Project description:Treponema denticola is a major pathogen in periodontal disease, frequently isolated from lesions of chronic periodontitis. The ability to adopt its environment is believed to be important for T. denticola in colonizing and proliferating in the gingival crevice. T. denticola use serum as a major nutrient source in the gingival crevices, suggesting that this microorganism utilize serum components to proliferate in gingival crevice. The purpose of this study was to identify T. denticola serum utilization genes. Precultured T. denticola were suspended in tryptone-yeast extract-gelatin-volatile fatty acids medium containing 0, 1 and 10% serum, and incubated anaerobically for 17 h. Total RNA was isolated and T. denticola gene expression was compared by microarray and reverse transcription-polymerase chain reaction. In serum-depleted conditions, the expression of a potential hydroxylamine reductase, several ABC transporters, and phosphoenolpyruvate synthase were increased, while methyl-accepting chemotaxis proteins and a transcriptional regulator were decreased. The results suggest that T. denticola may uptake serum components via ABC transporters. Decreased dmcA expression with decreased serum concentration suggests its involvement in chemotaxis toward serum-rich environments.
Project description:Cannabidiol (CBD) actions in the brain are largely complex and not fully understood. One important brain structure that is a target for CBD, and for which health-beneficial CBD consequences can be seen is the hypothalamus. Given the fundamental role of gene expression in the hypothalamus's regulatory functions, this study undertook the analysis of changes arising in hypothalamic cells’ transcriptomes following various CBD treatments. For this purpose, we used a hypothalamic cellular model, namely we employ adult-derived mHypoA-2/12 mouse cell lines. In the course of the study, the neural cells were treated with different CBD doses (ranging from 0.325 to 3 µM) and vehicle as a control for 6 and 24 hours. The experiment setup allowed us to evaluate both the dosage and time-dependent effect of CBD on hypothalamic cells, particularly on their viability, apoptosis, and transcriptome profile.
Project description:Treponema denticola has been strongly implicated in the pathogenesis of chronic periodontitis. Previously, we reported increased expression of TDE_0259 (oxtR), which exhibits similarities with the multiple-antibiotic resistance regulator (marR)-like gene in the bacteriocin ABC transporter gene-deficient mutant, which altered susceptibility to antimicrobial agents. Therefore, we investigated the function of oxtR in T. denticola using an oxtR-deficient mutant. We observed increased OxtR expression upon oxygen exposure. The growth rate of the bacterium was unaffected by the inactivation of oxtR. We observed a relative increase in the expression of genes associated with iron-sulfur cluster-binding proteins, flavodoxin, oligopeptide/dipeptide ABC transporters, heat shock proteins, DNA helicase, iron compounds, ABC transporters, and lipoproteins. The oxtR mutant exhibited an increased minimum inhibitory concentration against ofloxacin. In addition, the mutant showed a slightly faster growth rate than that of the wild type under oxygen stress. Our findings also suggested that OxtR may regulate the operon by binding to a potential promoter region. The oxygen-sensing regulator oxtR plays a role in regulating the expression of a potential ferredoxin, which may contribute to the response to oxygen-induced stress in T. denticola.
Project description:Porphyromonas gingivalis and Treponema denticola are periodontalpathogens that are associated with the severity and progression of periodontal diseases. This study investigates the gene expression of Porphyromonas gingivalis during co-culture with Treponema denticola
Project description:Porphyromonas gingivalis and Treponema denticola are periodontalpathogens that are associated with the severity and progression of periodontal diseases. this study investigates the gene expression of Treponema denticola during co-culture with Porphyromonas gingivalis.
Project description:Primary glioma stem cells cultured as neurospheres in NBL media with growth factors were subjected to treatment with the non-toxic, non-psychoactive cannabis compound cannabidiol (CBD). Control and CBD- treated cultures were used to generate RNA used to hybridize on Affymetrix DNA arrays Fluorescence intensities data were RMA-normalized using Partek software.
Project description:Background: Treponema denticola is strongly associated with the development of periodontal disease. Both synergistic and antagonistic effects are observed among bacterial species in the process of biofilm formation. Bacteriocin-related genes have not yet been fully characterized in periodontopathic bacteria. The aim of this study was to detect and characterize bacteriocin-associated proteins in T. denticola. Methods: The whole genome sequence of T. denticola ATCC 35405 was screened with a Streptococcus mutans bacteriocin immunity protein (ImmA/Bip) sequence. The prevalence of homologous genes in T. denticola strains was then investigated by Southern blotting. Expression of the genes was evaluated by qRT-PCR. Results: In the genome sequence of T. denticola, an amino acid sequence coded by open reading frame TDE_0719 showed 26% identity with the S. mutans ImmA. Furthermore, two protein sequences coded by TDE_0425 and TDE_2431 in T. denticola ATCC 35405 showed ~40% identity with that coded by TDE0719. Therefore, TDE_0425, TDE_0719, and TDE_2431 were designated as tepA1, A2, and A3, respectively. Open reading frames showing similarity to the HlyD family of secretion proteins were detected downstream of tepA1, A2, and A3. They were designated as tepB1, B2, and B3, respectively. A gene harboring a bacteriocin-like signal sequence was detected upstream of tepA1. The prevalence of tepA1 and A2 differed among Treponema species. Susceptibility to chloramphenicol and ofloxiacin was slightly decreased in a tepA2 mutant while that to kanamycin was increased. Expression of tepA3-B3 was increased in the tepA2 mutant. Conclusion: These results indicate that T. denticola ATCC 35405 has three potential bacteriocin export proteins and that the presence of these genes differs among the Treponema strains. These proteins may be involved in resistance to chloramphenicol.