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Objective: To assess the effect of daily consumption of erythritol, xylitol, and sorbitol candies on caries development in mixed dentition during a 3-year intervention and 3 years after the intervention.Methods: 485 Estonian first- and second-grade primary school children participated. Children were randomly allocated to an erythritol, xylitol, or sorbitol (control) group. Polyol-containing candies were administered on school days with a daily polyol consumption of 3 × 2.5 g. Yearly, caries development was assessed by calibrated dentists using the ICDAS criteria. Six years after initiation of the study and 3 years after cessation of daily polyol consumption, 420 participants were re-examined to identify potential long-term effects of polyol consumption. Survival curves were generated at the end of the intervention period and 3 years after intervention. The model included age of the subjects, schools, tooth surface ages and years of surface exposure to intervention. ICDAS scoring system-based events included enamel/dentin caries development, dentin caries development, increase in caries score, and dentist intervention.Results: At the end of the intervention, time to enamel/dentin caries development, dentin caries development, increase in caries score, and dentist intervention were significantly longer in the erythritol group as compared to the sorbitol group. Except for increase in caries score, all effects persisted 3 years after cessation of daily polyol consumption.Conclusions: A caries-preventive effect of 3-year erythritol consumption as compared to sorbitol was established in children with mixed dentition. The effect persisted up to 3 years after the end of the intervention. [ABSTRACT FROM AUTHOR]

Objective: The aim of this study was to test the efficacy of long-term, daily intake of erythritol and xylitol candy, compared with sorbitol candy, on the development of enamel and dentin caries lesions. Methods: The study was a double-blind randomized controlled prospective clinical trial. Altogether 485 primary school children, first- and second-graders at baseline, from southeastern Estonia participated in this 3-year intervention. Each child consumed four erythritol, xylitol or sorbitol (control) candies three times per school day. The daily intake of polyol was about 7.5 g. The International Caries Detection and Assessment System (ICDAS) was used in the clinical examinations by four calibrated examiners at baseline and at 12, 24 and 36 months. Results: The annual examination analyses and the follow-up analyses confirmed that the number of dentin caries teeth and surfaces at 24 months follow-up and surfaces at 36 months follow-up was significantly lower in the mixed dentition in the erythritol group than in the xylitol or control group. Time of enamel/dentin caries lesions to develop and of dentin caries lesions to progress was significantly longer in the erythritol group compared to the sorbitol and xylitol groups. Also the increase in caries score was lower in the erythritol group than in the other groups. Conclusions: In the follow-up examinations, a lower number of dentin caries teeth and surfaces was found in the erythritol group than in the xylitol or control groups. Time to the development of caries lesions was longest in the erythritol group. Trial registration: ClinicalTrials.gov Identifier NCT01062633. © 2014 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Addition of common dietary carbohydrates to Millipore-treated human whole saliva either enhances or inhibits the formation of salivary precipitates, some carbohydrates showing no effect. The purpose of this study was to investigate the precipitation conditions more thoroughly and to elucidate the chemical nature of the precipitates formed. D-Xylose either enhanced precipitation (in long-term incubations) or had no appreciable effect (in 10 minute incubations). Other aldo- and keto-sugars and disaccharides (maltose, sucrose, lactose) generally enhanced precipitation, whereas all polyols (xylitol, D-sorbitol, mannitol, and maltitol) retarded the formation of turbidity in saliva. Xylitol inhibited formation of precipitates also in the presence of D-xylose, dextrans, and starch. Fast protein liquid chromatography (FPLC) of EDTA-soluble pellets obtained by centrifugation of the precipitates produced two major protein fractions (I and II) with a molecular weight of 112,000 and 46,000, respectively. The carbohydrates exerted a selective effect on the relative size of I and II in that polyol incubations resulted in a I to II ratio of 1:3, whereas control incubations (without added sugars) and incubations with other carbohydrates gave ratios of 1:6 to 1:10. Both peaks contained large amounts of acidic amino acids, proline, and glycine. The saliva precipitates contained a substantial portion of a crystalline phase that had the crystal structure of apatite, the individual crystallites being extremely small (less than 1 micron) with a Ca:P ratio of 1.46. The carbohydrates had a similar effect on the overall inorganic composition of the precipitates, but they had a clearly selective effect on the rate of formation of precipitates and on the relative amount of coprecipitating salivary proteins. This selectivity indicates that these carbohydrates, when consumed habitually, may exert different effects on the precipitation of Ca-salts at mineral-deficient enamel and dentine sites. [ABSTRACT FROM AUTHOR]

The proteolytic profile of Treponema vincentii ATCC 35580 was studied using PZ- PLGPA (phenylazobenzyloxycarbonyl-L-prolyl-L-leucylglycyl-L-prolyl-D-arginine; a substrate of bacterial collagenases) and amino acid 2-naphthylamides (2NA) as substrates. The cell extracts showed high activity toward PZ-PLGPA, Nα-L-arginyl-2NA and Nγ-L-glutamyl-2NA. Gel permeation chromatography revealed 2 major endopeptidases (I and II) hydrolyzing PZ-PLGPA, the molecular weights of which were 75,000 and 23,000, respectively. Enzyme I was stable enough for subsequent fast protein liquid chromatography on an anion exchange column. The enzyme had a broad pH optimum of 6.5 to 7.5 with PZ-PLGPA as substrate, hydrolyzed gelatin and was moderately inhibited by metal chelators, but was very sensitive to p-chloromercuribenzoic acid (PCMB). Enzyme II with a pH optimum of 7 to 8 was more labile, quite sensitive to PCMB and moderately inhibited by chelators. A high-molecular weight arginine aminopeptidase (mol wt. > 200,000) was sensitive to PCMB and showed a value of 0.55 mM for Km in the hydrolysis of Nα-L-arginyl-2NA. The hydrolysis of PZ-PLGPA and gelatin suggests that this organism may contain collagenolytic proteinases. Because the insoluble proteinase substrate Azocoll was not hydro1yzed, these enzymes may be active on soluble collagenous substances only. T. vincentii ATCC 35580 typifies an organism rich in PZ-PLGPA-endopeptidase, argimne aimnopeptidase and γ-glutamylpeptidase activity. [ABSTRACT FROM AUTHOR]

A previous clinical trial showed that long-term use of saliva-stimulating polyol (xylitol and sorbitol) chewing gums was associated with arrest of dental caries in young subjects. After a 20-22-month intervention (when the subjects were 8 years old), a total of 23 primary teeth with extensive dentin caries lesions whose surface in clinical examination was found to be totally rehardened (remineralized) could be removed because the teeth were near their physiologic exfoliation time. These teeth were subjected to histologic, microhardness, and electron microscopic tests. The majority of the specimens had been remineralized from the surface by a non-cellular-mediated process within the remaining collapsed, organic extracellular matrix associated with the remaining dentinal surface. Many of the underlying dentinal tubules were filled with a matrix that had been subsequently mineralized. Dental microanalyses showed that the topmost (outer) 20-μm-thick rehardened layer of the lesions exhibited the highest Ca:P ratio, which leveled off at a depth of approximately 150 μm. The rehardened surface layer (normally <0.1 mm in thickness) was significantly (P < 0.001) harder than sound dentin and nearly as hard as sound enamel. Although the main source of the mineral present in the rehardened layer was most likely of salivary origin, some extracellular remineralization was probably mediated by odontoblasts. The results complete the clinical diagnoses of the original trial and suggest that regular use of polyol chewing gums may induce changes in dentin caries lesions, which in histologic and physiochemical studies show typical characteristics of rehardening and mineralization. [ABSTRACT FROM AUTHOR]

Several studies indicate that xylitol (X) consumption is associated with certain biochemical changes in dental plaque and whole saliva. In making X-containing saliva stimulants more cost-effective and palatable, manufacturers may use maltitol syrup (MS, which normally contains some sorbitol and higher polyols) or polydextrose (PD, a polysaccharide molecule with a mass >22 kDa) as bulking agents. Combinations of X with MS and PD have not been tested regarding their salivary effects. One hundred and eighty-eight young subjects (mean age, 22 years) of both sexes were divided into three groups of equal size for a 4-month study. The subjects in one group used X-MS dragées (in 7 daily episodes; 8 g X per day), while the subjects in another group used X-PD dragées in as many daily episodes (8 g X per day). Subjects in the third (comparison) group did not receive saliva stimulants. Paraffin-stimulated whole saliva samples were collected at baseline, after 2 months, and at endpoint. The usage of X-MS was associated with a significant (P < 0.05) reduction in the salivary sucrase activity. After 4 months, the activity of enzymes hydrolyzing Nα-benzoyl-DL-arginyl-p- nitroaniline was significantly reduced in all groups, while the levels of free sialic acid were reduced in group X-PD only (P < 0.05). These salivary changes most likely reflected microbial shifts in the oral cavity and suggest that information from saliva studies may be of avail when deciding which bulking agents should be used in xylitol-based saliva stimulants. [ABSTRACT FROM AUTHOR]

The cell extracts of two human oral strains (FN2 and FN3) of Fusobacterium nucleatum displayed exceptionally high γ-glutamylpeptidase activity as determined with N-γ- l-glutamyl-2-naphthylamine as substrate. This activity was so dominant that the hydrolysis of other N-aminoacyl-2-naphthylamines progressed at a rate <10% of the former. Two major enzymes (I and II) were partially purified from FN2. I had a molecular weight of 115,000 and did not hydrolyze γ-glutamylcysteinylglycine (glutathione). II had a molecular weight of 70,000 and rapidly liberated only glutamic acid from glutathione. Strain FN3 contained several enzymes hydrolyzing γ-glu-2NA. Direct anion exchange chromatography of FN3 cell extracts separated one enzyme that liberated both glutamic acid and glycine from glutathione, one that was inactive against glutathione (but hydrolyzed γ-glu-2NA), and one that liberated only glutamic acid. Although γ-glu-2NA was a good synthetic substrate, glutathione was hydrolyzed at least 500 times faster by an enzyme present in both strains. These results indicate that the presence of γ-glutamylpeptidase activity is very characteristic of these F. nucleatum strains. [ABSTRACT FROM AUTHOR]

Cell extracts prepared from several oral treponemes isolated from the subgingival plaque of periodontitis patients showed high enzyme activity toward phenylazobenzyl-oxycarbonyl- l-prolyl- l-leucylglycyl- l-prolyl- d-arginine (a compound used as a substrate for microbial collagenases). One major enzyme hydrolyzing this substrate at the Leu-Gly bond only was partially purified from an unspeciated treponeme (strain US), Treponema denticola ATCC 35405, and 29 different clinical isolates of T. denticola. The Treponema US enzyme also hydrolyzed furylacryloyl- l-leucylglycyl- l-prolyl- l-alanine (another substrate of bacterial collagenases) at the Leu-Gly bond. This enzyme also hydrolyzed various collagens and collagen-derived peptides. These treponemal proteases were sensitive to metal chelators and p-chloromercury compounds. The results indicate that human oral treponemes contain enzymes that readily hydrolyze in chromogenic protease substrates the Leu-Gly bond only that is the cleavage site of these substrates also by 'true' microbial collagenases. [ABSTRACT FROM AUTHOR]

Treponema denticola ATCC 35405, a human oral spirochete associated with periodontal disease, was shown to contain three enzymes (I, II, and III) with proline iminopeptidase activity. II and III were considered to be true iminopeptidases, whereas enzyme I was found to be a benzoylarginine peptidase with iminopeptidase activity. Enzyme III, the dominant proline iminopeptidase of T. denticola in terms of its activity toward N- l-prolyl-2-naphthylamine, was considered to be a sulfhydryl peptidase: 0.167 μ M p-chloromercuribenzoic acid totally inactivated the enzyme, and 1.0 m M dithiothreitol restored 92% of activity. The activity of this enzyme was not affected by metal chelators. Chemical modification of enzyme III suggests that tyrosyl (or histidyl) and carboxyl groups may be necessary for its activity. The hydrolysis of N- l-prolyl-2-naphthylamine was found to be very characteristic of T. denticola ATCC 35405; out of 24 different N- l-aminoacyl-2-naphthylamines tested, only the proline derivative was hydrolyzed at a high rate. The substrate specificity of the enzymes discovered indicates that they may be important for the nutrition of T. denticola. The iminopeptidase activity may be related to the pathogenicity of this organism in periodontal disease. [ABSTRACT FROM AUTHOR]

Seven clinical isolates and the ATCC strain 35405 of Treponema denticola, obtained from human periodontal pockets, were studied for peptidase activity with several chromogenic compounds as substrates. The cell sonicates of all strains hydrolyzed phenylazobenzyloxycarbonyl- l-prolyl- l-leucyl-glycyl- l-prolyl- d-arginine (a collagenase substrate), azocasein, and the 2-naphthylamines of l-proline, l-hydroxyproline, l-pyrrolidine, and benzoyl- l-arginine, but the rates of hydrolysis varied considerably from strain to strain. Fast protein liquid chromatography on gel and anion exchange columns revealed further biochemical differences between the strains. The ATCC strain consistently produced several proline iminopeptidases, whereas four of the clinical isolates yielded high and three yielded low iminopeptidase activity. The ATCC strain and six clinical isolates displayed high benzoylarginine peptidase activity. The use of N- l-prolyl-2-naphthylamine as substrate revealed more differences between the strains than other substrates. The substrate specificity of the enzymes discovered suggests that they may be important for the nutrition of the organism or in the protection of the organism against chemical defense factors present in the gingival pocket. [ABSTRACT FROM AUTHOR]

Background: Oral microbiome has significant impact on both oral and general health. Polyols have been promoted as sugar substitutes in prevention of oral diseases. We aimed to reveal the effect of candies containing erythritol, xylitol or control (sorbitol) on salivary microbiome. Methods: Ninety children (11.3 ± 0.6 years) consumed candies during 3 years. Microbial communities were profiled using Illumina HiSeq 2000 sequencing and real-time PCR. Results: The dominant phyla in saliva were Firmicutes (39.1%), Proteobacteria (26.1%), Bacteroidetes (14.7%), Actinobacteria (12%) and Fusobacteria (6%). The microbiome of erythritol group significantly differed from that of the other groups. Both erythritol and xylitol reduced the number of observed bacterial phylotypes in comparison to the control group. The relative abundance of the genera Veillonella, Streptococcus and Fusobacterium were higher while that of Bergeyella lower after erythritol intervention when comparing with control. The lowest prevalence of caries-related mutans streptococci corresponded with the lowest clinical caries markers in the erythritol group. Conclusions: Daily consumption of erythritol, xylitol or control candies has a specific influence on the salivary microbiome composition in schoolchildren. Erythritol is associated with the lowest prevalence of caries-related mutans streptococci and the lowest levels of clinical caries experience. Trial registration: ClinicalTrials.gov Identifier NCT01062633. [ABSTRACT FROM AUTHOR]