties and their transformation into practical forms present challenges. This difficulty often arises from the heterogeneity in cell density, which is influenced by the presence of N-acyl-homoserine lactones (AHLs) serving as quorum sensing signaling molecules during the biosynthesis of BNC biofilms. In this study, we employed surface characterization methodologies including scanning electron microscopy, energy-dispersive spectroscopy, diffuse reflectance infrared Fourier transform spectroscopy, and atomic force microscopy to characterize BNC biofilms derived from growth media supplemented with varying concentrations of distinct N-acyl-homoserine lactone signaling molecules. The data obtained through these analytical techniques elucidated that the morphological properties of the BNC biofilms were influenced by the specific AHLs, signaling molecules, introduced into the growth media. These findings lay the groundwork for future exploration of leveraging synthetic biology and biomimetic methods for tailoring BNC with predetermined morphological properties.

Small RNAs direct attack and defense mechanisms in a quorum sensing phage and its host

Many, if not all, bacteria use quorum sensing (QS) to control collective behaviors, and more recently, QS has also been discovered in bacteriophages (phages). Phages can produce communication molecules of their own, or "listen in" on the host's communication processes, to switch between lytic and lysogenic modes of infection. Here, we study the interaction of Vibrio cholerae with the lysogenic phage VP882, which is activated by the QS molecule DPO. We discover that induction of VP882 results in the binding of phage transcripts to the major RNA chaperone Hfq, which in turn outcompetes and downregulates host-encoded small RNAs (sRNAs). VP882 itself also encodes Hfq-binding sRNAs, and we demonstrate that one of these sRNAs, named VpdS, promotes phage replication by regulating host and phage mRNA levels. We further show that host-encoded sRNAs can antagonize phage replication by downregulating phage mRNA expression and thus might be part of the host's phage defense arsenal.