ARTICLE | doi:10.20944/preprints202305.1041.v1
Subject: Biology And Life Sciences, Biochemistry And Molecular Biology Keywords: Microbial physiology; Quorum Sensing; Quorum Quenching; Pseudomonas aeruginosa
Online: 15 May 2023 (12:13:55 CEST)
After a time away from the classrooms and laboratories due to the global pandemic, the return to the teaching activities during the semester represented a challenge to both teachers and students. Our particular situation in a Microbial Physiology course was the necessity of imparting in a shorter time, laboratory practices that usually take longer. This article describes a two-week long laboratory exercise that covers several concepts in an interrelated way: conjugation as a gene transfer mechanism, regulation of microbial physiology, production of secondary metabolites, degradation of macromolecules and biofilm formation. Utilizing a Quorum Quenching (QQ) strategy, the Quorum Sensing (QS) system of Pseudomonas aeruginosa is first attenuated. Then, phenotypes regulated by QS are evidenced. QS is a regulatory mechanism of the microbial physiology that relays on signal molecules. QS is related in P. aeruginosa to several virulence factors, some of which are exploited in the laboratory practices presented in this work. QQ is phenomenon by which QS is interrupted or attenuated. We utilized a QQ approach based on the enzymatic degradation of the P. aeruginosa QS signals in order to put in evidence QS-regulated traits that are relevant for our Microbial Physiology course.
Subject: Biology And Life Sciences, Immunology And Microbiology Keywords: Quorum sensing; Quorum quenching; AHL molecules; 16S rRNA
Online: 22 September 2020 (09:43:55 CEST)
N-Acyl-homoserine lactones, the Quorum sensing signaling molecules predominantly found in gram-negative bacteria, which regulate several bacterial genes including virulence and antibiotic resistant genes. The study was aimed to identify and characterize QS and QQ bacteria from different samples. 5 samples with different ecological background were collected from soil and 10 samples from hospital setup. 31 different bacteria were isolated with either QS or QQ activities all together. CV026 and A136 biosensor strains were used for the detection of QS and QQ positive strains. QS activity was observed by cross streaking test of bacteria against CV026, it was affirmed that 13 isolates from the soil and 5 from hospital equipment’s showed positive QS activity. QQ activity of each isolate was tested by well diffusion assay, C6-HSL and C12-HSL were our candidate AHL molecules. The AHL molecule degradation was detected in 4 isolates of soil and none from the samples obtained from hospital setup. The total of 6 strongly positive QS and QQ isolates were identified and selected for 16S rRNA gene sequencing. The phylogenetic analysis revealed that these isolates were closely related to Pseudomonas, Bacillus and Exiguobacterium genera. In contrast, 1 Gram positive bacterial isolate was also purified with QS potential.
ARTICLE | doi:10.20944/preprints201807.0472.v1
Subject: Biology And Life Sciences, Immunology And Microbiology Keywords: chitosan; quorum sensing; antibacterial activity; quorum sensing inhibition
Online: 25 July 2018 (08:32:31 CEST)
New approaches to deal with drug-resistant pathogenic bacteria are urgent. We studied the antibacterial effect of chitosans against an E. coli quorum sensing biosensor reporter strain, and selected a non-toxic chitosan to evaluate its QS inhibition activity and its effect on bacterial aggregation. To this end, chitosans of varying DA (12 to 69%) and Mw (29 to 288 KDa) were studied. Only chitosans of low DA (~12%) inhibited the bacterial growth, regardless of the Mw. Chitosan MDP DA30 (DA 42% and Mw 115 kDa) was selected for further QS inhibition and SEM imaging studies. MDP DA30 chitosan exhibited QS inhibition activity in an inverse dose-dependent manner (≤12.5 µg/mL). SEM images revealed that this chitosan, when added at low concentration (≤30.6 µg/mL), induced substantial bacterial aggregation, whereas at high concentration (234.3 µg/mL), it did not. Aggregation explains the QS inhibition activity as the consequence of retardation of the diffusion of AHL.
REVIEW | doi:10.20944/preprints202305.0976.v1
Subject: Biology And Life Sciences, Immunology And Microbiology Keywords: quorum sensing; quorum sensing inhibitors; Chromobacterium violaceum; plant extracts
Online: 15 May 2023 (05:07:34 CEST)
In the new antibiotic era, the exponential increase of multiresistant bacterial strains become the main global health problem. Many researchers focused their efforts to explore novel or combined strategies for combating bacterial resistance. The good knowledge of molecular mechanisms of resistance and bacterial virulence factors as key targets gives us a good scenario to resolve the problem. One particularly attractive and promising way is to attack the main regulatory “network” of bacterial virulence determinants known as Quorum sensing (QS). The inhibition of QS signals will be a novel way for screening more effective Quorum sensing inhibitors (QSIs) and will put a key role in next-generation antimicrobials in the resistance battle. This determined the aim of the present review: comprehensive clarification of the regulatory mechanisms of quorum-sensing signaling pathways in Chromobacterium violaceum and discovery of potential plant quorum sensing inhibitors.
ARTICLE | doi:10.20944/preprints202111.0105.v1
Subject: Chemistry And Materials Science, Medicinal Chemistry Keywords: quorum sensing; furanones; biofilm
Online: 4 November 2021 (16:18:19 CET)
Clinical evidence has shown that bacterial infections are more difficult to eradicate when form-ing a biofilm aggregate than when are produced by bacteria in planktonic form. Therefore, com-pounds that inhibit biofilm formation could be used against severe infections. It has been re-ported that bromo 2-(5H) furanones inhibited biofilm formation by their anti-quorum sensing properties. To determine if the 2-(5H) furanone moiety is essential to induce inhibition of biofilm formation, we evaluated ten halogen 2-(5H) furanones derivates previously synthesized. Besides evaluating the inhibition of biofilm formation, we assessed pyocyanin production, swarming motility, and transcription of essential QS genes: rsaL, rhlA, pqsA and phz1 genes. Our results showed that although three bromo-furan-2(5H)-one-type derivatives (A1-A3) and two bromo-4-(phenylamino)-furan-2(5H)-one-type compounds (B2 and B6) inhibited the biofilm formation in both P. aeruginosa PA14 (reference) and PA64 (drug-resistant) strains only the furanones A1-A3 were efficient to inhibit QSS.
ARTICLE | doi:10.20944/preprints202304.0914.v1
Subject: Biology And Life Sciences, Immunology And Microbiology Keywords: Metformin; Quorum sensing; Quorum quenching; Pseudomonas aeruginosa; Virulence; Drug repurposing; Anti-virulence therapy; Biofilm inhibition
Online: 25 April 2023 (09:54:20 CEST)
Metformin (MeT) is an FDA-approved drug with a myriad of health benefits. Besides being used as an anti-diabetic drug, MeT is also effective against various cancers, liver-, cardiovascular-, and renal diseases. It has also been proven to demonstrate anti-ageing and neuroprotective effects. This study was undertaken to examine its unique potential as an anti-virulence drug against an opportunistic bacterial pathogen, Pseudomonas aeruginosa. Due to the menace of multidrug resistance in pathogenic microorganisms, many novel or repurposed drugs with anti-virulence prospects are emerging as next-generation therapies with the aim to overshadow the application of existing antimicrobial regimens. The quorum sensing (QS) mechanisms of P. aeruginosa are an attractive drug target for attenuating bacterial virulence. In this context, the anti-QS potential of MeT was scrutinized using biosensor assays. MeT was comprehensively evaluated for its effects on different motility phenotypes, virulence factor production (phenotypic and genotypic expression) along with biofilm development in P. aeruginosa in vitro. At sub-lethal concentrations, MeT displayed prolific quorum quenching (QQ) ability and remarkably inhibited AHL biosynthesis in P. aeruginosa. Moreover, MeT (1/8 MIC) effectively downregulated the expression levels of various QS- and virulence genes in P. aeruginosa, which coincided with a notable reduction in the levels of alginate, hemolysin, pyocyanin, pyochelin, elastase, and protease production. In silico analysis through molecular docking also predicted strong associations between the QS receptors of P. aeruginosa and MeT. MeT also compromised the motility phenotypes and successfully abrogated biofilm formation by inhibiting EPS production in P. aeruginosa. Hence, the QQ, anti-virulence, and anti-fouling potential of MeT was elucidated for the first time against P. aeruginosa.
ARTICLE | doi:10.20944/preprints202208.0050.v1
Subject: Physical Sciences, Applied Physics Keywords: quorum sensing; resistance random network; complex networks
Online: 2 August 2022 (08:21:25 CEST)
We propose a model for bacterial Quorum Sensing based on an auxiliary electrostatic-like interaction originating from a fictitious electrical charge that represents bacteria activity. A cooperative mechanism for charge/activity exchange is introduced to implement chemotaxis and replication. The bacteria system is thus represented by means of a complex resistor network where link resistances take into account the allowed activity-flow among individuals. By explicit spatial stochastic simulations, we show that the model exhibits different quasi-realistic behaviors from colony formation to biofilm aggregation. The electrical signal associated with Quorum Sensing is analyzed in space and time and provides useful information about the colony dynamics. In particular, we analyze the transition between the planktonic and the colony phases as the intensity of Quorum Sensing is varied.
ARTICLE | doi:10.20944/preprints202008.0259.v1
Subject: Medicine And Pharmacology, Dentistry And Oral Surgery Keywords: DMTU; Multispecies biofilms; Porphyromonas gingivalis; Quorum sensing
Online: 11 August 2020 (08:11:20 CEST)
Imbalance of homeostasis between the microbial communities and the host system leads to dysbiosis in oral micro flora. DMTU (1,3-di-m-tolyl-urea), is a biocompatible compound that was shown to inhibit Streptococcus mutansbiofilms by inhibiting its communication system (quorum sensing). Here, we hypothesized that DMTU is able to inhibit multispecies biofilms. We developed a multispecies oral biofilm model comprising an early colonizer Streptococcus gordonii, a bridge colonizer Fusobacterium nucleatum, and late colonizers Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans. We performed comprehensive investigations to demonstrate the effect of DMTU on planktonic cells and biofilms. Our findings showed that DMTU inhibits and disrupts multispecies biofilms without bactericidal effects. Mechanistic studies revealed significant down regulation of biofilm and virulence related genes in P. gingivalis. Taken together, our study highlights the potential of DMTU to inhibit polymicrobial biofilm communities and their virulence.
ARTICLE | doi:10.20944/preprints201912.0398.v1
Subject: Chemistry And Materials Science, Organic Chemistry Keywords: Cladosporium sp.; altertoxins; quorum sensing inhibitory activity
Online: 31 December 2019 (02:31:16 CET)
Five new perylenequinone derivatives, altertoxins VIII-XII (1-5), as well as one known compound cladosporol I (6), were isolated from the fermentation broth of Cladosporium sp. KFD33 from a blood cockle from Haikou Bay, China. Their structures were determined based on spectroscopic methods and ECD spectra analysis along with quantum ECD calculations. Compounds 1-6 exhibited quorum sensing inhibitory activities against Chromobacterium violaceum CV026 with MIC values of 30, 30, 20, 30, 20 and 30 μg/well, respectively.
ARTICLE | doi:10.20944/preprints202307.0402.v1
Subject: Biology And Life Sciences, Immunology And Microbiology Keywords: oral biofilm; caries; quorum sensing; quorum quenching; acylhomoserine lactones; Streptococcus; 16S community profiling; caries-free children; caries-active children
Online: 6 July 2023 (10:25:25 CEST)
Caries are lesions caused by acidic compounds derived from the metabolism of bacteria such as streptococci. It is not merely the presence of acidogenic bacteria that causes caries but an imbalance in the oral biofilm microhabitat and its behavior. One factor underlying biofilm formation is quorum sensing (QS), that in gram-positives relies on peptidic molecules, whereas in gram-negatives is driven by small diffusible signals. Strategies based on QS inhibition have been proposed as alter-natives to antimicrobial therapies. Here, we investigate the antibiofilm potential of the lactonase enzyme Aii20J, previously reported as effective against biofilms of periodontal disease origin. We generated in vitro polymicrobial biofilms dominated by gram-positive taxa using supragingival samples from caries-free and caries-active children. The effect of Aii20J on the biofilms was evalu-ated regarding its biomass-reducing ability, as assessed with crystal violet assay, and its effects on the bacterial composition of the polymicrobial biofilms, as assessed by 16S community profiling. We describe significant biomass reductions upon Aii20J exposure without significant changes in bac-terial composition at the genus level. Our results support the use of Aii20J to prevent oral biofilm formation while highlighting the influence of the polymicrobial environment on bacterial commu-nities and their response to antibiofilm treatments.
REVIEW | doi:10.20944/preprints202306.0051.v1
Subject: Biology And Life Sciences, Biology And Biotechnology Keywords: green synthesis; MOFs; amyloid proteins; prionase; mycotoxins; growth inhibition; quorum sensing; quorum quenching; lactamases; biofilms; chitinases; lactonases; His6-OPH
Online: 1 June 2023 (07:31:58 CEST)
Active research of metal-containing compounds and enzymes as effective antifungal agents is currently noted. The interest in metals is due to the wide variety of ligands that can be used for metals, including chemically synthesized and naturally obtained variants as a result of the so-called "green synthesis". The main mechanism of antifungal action of metals is the triggering of generation and accumulation of reactive oxygen species (ROS). Further action of ROS on various biomolecules is nonspecific. This review highlights various hydrolytic enzymes (glucanases and proteases) that affect the structural elements of fungal cells (cell walls, membranes), fungal quorum sensing molecules, fungal own protective agents (mycotoxins and antibiotics), proteins responsible for the adhesion and formation of stable highly concentrated populations in the form of biofilms. A wide range of the substrates for enzymes allows the use of various mechanisms of their antifungal actions. The prospects of combining two different types of antifungal agents (metals and enzymes) for mycelial fungi and yeast cells are discussed in this review. Special attention is paid to the possible influence of metals on activity of the enzymes and the possible effects of proteins on antifungal activity of metal-containing compounds.
ARTICLE | doi:10.20944/preprints201912.0184.v1
Subject: Chemistry And Materials Science, Medicinal Chemistry Keywords: sponge; quorum sensing; quorum sensing inhibition; N-acyl homoserine lactone; Sarcotragus spinosulus; 3-Br-N-methyltyramine; 5,6-dibromo-N,N-dimethyltryptamine
Online: 13 December 2019 (12:12:54 CET)
Marine sponges, a well documented prolific source of natural products, harbors numerous microbial communities believed to possess N-acyl homoserine lactones (AHLs) mediated Quorum sensing (QS) as one of the mechanisms of interaction. Bacteria and eukaryotic organisms are known to produce molecules that can interfere with QS signaling, thus affecting microbial genetic regulation and function. In the present study, we established the potential for production of both QS signal molecules as well as QS interfering molecules (QSI) in the same sponge species Sarcotragus spinosulus. A total of eighteen saturated acyl chain AHLs were identified along with six putative unsaturated acyl chain AHLs. Bioassay guided purification led to the isolation of two brominated metabolites with QS-interfering activity. The structures of these compounds were elucidated by comparative spectral analysis of 1HNMR and HR-MS data and was identified as 3-Br-N-methyltyramine (1) and 5,6-dibromo-N,N-dimethyltryptamine (2). The QSI activity of compounds 1 and 2 were evaluated using reporter gene assays for long- and short-chain signals (E. coli pSB1075 and E. coli pSB401) and was confirmed by measuring dose dependent inhibition of proteolytic activity and pyocyanin production in P. aeruginosa PAO1. The obtained results showed the co-existence of QS and QSI in S. spinosulus, a complex network which may mediate the orchestrated function of the microbiome within the sponge holobiont.
ARTICLE | doi:10.20944/preprints201908.0096.v1
Subject: Biology And Life Sciences, Immunology And Microbiology Keywords: AgrA protein; biofilm; MRSA; quorum sensing; SarA protein; swarming
Online: 7 August 2019 (10:29:04 CEST)
Background: Staphylococcus aureus (S. aureus) is an opportunistic pathogen and a predominant cause of life-threatening nosocomial infections. Drug resistance in S. aureus is attributed to production of biofilm, which is controlled largely by bacterial quorum sensing (QS) systems. Methodology: In vitro analysis of biofilm inhibition assay was performed using crystal violet staining assay, swarming motility, light microscopy and growth curve analyses. Identification of the major constituents of I. verum fruit extract was performed by GC-MS. Ligand-protein interaction was analyzed by molecular docking investigations. Results: The methanol extract of I. verum inhibited the growth of MRSA at the concentration of 4.8 mg/ml. At the sub-inhibitory concentration (2.4mg/ml), the extract showed significant reduction in biofilmogenesis. Light microscopy analysis confirmed the antibiofilm activity as well as the efficacy in disturbing biofilm architecture. A reduced swarming motility was observed at the lowest concentration of 2.4mg/ml. GC-MS analysis revealed anethol (AL) as the major constituent. The molecular docking analysis attributes the antibiofilm activity to an active ligand AL, which strongly interacted with the active site residues of AgrA and SarA proteins of S. aureus. Conclusion: We report the activities of I. verum to be immensely interfering with QS system and biofilm formation in MRSA.
REVIEW | doi:10.20944/preprints202105.0572.v1
Subject: Biology And Life Sciences, Immunology And Microbiology Keywords: microalgae; marine bacteria; quorum sensing signals; alkyl quinolones; microbial loop
Online: 24 May 2021 (13:34:11 CEST)
Quorum sensing (QS) describes a process by which bacteria can sense the local cell density of their own species, thus enabling them to coordinate gene expression and physiological processes on a community-wide scale. Small molecules called autoinducers or QS signals, which act as intraspecies signals, mediate quorum sensing. As our knowledge of QS has progressed, so too has our understanding of the structural diversity of QS signals, along with the diversity of bacteria conducting QS and the range of ecosystems in which QS takes place. It is now also clear that QS signals are more than just intraspecies signals. QS signals mediate interactions between species of prokaryotes, and between prokaryotes and eukaryotes. In recent years, our understanding of QS signals as mediators of algae–bacteria interactions has advanced such that we are beginning to develop a mechanistic understanding of their effects. This review will summarize the recent efforts to understand how different classes of QS signals contribute to the interactions between planktonic microalgae and bacteria in our oceans, primarily N-acyl-homoserine lactones, their degradation products tetramic acids, and 2-alkyl-4-quinolones. In particular, this review will discuss the ways in which QS signals alter microalgae growth and metabolism, namely as direct effectors of photosynthesis, regulators of the cell cycle, and as modulators of other algicidal mechanisms. Furthermore, the contribution of QS signals to nutrient acquisition is discussed, and finally how microalgae can modulate these small molecules to dampen their effects.
ARTICLE | doi:10.20944/preprints202103.0552.v1
Subject: Biology And Life Sciences, Anatomy And Physiology Keywords: Huma Serum Albumin; Acinetobacter baumannii; quorum sensing; iron; human fluids.
Online: 22 March 2021 (15:49:16 CET)
Acinetobacter baumannii is a nosocomial pathogen capable of causing serious infections associated with high rates of morbidity and mortality. Due to its antimicrobial drug resistance profile, A. baumannii is categorized as an urgent priority pathogen by the Centers for Disease Control and Prevention in the United States and priority group 1 critical microorganism by the World Health Organization. Understanding how A. baumannii adapts to different host environments may provide critical insights into strategically targeting this pathogen with novel antimicrobial and biological therapeutics. Exposure to human fluids was previously shown to alter the gene expression profile of a highly drug susceptible A. baumannii strain A118 leading to persistence and survival of this pathogen. Herein, we explore the impact of human pleural fluid (HPF) and human serum albumin (HSA) on the gene expression profile of a highly multi-drug resistant strain of A. baumannii AB5075. Differential expression was observed for ~30 genes, whose products are involved in quorum sensing, quorum quenching, iron acquisition, fatty acid metabolism, biofilm formation, secretion systems and type IV pilus formation. Phenotypic and further transcriptomic analysis using quantitative RT-PCR confirmed RNA-seq data and pointed out a distinctive role of HSA as the molecule involved in A. baumannii response.
REVIEW | doi:10.20944/preprints202008.0379.v2
Subject: Biology And Life Sciences, Cell And Developmental Biology Keywords: hormone; quorum sensing; receptor; bacteria; fungi; metazoa; plants; microbiota; evolution
Online: 25 September 2020 (10:40:10 CEST)
Cellular communications play pivotal roles in multi-cellular species, but they do so also in uni-cellular species. Moreover, cells communicate with each other not only within the same individual but also with cells in other individuals belonging to the same or other species. These communications occur between two unicellular species, two multicellular species, or between unicellular and multicellular species. The molecular mechanisms involved exhibit diversity and specificity, but they share common basic features which allow common pathways of communication between different, and sometimes very different species. These interactions have been made possible by the high degree of conservation of the basic molecular mechanisms of interaction of many ligand-receptor pairs in evolutionary remote species. These inter-species cellular communications played crucial roles during Evolution and must have been positively selected, particularly when collectively beneficial in hostile environments. We think that communications between cells did not arise after their emergence but was part of the very nature of first cells. Synchronization of populations of non-living protocells through chemical communications may have been a mandatory step towards their emergence as populations of living cells and explain the large commonality of cell communication mechanisms among microorganisms, plants, and animals.
ARTICLE | doi:10.20944/preprints201910.0074.v1
Subject: Medicine And Pharmacology, Pharmacology And Toxicology Keywords: Pseudomonas aeruginosa; Quorum sensing; Virtual screening; E-pharmacophore; Drug discovery.
Online: 7 October 2019 (12:33:33 CEST)
Pseudomonas aeruginosa is an emerging opportunistic pathogen responsible for cystic fibrosis and nosocomial infections. In addition, empirical treatments are become inefficient due to their multiple-antibiotic resistance and extensive colonizing ability. Quorum sensing (QS) plays a vital role in the regulation of virulence factors in P. aeruginosa. Attenuation of virulence by QS inhibition could be an alternative and effective approach to control infections. Therefore, we sought to discover new QS inhibitors (QSIs) against LasR receptor in P. aeruginosa using chemoinformatics. Initially, a structure-based high-throughput virtual screening was performed using the LasR active site that identified 61404 relevant molecules. E-pharmacophore (ADAHH) screening of these molecules rendered 72 QSI candidates. In standard-precision docking, only 7 compounds were found as potential QSIs due to their higher binding affinity to LasR receptor (-7.53 to -10.32 kcal/mol compared to -7.43 kcal/mol of native ligands). The ADMET properties of these compounds were suitable to be QSIs. Later, extra-precision docking and binding energy calculation suggested ZINC19765885 and ZINC72387263 as the most promising QSIs. The dynamic simulation of the docked complexes showed good binding stability and molecular interactions. The current study suggested that these two compounds could be used in P. aeruginosa QS inhibition to combat bacterial infections.
ARTICLE | doi:10.20944/preprints202207.0425.v1
Subject: Biology And Life Sciences, Immunology And Microbiology Keywords: quorum sensing; autoinducer; organic plant extracts; biofilms; swimming motility; swarming motility
Online: 27 July 2022 (15:07:29 CEST)
Quorum sensing (QS) is a form of intra- and inter-species communication system which is employed by bacteria to regulate their collective behavior in a cell population-dependent manner. QS has been implicated in the virulence of several pathogenic bacteria. This work aimed to investigate the anti-quorum sensing (anti-QS) potential of ethanolic extracts of eight aromatic plants of Cyprus namely, Origanum vulgare subsp. hirtum, Rosmarinus officinalis, Salvia officinalis, Lavendula spp., Calendula officinalis, Melissa officinalis, Sideritis cypria, and Aloysia citriodora. We initially assess the effects of the extracts on autoinducer 2 (AI-2) signaling activity, using Vibrio harveyi BB170 as a reported strain. We subsequently assess the effect of the ethanolic extracts on QS-related processes including biofilm formation and swarming and swimming motilities of Escherichia coli MG1655. Of the tested ethanolic extracts those of Origanum vulgare subsp. hirtum, Rosmarinus officinalis, and Salvia officinalis were the most potent AI-2 signaling inhibitors while the extracts from the other plants exhibited low to moderate inhibitory activity. The three ethanolic extracts also inhibited the biofilm formation (>60%) of E. coli MG1655, as well as its swimming and swarming motility in a concentration-dependent manner. These extracts may consider true anti-QS inhibitors because they disrupt QS-related activities of E. coli MG1655 without affecting bacterial growth. The results suggest that plants from the unexplored flora of Cyprus could serve as a source to identify novel anti-QS inhibitors to treat infectious diseases caused by pathogens resistant to antibiotics
Subject: Chemistry And Materials Science, Medicinal Chemistry Keywords: azolines synthesis; quorum sensing; Chromobacterium violaceum CV026; molecular docking; molecular dynamics
Online: 6 May 2019 (08:59:08 CEST)
The increasingly common occurrence of antibiotic-resistant bacteria has become an urgent public health issue. There are currently some infections without any effective treatment, which require new therapeutic strategies. An attractive alternative is the design of compounds capable of disrupting bacterial communication known as quorum sensing (QS). In gram-negative bacteria, such communication is regulated by acyl-homoserine lactones (AHLs). QS allows bacteria to proliferate before expressing virulence factors. Our group previously reported that hexyloxy phenyl imidazoline (9) demonstrated 71% inhibitory activity of QS at 100 µM (IC50=90.9 µM) in Chomobacterium violaceum, a gram-negative bacterium. The aim of the present study was to take 9 as a lead compound to design and synthesize three 2-imidazolines (13-15) and three 2-oxazolines (16-18), to be evaluated as quorum sensing inhibitors on C. violaceum CV026. We were looking for compounds with a higher affinity towards the Cvi receptor of this bacterium and the ability to inhibit QS. The binding mode of the test compounds on the Cvi receptor was explored with docking studies and molecular dynamics. It was found that 8-pentyloxyphenyl-2-imidazoline 13 reduced the production of violacein (IC50=56.38 µM) without affecting bacterial growth, suggesting inhibition of quorum sensing. Indeed, compound 13 is apparently one of the best QS inhibitors known to date. Molecular docking revealed the affinity of compound 13 for the orthosteric site of the N-hexanoyl homoserine lactone (C6-AHL) on the CviR protein. Ten aminoacid residues in the active site of C6-AHL interacted with 13, and 7 of these are the same as those interacting with AHL. Contrarily, 8-octyloxyphenyl-2-imidazoline 14, 8-decyloxyphenyl-2-imidazoline 15 and 9-decyloxyphenyl-2-oxazoline 18 bound only to an allosteric site and thus did not compete with C6-AHL for the orthosteric site.
ARTICLE | doi:10.20944/preprints202306.0628.v1
Subject: Biology And Life Sciences, Immunology And Microbiology Keywords: Staphylococcus aureus; MRSA; mBTL; mBTL-ChNPs; antibiotic resistance; quorum sensing; virulence; biofilm
Online: 8 June 2023 (10:36:45 CEST)
Of late, the focus has been shifted towards quorum sensing inhibitors which reduce bacterial virulence and lower the probability of resistance and refining infections. In this work, meta-bromo-thiolactone (mBTL), a potent quorum and virulence inhibitor against Staphylococcus aureus and MRSA phenotypes, was formulated in chitosan nanoparticles (ChNPs) using ionic gelation method. mBTL-loaded-ChNPs were characterized for particle size, polydispersity index, zeta potential. Morphology was visualized by Transmission Electron Microscopy (TEM), drug release profile and antibiofilm analysis using Confocal Laser Scanning Microscope (CLSM) and Scanning Electron Microscopy (SEM) were performed. Synthesized mBTL-loaded-CNPs showed homogenized nano-size particles ranging from 158+1.3 to 284+5.6 nm with spherical particles that exhibited sustainable release profile over 48 hr at 37 °C. These findings revealed successful preparation of mBTL-loaded-ChNPs, that further showed effective antibiofilm activity at MIC50 (0.5 mg/mL) where all strains displayed reduced biofilm formation compared to untreated strains. CLSM results showed a significant reduction in the number of viable cells, indicating the effectiveness of m-BTL as an antibacterial agent. SEM analysis permitted visualization of biofilm structure in relation to the spatial localization of important biofilm matrix components, the formed biofilms were clearly distinguished in the SEM images. Bacterial cells in the control group were enclosed in thick biofilms. In contrast, there was a considerable reduction in biofilm production when mBTL was present, where bacterial cells seemed less ordered and more scattered with no detectable biofilms. In conclusion, mBTL-loaded-ChNPs is a potential alternative treatment to overcome antimicrobial resistance and condensed MRSA infections.
ARTICLE | doi:10.20944/preprints202005.0267.v1
Subject: Computer Science And Mathematics, Computer Science Keywords: Blockchain; ÐApp; UML; conceptual modelling; Ethereum; Smart Contract; Solidity; Quorum; middleware; Clockchain
Online: 16 May 2020 (16:29:58 CEST)
Blockchain decentralized applications (ÐApps) are applications which run on Blockchains nodes. Thus, in order to interact directly with this sort of applications, users need to have a blockchain address, wallet and knowledge about how to make transactions in order to interact with ÐApps. Therefore, the knowledge required to use a ÐApp can easily make users to desist when trying to interact with them. In order to tackle this issue, we propose a software architecture that will be located in the middle of the user and the ÐApp, thus making users initially unaware that they are interacting with a ÐApp. This is achieved by analyzing the relationship between ÐApps and Apps by using UML modelling. Next, based in the previous analysis, we created a middleware for users to interact with a ÐApp in the same manner the do with a traditional web app, i.e. by using usernames, passwords and UI elements instead of addresses, private keys or transactions. Finally, in order to put the developed middleware into practice, we developed a ÐApp that makes use of it. This ÐApp registers the time control of workers from companies by using Blockchain to store the data in a secure and non-modifiable manner.
REVIEW | doi:10.20944/preprints202306.0101.v1
Subject: Medicine And Pharmacology, Dentistry And Oral Surgery Keywords: Quorum sensing; Autoinducer-2 (AI-2); Acetyl homoserine lactones (AHL); Periodontal diseases; Oral Biofilm
Online: 1 June 2023 (13:54:31 CEST)
Oral diseases remain a major health problem worldwide, with the World Health Organisation reporting that oral health neglect affects almost half of the world’s population. Quorum sensing molecules (QSMs) influence oral biofilms in various ways with knowledge of direct oral QSM-host interaction being limited, and such studies could provide more insight into the cross-kingdom communication occurring during oral disease development. This review aims to explore the literature on oral QSM-host interaction and to highlight areas of advancement in this field. QSM CSP-1 produced by Gram-positive oral bacteria was found to interact with T2R receptors, activating NF-kB signalling and leading to remodeling of the cytoskeleton. AI-2 in the oral cavity was found in various studies to elicit an inflammatory response in specific oral cells. Overall AHL detection methods remain an area for development, through which greater understanding of oral QSMs’ influence on host cells can be achieved. In conclusion, from the current literature it can be inferred that that QSMs are utilised by host cells to detect bacterial presence and in the majority of cases elicit an immune response towards the environmental QSMs. This may provide a base to target QSMs as novel treatment of oral diseases.
REVIEW | doi:10.20944/preprints202307.0847.v1
Subject: Environmental And Earth Sciences, Environmental Science Keywords: quorum sensing; microbial communication; biofilm formation; biogeochemical cycling; biotechnological applications; marine environment; deep‐sea environment
Online: 13 July 2023 (05:26:58 CEST)
Quorum sensing (QS), a well-established phenomenon in microorganisms, involves the communication between cells through chemical signals, which is dependent on cell density. Extensive research has been conducted on this microbial ability, encompassing the early stages of understanding QS to the latest advancements in the identification and characterization of its mechanisms. This minireview comprehensively examines the role of QS in various aspects, including biofilm formation, virulence in pathogenic bacteria such as Helicobacter pylori and Pseudomonas sp., as well as its influence on biogeochemical cycling in deep-sea environments. Furthermore, future progress in the field will be achieved by combining state-of-the-art methods for observing QS in the deep sea with a deeper understanding of the underlying processes, which will facilitate the engineering of microorganisms for improved degradation of persistent environmental pollutants and other biotechnological applications.
REVIEW | doi:10.20944/preprints202308.1824.v1
Subject: Biology And Life Sciences, Life Sciences Keywords: antifungal peptides; enzyme-like activity; growth inhibition; antifungal activity; fungal metabolites; quorum response molecules; combined antifungals
Online: 28 August 2023 (09:44:34 CEST)
Biomimetics, which are similar to natural compounds that play an important role in the metabolism, manifestation of functional activity and reproduction of various fungi, have a pronounced attrac-tion in the current search for new effective antifungals. Actual trends in the development of this area of research indicate that unnatural amino acids can be used as such biomimetics, including those containing halogen atoms; compounds similar to nitrogenous bases embedded in the nucleic acids synthesized by fungi; peptides imitating fungal analogues; molecules similar to natural sub-strates of numerous fungal enzymes and Quorum Sensing signaling molecules of fungi and yeast; etc. Most part of the review is devoted to the analysis of semi-synthetic and synthetic antifungal peptides and their targets of action. This review is aimed at combining and systematizing the cur-rent scientific information accumulating in this area of research, developing various antifungals with an assessment of the effectiveness of the created biomimetics and the possibility of combining them with other antimicrobial substances to reduce cell resistance and improve antifungal effects.
ARTICLE | doi:10.20944/preprints202307.0765.v1
Subject: Medicine And Pharmacology, Pharmacy Keywords: 4-amino-7-chloroquinoline; 1,3,4-oxadiazole; anthranilic acid; synthesis; quorum sensing; anti-biofilm; P. aeruginosa; PQS
Online: 12 July 2023 (21:25:14 CEST)
Inhibition of quorum sensing (QS)-dependent biofilm formation and virulence is extensively investigated as a novel approach to combat bacterial pathogens by impeding their ability to cause antibiotic-tolerant diseases. Such strategies are considered a promising alternative to conventional antibacterial therapy since interrupting the central cell-to-cell communication system imposes less selective pressure on the target pathogen. In this study, novel hybrid compounds composed of anthranilic acids substituted with halogens at different positions of the phenyl ring, and 4-(2-aminoethyl/4-aminobuthyl)amino-7-chloroquinoline linked via 1,3,4-oxadiazole were synthesized using standard procedures. Their anti-QS activities were evaluated using Chromobacterium violaceum ATCC31532 (anti-biofilm and bactericidal activities) and Pseudomonas aeruginosa PAO1 (anti-biofilm/-virulence activities). The results showed that compounds 15–19 and 23 inhibited the production of violacein, the QS-inducible pigment, in C. violaceum to a similar extent as the model QS inhibitor - quercetin (83.5–90%). Compound 15 exhibited the strongest effect on P. aeruginosa in the anti-biofilm screening, reducing its ability to form biofilm by almost 50%, while compounds 16 and 19 were able to reduce the biofilm formation by approximately 30%. However, compound 23 did not demonstrate significant anti-biofilm activity and only inhibited pyocyanin production in P. aeruginosa. In conclusion, this study suggests that 1,3,4-oxadiazoles 15, 16, and 19 are the most promising compounds for future research as novel QS inhibitors against Gram-negative bacteria equipped with different QS signaling pathways.
ARTICLE | doi:10.20944/preprints202308.1409.v1
Subject: Medicine And Pharmacology, Dentistry And Oral Surgery Keywords: periodontal diseases; Porphyromonas gingivalis; plant extract; Symphytum Officinale; Panax Ginseng; metronidazole; acylated homoserine lactones; biofilm; quorum sensing
Online: 21 August 2023 (08:56:14 CEST)
Background: The biofilm-forming bacteria Porphyromonas gingivalis (P. gingivalis) is primarily responsible for periodontal disorders, which continue to be a global health issue. Despite the widespread use of antibiotics like metronidazole, other potent therapeutic a crucial quorum sensing molecule implicated in the formation of P. gingivalis biofilm. Approaches are now being explored due to the growing challenge of antibiotic resistance. Objective: In particular, chemicals from Symphytum Officinale (S) and Panax Ginseng (G) plants will be examined in this study to assess if they have the ability to inhibit biofilm and to prevent the development of acylated homoserine lactones (AHLs), a quorum sensing molecule. Materials and Methods: With Metronidazole serving as a control, the antibacterial effects of these substances (Symphytum Officinale and Panax Ginseng) were examined against a standard strain and a clinical isolate of P. gingivalis. Dilutions of these plant extracts were used either alone (G+S) or in conjunction with metronidazole (G+S+F), to assess their antibacterial activity using antibacterial susceptibility test (inhibition zone), biofilm inhibition and disruption assay (optical density) and quorum sensing inhibition assay (AHL). Results: The combinations of Symphytum officinale, Panax Ginseng and metronidazole (S+G+F) showed the maximum effectiveness with highest zone of inhibition (25.000±0.001 mm) and biofilm inhibition (98.46%), and were comparable to G+S (inhibition zone of 24.341±0.593 mm and biofilm inhibition of 97.76%), and significantly different in the degree of biofilm inhibition with different treatment scenarios. Additionally, the plant's extracts and combinations are specific concentrations had a significant effect on the suppression of the generation of AHL (p < 0.05). Conclusion: According to preliminary research, these plant-derived substances, especially when paired with metronidazole, significantly inhibited the growth of P. gingivalis biofilm providing valuable information for the development of innovative therapeutic approaches for periodontitis and other biofilm-associated illnesses.
REVIEW | doi:10.20944/preprints202105.0131.v1
Subject: Biology And Life Sciences, Anatomy And Physiology Keywords: rhizobia; legume; rhizobium-legume symbiosis; exopolysaccharide synthesis; quorum sensing (QS); nodulation; nod regulon; flavonoids; RosR/MucR; SyrM
Online: 7 May 2021 (09:29:45 CEST)
Abstract: Rhizobia are soil proteobacteria able to engage in a nitrogen-fixing symbiotic interaction with legumes which involves root rhizobial infection and bacterial invasion of new organs formed by the plant in response to the presence of appropriate bacterial partners. This interaction relies on a complex molecular dialogue between both symbionts. Bacterial N-acetyl-glucosamine oligomers called Nod factors are indispensable in most cases for early steps of the symbiotic interaction. In addition, different rhizobial surface polysaccharides, such as exopolysaccharides (EPS), may also be symbiotically relevant. EPS are acidic polysaccharides located out of the cell with little or no cell association that carry out important roles both in free-life and in symbiosis. EPS production is very complexly modulated and, frequently, co-regulated with that of Nod factors, but the type of co-regulation varies depending on the rhizobial strain. Many studies point out a signalling role of EPS-derived oligosaccharides in root infection and nodule invasion but, in certain symbiotic couples, EPS can be dispensable for a successful interaction. In summary, the complex regulation of the production of rhizobial EPS varies in different rhizobia and the relevance of this polysaccharide in symbiosis with legumes depends on the specific interacting couple.
REVIEW | doi:10.20944/preprints202203.0168.v1
Subject: Biology And Life Sciences, Immunology And Microbiology Keywords: cAMP signaling; quorum sensing; alternative splicing; lipid signaling; MAPK cascade; multistep phosphorelay; pheromone signaling; glucose signaling; light signaling
Online: 11 March 2022 (10:15:46 CET)
Biochemical signaling is the key mechanism to coordinate a living organism in all aspects of its life. It is still enigmatic how exactly cells and organisms deal with environmental signals and irritations precisely because of the limited number of signaling proteins and a multitude of transitions inside and outside the cell. Many components of signaling pathways are functionally pleiotropic, which means they have several functions. A single stimulus often activates multiple effectors, a distinct effector can be activated by numerous stimuli and signals triggered by different stimuli are often transduced via shared network components. This very compact and concise review sheds light on the most important molecular mechanisms of cellular signaling in fungi.
ARTICLE | doi:10.20944/preprints201912.0399.v1
Subject: Biology And Life Sciences, Immunology And Microbiology Keywords: natural product; actinobacteria; quorum sensing inhibition (QSI); biosynthetic gene clusters (BGCs); global natural product social networking (GNPS); cyclic dipeptides (2,5-diketopiperazines, DKPs); LC-HRMS
Online: 31 December 2019 (02:59:59 CET)
Streptomyces, being one of the most promising genera due to its ability to synthesize a variety of bioactive secondary metabolites of pharmaceutical interest, here studied in relation to its genomic and metabolomic potential. Coinciding with the increase in sequenced data, mining of bacterial genomes for biosynthetic gene clusters (BGCs) has become a routine component of natural product discovery. Herein, we describe the isolation and characterization of a Streptomyces tendae VITAKN with quorum sensing inhibitory activity (QSI) that was isolated from southern coastal parts of India. The nearly complete genome consists of 8,621,231bp with a GC content of 72.2%. Utilizing the BiG-SCAPE-CORASON platform, a sequence similarity network predicted from this strain was evaluated through sequence similarity analysis with the MIBiG database and existing 3,365 BGCs predicted by antiSMASH analysis of publicly available complete Streptomyces genomes. Crude extract analyzed on LC-HRMS/MS and Global Natural Product Social Molecular Networking (GNPS) online workflow using dereplication resulted in the identification of cyclic dipeptides (2,5-diketopiperazines, DKPs) in the extract, which are known to possess QSI activity. Our results highlight the potential use of genomic mining coupled with LC-HRMS/MS and bionformatic tools (GNPS) as a potent approach for metabolome studies in discovering novel QSI lead compounds. This study also provides the biosynthetic diversity of these BGCs and an assessment of the predicted chemical space yet to be discovered.