scholarly journals Ibuprofen-mediated potential inhibition of biofilm development and quorum sensing inPseudomonas aeruginosa

2019 ◽  
Author(s):  
Lu Dai ◽  
Tian-qi Wu ◽  
Zhong-hui Wang ◽  
Ruoyu Yuan ◽  
Ye Ding ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Quorum Sensing ◽  
Virulence Factors ◽  
Mass Spectrometry Analysis ◽  
Significant Suppression ◽  
Spectrometry Analysis ◽  
Acridine Orange Staining ◽  
Drug Concentrations ◽  
Associated Proteins ◽  
Biofilm Development

AbstractPseudomonas aeruginosais one of the leading causes of opportunistic and hospital-acquired infections worldwide. The infection withP. aeruginosais frequently linked with clinical treatment difficulties given drug resistance and abuse of antibiotics. Ibuprofen, a widely used non-steroidal anti-inflammatory drug, has been previously reported to exert antimicrobial activity, although the specific mechanism of its action requires additional investigation. Given the regulation effects on quorum sensing (QS), we hypothesized that inhibition ofP. aeruginosawith ibuprofen is linked with the QS systems. First, we assessed the action of ibuprofen inP. aeruginosaby measuring CFU. The antimicrobial activity of ibuprofen was evaluated by crystal violent staining and acridine orange staining at various drug concentrations (0, 50, 75, and 100 μg/mL). Moreover, the effect of ibuprofen on different QS virulence factors, such as pyocyanin, elastase, protease, and rhamnolipids, was assessed revealing a concentration-dependent decrease (P<0.05). The effect of ibuprofen was confirmed by liquid chromatography/mass spectrometry analysis of 3-oxo-C12-HSL and C4-HSL production. In addition, qRT-PCR results identified significant suppression of Las and Rhl gene expression after 18 hours of treatment with ibuprofen (P<0.05), with the most significant suppression observed at the concentration of 75 μg/mL. Functional complementation with exogenous 3-oxo-C12-HSL and C4-HSL suggested that C4-HSL can recover the production of virulence factors and biofilm formation inP. aeruginosa.Molecular docking of ibuprofen with QS-associated proteins revealed high binding affinity. In summary, the results suggest that ibuprofen is a candidate drug for the treatment of clinical infections withP. aeruginosa.

Genes Encoding Bacteriocins and Their Expression and Potential Virulence Factors of Enterococci Isolated from Wood Pigeons (Columba palumbus)

2006 ◽  
Vol 69 (3) ◽  
pp. 520-531 ◽  
Author(s):  
MARÍA MARTÍN ◽  
JORGE GUTIÉRREZ ◽  
RAQUEL CRIADO ◽  
CARMEN HERRANZ ◽  
LUIS M. CINTAS ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Virulence Factors ◽  
Virulence Genes ◽  
Pcr Amplification ◽  
Virulence Gene ◽  
Antagonistic Activity ◽  
Mass Spectrometry Analysis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virulence Determinants ◽  
Spectrometry Analysis

Samples of the intestinal content and carcasses of wood pigeons (Columba palumbus) were evaluated for enterococci with antimicrobial activity. Enterococcus faecium comprised the largest enterococcal species with antagonistic activity, followed by Enterococcus faecalis and Enterococcus columbae. PCR amplification of genes coding bacteriocins and determination of their nucleotide sequence, and the use of specific antipeptide bacteriocin antibodies and a noncompetitive indirect enzyme-linked immunosorbent assay, permitted characterization of enterococci coding that described bacteriocins and their expression. The efaAfm determinant was the only virulence gene detected in E. faecium, whereas E. faecalis showed a larger number of virulence determinants, and E. columbae did not carry any of the virulence genes examined. Although all E. faecalis isolates manifested a potent direct antimicrobial activity, no activity was detected in supernatants of producer cells. Purification of the antagonistic activity of E. columbae PLCH2 showed multiple chromatographic fragments after matrix-assisted laser desorption–ionization time-of-flight mass spectrometry analysis, suggesting the active peptide(s) had not yet purified to homogeneity. Bacteriocinogenic E. faecium and E. columbae isolates may be considered hygienic for production of enterocins and potentially safe due to their low incidence of potential virulence genes and susceptibility of most relevant clinical antibiotics. However, the presence among the enterococci of E. faecalis strains with a potent antagonistic activity and multiple virulence factors is an issue that must be considered further.

scholarly journals The Oligopeptide Transport System Is Essential for the Development of Natural Competence in Streptococcus thermophilus Strain LMD-9

2009 ◽  
Vol 191 (14) ◽  
pp. 4647-4655 ◽  
Author(s):  
Rozenn Gardan ◽  
Colette Besset ◽  
Alain Guillot ◽  
Christophe Gitton ◽  
Véronique Monnet
Keyword(s):  
Quorum Sensing ◽  
Streptococcus Thermophilus ◽  
Defined Medium ◽  
Mass Spectrometry Analysis ◽  
Transport Systems ◽  
Label Free ◽  
Spectrometry Analysis ◽  
Proteomic Approach ◽  
Liquid Chromatography Tandem Mass ◽  
Oligopeptide Transport

ABSTRACT In gram-positive bacteria, oligopeptide transport systems, called Opp or Ami, play a role in nutrition but are also involved in the internalization of signaling peptides that take part in the functioning of quorum-sensing pathways. Our objective was to reveal functions that are controlled by Ami via quorum-sensing mechanisms in Streptococcus thermophilus, a nonpathogenic bacterium widely used in dairy technology in association with other bacteria. Using a label-free proteomic approach combining one-dimensional electrophoresis with liquid chromatography-tandem mass spectrometry analysis, we compared the proteome of the S. thermophilus LMD-9 to that of a mutant deleted for the subunits C, D, and E of the ami operon. Both strains were grown in a chemically defined medium (CDM) without peptides. We focused our attention on proteins that were no more detected in the ami deletion mutant. In addition to the three subunits of the Ami transporter, 17 proteins fulfilled this criterion and, among them, 7 were similar to proteins that have been identified as essential for transformation in S. pneumoniae. These results led us to find a condition of growth, the early exponential state in CDM, that allows natural transformation in S. thermophilus LMD-9 to turn on spontaneously. Cells were not competent in M17 rich medium. Furthermore, we demonstrated that the Ami transporter controls the triggering of the competence state through the control of the transcription of comX, itself controlling the transcription of late competence genes. We also showed that one of the two oligopeptide-binding proteins of strain LMD-9 plays the predominant role in the control of competence.

scholarly journals Two Homologous Agr-Like Quorum-Sensing Systems Cooperatively Control Adherence, Cell Morphology, and Cell Viability Properties in Lactobacillus plantarum WCFS1

2008 ◽  
Vol 190 (23) ◽  
pp. 7655-7665 ◽  
Author(s):  
Toshio Fujii ◽  
Colin Ingham ◽  
Jiro Nakayama ◽  
Marke Beerthuyzen ◽  
Ryoko Kunuki ◽  
...  
Keyword(s):  
Cell Wall ◽  
Quorum Sensing ◽  
Lactobacillus Plantarum ◽  
Cell Morphology ◽  
Response Regulator ◽  
Regulatory System ◽  
Mass Spectrometry Analysis ◽  
Cell Wall Polysaccharide ◽  
Transcription Analysis ◽  
Spectrometry Analysis

ABSTRACT A two-component regulatory system of Lactobacillus plantarum, encoded by genes designated lamK and lamR (hpk10 and rrp10), was studied. The lamK and lamR genes encode proteins which are highly homologous to the quorum-sensing histidine kinase LamC and the response regulator LamA, respectively. Transcription analysis of the lamKR operon and the lamBDCA operon and liquid chromatography-mass spectrometry analysis of production of the LamD558 autoinducing peptide were performed for ΔlamA, ΔlamR, ΔlamA ΔlamR deletion mutants and a wild-type strain. The results suggested that lamA and lamR are cooperating genes. In addition, typical phenotypes of the ΔlamA mutant, such as reduced adherence to glass surfaces and filamentous cell morphology, were enhanced in the ΔlamA ΔlamR mutant. Microarray analysis suggested that the same cell wall polysaccharide synthesis genes, stress response-related genes, and cell wall protein-encoding genes were affected in the ΔlamA and ΔlamA ΔlamR mutants. However, the regulation ratio was more significant for the ΔlamA ΔlamR mutant, indicating the cooperative effect of LamA and LamR.

scholarly journals GAS CHROMATOGRAPHY-MASS SPECTROMETRY ANALYSIS AND IN VITRO ANTIMICROBIAL SCREENING OF WEDELIA GLAUCA (ORTEGA) O. HOFFM. EX HICKEN

2017 ◽  
Vol 10 (11) ◽  
pp. 109
Author(s):  
Krishnavignesh L Krishnavignesh ◽  
Mahalakshmipriya A ◽  
Ramesh M
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Antimicrobial Activity ◽  
Mass Spectrometry Analysis ◽  
Gas Chromatography Mass Spectrometry ◽  
Diffusion Method ◽  
Spectrometry Analysis ◽  
Chromatography Mass Spectrometry ◽  
Ms Analysis

  Objective: Continued resistance toward the antibiotics urges us to explore newer antibiotics. Plants are being the safer source of antibiotics with lesser or no side effects. This study was designed to study the presence of phytochemical constituents and antibacterial activity of leaf and flower extracts of Wedelia glauca against urinary tract infection causing pathogens.Methods: The plant leaves were extracted with five different solvents based on the polarity. The extraction was done using soxhalation. Antimicrobial activity was determined by agar well diffusion method for both the sample and standard. The acetone plant extract was subjected to gas chromatography-mass spectrometry (GC-MS) analysis for screening phytoconstituents.Results: Preliminary phytochemical screening revealed the presence of diverse phytoconstituents in the plant. The different extracts exhibited a considerable antimicrobial potential. Among the solvents used acetone extract showed comparably better antimicrobial activity with 100% of inhibition rate with the maximum zone of inhibition of 1.6±0.77 mm against Staphylococcus sp. and Aspergillus sp. at the concentration of 5 mg. GC-MS analysis provided 8 major peaks which revealed the existence of a variety of bioactive compounds which may attribute to the efficacy of the plant.Conclusion: W. glauca leaf and flower extracts displayed a broad spectrum of antibacterial and antifungal activity and can be considered as a potential source of newer antibiotic compounds.

scholarly journals Proteomic Identification of an Endogenous Synaptic SUMOylome in the Developing Rat Brain

2021 ◽  
Vol 14 ◽  
Author(s):  
Marie Pronot ◽  
Félicie Kieffer ◽  
Anne-Sophie Gay ◽  
Delphine Debayle ◽  
Raphaël Forquet ◽  
...  
Keyword(s):  
Subcellular Fractionation ◽  
Mass Spectrometry Analysis ◽  
Mammalian Brain ◽  
Synaptic Organization ◽  
Post Translational Modifications ◽  
Spectrometry Analysis ◽  
Functional Changes ◽  
Neuronal Communication ◽  
Associated Proteins ◽  
And Function

Synapses are highly specialized structures that interconnect neurons to form functional networks dedicated to neuronal communication. During brain development, synapses undergo activity-dependent rearrangements leading to both structural and functional changes. Many molecular processes are involved in this regulation, including post-translational modifications by the Small Ubiquitin-like MOdifier SUMO. To get a wider view of the panel of endogenous synaptic SUMO-modified proteins in the mammalian brain, we combined subcellular fractionation of rat brains at the post-natal day 14 with denaturing immunoprecipitation using SUMO2/3 antibodies and tandem mass spectrometry analysis. Our screening identified 803 candidate SUMO2/3 targets, which represents about 18% of the synaptic proteome. Our dataset includes neurotransmitter receptors, transporters, adhesion molecules, scaffolding proteins as well as vesicular trafficking and cytoskeleton-associated proteins, defining SUMO2/3 as a central regulator of the synaptic organization and function.

scholarly journals Trp-Containing Antibacterial Peptides Impair Quorum Sensing and Biofilm Development in Multidrug-Resistant Pseudomonas aeruginosa and Exhibit Synergistic Effects With Antibiotics

2021 ◽  
Vol 12 ◽  
Author(s):  
Dejing Shang ◽  
Xue Han ◽  
Wanying Du ◽  
Zhiru Kou ◽  
Fengquan Jiang
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Antibacterial Activity ◽  
Quorum Sensing ◽  
Virulence Factors ◽  
Biofilm Formation ◽  
Efflux Pump ◽  
Synergistic Effects ◽  
Multidrug Resistant ◽  
Biofilm Development ◽  
Antibiotic Efflux

Pseudomonas aeruginosa uses quorum sensing (QS) to control virulence, biofilm formation and antibiotic efflux pump expression. The development of effective small molecules targeting the QS system and biofilm formation represents a novel attractive strategy. In this present study, the effects of a series of Trp-containing peptides on the QS-regulated virulence and biofilm development of multidrug-resistant P. aeruginosa, as well as their synergistic antibacterial activity with three classes of traditional chemical antibiotics were investigated. The results showed that Trp-containing peptides at low concentrations reduced the production of QS-regulated virulence factors by downregulating the gene expression of both the las and rhl systems in the strain MRPA0108. Biofilm formation was inhibited in a concentration-dependent manner, which was associated with extracellular polysaccharide production inhibition by downregulating pelA, algD, and pslA transcription. These changes correlated with alterations in the extracellular production of pseudomonal virulence factors and swarming motility. In addition, the combination of Trp-containing peptides at low concentration with the antibiotics ceftazidime and piperacillin provided synergistic effects. Notably, L11W and L12W showed the highest synergy with ceftazidime and piperacillin. A mechanistic study demonstrated that the Trp-containing peptides, especially L12W, significantly decreased β-lactamase activity and expression of efflux pump genes OprM, MexX, and MexA, resulting in a reduction in antibiotic efflux from MRPA0108 cells and thus increasing the antibacterial activity of these antibiotics against MRPA0108.

scholarly journals Enterobacter cancerogenus produces short chain N-3-oxo-acylhomoserine lactones quorum sensing molecules

2015 ◽  
Author(s):  
Kok-Gan Chan ◽  
Wen-Si Tan
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
High Resolution ◽  
Quorum Sensing ◽  
Homoserine Lactone ◽  
Mass Spectrometry Analysis ◽  
Liquid Chromatography Mass Spectrometry ◽  
Spectrometry Analysis ◽  
Synthesis Activity

Enterobacter cancerogenus strain M004 genome size is 5.67 Mb. Here, its luxI homologue, designated as ecnI which is ecnI gene (633 bp) was cloned and overexpressed. Its AHL synthesis activity was verified using the high-resolution liquid chromatography-mass spectrometry analysis revealed the production of N-(3-oxo-hexanoyl)-L-homoserine lactone (3-oxo-C6-HSL) and N-(3-oxo-hexanoyl)-L-homoserine lactone (3-oxo-C8-HSL). The cloning and characterization of luxI homologue of E. cancerogenus strain M004 was firstly reported here.

scholarly journals Identification of Extracellular N-Acylhomoserine Lactone Acylase from a Streptomyces sp. and Its Application to Quorum Quenching

2005 ◽  
Vol 71 (5) ◽  
pp. 2632-2641 ◽  
Author(s):  
Sun-Yang Park ◽  
Hye-Ok Kang ◽  
Hak-Sun Jang ◽  
Jung-Kee Lee ◽  
Bon-Tag Koo ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Pathogenic Bacteria ◽  
Quorum Quenching ◽  
Homoserine Lactone ◽  
Amino Acid Sequences ◽  
Amide Bond ◽  
Mass Spectrometry Analysis ◽  
Penicillin G ◽  
Spectrometry Analysis ◽  
Acyl Chains

ABSTRACT N-Acylhomoserine lactones (AHLs) play an important role in regulating virulence factors in pathogenic bacteria. Recently, the enzymatic inactivation of AHLs, which can be used as antibacterial targets, has been identified in several soil bacteria. In this study, strain M664, identified as a Streptomyces sp., was found to secrete an AHL-degrading enzyme into a culture medium. The ahlM gene for AHL degradation from Streptomyces sp. strain M664 was cloned, expressed heterologously in Streptomyces lividans, and purified. The enzyme was found to be a heterodimeric protein with subunits of approximately 60 kDa and 23 kDa. A comparison of AhlM with known AHL-acylases, Ralstonia strain XJ12B AiiD and Pseudomonas aeruginosa PAO1 PvdQ, revealed 35% and 32% identities in the deduced amino acid sequences, respectively. However, AhlM was most similar to the cyclic lipopeptide acylase from Streptomyces sp. strain FERM BP-5809, exhibiting 93% identity. A mass spectrometry analysis demonstrated that AhlM hydrolyzed the amide bond of AHL, releasing homoserine lactone. AhlM exhibited a higher deacylation activity toward AHLs with long acyl chains rather than short acyl chains. Interestingly, AhlM was also found to be capable of degrading penicillin G by deacylation, showing that AhlM has a broad substrate specificity. The addition of AhlM to the growth medium reduced the accumulation of AHLs and decreased the production of virulence factors, including elastase, total protease, and LasA, in P. aeruginosa. Accordingly, these results suggest that AHL-acylase, AhlM could be effectively applied to the control of AHL-mediated pathogenicity.

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