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Bacterial growth is closely tied to nutrient availability, with cells proliferating exponentially under favorable conditions and entering a stationary phase when resources become scarce. This transition is mediated by a regulatory mechanism known as the stringent response, which allows bacteria to adapt to nutrient deprivation by modulating gene expression and metabolic activity.During nutrient scarcity, intracellular amino acid levels decline. It results in the accumulation of uncharged tRNAs...

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Live Cell Calcium Imaging Combined with siRNA Mediated Gene Silencing Identifies Ca2+ Leak Channels in the ER Membrane and their Regulatory Mechanisms
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Transcriptome analysis of calcium hydroxide tolerance in Enterococcus faecalis.

Zhibo Xu1,2, Haonan Ma3, Xinmiao Jiang1

  • 1Department of Endodontics, Hospital of Stomatology, Jilin University, Changchun, China.

Frontiers in Microbiology
|May 15, 2025
PubMed
Summary

Repeated exposure to calcium hydroxide (Ca(OH)2) increases Enterococcus faecalis survival and biofilm formation. RNA sequencing reveals significant gene expression changes, indicating complex signaling pathways contribute to Ca(OH)2 tolerance in this bacterium.

Keywords:
Enterococcus faecaliscalcium hydroxidepersistent apical periodontitistolerancetranscriptomics

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Dental Research

Background:

  • Calcium hydroxide (Ca(OH)2) is a common root canal disinfectant.
  • Enterococcus faecalis exhibits tolerance to Ca(OH)2, limiting its disinfectant efficacy.
  • Understanding the mechanisms of E. faecalis tolerance is crucial for effective endodontic treatment.

Purpose of the Study:

  • To investigate the mechanism of Enterococcus faecalis tolerance to repeated calcium hydroxide exposure.
  • To identify molecular pathways involved in Ca(OH)2 tolerance in E. faecalis.

Main Methods:

  • Enterococcus faecalis was subjected to 10 successive passages with Ca(OH)2.
  • Bacterial survival rates and biofilm formation (crystal violet staining) were assessed.
  • RNA sequencing and bioinformatics analysis were performed on Ca(OH)2-tolerant strains.

Main Results:

  • Progressive increase in E. faecalis survival rate after repeated Ca(OH)2 exposure.
  • Ca(OH)2-tolerant strains exhibited enhanced biofilm formation compared to standard strains.
  • RNA sequencing identified 683 differentially expressed genes, implicating pathways in carbohydrate metabolism, quorum sensing, and two-component systems.

Conclusions:

  • Prolonged Ca(OH)2 exposure induces tolerance in E. faecalis.
  • Tolerance mechanisms involve alterations in multiple interconnected signaling pathways.
  • Findings provide a basis for developing strategies against E. faecalis endodontic infections.