For human RAG2 expression, the following primers were used: 5′-CAC AGT CAT AGT GGG CAG TCA-3′ and 5′-TGA TGG TAC GTA GAT TTT TGT CTG A-3′. Quantification of the transcript was performed by real-time PCR on an ABI Prism 7000 light cycler (Applied Biosystems, Zug, Switzerland) using SYBR green PCR MasterMix (Fermentas). Ct values were normalized against GAPDH and fold induction was calculated as . We thank Professor Small molecule library mouse Andera Biondi and Dr. Grazia Fazio, Centro Ricerca Tettamanti, Clinica Pediatrica Universitá di Milano-Bicocca, Ospedale San Gerardo, Monza, MI, Italy for providing
the human BM samples. We thank Professors Rod Ceredig and David Nemazee for critical reading of the manuscript. Antonius G. Rolink is the holder of the chair in Immunology endowed by F. Hoffmann-La Roche Ltd., Basel. This work was supported by a grant from the Swiss National Science Foundation to A. G. R. Conflict of interest: The authors declare no financial or commercial conflict of interest.
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“DGGE of 16S rDNA is one of the most frequently used methods to study microbial communities. In this study, the DGGE profiles of different 16S rDNA regions of the periodontal pathogens Porphyromonas gingivalis, Fusobacterium nucleatum, and Prevotella nigrescens were investigated. The results suggested that V3-V5 and V6-V8 fragments may be suitable for community analysis of subgingival bacteria. Further analysis of subgingival samples with V3-V5 and V6-V8 regions as target fragments suggested that, in chronic periodontitis, PR-171 supplier re-colonization by periodontal bacteria with a population very similar to the baseline may occur by 6 weeks after mechanical debridement. Periodontal infection is initiated by invasive periodontal pathogens in subgingival plaque biofilm.
The first step in periodontal therapy is to alter or eliminate the bacterial communities PtdIns(3,4)P2 responsible for the infection (1). Mechanical debridement significantly improves clinical parameters and is necessary for successful periodontal treatment. However, the data from studies of the effects of periodontal therapy on the subgingival microbiota are confusing (2, 3). So far, several 16S rDNA-based methods have been used for analysis of the effect of mechanical debridement on subgingival bacterial communities. Species-specific regions in 16S rDNA have been used to design primers for PCR analysis to identify unique periodontal pathogens such as Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola (4–6). In addition, PCR primers designed by conserved sequences of 16S rDNA have also been used for amplification of 16S rDNA fragments from all bacterial species found in periodontal pockets. Further separation and analysis of PCR amplicons can profile the bacterial communities of subgingival plaque and elucidate microbial population dynamics (7–9).