Salivary-free fluoride ion concentration measured using a flow-injection analysis device and oral environment in 4-6-year-old children.

Although fluoride (F) products are widely used for caries prevention, the safest and most effective modes of application, in particular for young children, remain to be elucidated. The limitations associated with the detection of ultra-low F ion concentrations are the major obstacles in accurately assessing the salivary F ion concentrations in children. This study aimed to measure accurate salivary-free F ion concentrations in children using a flow-injection analysis device and highlight the conditions or substances that influence changes in salivary content. Subjects were 4-6-year-old children, and we statistically compared the data involving the number of decayed, missing, or filled surfaces (dmfs), the levels of Mutans streptococci (MS) and Lactobacilli (LB) cariogenic bacteria, and oral hygiene habits. The information on the latter was obtained using a parent/guardian questionnaire. The average free F ion concentration measured was 0.421 ± 0.158 µmol/L (0.008 ± 0.003 ppm), which was considerably lower than that obtained in previous studies using the conventional F electrode method. No significantly different correlations were seen between salivary-free F ion concentrations and dmfs, MS and LB levels. With regard to salivary-free F ion concentrations and oral hygiene habits, only finishing brush of subjects' teeth by guardians showed a significant difference. In summary, the frequency of brushing was shown to correlate with free F ion concentration in saliva of children. Further studies are needed to circumstantially evaluate some other substances in saliva and oral hygiene habits.

Contribution of phosphoglucosamine mutase to the resistance of Streptococcus gordonii DL1 to polymorphonuclear leukocyte killing.

Phosphoglucosamine mutase (GlmM; EC 5.4.2.10) catalyzes the interconversion of glucosamine-6-phosphate to glucosamine-1-phosphate, an essential step in the biosynthetic pathway leading to the formation of the peptidoglycan precursor uridine 5'-diphospho-N-acetylglucosamine. We have recently identified the gene (glmM) encoding the enzyme of Streptococcus gordonii, an early colonizer on the human tooth and an important cause of infective endocarditis, and indicated that the glmM mutation in S. gordonii appears to influence bacterial cell growth, morphology, and sensitivity to penicillins. In the present study, we assessed whether the glmM mutation also affects escape from polymorphonuclear leukocyte (PMN)-dependent killing. Although no differences in attachment to human PMNs were observed between the glmM mutant and the wild-type S. gordonii, the glmM mutation resulted in increased sensitivity to PMN-dependent killing. Compared with the wild type, the glmM mutant induced increased superoxide anion production and lysozyme release by PMNs. Moreover, the glmM mutant is more sensitive to lysozyme, indicating that the GlmM may be required for synthesis of firm peptidoglycans for resistance to bacterial cell lysis. These findings suggest that the GlmM contributes to the resistance of S. gordonii to PMN-dependent killing. Enzymes such as GlmM could be novel drug targets for this organism.

Identification of the Streptococcus gordonii glmM gene encoding phosphoglucosamine mutase and its role in bacterial cell morphology, biofilm formation, and sensitivity to antibiotics.

Phosphoglucosamine mutase (EC 5.4.2.10) catalyzes the interconversion of glucosamine-6-phosphate into glucosamine-1-phosphate, an essential step in the biosynthetic pathway leading to the formation of peptidoglycan precursor uridine 5'-diphospho-N-acetylglucosamine. The gene (glmM) of Escherichia coli encoding the enzyme has been identified previously. We have now identified a glmM homolog in Streptococcus gordonii, an early colonizer on the human tooth and an important cause of infective endocarditis, and have confirmed that the gene encodes phosphoglucosamine mutase by assaying the enzymatic activity of the recombinant GlmM protein. Insertional glmM mutant of S. gordonii did not produce GlmM, and had a growth rate that was approximately half that of the wild type. Morphological analyses clearly indicated that the glmM mutation causes marked elongation of the streptococcal chains, enlargement of bacterial cells, and increased roughness of the bacterial cell surface. Furthermore, the glmM mutation reduces biofilm formation and increases sensitivity to penicillins relative to wild type. All of these phenotypic changes were also observed in a glmM deletion mutant, and were restored by the complementation with plasmid-borne glmM. These results suggest that, in S. gordonii, mutations in glmM appear to influence bacterial cell growth and morphology, biofilm formation, and sensitivity to penicillins.