O176 Effect of Fe and glucose on the glycolysis and gluconeogenesis in biofilm-associated Staphylococcus epidermidis

Massonet, Caroline; Pintens, Valerie; Merckx, Roel; Eldere, J. Van

doi:10.1016/s0924-8579(07)70116-4

Cited by 4 publications

(8 citation statements)

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“…The amplicon was ligated into the pGEM-T Easy vector (Promega, Madison, WI), yielding pGEMsesC. Pure plasmid pGEMsesC was prepared and quantified as described previously (18,23). Standard dilutions of a known quantity of pGEMsesC were used in real-time PCR.…”

Section: Methodsmentioning

confidence: 99%

“…modifications were incorporated into the anesthesia and euthanasia protocols for rats (18,23) in accordance with guidelines of the animal ethics committee. In vivo and in vitro extraction of DNA and RNA from sessile and planktonic bacteria and reverse transcription were performed as described previously (18,23,29,31).…”

Section: Vol 77 2009 Inhibition Of S Epidermidis Biofilm Formationmentioning

confidence: 99%

“…In vivo and in vitro extraction of DNA and RNA from sessile and planktonic bacteria and reverse transcription were performed as described previously (18,23,29,31).…”

Section: Vol 77 2009 Inhibition Of S Epidermidis Biofilm Formationmentioning

confidence: 99%

“…Using the TaqMan primers sesC-TF and sesC-TR and a dual-label probe (5Ј 6-carboxyfluorescein and 3Ј 6-carboxytetramethylrhodamine), sesC-P quantification of both cDNA and gDNA was performed as described previously (18,23,29,31). In each run, a standard dilution of the plasmid (pGEMsesC) with a known quantity that allowed gene quantification and a negative control (distilled water) were included.…”

Section: Vol 77 2009 Inhibition Of S Epidermidis Biofilm Formationmentioning

confidence: 99%

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Inhibition of Staphylococcus epidermidis Biofilm Formation by Rabbit Polyclonal Antibodies against the SesC Protein

et al. 2009

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Several well-studied proteins with defined roles in Staphylococcus epidermidis biofilm formation are LPXTG motif-containing proteins. Here, we investigate the possible use of the LPXTG motif-containing protein SesC (S. epidermidis surface protein C; accession no. NP_765787) as a target for antibodies to prevent biofilm formation. In vitro and in a in vivo rat model of catheter infection, gene and protein expression analysis showed that SesC is expressed more strongly in biofilm-associated cells than in planktonic cells and is expressed particularly during the late phase of in vivo biofilm formation. Polyclonal rabbit antibodies raised against SesC reduced the fibrinogen-binding ability of S. epidermidis RP62A and Staphylococcus aureus RN4220 transformants expressing SesC, inhibited in vitro biofilm formation by S. epidermidis strains 10b and 1457, and significantly reduced the numbers of bacteria in a 1-day-old in vivo biofilm (P < 0.001, one-way analysis of variance). Our findings revealed that SesC is a promising target for prevention and treatment of S. epidermidis biofilms because it affects both the primary attachment and biofilm accumulation phases. The precise role of SesC in biofilm formation remains to be identified.

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Section: Methodsmentioning

confidence: 99%

Section: Vol 77 2009 Inhibition Of S Epidermidis Biofilm Formationmentioning

confidence: 99%

“…In vivo and in vitro extraction of DNA and RNA from sessile and planktonic bacteria and reverse transcription were performed as described previously (18,23,29,31).…”

Section: Vol 77 2009 Inhibition Of S Epidermidis Biofilm Formationmentioning

confidence: 99%

Section: Vol 77 2009 Inhibition Of S Epidermidis Biofilm Formationmentioning

confidence: 99%

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Inhibition of Staphylococcus epidermidis Biofilm Formation by Rabbit Polyclonal Antibodies against the SesC Protein

et al. 2009

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“…It seems as if intracellular R. equi cells manage to readily acquire iron in sufficient quantities, possibly through a constitutively expressed iron acquisition system (28) or through the baseline expression of the iup genes or another inducible iron acquisition system. In summary, this study shows that R. equi cells survive for at least 24 h in activated macrophages, where they are able to establish their unusual compartment.…”

Section: Discussionmentioning

confidence: 99%

Nitric Oxide-Mediated Intracellular Growth Restriction of Pathogenic Rhodococcus equi Can Be Prevented by Iron

et al. 2011

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Rhodococcus equi is an intracellular pathogen which causes pneumonia in young horses and in immunocompromised humans. R. equi arrests phagosome maturation in macrophages at a prephagolysosome stage and grows inside a privileged compartment. Here, we show that, in murine macrophages activated with gamma interferon and lipopolysaccharide, R. equi does not multiply but stays viable for at least 24 h. Whereas infection control of other intracellular pathogens by activated macrophages is executed by enhanced phagosome acidification or phagolysosome formation, by autophagy or by the interferoninducible GTPase Irgm1, none of these mechanisms seems to control R. equi infection. Growth control by macrophage activation is fully mimicked by treatment of resting macrophages with nitric oxide donors, and inhibition of bacterial multiplication by either activation or nitric oxide donors is annihilated by cotreatment of infected macrophages with ferrous sulfate. Transcriptional analysis of the R. equi ironregulated gene iupT demonstrates that intracellular R. equi encounters iron stress in activated, but not in resting, macrophages and that this stress is relieved by extracellular addition of ferrous sulfate. Our results suggest that nitric oxide is central to the restriction of bacterial access to iron in activated macrophages.

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Candida albicans biofilm formation in a new in vivo rat model

et al. 2010

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Device-associated microbial growth, including Candida biofilms, represents more than half of all human microbial infections and, despite a relatively small risk of implant-associated diseases, this type of infection usually leads to high morbidity, increased health-care costs and prolonged antimicrobial therapy. Animal models are needed to elucidate the complex host-pathogen interactions that occur during the development of attached and structured biofilm populations. We describe here a new in vivo model to study Candida biofilm, based on the avascular implantation of small catheters in rats. Polyurethane biomaterials challenged with Candida cells were placed underneath the skin of immunosuppressed animals following only minor surgery. The model allowed the study of up to ten biofilms at once, and the recovery of mature biofilms from 2 days after implantation. The adhering inoculum was adjusted to the standard threshold of positive diagnosis of fungal infection in materials recovered from patients. Wild-type biofilms were mainly formed of hyphal cells, and they were unevenly distributed across the catheter length as observed in infected materials in clinical cases. The hyphal multilayered structure of the biofilms of wild-type strains was observed by confocal microscopy and compared to the monolayer of yeast or hyphal cells of two well-known biofilm-deficient strains, efg1D/efg1D cph1D/cph1D and bcr1D/ bcr1D, respectively. The subcutaneous Candida biofilm model relies on the use of implanted catheters with accessible, fast and minor surgery to the animals. This model can be used to characterize the ability of antimicrobial agents to eliminate biofilms, and to evaluate the prophylactic effect of antifungal drugs and biomaterial coatings.

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O176 Effect of Fe and glucose on the glycolysis and gluconeogenesis in biofilm-associated Staphylococcus epidermidis

Cited by 4 publications

References 0 publications

Inhibition of Staphylococcus epidermidis Biofilm Formation by Rabbit Polyclonal Antibodies against the SesC Protein

Inhibition of Staphylococcus epidermidis Biofilm Formation by Rabbit Polyclonal Antibodies against the SesC Protein

Nitric Oxide-Mediated Intracellular Growth Restriction of Pathogenic Rhodococcus equi Can Be Prevented by Iron

Candida albicans biofilm formation in a new in vivo rat model

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