Barna da Siena, 'The Resurrection of Lazarus'

Davies, Damian Walford

doi:10.1093/english/51.200.181

Cited by 2 publications

(3 citation statements)

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“…Other similar techniques such as methylation-sensitive arbitrarily primed PCR (MS AP-PCR) (Gonzalgo et al, 1997) and methylated CpG island amplification (MCA) (Toyota et al, 1999) were developed about the same time in the late 1990s. Subsequent modifications to these techniques have recently been reviewed by Davies (2002) and Liang et al (2002). These methodologies hold the promise of providing a rapid and efficient way to screen for changes in methylation patterns in genomic DNA and to isolate specific sequences associated with these changes, in a nonselective manner.…”

Section: Discussionmentioning

confidence: 99%

“…False positives are a major problem, resulting in the low yield of MSRF and similar techniques (Davies, 2002;Liang et al, 2002). In addition to the possibility of incomplete restriction digestion, this problem is compounded by the heterogeneity of tumor tissues that commonly comprise mixtures of benign and cancerous tissues expressing varying levels of the differentially methylated genes.…”

Section: Discussionmentioning

confidence: 99%

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PMP24, a gene identified by MSRF, undergoes DNA hypermethylation-associated gene silencing during cancer progression in an LNCaP model

2004

Oncogene

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Transcriptional silencing of antitumor genes via CpG island methylation could be a mechanism mediating prostate cancer (PCa) progression from an androgensensitive (AS) to an androgen-insensitive (AI) state. We have used the methylation-sensitive restriction fingerprinting (MSRF) technique to identify novel CpG-rich sequences that are differentially methylated between the genome of the AS PCa cell line LNCaP and that of an AI subline LNCaP CS generated by maintaining LNCaP in medium with charcoal-stripped (CS) serum for over 30 passages. One such sequence identified was located on a 5 0 CpG island that was found to span part of the promoter, exon 1, and part of intron 1 of the peroxisomal membrane protein 24 kDa (PMP24) gene. Using semiquantitative RT-PCR and bisulfite genomic sequencing, we established an inverse relationship between mRNA expression and methylation of the 5 0 CpG island of PMP24. PMP24 mRNA was absent in LNCaP CS and the androgen receptor-negative PC-3 cell line; both exhibited dense methylation in the said CpG island. In contrast, PMP24 mRNA was expressed in LNCaP and normal prostatic epithelial cells (NPrECs) whose PMP24 5 0 CpG island remained unmethylated. Treatment of LNCaP CS and PC-3 with the demethylating agent 5-aza-2 0 -deoxycytidine (5-AZAdC) reactivated PMP24 mRNA expression. Transient transfection of PMP24 into LNCaP CS and PC-3 cells induced a significant reduction in cell growth and soft-agar colony formation potential, suggesting that PMP24 gene product has antitumor properties. These results demonstrate the utility of MSRF in the identification of novel, differentially methylated DNA sequences in the genome and suggest that hypermethylation-mediated silencing of PMP24 is an epigenetic event involved in PCa progression to androgen independence.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

PMP24, a gene identified by MSRF, undergoes DNA hypermethylation-associated gene silencing during cancer progression in an LNCaP model

2004

Oncogene

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“…~453,000 cases of CDI were reported in the United States in 2011 [1] and there have been increasing reports of CDI from different parts of the world [2][3][4]. Treatment of CDI is complicated by the fact that 20-36% of cases experience recurrence, relapsing after completion of initial treatment [5].…”

Section: Introductionmentioning

confidence: 99%

Clostridioides difficileLuxS mediates inter-bacterial interactions within biofilms

Slater

Frost

Jossi

et al. 2018

Preprint

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The anaerobic gut pathogen, Clostridioides difficile, forms adherent biofilms that may play an important role in recurrent C. difficile infections. The mechanisms underlying C. difficile community formation and inter-bacterial interactions are nevertheless poorly understood. C. difficile produces AI-2, a quorum sensing molecule that modulates biofilm formation across many bacterial species. We found that a strain defective in LuxS, the enzyme that mediates AI-2 production, is defective in biofilm development in vitro. Transcriptomic analyses of biofilms formed by wild type (WT) and luxS mutant (luxS) strains revealed a downregulation of prophage loci in the luxS mutant biofilms compared to the WT. Detection of phages and eDNA within biofilms may suggest that DNA release by phage-mediated cell lysis contributes to C. difficile biofilm formation. In order to understand if LuxS mediates C. difficile crosstalk with other gut species, C. difficile interactions with a common gut bacterium, Bacteroides fragilis, were studied. We demonstrate that C. difficile growth is significantly reduced when co-cultured with B. fragilis in mixed biofilms. Interestingly, the absence of C.difficile LuxS alleviates the B. fragilis mediated growth inhibition. Dual species RNAsequencing analyses from single and mixed biofilms revealed differential modulation of distinct metabolic pathways for C. difficile WT, luxS and B. fragilis upon co-culture, indicating that AI-2 may be involved in induction of selective metabolic responses in B. fragilis. Overall, our data suggest that C. difficile LuxS/AI-2 utilises different mechanisms to mediate formation of single and mixed species communities.

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Barna da Siena, 'The Resurrection of Lazarus'

Cited by 2 publications

References 0 publications

PMP24, a gene identified by MSRF, undergoes DNA hypermethylation-associated gene silencing during cancer progression in an LNCaP model

PMP24, a gene identified by MSRF, undergoes DNA hypermethylation-associated gene silencing during cancer progression in an LNCaP model

Clostridioides difficileLuxS mediates inter-bacterial interactions within biofilms

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