Streptococcus mutans glutamate binding protein (GlnH) as antigen target for a mucosal anti-caries vaccine.

BACKGROUND: In recent years, several studies have demonstrated that bacterial ABC transporters present relevant antigen targets for the development of vaccines against bacteria such as Streptococcus pneumoniae and Enterococcus faecalis. In Streptococcus mutans, the glutamate transporter operon (glnH), encoding an ABC transporter, is associated with acid tolerance and represents an important virulence-associated factor for the development of dental caries. RESULTS: In this study, we generated a recombinant form of the S. mutans GlnH protein (rGlnH) in Bacillus subtilis. Mice immunized with this protein antigen elicited strong antigen-specific antibody responses after sublingual administration of a vaccine formulation containing a mucosal adjuvant, a non-toxic derivative of the heat-labile toxin (LTK63) originally produced by enterotoxigenic Escherichia coli (ETEC) strains. Serum anti-rGlnH antibodies reduced adhesion of S. mutans to the oral cavity of naïve mice. Moreover, mice actively immunized with rGlnH were partially protected from oral colonization after exposure to the S. mutans NG8 strain. CONCLUSIONS: Our results indicate that S. mutans rGlnH is a potential target antigen capable of inducing specific and protective antibody responses after immunization. Overall, these observations raise the prospect of the development of mucosal anti-caries vaccines.

Total fluoride intake and excretion in children up to 4 years of age living in fluoridated and non-fluoridated areas.

Fractional fluoride retention is important during the early years of life when considering the risk of development of dental fluorosis. This study aimed to measure fractional fluoride retention in young children. The objectives were to investigate the relationships between fractional fluoride retention and total daily fluoride intake, age, and body mass index (BMI). Twenty-nine healthy children, up to 4 yr of age, participated; 14 lived in a fluoridated area (0.64 µg ml(-1) of fluoride in drinking water) and 15 lived in a non-fluoridated area (0.04 µg ml(-1) of fluoride in drinking water). The total daily fluoride intake of each child was calculated from the daily dietary fluoride intake and toothpaste ingestion (if fluoride toothpaste was used). Total daily fluoride excretion was measured by collecting voided urine and faeces over a 24-h period, and fractional fluoride retention was calculated by dividing the amount of fluoride retained in the body (total daily fluoride intake minus total daily fluoride excretion) by the total daily fluoride intake. Nine children were excluded from data analysis because of suspected invalid samples. Mean (range) fractional fluoride retention for the remaining 20 children was 0.61 (0.06-0.98). There were no statistically significant correlations between fractional fluoride retention and either age or BMI. However, fractional fluoride retention was correlated with total daily fluoride intake: fractional fluoride retention = 1 - exp (-C × total daily fluoride intake), where C = 28.75 (95% CI = 19.75-37.75). The wide variation in fluoride retention in young children could have important implications when recommendations for fluoride use are being considered.

Physical exercise ameliorates the toxic effect of fluoride on the insulin-glucose system.

Daily intake of water with fluoride concentrations >1.5 mg/l produces insulin resistance (IR). On the other hand, physical activity increases insulin sensitivity in the muscle. Therefore, the aim of this study was to evaluate the effect of physical activity on IR in rats treated with sodium fluoride (NaF) in drinking water. Sprague-Dawley rats were divided into three groups (n=10/group): Control (drinking water without NaF), NaF (drinking water with NaF 15 mg/l for 30 days), and Exercise (daily running on a treadmill for 60 min at 2.25 m/min and drinking water with NaF 15 mg/l for 30 days). IR was evaluated with the homeostasis model assessment-IR (HOMA-IR) index using fasting plasma levels of glucose and insulin. IR increased in rats treated with 15 mg/l NaF in drinking water. A decrease in IR was observed in rats that performed physical activity and drank water with 15 mg/l NaF; the Exercise group also showed an increase in the amounts of bone fluoride. The variation in the HOMA-IR values could be the consequence of variation in the sensitivity of tissues to insulin or decrease in plasma fluoride levels due to bone fluoride intake. These findings indicate that the performance of daily physical activity could reduce the negative effects of the chronic ingestion of NaF on glucose homeostasis.

Molecular mechanisms of fluoride toxicity.

Halfway through the twentieth century, fluoride piqued the interest of toxicologists due to its deleterious effects at high concentrations in human populations suffering from fluorosis and in in vivo experimental models. Until the 1990s, the toxicity of fluoride was largely ignored due to its "good reputation" for preventing caries via topical application and in dental toothpastes. However, in the last decade, interest in its undesirable effects has resurfaced due to the awareness that this element interacts with cellular systems even at low doses. In recent years, several investigations demonstrated that fluoride can induce oxidative stress and modulate intracellular redox homeostasis, lipid peroxidation and protein carbonyl content, as well as alter gene expression and cause apoptosis. Genes modulated by fluoride include those related to the stress response, metabolic enzymes, the cell cycle, cell-cell communications and signal transduction. The primary purpose of this review is to examine recent findings from our group and others that focus on the molecular mechanisms of the action of inorganic fluoride in several cellular processes with respect to potential physiological and toxicological implications. This review presents an overview of the current research on the molecular aspects of fluoride exposure with emphasis on biological targets and their possible mechanisms of involvement in fluoride cytotoxicity. The goal of this review is to enhance understanding of the mechanisms by which fluoride affects cells, with an emphasis on tissue-specific events in humans.

Fingernail may not be a reliable biomarker of fluoride body burden from dentifrice.

Fingernail has been suggested as a biomarker of fluoride (F) body burden, but there is no consensus if it would be a reliable indicator of F exposure from dentifrice. Therefore, the present study was conducted to investigate if fingernails would have sensitivity to detect F exposure from dentifrice in young children. Twenty-three 1-3-year-old children living in the city of Piracicaba (0.72 ppm F in water), Brazil, were enrolled in two phases of different F exposure: in phase A (1st to 11th week), they were exposed to the combination of F from diet (solids and liquids) and dentifrice (1,500 microg F/g as MFP), and in phase B (12th to 29th week), only to F from diet (the use of F dentifrice was interrupted). Fingernails were weekly clipped during 35 weeks for F determination. F intake from diet and dentifrice in each phase was also determined. Both analyses were made with ion-specific electrode. F intake (Mean +/- SD) was significantly higher (p<0.01) when the children were exposed to F from diet+dentifrice than only to F from diet (0.086 +/- 0.032 and 0.040 +/- 0.009 mg F/day/kg body weight, respectively). However, F concentrations in nails collected during the whole experimental period of 35 weeks presented great variation with no trend of decreasing after F dentifrice intake interruption. The findings suggest that fingernail may not be a reliable F biomarker of body burden from dentifrice.

Kinetics of monofluorophosphate hydrolysis in a bacterial test plaque in situ.

Models to evaluate the anticaries potential of fluoride (F) formulations containing monofluorophosphate (MFP) should consider the release of F ion to the oral environment by its enzymatic hydrolysis. This was tested in situ, using a test plaque of a strain of Streptococcus mutans which presents high MFPase activity at pH 5.0. The test plaque was exposed to non-F or MFP (1,450 microg F/g) dentifrices and the fluid phase of the plaque was analyzed after 15, 30, 45 and 75 min. MFP concentration in the plaque fluid decreased over time after exposure to MFP dentifrice, but F ion reached 134.9 +/- 32.0 microM at 15 min and decreased significantly only at 75 min, suggesting continuous MFP hydrolysis by the test plaque.

Fingernail may not be a reliable biomarker of fluoride body burden from dentifrice

Fingernail has been suggested as a biomarker of fluoride (F) body burden, but there is no consensus if it would be a reliable indicator of F exposure from dentifrice. Therefore, the present study was conducted to investigate if fingernails would have sensitivity to detect F exposure from dentifrice in young children. Twenty-three 1-3-year-old children living in the city of Piracicaba (0.72 ppm F in water), Brazil, were enrolled in two phases of different F exposure: in phase A (1st to 11th week), they were exposed to the combination of F from diet (solids and liquids) and dentifrice (1,500 µg F/g as MFP), and in phase B (12th to 29th week), only to F from diet (the use of F dentifrice was interrupted). Fingernails were weekly clipped during 35 weeks for F determination. F intake from diet and dentifrice in each phase was also determined. Both analyses were made with ion-specific electrode. F intake (Mean ± SD) was significantly higher (p<0.01) when the children were exposed to F from diet+dentifrice than only to F from diet (0.086 ± 0.032 and 0.040 ± 0.009 mg F/day/kg body weight, respectively). However, F concentrations in nails collected during the whole experimental period of 35 weeks presented great variation with no trend of decreasing after F dentifrice intake interruption. The findings suggest that fingernail may not be a reliable F biomarker of body burden from dentifrice.

As unhas têm sido consideradas um biomarcador para a exposição ao flúor (F), mas não há consenso se é um indicador confiável para exposição ao F a partir do dentifrício. Vinte e três crianças, com idade entre 1 a 3 anos, moradoras de Piracicaba (0,72 ppm F na água), Brasil, foram submetidas a duas fases de diferentes exposição ao F: fase A (1a a 11a semanas), as crianças foram expostas à combinação de F a partir da dieta (sólidos e líquidos) e dentifrício (1500 µg F/g como MFP); e na fase B (12ª a 29ª semanas), apenas ao F da dieta, uma vez que usaram dentifrício não fluoretado. As unhas das mãos foram coletadas semanalmente durante 35 semanas para determinação de F. A exposição ao F a partir da dieta e dentifrício foi também determinada. Ambas análises foram feitas com eletrodo específico para F. A exposição ao F foi significativamente maior (p<0,001) quando as crianças foram expostas ao F da dieta + dentifrício que ao F da dieta (0,086 ± 0,032 e 0,040 ± 0,009 mg F/kg corpóreo/dia, respectivamente). Entretanto, a concentração de F nas unhas coletadas durante todo o período experimental não diminuiu após a interrupção da ingestão do F a partir do dentifrício. Os resultados sugerem que as unhas das mãos não são um biomarcador confiável para refletir a exposição ao F pelo dentifrício.