The purpose of this work was to explain how the caries-preventive agent xylitol interferes with the growth of Streptococcus mutans. It was found that the xylitol-sensitive strain of S. mutans 27352 (serotype g) and LG1 (serotype c) took up 14C-xylitol when the labelled pentitol was added to cells growing at the expense of glucose. Uptake of xylitol by growing cells of S. mutans 27352 XR and LG1 XR, two xylitol-insensitive spontaneous mutants, and of S. mutans GS5-2, which was also insensitive to xylitol, was practically inexistent under the same conditions. Alkaline phosphatase treatment followed by enzymatic analysis and thin-layer chromatography revealed that the accumulated product was xylitol phosphate. Intracellular concentrations of 5–7 mM for resting cells and of up to 60 mM for growing cells were calculated. Xylitol was phosphorylated at the expense of phosphoenolpyruvate by toluenized cells of S. mutans LG1, but not by toluenized cells of GS5–2 and S. mutans LG1 XR. The phosphorylation of xylitol was dependent on phosphoenolpyruvate and required the presence of both soluble and membrane cellular fractions in the reaction mixture. This indicated that xylitol was transported and phosphorylated by a phosphoenolpyruvate: sugar phosphotransferase system. The phosphoenolpyruvate-dependent phosphorylation by isolated membranes of S. mutans LG1 in the presence of the soluble fraction was inhibited by fructose but not by glucose, mannose or galactose. Measurement of phosphoenolpyruvate: phosphotransferase activities in isolated membrane revealed that strain 27352 and LG1 had activities for fructose and xylitol; membrane from 27352 XR and LG1 XR had very little activity for fructose and xylitol. It was concluded that xylitol was transported and phosphorylated by a constitutive phosphoenolpyruvate:fructose phosphotransferase system in S. mutans. The data suggested that xylitol toxicity in S. mutans is caused by the intracellular accumulation of xylitol phosphate.
Since the exposure of mutans streptococci to xylitol is known to select for xylitol-resistant (XR) natural mutants, the occurrence and long-term survival of such xylitol-resistant strains was evaluated in a cross-sectional sampling of participants of the Ylivieska xylitol study four years after the original two-year experimental period. Paraffin-stimulated whole saliva was first collected, and then plaque was collected and pooled. The salivary and dental plaque mutans streptococci were enumerated after growth on TSY20B agar. The proportion of XR strains was determined by autoradiography with 14C-xylitol. A strong and significant correlation (r = 0.645 and p = 0.005) between the number of mutans streptococci in saliva and in dental plaque was observed in non-consumers of xylitol. Such a correlation totally disappeared (r = 0.098 and p = 0.612) in xylitol-exposed consumers (habitual and former xylitol-consumers). The proportion of the salivary XR mutants (35%) in non-consumers (n = 16) was significantly lower than in the xylitol-exposed consumers (79%) (n = 27), (p = 0.0001) or in former consumers (75%) (n = 13), (p = 0.0008) or in the habitual consumers (83%) (n = 14), (p = 0.004). The proportion of XR mutants in dental plaque was, on the average, much lower than in the corresponding saliva. The proportion of XR in the plaque of xylitol non-consumers was half of that of the xylitol-exposed group, but the difference was not statistically significant.(ABSTRACT TRUNCATED AT 250 WORDS)
The effect of long-term consumption of refined xylitol on the natural populations of S. mutans in the human oral cavity has been investigated. Fifty-four S. mutans strains were isolated from adults and children who had been consuming commercial food products containing xylitol for a period of from 1 1/2 to 10 years. Twenty isolates were also obtained from control subjects who had never consumed xylitol-containing commercial food products. The inhibitory effect of xylitol on the isolated strains was determined by monitoring growth on glucose in the presence or absence of xylitol. This was used to define the sensitivity of each isolate to xylitol. Phosphoenolpyruvate:sugar phosphotransferase (PEP-PTS) activities were measured by means of the soluble and membrane fractions prepared from strains from both study populations. It was found that 87% of the fresh isolates from xylitol consumers were xylitol-resistant (XR), compared with only 10% of the strains isolated from the control subjects. The XR strains had low constitutive fructose PTS activity and very low xylitol-phosphorylating capacity. The xylitol-sensitive (XS) strains, however, had much higher levels of constitutive fructose PTS activity and phosphorylated xylitol 16 times more rapidly than did the XR strains. Evidence for the phosphorylation of xylitol by a fructose PEP-PTS in the XS strains was obtained. The growth inhibition by the intracellular accumulation of non-metabolizable toxic xylitol phosphate and its prevention by the presence of fructose are discussed.
We investigated the effect of xylitol on the growth of different oral bacteria in the presence of glucose. Xylitol inhibited the growth of all but one of ten strains of S. mutans and failed to inhibit the growth of the lactobacilli, actinomycetes, and other streptococci tested except S. sanguis 10556, which was slightly inhibited. However, the rate of acid production of the sensitive S. mutans strains was not equally affected by xylitol. These data, obtained with pure cultures of acidogenic oral bacteria, may explain the lack of an in vitro inhibitory effect of xylitol on dental plaque samples.
Several factors affecting the amount of fluoride ingested during toothbrushing by 2- to 7-year-old children were investigated. The specific purpose of this study was to determine the contribution of age, the amount of dentifrice used, and rinsing after brushing to the variation in the ingestion of fluoride dentifrice. Four hundred and five children brushed their teeth in front of a portable sink. The tubes of dentifrice in gel (0.24% NaF) were weighed before and after use to determine the amount of toothpaste used. The fluoride content of the collected liquids was determined with a fluoride-ion-specific electrode. The amount of fluoride ingested was derived by determining the difference between the amounts used and recovered. The amount of dentifrice used, the age, and the rinsing habits, entered in a multiple regression model, explained up to 66 percent of the total variation in the amount of fluoride ingested. The amount of dentifrice used accounted by itself for 60 percent of the total variation. Therefore, these results indicate that the quantity of dentifrice used was the most important factor affecting the ingestion of fluoride through toothbrushing by young children.
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