scholarly journals Inhibitory Effects of Ethyl Gallate on Streptococcus mutans Biofilm Formation by Optical Profilometry and Gene Expression Analysis

Molecules ◽  
2019 ◽  
Vol 24 (3) ◽  
pp. 529 ◽  
Author(s):  
Vika Gabe ◽  
Tomas Kacergius ◽  
Saleh Abu-Lafi ◽  
Povilas Kalesinskas ◽  
Mahmud Masalha ◽  
...  
Keyword(s):  
Gene Expression ◽  
Biofilm Formation ◽  
Fold Change ◽  
Solid Surfaces ◽  
Dependent Manner ◽  
Ethyl Gallate ◽  
Optical Profilometry ◽  
Expression Change ◽  
Significant Gene ◽  
Glass Surfaces

This study aimed to test the effectiveness of ethyl gallate (EG) against S. mutans biofilm formation on solid surfaces (polystyrene, glass) and acidogenicity, and to examine the effect on expression of related genes. The biofilm that is formed by S. mutans bacteria was evaluated using colorimetric assay and optical profilometry, while the pH of the biofilm growth medium was measured with microelectrode. The expression of genes encoding glucan binding protein B (gbpB), glucosyltranferases B, -C, -D (gtfB, -C, -D) and F-ATPase (atpD, atpF) was assessed using a quantitative reverse transcription-polymerase chain reaction (RT-qPCR). It was revealed that all of the EG concentrations significantly suppressed S. mutans biofilm build-up on polystyrene and glass surfaces, and inhibited acidogenicity, in a dose-dependent manner, compared to the activity of untreated bacteria (p < 0.05). The highest concentration of EG (3.53 mM) reduced biofilm formation on polystyrene and glass surfaces by 68% and more than 91%, respectively, and prevented a decrease in pH levels by 95%. The RT-qPCR data demonstrate that the biofilm-producing bacteria treated with EG underwent significant gene expression changes involving the gtfC (a 98.6 increase in fold change), gtfB gene (a 47.5 increase in fold change) and the gbpB gene (a 13.8 increase in fold change). However, for the other genes tested (gtfD, atpD and atpF), the EG treatments did not produce significant expression change compared to the control. EG produced significant gene expression change in three genes—gtfC, gtfB, and gbpB; it has the capacity to inhibit S. mutans biofilm formation on solid surfaces (polystyrene, glass), as well as acidogenicity. Therefore, EG might be used as an antibiofilm and/or anticaries agent for oral formulations in order to reduce the prevalence of dental caries.

scholarly journals Suppressive Effects of Octyl Gallate on Streptococcus mutans Biofilm Formation, Acidogenicity, and Gene Expression

Molecules ◽  
2019 ◽  
Vol 24 (17) ◽  
pp. 3170 ◽  
Author(s):  
Vika Gabe ◽  
Tomas Kacergius ◽  
Saleh Abu-Lafi ◽  
Mouhammad Zeidan ◽  
Basheer Abu-Farich ◽  
...  
Keyword(s):  
Gene Expression ◽  
Dental Caries ◽  
Streptococcus Mutans ◽  
Biofilm Formation ◽  
Colorimetric Method ◽  
Solid Surfaces ◽  
Dependent Manner ◽  
Oral Diseases ◽  
Expression Change ◽  
Biofilm Development

The accumulation of biofilm by Streptococcus mutans bacteria on hard tooth tissues leads to dental caries, which remains one of the most prevalent oral diseases. Hence, the development of new antibiofilm agents is of critical importance. The current study reports the results from testing the effectiveness of octyl gallate (C8-OG) against: (1) S. mutans biofilm formation on solid surfaces (polystyrene, glass), (2) acidogenicity, (3) and the expression of biofilm-related genes. The amount of biofilm formed by S. mutans bacteria was evaluated using the colorimetric method and optical profilometry. The pH of the biofilm growth medium was measured with microelectrode. A quantitative reverse transcription-polymerase chain reaction (RT-qPCR) was used to assess the expression of genes encoding glucan binding protein B (gbpB), glucosyltransferases B, -C, -D (gtfB, -C, -D), and the F-ATPase β subunit of the F1 protein (atpD). The results show that C8-OG significantly diminished biofilm formation by exposed S. mutans on solid surfaces and suppressed acidogenicity in a dose-dependent manner, compared to unexposed bacteria (p < 0.05). The C8-OG concentration of 100.24 µM inhibited S. mutans biofilm development on solid surfaces by 100% and prevented a decrease in pH levels by 99%. In addition, the RT-qPCR data demonstrate that the biofilm-producing bacteria treated with C8-OG underwent a significant reduction in gene expression in the case of the four genes under study (gbpB, gtfC, gtfD, and atpD), and there was a slight decrease in expression of the gtfB gene. However, C8-OG treatments did not produce significant expression change compared to the control for the planktonic cells, although there was a significant increase for the atpD gene. Therefore, C8-OG might be a potent antibiofilm and/or anticaries agent for oral formulations that aim to reduce the prevalence of dental caries.

scholarly journals Lauryl Gallate Activity and Streptococcus mutans: Its Effects on Biofilm Formation, Acidogenicity and Gene Expression

Molecules ◽  
2020 ◽  
Vol 25 (16) ◽  
pp. 3685
Author(s):  
Vika Gabe ◽  
Mouhammad Zeidan ◽  
Tomas Kacergius ◽  
Maksim Bratchikov ◽  
Mizied Falah ◽  
...  
Keyword(s):  
Gene Expression ◽  
Streptococcus Mutans ◽  
Biofilm Formation ◽  
Solid Surfaces ◽  
Dependent Manner ◽  
Ethyl Gallate ◽  
Planktonic Cells ◽  
Biofilm Growth ◽  
Biofilm Development ◽  
Lauryl Gallate

Streptococcus mutans bacterium is implicated in the pathogenesis of dental caries due to the production of biofilm and organic acids from dietary sucrose. Despite the availability of various means of prophylaxis, caries still has a high worldwide prevalence. Therefore, it is important to find new pharmaceuticals to inhibit S. mutans biofilm formation and acidogenicity. The aim of the current study was to evaluate the activity of lauryl gallate (dodecyl gallate) against S. mutans acidogenicity, the expression of biofilm-associated genes, and biofilm development on solid surfaces (polystyrene, glass). The biofilm quantities produced by S. mutans bacteria were assessed using colorimetric and optical profilometry techniques. Acidogenicity was evaluated by measuring the pH of the biofilm growth medium with microelectrode. Assessment of the expression of gene coding for glucan-binding protein B (gbpB), glucosyltranferases B, -C, -D (gtfB, -C, -D), and the F-ATPase β subunit of F1 protein (atpD) was carried out using a quantitative reverse transcription-polymerase chain reaction (RT-qPCR). The results demonstrate the capacity of lauryl gallate to significantly inhibit S. mutans acidogenicity and biofilm development on solid surfaces, in a dose-dependent manner, compared to untreated bacteria (p < 0.05). The highest activity of lauryl gallate occurred at a concentration of 98.98 µM, at which it suppressed biofilm formation by 100% and lowered pH levels by 98%. The effect of lauryl gallate treatment on gene expression changes, as demonstrated by our RT-qPCR data, was limited to the gtfD gene only, was a significant (48%) decrease in gene expression, obtained for the biofilm-producing bacteria, while a 300% increase in fold change for the same gene occurred in the planktonic cells. It is important to note that in previous studies we showed a broader effect of related derivatives. However, a similar magnitude of difference in effects between biofilm and planktonic cells for the atpD gene was obtained after treatment with octyl gallate and reverse magnitude for the same gene after treatment with ethyl gallate. Therefore, to ascertain the possible direct or indirect effects of lauryl gallate, as well as octyl gallate and ethyl gallate, more research is needed to examine the effects on the amount of enzymes and on the enzymatic activity of the products of the affected genes that are involved in the production and maintenance of biofilm by S. mutans.

An Alanine-Rich Peptide Attenuates Quorum Sensing-Regulated Virulence and Biofilm Formation in Staphylococcus aureus

2019 ◽  
Vol 102 (4) ◽  
pp. 1228-1234 ◽  
Author(s):  
Raid Al Akeel ◽  
Ayesha Mateen ◽  
Rabbani Syed
Keyword(s):  
Gene Expression ◽  
Staphylococcus Aureus ◽  
Quorum Sensing ◽  
Biofilm Formation ◽  
Bacterial Attachment ◽  
The Body ◽  
Dependent Manner ◽  
Virulence Determinants ◽  
Microarray Gene Expression ◽  
Concentration Dependent Manner

Abstract Background: Alanine-rich proteins/peptides (ARP), with bioactivity of up to 20 amino acid residues, can be observed by the body easily during gastrointestinal digestion. Objective: Populus trichocarpa extract’s capability to attenuate quorum sensing-regulated virulence and biofilm formation in Staphylococcus aureus is described. Methods: PT13, an ARP obtained from P. trichocarpa, was tested for its activity against S. aureus using the broth microdilution test; a crystal-violet biofilm assay was performed under a scanning electron microscope. The production of various virulence factors was estimated with PT13 treatment. Microarray gene expression profiling of PT13-treated S. aureus was conducted and compared with an untreated control. Exopolysaccharides (EPS) was estimated to observe the PT13 inhibition activity. Results: PT13 was antimicrobial toward S. aureus at different concentrations and showed a similar growth rate in the presence and absence of PT13 at concentrations ≤8 μg/mL. Biofilm production was interrupted even at low concentrations, and biofilm-related genes were down-regulated when exposed to PT13. The genes encoding cell adhesion and bacterial attachment protein were the major genes suppressed by PT13. In addition, hemolysins, clumping activity, and EPS production of S. aureus decreased after treatment in a concentration-dependent manner. Conclusions: A long-chain PT13 with effective actions that, even at low concentration levels, not only regulated the gene expression in the producer organism but also blocked the virulence gene expression in this Gram-positive human pathogen is described. Highlights: We identified a PT13 as a potential antivirulence agent that regulated production of bacterial virulence determinants (e.g., toxins, enzymes and biofilm), downwards and it may be a promising anti-virulence agent to be further developed as an anti-infective agent.

scholarly journals Cigarette smoke alters the transcriptome of non-involved lung tissue in lung adenocarcinoma patients

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Giulia Pintarelli ◽  
Sara Noci ◽  
Davide Maspero ◽  
Angela Pettinicchio ◽  
Matteo Dugo ◽  
...  
Keyword(s):  
Gene Expression ◽  
Lung Adenocarcinoma ◽  
Cigarette Smoke ◽  
Lung Tissue ◽  
Gene Networks ◽  
Lung Parenchyma ◽  
Fold Change ◽  
Differentially Expressed ◽  
Significant Gene ◽  
Never Smokers

Abstract Alterations in the gene expression of organs in contact with the environment may signal exposure to toxins. To identify genes in lung tissue whose expression levels are altered by cigarette smoking, we compared the transcriptomes of lung tissue between 118 ever smokers and 58 never smokers. In all cases, the tissue studied was non-involved lung tissue obtained at lobectomy from patients with lung adenocarcinoma. Of the 17,097 genes analyzed, 357 were differentially expressed between ever smokers and never smokers (FDR < 0.05), including 290 genes that were up-regulated and 67 down-regulated in ever smokers. For 85 genes, the absolute value of the fold change was ≥2. The gene with the smallest FDR was MYO1A (FDR = 6.9 × 10−4) while the gene with the largest difference between groups was FGG (fold change = 31.60). Overall, 100 of the genes identified in this study (38.6%) had previously been found to associate with smoking in at least one of four previously reported datasets of non-involved lung tissue. Seven genes (KMO, CD1A, SPINK5, TREM2, CYBB, DNASE2B, FGG) were differentially expressed between ever and never smokers in all five datasets, with concordant higher expression in ever smokers. Smoking-induced up-regulation of six of these genes was also observed in a transcription dataset from lung tissue of non-cancer patients. Among the three most significant gene networks, two are involved in immunity and inflammation and one in cell death. Overall, this study shows that the lung parenchyma transcriptome of smokers has altered gene expression and that these alterations are reproducible in different series of smokers across countries. Moreover, this study identified a seven-gene panel that reflects lung tissue exposure to cigarette smoke.

Abstract 174: Regulation of Post-Stroke Gene Expression and Pathophysiology by an Enhancer RNA

Stroke ◽  
2020 ◽  
Vol 51 (Suppl_1) ◽  
Author(s):  
Sunil Bhattarai ◽  
Aparna Akella ◽  
Asad Ghani ◽  
Atish Gandhi ◽  
Ashutosh A Dharap
Keyword(s):  
Gene Expression ◽  
Group Versus ◽  
Fold Change ◽  
Luciferase Reporter ◽  
Mitochondrial Activity ◽  
Cell Fractionation ◽  
Dependent Manner ◽  
Post Stroke ◽  
Genome Wide ◽  
Absolute Fold Change

Enhancer elements orchestrate genomic regulatory interactions to facilitate gene expression and phenotypic changes in development and disease. Recent studies have shown that mammalian enhancers are transcribed into noncoding enhancer RNAs (eRNAs) in a stimulus-dependent manner and some of these eRNAs have central roles in enhancer-mediated gene induction. Due to the very recent discovery of eRNAs, their expression and functions in cerebral ischemia are virtually unknown. Recently, we applied a combination of genome-wide H3K27ac ChIP-seq and genome-wide RNA-seq in ipsilateral cortices from adult male C57BL6/N mice following a 1 h middle cerebral artery occlusion (or sham surgery) and 6 h of reperfusion (n=6/group) to identify enhancer elements and stroke-induced eRNAs. We identified 77 eRNAs that were significantly upregulated (absolute fold-change ≥2.0; q<0.05) in stroke as compared to sham, of which 55 were exclusively expressed in stroke. All 77 eRNAs were novel, unspliced transcripts. Using luciferase reporter assays, we confirmed the enhancer capabilities of multiple eRNA-encoding genomic loci, and using cell fractionation experiments we determined that the eRNAs are preferentially localized to the chromatin, consistent with their eRNA status. Knockdown of one such eRNA in the mouse brain using antisense oligos, followed by a 1 h MCAO and 6 h of reperfusion, resulted in 49 downregulated genes and 106 upregulated genes (absolute fold-change ≥2.0; q<0.05) versus negative controls. The downregulated subset was enriched in several important genes with roles in mitochondrial activity, receptor signaling, ion channels and calcium homeostasis, and the upregulated subset was enriched in genes involved in inflammation and apoptosis. These changes were associated with worsened neurological scores and an 11.9% increase in infarct volume in the eRNA knockdown group versus negative control (p=0.004). This suggests an innate neuroprotective role for the eRNA during the stroke injury. Together, our study presents the first genome-wide identification of stroke-induced eRNAs in the adult cerebral cortex and illuminates the functional importance of one such eRNA in modulating post-stroke cortical gene expression and pathophysiology.

scholarly journals Relative Abundances ofCandida albicansandCandida glabratainIn VitroCoculture Biofilms Impact Biofilm Structure and Formation

2018 ◽  
Vol 84 (8) ◽  
pp. e02769-17 ◽  
Author(s):  
Michelle L. Olson ◽  
Arul Jayaraman ◽  
Katy C. Kao
Keyword(s):  
Gene Expression ◽  
Candida Albicans ◽  
Biofilm Formation ◽  
Candida Glabrata ◽  
Interspecific Interactions ◽  
Host Immune System ◽  
Dependent Manner ◽  
Important Species ◽  
Content Type ◽  
Content Ratio

ABSTRACTCandidais a member of the normal human microbiota and often resides on mucosal surfaces such as the oral cavity or the gastrointestinal tract. In addition to their commensality,Candidaspecies can opportunistically become pathogenic if the host microbiota is disrupted or if the host immune system becomes compromised. An important factor forCandidapathogenesis is its ability to form biofilm communities. The two most medically important species—Candida albicansandCandida glabrata—are often coisolated from infection sites, suggesting the importance ofCandidacoculture biofilms. In this work, we report that biofilm formation of the coculture population depends on the relative ratio of starting cell concentrations ofC. albicansandC. glabrata. When using a starting ratio ofC. albicanstoC. glabrataof 1:3, ∼6.5- and ∼2.5-fold increases in biofilm biomass were observed relative to those of aC. albicansmonoculture and aC. albicans/C. glabrataratio of 1:1, respectively. Confocal microscopy analysis revealed the heterogeneity and complex structures composed of longC. albicanshyphae andC. glabratacell clusters in the coculture biofilms, and reverse transcription-quantitative PCR (qRT-PCR) studies showed increases in the relative expression of theHWP1andALS3adhesion genes in theC. albicans/C. glabrata1:3 biofilm compared to that in theC. albicansmonoculture biofilm. Additionally, only the 1:3C. albicans/C. glabratabiofilm demonstrated an increased resistance to the antifungal drug caspofungin. Overall, the results suggest that interspecific interactions between these two fungal pathogens increase biofilm formation and virulence-related gene expression in a coculture composition-dependent manner.IMPORTANCECandida albicansandCandida glabrataare often coisolated during infection, and the occurrence of coisolation increases with increasing inflammation, suggesting possible synergistic interactions between the twoCandidaspecies in pathogenesis. During the course of an infection, the prevalence of eachCandidaspecies may change over time due to differences in metabolism and in the resistance of each species to antifungal therapies. Therefore, it is necessary to understand the dynamics betweenC. albicansandC. glabratain coculture to develop better therapeutic strategies againstCandidainfections. Existingin vitrowork has focused on understanding how an equal-part culture ofC. albicansandC. glabrataimpacts biofilm formation and pathogenesis. What is not understood, and what is investigated in this work, is how the composition ofCandidaspecies in coculture impacts overall biofilm formation, virulence gene expression, and the therapeutic treatment of biofilms.

scholarly journals Inhibitory Effects ofChrysanthemum borealeEssential Oil on Biofilm Formation and Virulence Factor Expression ofStreptococcus mutans

2015 ◽  
Vol 2015 ◽  
pp. 1-11 ◽  
Author(s):  
Beom-Su Kim ◽  
Sun-Ju Park ◽  
Myung-Kon Kim ◽  
Young-Hoi Kim ◽  
Sang-Bong Lee ◽  
...  
Keyword(s):  
Gene Expression ◽  
Essential Oil ◽  
Virulence Factors ◽  
Bacterial Growth ◽  
Biofilm Formation ◽  
Acid Production ◽  
Acid Tolerance ◽  
Significant Inhibition ◽  
Pcr Analysis ◽  
Dependent Manner

The aim of the study was to evaluate the antibacterial activity of essential oil extracted fromChrysanthemum boreale(C. boreale) onStreptococcus mutans(S. mutans). To investigate anticariogenic properties, and bacterial growth, acid production, biofilm formation, bacterial adherence ofS. mutanswere evaluated. Then gene expression of several virulence factors was also evaluated.C. borealeessential oil exhibited significant inhibition of bacterial growth, adherence capacity, and acid production ofS. mutansat concentrations 0.1–0.5 mg/mL and 0.25–0.5 mg/mL, respectively. The safranin staining and scanning electron microscopy results showed that the biofilm formation was also inhibited. The result of live/dead staining showed the bactericidal effect. Furthermore, real-time PCR analysis showed that the gene expression of some virulence factors such asgtfB,gtfC,gtfD,gbpB,spaP,brpA,relA, andvicR ofS. mutanswas significantly decreased in a dose dependent manner. In GC and GC-MS analysis, seventy-two compounds were identified in the oil, representing 85.42% of the total oil. The major components were camphor (20.89%),β-caryophyllene (5.71%),α-thujone (5.46%), piperitone (5.27%),epi-sesquiphellandrene (5.16%),α-pinene (4.97%), 1,8-cineole (4.52%),β-pinene (4.45%), and camphene (4.19%). These results suggest thatC. borealeessential oil may inhibit growth, adhesion, acid tolerance, and biofilm formation ofS. mutansthrough the partial inhibition of several of these virulence factors.

scholarly journals Eucalyptol inhibits biofilm formation of Streptococcus pyogenes and its mediated virulence factors

2020 ◽  
Vol 69 (11) ◽  
pp. 1308-1318
Author(s):  
Karuppiah Vijayakumar ◽  
Vajravelu Manigandan ◽  
Danaraj Jeyapragash ◽  
Veeraiyan Bharathidasan ◽  
Balaiyan Anandharaj ◽  
...  
Keyword(s):  
Gene Expression ◽  
Streptococcus Pyogenes ◽  
Biofilm Formation ◽  
Type Species ◽  
Inhibitory Concentration ◽  
Antibiofilm Activity ◽  
Dependent Manner ◽  
Biofilm Inhibition ◽  
Content Type ◽  
Link Type

Introduction. Streptococcus pyogenes is a diverse virulent synthesis pathogen responsible for invasive systemic infections. Establishment of antibiotic resistance in the pathogen has produced a need for new antibiofilm agents to control the biofilm formation and reduce biofilm-associated resistance development. Aim. The present study investigates the in vitro antibiofilm activity of eucalyptol against S. pyogenes . Methodology. The antibiofilm potential of eucalyptol was assessed using a microdilution method and their biofilm inhibition efficacy was visualized by microscopic analysis. The biochemical assays were performed to assess the influence of eucalyptol on virulence productions. Real-time PCR analysis was performed to evaluate the expression profile of the virulence genes. Results. Eucalyptol showed significant antibiofilm potential in a dose-dependent manner without affecting bacterial growth. Eucalyptol at 300 µg ml−1 (biofilm inhibitory concentration) significantly inhibited the initial stage of biofilm formation in S. pyogenes . However, eucalyptol failed to diminish the mature biofilms of S. pyogenes at biofilm inhibitory concentration and it effectively reduced the biofilm formation on stainless steel, titanium, and silicone surfaces. The biochemical assay results revealed that eucalyptol greatly affects the cell-surface hydrophobicity, auto-aggregation, extracellular protease, haemolysis and hyaluronic acid synthesis. Further, the gene-expression analysis results showed significant downregulation of virulence gene expression upon eucalyptol treatment. Conclusion. The present study suggests that eucalyptol applies its antibiofilm assets by intruding the initial biofilm formation of S. pyogenes . Supplementary studies are needed to understand the mode of action involved in biofilm inhibition.

Sign in / Sign up

Export Citation Format

Share Document