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An extension of the uniform invariance principle for ordinary differential equations with finite delay is developed. The uniform invariance principle allows the derivative of the auxiliary scalar function to be positive in some bounded sets of the state space while the classical invariance principle assumes that . As a consequence, the uniform invariance principle can deal with a larger class of problems. The main difficulty to prove an invariance principle for functional differential equations is the fact that flows are defined on an infinite dimensional space and, in such spaces, bounded solutions may not be precompact. This difficulty is overcome by imposing the vector field taking bounded sets into bounded sets.

The existence of asymptotically almost periodic mild solutions for a class of abstract partial neutral integro-differential equations with unbounded delay is studied.

Background: The aim was to evaluate whether aspartame regular mouthwash prior to erosive challenges with citric or hydrochloric acids would be able to prevent erosive enamel wear., Material and Methods: This randomized, single blind in situ study was conducted with 3 crossover phases of 5 days. Polished bovine enamel blocks (n=252) were randomly divided among 6 groups/ 3 phases/ 21 volunteers. The groups under study were: aspartame solution (0.024% of aspartame in deionized water - experimental group), deionized water (negative-control) and stannous-containing solution (Elmex® Erosion Protection Dental Rinse; positive-control); subjected to erosion on citric acid or hydrochloric acid. Four times per day the volunteers rinsed the intraoral appliance with the respective solutions ( in situ ) prior to immersion of half of the appliance in 0.05M citric acid and the other half in 0.01M hydrochloric acid for 120 seconds (extraoral). The response variable was enamel loss by profilometry. Data were analyzed by ANOVA and Tukey's test ( p <0.05)., Results: No difference on enamel loss was found between aspartame solution and deionized water. Stannous-solution resulted in less enamel loss compared to deionized water. Hydrochloric acid resulted in higher enamel loss than citric acid., Conclusions: In this model, aspartame was not able to prevent erosive tooth wear against citric or hydrochloric acids. Key words: Dental erosion, aspartame, stannous fluoride, citric acid, hydrochloric acid., Competing Interests: Conflicts of interest The authors declare that they have no conflict of interest., (Copyright: © 2020 Medicina Oral S.L.)

Background: The aim of this study was evaluated the eroded enamel rehardening potential using upper palatal and lower buccal removable appliances in different times of salivary exposure (30 min, 1h, 2h, 12h) after a single erosive challenge event., Material and Methods: After initial surface hardness evaluation, bovine enamel blocks were eroded in vitro (0.01 M hydrochloric acid, pH 2.3, 30 seconds), selected (n = 160) and randomly assigned to the two appliance designs and twenty volunteers. Four enamel blocks were inserted in each removable appliance. On the in situ phase, the volunteers were instructed to use the upper palatal and lower buccal appliances simultaneously for 12 nonconsecutive hours. After each predetermined period of time of salivary exposure, the enamel blocks were removed from the appliances for immediate evaluation of surface hardness, enabling percentage of surface hardness recovery calculation (%SHR). The data were analyzed using two-way ANOVA and Tukey's test (α=5%)., Results: The results showed no difference in the degree of enamel rehardening by the upper palatal or lower buccal appliances ( p >0.0001). Regarding the time of use of the appliances, it was demonstrated that 30 minutes (upper = 21.12%, lower = 19.84%) and 1 hour (upper = 35.69%, lower = 30.50%) promoted lower hardness recovery than two hours (upper = 44.65%, lower 40.80%) of salivary exposure ( p <0.0001). The use of 12 hours (upper = 49.33%, lower = 49.00%), including the sleeping time of the volunteers did not increase the %SHR., Conclusions: The location of the appliance does not influence the re-hardening ability of saliva and the use of intraoral appliances for 2 hours seems to be appropriate for partial rehardening of the softened enamel surface. Key words: Tooth erosion, in situ, saliva, tooth remineralization., Competing Interests: Conflict of interest statement:The authors report no conflicts of interest in this study., (Copyright: © 2019 Medicina Oral S.L.)

Introduction: The increasing reports of vancomycin-resistant Staphylococcus strains (VRS) haves caused concern worldwide, from the laboratory detection to patient management. This study aimed to identify the occurrence of VRS strains among healthcare professionals from a university hospital., Methods: A total of 102 Staphylococcus sp. isolates from healthcare professionals, obtained in a previous study were evaluated according to standard techniques for VRS detection., Results: After screening inoculation of plates containing 6µg/ml of vancomycin, 19 resistant isolates were identified. The susceptibility profile to other antimicrobials revealed 18 multidrug resistant isolates. The minimum inhibitory concentration (MIC) was determined by E-test and broth microdilution. According to E-tests, of 19 isolates grown in BHI-V6, four isolates presented MIC ≥ 128 µg/ml, seven with MIC ranging from 4 to 8 µg/ml, and eight with MIC ≤ 2µg/ml. By broth microdilution, 14 isolates presented MIC ≤ 2 µg/ml and five with MIC ≥ 16µg/ml. The presence of the gene vanA was determined by PCR in the five resistant isolates, and this gene was detected in one of the strains. Furthermore, among the 19 strains, the gene mecA was found in 13 (39,4%) isolates, including the strain carrying the gene vanA., Conclusions: Based on these results, we highlight the presence of one strain carrying both vanA and the mecA genes, as well as multidrug-resistant strains colonizing healthcare professionals, and their importance as potential vectors to spread strains carrying resistance genes in the hospital environment.

Objective: To phenotypically evaluate biofilm production by Pseudomonas aeruginosa clinically isolated from patients with ventilator-associated pneumonia., Methods: Twenty clinical isolates of P. aeruginosa were analyzed, 19 of which were from clinical samples of tracheal aspirate, and one was from a bronchoalveolar lavage sample. The evaluation of the capacity of P. aeruginosa to produce biofilm was verified using two techniques, one qualitative and the other quantitative., Results: The qualitative technique showed that only 15% of the isolates were considered biofilm producers, while the quantitative technique showed that 75% of the isolates were biofilm producers. The biofilm isolates presented the following susceptibility profile: 53.3% were multidrug-resistant, and 46.7% were multidrug-sensitive., Conclusion: The quantitative technique was more effective than the qualitative technique for the detection of biofilm production. For the bacterial population analyzed, biofilm production was independent of the susceptibility profile of the bacteria, demonstrating that the therapeutic failure could be related to biofilm production, as it prevented the destruction of the bacteria present in this structure, causing complications of pneumonia associated with mechanical ventilation, including extrapulmonary infections, and making it difficult to treat the infection.

Objectives: The present study evaluated fluoride (F) and calcium (Ca) concentrations in the biofilm fluid formed in situ under cariogenic challenge after using F dentifrices supplemented or not with sodium trimetaphosphate (TMP) or calcium glycerophosphate (CaGP)., Methods: Volunteers (n = 12) were randomly divided into 5 groups according to the toothpastes used: placebo (without F, CaGP or TMP), 1100 ppm F (1100F) and low-fluoride dentifrice (LFD, 550 ppm F) with no supplementation (550F) or supplemented with 1 % TMP (550F-TMP) or 0.25 % CaGP (550F-CaGP). In each phase, volunteers wore palatal appliances containing 4 bovine enamel blocks. Cariogenic challenge was performed with 30 % sucrose solution, 6 times/day. On the morning of the eigth day, biofilm samples were collected 12 h and 1 h after brushing and cariogenic challenge. F and Ca analyses in the biofilm fluid were performed with the inverted electrode after buffering with TISAB III and using the Arsenazo III method, respectively. Data were submitted to two-way ANOVA (repeated measures) and Student-Newman-Keuls test (p < 0.05)., Results: A dose-response relationship was verified between F concentrations in the dentifrices and in the biofilm fluid. Significant differences were observed among placebo, 550F, and 1100F only 1 h after brushing, without statistical differences among 550F, 550F-TMP, and 550F-CaGP. No defined trend was observed among the groups regarding Ca concentrations, with the highest values seen for placebo and 550F-CaGP., Conclusion: The anticaries effect of LFDs supplemented with CaGP or TMP cannot be related to an increased availability of F and Ca in the biofilm fluid., Clinical Significance: The better performance of LFDs containing CaGP or TMP shown in previous studies should be attributed to their ability to interact with tooth enamel and with the biofilm, rather to their effect on the biofilm fluid.

Introduction: Information on the prevalence of lead exposure is essential to formulate efficient public health policies. Developed countries have implemented successful public policies for the prevention and control of lead poisoning. In the United States, Canada, Japan and the European Union, for instance, periodically repeated prevalence studies show that blood lead levels (BLLs) in children have decreased overall. Although BLL of Latino children in the U.S. have also dropped in recent years, the geometric mean remains higher than that of white children. Little is known about lead exposure in children in Latin America and the Caribbean (LAC). In this review, we responded to two questions: What is currently known about lead sources and levels in children in LAC? Are there public policies to prevent children's exposure to lead in LAC?, Method: We conducted a literature review covering the period from January 2000 to March 2014 in the PubMed and Lilacs databases to obtain English, Portuguese and Spanish language studies reporting the prevalence of BLLs in children aged 0-18years living in LAC countries. No specific analytical method was selected, and given the scarcity of data, the study was highly inclusive., Results: Fifty-six papers were selected from 16 different LAC countries. The children's BLLs found in this review are high (≥10μg/dL) compared to BLLs for the same age group in the U. S. However, most studies reported an association with some type of "lead hot spot", in which children can be exposed to lead levels similar to those of occupational settings. Only Peru and Mexico reported BLLs in children from population-based studies., Conclusions: Most BLLs prevalence studies carried out in LAC were in areas with known emission sources. The percentage of children at risk of lead poisoning in LAC is unknown, and probably underestimated. Thus, there is an urgent need to establish public health policies to quantify and prevent lead poisoning, specifically by prioritizing the identification and control of "hot spots"., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Low level laser therapy (LLLT) has been shown to stimulate bone cell metabolism but their impact on the matrix metalloproteinase (MMP) expression and activity is little explored. This study evaluated the influence of LLLT at two different wavelengths, red and infrared, on MC3T3-E1 preosteoblast viability, alkaline phosphatase (ALP) and MMP-2 and -9 activities. To accomplish this, MC3T3-E1 cells were irradiated with a punctual application of either red (660nm; InGaAIP active medium) or infrared (780nm; GaAlAs active medium) lasers both at a potency of 20mW, energy dose of 0.08 or 0.16J, and energy density of 1.9J/cm 2 or 3.8J/cm 2 , respectively. The control group received no irradiation. Cellular viability, ALP and MMP-2 and -9 activities were assessed by MTT assay, enzymatic activity and zymography, respectively, at 24, 48 and 72h. The treatment of cells with both red and infrared lasers significantly increased the cellular viability compared to the non-irradiated control group at 24 and 48h. The ALP activity was also up modulated in infrared groups at 24 and 72h, depending on the energy densities. In addition, the irradiation with red laser at the energy density of 1.9J/cm 2 promoted an enhancement of MMP-2 activity at 48 and 72h. However, no differences were observed for the MMP-9 activity. In conclusion, when used at these specific parameters, LLL modulates both preosteoblast viability and differentiation highlighted by the increased ALP and MMP-2 activities induced by irradiation., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The objective of this study was to evaluate markers of oxidative stress in the brains of rats exposed to lead acetate (Pb(C 2 H 3 O 2 ) 2 ), either associated or not associated with ferrous sulfate (FeSO 4 ). A total of 36 weaning rats (Rattus norvegicus) were divided into 6 groups of six animals and exposed to lead acetate for six weeks. In the control group (control), the animals received deionized water. The Pb260 and Pb260 + Fe received 260 µM lead acetate, and the Pb1050 and Pb1050 + Fe received 1050 µM lead acetate. The Pb260 + Fe and Pb1050 + Fe were supplemented with 20 mg of ferrous sulfate/Kg body weight every 2 days. Group Fe received deionized water and ferrous sulfate. The rat brains were collected to analyze the enzymatic activity of catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), and the concentration of reduced glutathione (GSH), lipid peroxidation (TBARS), and total antioxidant substance (TAS) (DPPH • technique). The activity of SOD and GPx in the experimental groups decreased compared to the control, together with the concentration of GSH (p < 0.05). For CAT analysis, SOD tended to increase in concentration in the experimental groups without a concomitant exposure to FeSO 4 , whereas GPx showed a slight tendency to increase in activity compared to the control. For TAS-DPPH • , there was a decrease in the experimental groups (p < 0.05). According to the results, SOD, GPx, and GSH were affected by lead acetate and exposure to ferrous sulfate changed this dynamic. However, further studies are needed to verify whether ferrous sulfate acts as a protectant against the toxic effects of lead. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 813-822, 2017., (© 2016 Wiley Periodicals, Inc.)

Abnormalities in lungs caused by emphysema might alter their response to sepsis and the occurrence of acute lung injury (ALI). This study compared the extension of ALI in response to intraperitoneal lipopolysaccharide (LPS) injection in Wistar rats with and without emphysema induced by elastase. Adult male Wistar rats were randomized into four groups: control, emphysema without sepsis, normal lung with sepsis and emphysema with sepsis. Sepsis was induced, and 24 h later the rats were euthanised. The following analysis was performed: blood gas measurements, bronchoalveolar lavage (BAL), lung permeability and histology. Animals that received LPS showed significant increase in a lung injury scoring system, inflammatory cells in bronchoalveolar lavage (BAL) and IL-6, TNF-α and CXCL2 mRNA expression in lung tissue. Animals with emphysema and sepsis showed increased alveolocapillary membrane permeability, demonstrated by higher BAL/serum albumin ratio. In conclusion, the presence of emphysema induced by elastase increases the inflammatory response in the lungs to a systemic stimulus, represented in this model by the intraperitoneal injection of LPS., (© 2016 The Authors. International Journal of Experimental Pathology © 2016 International Journal of Experimental Pathology.)

Osteoporosis is a systemic disease characterized by bone degradation and decreased bone mass that promotes increased bone fragility and eventual fracture risk. Studies have investigated the use of sodium fluoride (NaF) for the treatment of osteoporosis. However, fluoride can alter glucose homeostasis. The aim of this study was to evaluate the effects of NaF intake (50 mg/L) from water on the following parameters of ovariectomized (OVX) rats: (1) tyrosine phosphorylation status of insulin receptor substrate (pp185 (IRS-1/IRS-2)) in white adipose tissue; (2) insulin sensitivity; (3) plasma concentrations of glucose, insulin, total cholesterol, triglyceride, TNF-α, IL-6, osteocalcin, calcium, and fluoride; (4) bone density and biomechanical properties in the tibia; and (5) tibia histomorphometric analysis. Fifty-two Wistar rats (2 months old) were ovariectomized and distributed into two groups: control group (OVX-C) and NaF group (OVX-F), which was subjected to treatment with NaF (50 mg/L) administered in drinking water for 42 days. The chronic treatment with NaF promoted (1) a decrease in pp185 (IRS-1/IRS-2) tyrosine phosphorylation status after insulin infusion in white adipose tissue and in insulin sensitivity; (2) an increase in the plasma concentration of insulin, fluoride, osteocalcin, calcium, triglyceride, VLDL-cholesterol, TNF-α, and IL-6; (3) a reduction in the trabecular width, bone area, stiffness, maximum strength, and tenacity; (4) no changes in body weight, food and water intake, plasma glucose, total cholesterol, HDL-cholesterol, LDL-cholesterol, bone mineral content, and bone mineral density. It was concluded that chronic treatment with NaF (50 mg/L) in OVX rats causes a decrease in insulin sensitivity, insulin signaling transduction, and biochemical, biomechanical, and histomorphometric bone parameters.

Background: Here, we evaluated the relationship of diet and F-induced oxidative stress to lipid metabolism in the liver of rats eating normocaloric or hypercaloric diets for two time periods (20 or 60 days)., Methods: Seventy-two 21-day-old Wistar rats were divided into 2 groups (n = 36) based on the type of diet they were eating; each of these groups was then further divided into another two groups (n = 18) based on the time periods of either 20 or 60 days, for a total of four groups. Each of these was divided into 3 subgroups (n = 6 animals/subgroup), dependent on the dose of F administered in the drinking water (0 mg/L(control), 15 mg/L or 50 mg/L). After the experimental period, blood samples and the liver were collected. Plasma samples were analyzed for HDL, cholesterol and triglycerides. Western blots were performed to probe for GRP78, Erp29, SOD2, Apo-E and SREBP in hepatic tissues., Results: As expected,the expression of target proteins involved in oxidative stress increased in the F-treated groups, especially in liver tissue obtained from animals eating a hypercaloric diet. Most changes in the lipid levels and pathological conditions were seen earlier in the time period, at day 20. The morphometric analyses showed a reduction in steatosis in groups on ahypercaloric diet and treated with 50 mg F/L compared to the control, while no changes were obtained in normocaloric-fed rats. Accordingly, plasma TG was reduced in the F-treated group. The reduced expression of Apo-E in a time- and diet-dependent pattern may account for the particular decrease in steatosis in hypercaloric-fed F-treated rats., Conclusions: These results suggest that F changes liver lipid homeostasis, possibly because of the induction of oxidative stress, which seems to be higher in animals fed hypercaloric diets.

Introduction: Acidic dentifrices have been shown to be more effective than neutral ones against dental caries using in vitro, in situ, and clinical protocols., Objectives: Given the scarcity of studies assessing intraoral fluoride (F) retention after using such formulations, the present study evaluated the influence of pH and F concentration of dentifrices on F uptake by saliva, biofilm, and biofilm fluid., Methods: Volunteers (n = 22) were randomly assigned to dentifrices containing 0 (placebo), 550 (LFD, low-fluoride dentifrice), and 1100 ppm F (CD, conventional dentifrice) at pH 4.5 and 7.0 and brushed their teeth 3 times/day following a double-blind, crossover protocol. Saliva and biofilm samples were collected after 7 days of using the dentifrices, 1 and approximately 12 h after last brushing. F and calcium (Ca) analyses were performed with the inverted electrode after buffering with TISAB III and the Arsenazo III method, respectively. Data were analyzed by 3-way ANOVA, Tukey's HSD test, and Pearson's correlation coefficient (p < 0.05)., Results: F concentrations in biofilm fluid and whole biofilm 1 h after brushing with acidic F-toothpastes were higher than those related to neutral counterparts, although the differences were small and not significant; no increases were observed in salivary F concentrations influenced by dentifrice pH. Moreover, no definite trend was observed for Ca concentrations in these compartments., Conclusion: Dentifrice pH had some influence on F uptake by the biofilm fluid, having lesser or no impact on F uptake by the biofilm and saliva, respectively., Clinical Significance: Toothbrushing with acidic toothpastes leads to slight increases in F concentrations in the biofilm fluid when compared to neutral formulations, which may contribute to the higher anticaries effect of acidic formulations.

Objective: To evaluate the effects that administering dexamethasone before the induction of ventilator-induced lung injury (VILI) has on the temporal evolution of that injury., Methods: Wistar rats were allocated to one of three groups: pre-VILI administration of dexamethasone (dexamethasone group); pre-VILI administration of saline (control group); or ventilation only (sham group). The VILI was induced by ventilation at a high tidal volume. Animals in the dexamethasone and control groups were euthanized at 0, 4, 24, and 168 h after VILI induction. We analyzed arterial blood gases, lung edema, cell counts (total and differential) in the BAL fluid, and lung histology., Results: At 0, 4, and 24 h after VILI induction, acute lung injury (ALI) scores were higher in the control group than in the sham group (p < 0.05). Administration of dexamethasone prior to VILI induction decreased the severity of the lung injury. At 4 h and 24 h after induction, the ALI score in the dexamethasone group was not significantly different from that observed for the sham group and was lower than that observed for the control group (p < 0.05). Neutrophil counts in BAL fluid were increased in the control and dexamethasone groups, peaking at 4 h after VILI induction (p < 0.05). However, the neutrophil counts were lower in the dexamethasone group than in the control group at 4 h and 24 h after induction (p < 0.05). Pre-treatment with dexamethasone also prevented the post-induction oxygenation impairment seen in the control group., Conclusions: Administration of dexamethasone prior to VILI induction attenuates the effects of the injury in Wistar rats. The molecular mechanisms of such injury and the possible clinical role of corticosteroids in VILI have yet to be elucidated., Objetivo: Avaliar os efeitos da administração de dexametasona antes da indução de lesão pulmonar induzida por ventilação mecânica (LPIVM) na evolução temporal dessa lesão., Métodos: Ratos Wistar foram alocados em um dos três grupos: administração de dexametasona pré-LPIVM (grupo dexametasona); administração de salina pré-LPIVM (grupo controle); e somente ventilação (grupo sham). A LPIVM foi realizada por ventilação com volume corrente alto. Os animais dos grupos dexametasona e controle foram sacrificados em 0, 4, 24 e 168 h após LPIVM. Analisamos gasometria arterial, edema pulmonar, contagens de células (totais e diferenciais) no lavado broncoalveolar e histologia de tecido pulmonar., Resultados: Em 0, 4 e 24 h após LPIVM, os escores de lesão pulmonar aguda (LPA) foram maiores no grupo controle que no grupo sham (p < 0,05). A administração de dexametasona antes da LPIVM reduziu a gravidade da lesão pulmonar. Em 4 e 24 h após a indução, o escore de LPA no grupo dexametasona não foi significativamente diferente daquele observado no grupo sham e foi menor que o observado no grupo controle (p < 0,05). As contagens de neutrófilos no lavado broncoalveolar estavam aumentadas nos grupos controle e dexametasona, com pico em 4 h após LPIVM (p < 0,05). Entretanto, as contagens de neutrófilos foram menores no grupo dexametasona que no grupo controle em 4 e 24 h após LPIVM (p < 0,05). O pré-tratamento com dexametasona também impediu o comprometimento da oxigenação após a indução visto no grupo controle., Conclusões: A administração de dexametasona antes de LPIVM atenua os efeitos da lesão em ratos Wistar. Os mecanismos moleculares dessa lesão e o possível papel clínico dos corticosteroides na LPIVM ainda precisam ser elucidados.

Objective: To evaluate cytotoxicity and effect on protease activity of epigallocatechin-gallate extracted from experimental restorative dental copolymers in comparison to the control compound chlorhexidine., Methods: Copolymer disks were prepared from bis-GMA/TEGDMA (70/30 mol%) containing no compound (control) or 1% w/w of either epigallocatechin-gallate or chlorhexidine. MDPC-23 odontoblast-like cells were seeded with the copolymer extracts leached out into deionized water. Cell metabolic activity was quantified by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay at 24, 48, 72 h. Inhibition of protease activity by resin extracts was measured by a collagenolytic/genatinolytic enzyme activity assay and gelatin zymography. Data for MTT and protease inhibition were analyzed using two-way ANOVA followed by Tukey or Bonferroni post hoc tests (α=0.05)., Results: The MTT revealed that at 72 h, extracts from control (16.7%) and chlorhexidine (22.3%) copolymers induced significant reduction in cell metabolism (p<0.05). All copolymer extracts caused enzymatic inhibition in a dose dependent manner (p<0.01). Even when highly diluted, epigallocatechin-gallate extract had a significant antiproteolytic activity (p<0.05). Zymograms showed that all extracts reduced activity of MMP-2 and MMP-9 (pro- and active forms), with MMP-9 exhibiting the highest percentage inhibition revealed by densitometry., Conclusions: Epigallocatechin-gallate and chlorhexidine extracts did not exert cytotoxicity on evaluated cells when compared to control extracts. Both compounds retained antiproteolytic activity after extraction from a dental copolymer., Clinical Significance: Once extracted from a dental copolymer, epigallocatechin-gallate is not cytotoxic and retains antiproteolytic activity. These results may allow incorporation of epigallocatechin-gallate as a natural-safe alternative to chlorhexidine in functionalized restorative materials., (Copyright © 2016. Published by Elsevier Ltd.)

Objective: In this study, we investigated the differential pattern of protein expression in the liver of these mice to provide insights on why they have different responses to F., Material and Methods: Weanling male A/J and 129P3/J mice (n=10 from each strain) were pared and housed in metabolic cages with ad libitum access to low-F food and deionized water for 42 days. Liver proteome profiles were examined using nLC-MS/MS. Protein function was classified by GO biological process (Cluego v2.0.7 + Clupedia v1.0.8) and protein-protein interaction network was constructed (PSICQUIC, Cytoscape)., Results: Most proteins with fold change were increased in A/J mice. The functional category with the highest percentage of altered genes was oxidation-reduction process (20%). Subnetwork analysis revealed that proteins with fold change interacted with Disks large homolog 4 and Calcium-activated potassium channel subunit alpha-1. A/J mice had an increase in proteins related to energy flux and oxidative stress., Conclusion: This could be a possible explanation for the high susceptibility of these mice to the effects of F, since the exposure also induces oxidative stress., Competing Interests: STATEMENT The authors have declared no conflict of interest.

Introduction: There is a mechanism of macrolide resistance in Staphylococcus spp. which also affects the lincosamides and type B streptogramins characterizing the so-called MLSB resistance, whose expression can be constitutive (cMLSB) or inducible (iMLSB) and is encoded mainly by ermA and ermC genes. The cMLSB resistance is easily detected by susceptibility testing used in the laboratory routine, but iMLSB resistance is not. Therapy with clindamycin in cases of infection with isolated iMLSB resistance may fail., Objective: To characterize the phenotypic (occurrence of cMLSB and iMLSB phenotypes) and molecular (occurrence of ermA and ermC genes) profiles of MLSB resistance of clinical isolates of susceptible and methicillin-resistant Staphylococcus aureus and CNS (coagulase-negative Staphylococcus) from patients of a university hospital, in Pernambuco., Methods: The antimicrobial susceptibility of 103 isolates was determined by the disk diffusion technique in Mueller-Hinton agar followed by oxacillin screening. The iMLSB phenotype was detected by D test. Isolates with cMLSB and iMLSB phenotypes were subjected to polymerase chain reaction (PCR) for the detection of ermA and ermC genes., Results: The cMLSB and iMLSB phenotypes were respectively identified in 39 (37.9%) and five (4.9%) isolates. The iMLSB phenotype was found only in four (10.8%) methicillin-susceptible S. aureus and one (4.5%) methicillin-resistant S. aureus. In the 44 isolates subjected to PCR, four (9.1%) only ermA gene was detected, a lower frequency when compared to only ermC 17 (38.6%) gene and to one (2.3%) isolate presenting both genes., Conclusion: In the Staphylococcus spp. analyzed, the ermC gene was found more often than the ermA, although the iMLSB phenotype had been less frequent than the cMLSB. It was important to perform the D test for its detection to guide therapeutic approaches., (Copyright © 2016 Elsevier Editora Ltda. All rights reserved.)

Introduction: The benefits of prone position ventilation are well demonstrated in the severe forms of acute respiratory distress syndrome, but not in the milder forms. We investigated the effects of prone position on arterial blood gases, lung inflammation, and histology in an experimental mild acute lung injury (ALI) model., Methods: ALI was induced in Wistar rats by intraperitoneal Escherichia coli lipopolysaccharide (LPS, 5 mg/kg). After 24 h, the animals with PaO2/FIO2 between 200 and 300 mmHg were randomized into 2 groups: prone position (n = 6) and supine position (n = 6). Both groups were compared with a control group (n = 5) that was ventilated in the supine position. All of the groups were ventilated for 1 h with volume-controlled ventilation mode (tidal volume = 6 ml/kg, respiratory rate = 80 breaths/min, positive end-expiratory pressure = 5 cmH2O, inspired oxygen fraction = 1) RESULTS: Significantly higher lung injury scores were observed in the LPS-supine group compared to the LPS-prone and control groups (0.32 ± 0.03; 0.17 ± 0.03 and 0.13 ± 0.04, respectively) (p < 0.001), mainly due to a higher neutrophil infiltration level in the interstitial space and more proteinaceous debris that filled the airspaces. Similar differences were observed when the gravity-dependent lung regions and non-dependent lung regions were analyzed separately (p < 0.05). The BAL neutrophil content was also higher in the LPS-supine group compared to the LPS-prone and control groups (p < 0.05). There were no significant differences in the wet/dry ratio and gas exchange levels., Conclusions: In this experimental extrapulmonary mild ALI model, prone position ventilation for 1 h, when compared with supine position ventilation, was associated with lower lung inflammation and injury.

Background: Artificial lesions produced by different protocols might directly influence the response to different remineralising treatments. This study compared the response of different artificial caries-like enamel lesions to home-care and professional fluoride based-remineralising treatments in situ., Methods: The tested demineralising protocols were methylcellulose- MC gel, polyacrylic acid - PA gel, tetraethyl methylene diphosphanate - TEMDP solution, and acetate- Buffer solution. The lesions were remineralised using an in situ model, following a crossover and double blind design. Twelve subjects wore intra-oral appliances during 3 phases (3 d each): control (C) (saliva); home-care F(-) treatment (FD) (1,100 ppm F(-) dentifrice, 2x1 min/day); and professional (FVD) (22,600 ppm F(-) varnish) plus FD. The de-remineralisation was measured by transverse microradiography-TMR and hardness (surface hardness/cross-sectional hardness, SH/CSH, respectively)., Results: For SH, lesions produced by PA gel were the only one showing significant differences among the remineralising treatments (C x FD x FVD); while the TEMDP lesion were not responsive to any fluoride treatment (for both SH/CSH). For TMR, there were no differences among the remineralising treatments, regardless of the type of lesion. Generally, the most responsive lesions to fluoride were the less demineralised lesions (considering hardness: PA gel and Buffer)., Conclusions: The type of lesion has influence on the surface remineralisation degree induced by home-care and professional fluoride treatments using this in situ model.

Unlabelled: The use of gels and mouthrinses with MMP inhibitors (chlorhexidine, and green tea extract) was shown to prevent erosive wear. The aim of this study was to analyze the protective effect of toothpastes containing MMP inhibitors on dentine loss induced by erosion in vitro., Material and Methods: Five groups each containing 12 specimens of human root dentine were prepared. The specimens were subjected to 1 min erosion by immersion in a cola drink, 4 times a day, for 5 d. Each day, after the first and last erosive challenges, the specimens were brushed for 15 s with a slurry of dentifrice and water (1:3) containing placebo, 1,100 ppm fluoride, 0.61% green tea extract, 0.12% chlorhexidine or 0.004% chlorhexidine (commercial toothpaste). Between the acid challenges, the specimens were stored in artificial saliva with remineralizing potential until the next treatment. Dentine loss was determined using profilometry. Data were analyzed using one-way ANOVA after log transform (p<0.05)., Results: The mean wear values (μm) were as follows: placebo 1.83±0.53; 0.61% green tea extract 1.00±0.21; fluoride 1.27±0.43; 0.12% chlorhexidine 1.19±0.30; and 0.004% chlorhexidine 1.22±0.46. There was a significant difference in wear between placebo and all the treatment toothpastes, which did not differ from each other., Conclusion: The results suggest that toothpastes containing MMP inhibitors are as effective as those based on NaF in preventing dentine erosion and abrasion.

This study analysed in vitro the effect of milk against dental erosion, considering three factors: the type of milk (bovine whole/fat-free), the presence of different fluoride concentrations and the time of application (before/after erosive challenge). Bovine enamel (n = 15/group) and root dentine (n = 12/group) specimens were submitted to the following treatments: after the first erosive challenge - 0.9% NaCl solution (negative control), whole milk with 0, 2.5, 5.0 and 10.0 ppm F, fat-free milk with 0, 2.5, 5.0 and 10.0 ppm F, and 0.05% NaF solution (positive control); before the first erosive challenge - whole milk with 0, 2.5, 5.0 and 10.0 ppm F, fat-free milk with 0, 2.5, 5.0 and 10.0 ppm F, and 0.05% NaF solution (positive control). Specimens were submitted to demineralisation-remineralisation regimes 4 times/day for 5 days. The response variables were enamel and dentine loss (in micrometres). Data were analysed using Kruskal-Wallis/Dunn's test (p < 0.05). For enamel, whole milk containing 10 ppm F, applied before the erosive challenge, was the most protective treatment, but with no significant difference compared with the same treatment carried out after the erosive challenge. For dentine, whole fluoridated milk (all concentrations, after), fat-free 10 ppm F milk (after, before) and whole milk with or without F (except 2.5 ppm F, all before) significantly reduced dentine erosion. It seems that the presence of fluoride, especially at 10 ppm, is the most important factor in reducing dental erosion., (© 2016 S. Karger AG, Basel.)

This study investigated the isolated and combined effect of a single application of TiF4 or NaF varnish versus daily use of a solution containing a low concentration of TiF4/NaF against tooth erosion in vitro. A total of 90 bovine enamel and 108 root dentin samples were treated as follows: control (no treatment), solution containing TiF4/NaF (500 ppm F-, pH 4.4), NaF varnish (24,500 ppm F-, pH 5.0), TiF4 varnish (24,500 ppm F-, pH 1.0), TiF4 varnish + solution, and NaF varnish + solution. The erosive challenges were performed 4 × 90s/day (0.1% citric acid, pH 2.5) and, between them, the samples were immersed in artificial saliva. The tooth loss was measured using contact profilometry (after 7 days for dentin and after 7, 10, and 14 days for enamel). All treatments were effective in reducing tooth loss, except NaF varnish for enamel on day 7 (p < 0.0001). TiF4/NaF solution and TiF4 varnish did not differ with respect to enamel loss for 10 days; thereafter, TiF4 varnish lost its protective effect compared to TiF4/NaF solution. The combination of vehicles was more effective in reducing enamel loss than both varnishes on their own but not compared to the solution. For dentin, TiF4 varnish was more effective than NaF varnish, while TiF4/NaF solution and NaF varnish were similar. The combination of vehicles improved the protective effect only when compared to NaF varnish on its own (p < 0.0001). Both types of TiF4 applications, isolated or combined, were effective against tooth erosion, but some differences in their performance were seen between enamel and dentin., (© 2016 S. Karger AG, Basel.)

The mechanisms by which excessive ingestion of fluoride (F) during amelogenesis leads to dental fluorosis (DF) are still not precisely known. Inbred strains of mice vary in their susceptibility to develop DF, and therefore permit the investigation of underlying molecular events influencing DF severity. We employed a proteomic approach to characterize and evaluate changes in protein expression from secretory-stage and maturation-stage enamel in 2 strains of mice with different susceptibilities to DF (A/J, i.e. 'susceptible' and 129P3/J, i.e. 'resistant'). Weanling male and female susceptible and resistant mice fed a low-F diet were divided into 2 F-water treatment groups. They received water containing 0 (control) or 50 mg F/l for 6 weeks. Plasma and incisor enamel was analyzed for F content. For proteomic analysis, the enamel proteins extracted for each group were separated by 2-dimensional electrophoresis and subsequently characterized by liquid-chromatography electrospray-ionization quadrupole time-of-flight mass spectrometry. F data were analyzed by 2-way ANOVA and Bonferroni's test (p < 0.05). Resistant mice had significantly higher plasma and enamel F concentrations when compared with susceptible mice in the F-treated groups. The proteomic results for mice treated with 0 mg F/l revealed that during the secretory stage, resistant mice had a higher abundance of proteins than their susceptible counterparts, but this was reversed during the maturation stage. Treatment with F greatly increased the number of protein spots detected in both stages. Many proteins not previously described in enamel (e.g. type 1 collagen) as well as some uncharacterized proteins were identified. Our findings reveal new insights regarding amelogenesis and how genetic background and F affect this process., (© 2016 S. Karger AG, Basel.)

Objectives: This study characterized the proteome profile of the acquired pellicle formed in vivo on enamel. Changes in this proteome profile after exposure to lactic or citric acid were also evaluated., Methods: Volunteers (n=8) were subjected to dental prophylaxis. After 2 h to allow the formation of the acquired pellicle, the teeth were isolated with cotton rolls and 1 mL of citric acid (1%, pH 2.5) or lactic acid (0.1 M pH 4.8) or deionized water was gently applied with a pipette on the anterior teeth (both maxillary and mandibular) for 10 s. In sequence, the pellicle was collected with an electrode filter paper soaked in 3% citric acid. This procedure was repeated for two additional days following a crossover protocol. Proteins were subjected to reverse phase liquid chromatography coupled to mass spectrometry (nLC-ESI-MS/MS). MS/MS data were processed and submitted to Proteome Discoverer software. Searches were done using SWISS-PROT and TrEMBL databases for human proteins., Results: In total, seventy-two proteins were present in all groups and were submitted to quantitative analysis (SIEVE). Some of these proteins were increased more than two-fold after exposure to the acids. Among them, cystatin-B was increased 20- and 13-fold after exposure to citric and lactic acids, respectively. Additionally, some proteins were identified in only one of the groups (18, 5, and 11 proteins for deionized water, citric and lactic acids, respectively)., Conclusions: Our results open new insights regarding potentially acid-resistant proteins that could be added to dental products to prevent acidic dissolution of the teeth., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Objective: This in vitro study assessed the anti-erosive effect of experimental mouthrinses containing TiF4 and NaF on dentin erosive loss., Material and Methods: Bovine dentin specimens were randomly allocated into the groups (n=15): (1) SnCl2/NaF/AmF (Erosion Protection/GABA, pH 4.5, positive control); (2) experimental solution with 0.0815% TiF4(pH 2.5); (3) 0.105% NaF (pH 4.5); 4) 0.042% NaF+0.049% TiF4 (pH 4.4); (5) 0.063% NaF+0.036% TiF4 (pH 4.5); (6) no treatment (negative control). Each specimen was cyclically demineralized (Sprite Zero, pH 2.6, 4x90 s/day) and exposed to artificial saliva between the erosive challenges for 7 days. The treatment with the fluoride solutions was done 2x60 s/day, immediately after the first and the last erosive challenges of the day. Dentin erosive loss was measured by profilometry (μm). The data were analyzed using Kruskal Wallis/Dunn tests (p<0.05)., Results: Mouthrinses containing TiF4or Sn/F were able to show some protective effect against dentin erosive loss compared to negative control. The best anti-erosive effect was found for experimental solution containing 0.0815% TiF4 (100% reduction in dentin loss), followed by 0.042% NaF+0.049% TiF4 (58.3%), SnCl2/NaF/AmF (52%) and 0.063% NaF+0.036% TiF4 (40%). NaF solution (13.3%) did not significantly differ from control., Conclusion: The daily application of experimental mouthrinse containing TiF4 and NaF has the ability to reduce dentin erosion, as well as Erosion Protection and TiF4 alone.

This study aimed to identify metalloproteins that lose their metal ions in the presence of mercury (Hg) and bind to Hg in breast milk samples collected from the riverine population of the Madeira River, a tributary of the Amazon River. Initially, total Hg was determined from the hair of lactating women to identify individuals who were contaminated followed by a proteomic analysis of breast milk samples through two-dimensional polyacrylamide gel electrophoresis after acetone precipitation. The presence of Hg in the obtained protein spots was determined by cold vapor atomic absorption spectrometry and cold vapor atomic fluorescence spectrometry. These determinations indicated the presence of Hg in one protein spot, which was then characterized through electrospray ionization tandem mass spectrometry. Based on searches in the UniProt database, this protein spot was identified as lysozyme C.

Objectives: For the first time, this study characterized the proteome of the acquired pellicle formed on human dentine. The changes in this proteome after exposure to lactic or citric acid were also evaluated., Methods: Volunteers (n=9) wore a mandibular device containing 6 specimens of human root dentine. After the device remained in the volunteers' oral cavities for 10min or 2h to allow the formation of the acquired pellicle in situ, the specimens were immersed in citric acid (1%, pH 2.5) or lactic acid (0.1M, pH 4.8) or deionized water for 20s. In sequence, the pellicle was collected with an electrode filter paper soaked in 3% citric acid. This procedure was repeated for two additional days following a crossover protocol. After harvest, proteins were subjected to reverse phase liquid chromatography coupled to mass spectrometry (nLC-ESI-MS/MS). MS/MS data were processed and submitted to Proteome Discoverer software. Searches were done using SWISS-PROT and TrEMBL databases for human proteins., Results: In total, 223 distinct proteins were identified in the dentine acquired pellicle in each of the different conditions. Exposure to citric acid dramatically reduced the number of identified proteins. This did not occur for lactic acid. Acid-resistant proteins, such as mucins, were identified after pellicle was exposed to lactic or citric acid., Conclusions: These proteins could be related to protective effect of tooth homeostasis. Moreover, in the future, they could be candidates to the development of a supplemental therapy for the prevention and treatment of dental caries and dental erosion., Clinical Significance: This study indicates some acid-resistant proteins that could be used in dental products to prevent dental caries and erosion., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Objective: This study attempted to clarify the mechanisms of action of fluoridated acidic liquid dentifrices against dental caries., Design: In the in vitro leg, enamel specimens were submitted to a pH-cycling model, treated with distinct dentifrices (0, 550 μgF/g pH 4.5 and pH 7.0, 1100 or 5000 μgF/g pH 7.0) and analyzed using hardness. Alkali-soluble fluoride (F) deposition was quantified on pre-demineralized specimens treated with the dentifrices. In the clinical leg, 2-to-4-year-old children who had been using liquid dentifrices for 6 months (550 μgF/g pH 4.5 or pH 7.0 or 1100 μgF/g pH 7.0) had their plaque samples collected 5 and 60 min after the last brushing. Fluoride uptake in whole plaque was evaluated., Results: The reduction of the pH had a partial preventive effect on subsurface hardness loss only. [F] had a significant influence on the deposition of fluoride, surface and subsurface hardness loss. In vivo, the reduction of the pH was able to significantly increase plaque F uptake, leading to similar levels as those found for the neutral dentifrice containing twice [F]., Conclusion: The results obtained from in vitro studies whose design does not include the presence of dental plaque should be interpreted with caution., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The aims of this study were (1) to assess the amount of fluoride (F) released from varnishes containing calcium glycerophosphate (CaGP) and (2) to assess the effect of the experimental varnishes on in vitro demineralization. Six test groups using 5 varnishes: base varnish (no active ingredients); Duraphat® (2.26% NaF); Duofluorid® (5.63% NaF/CaF2); experimental varnish 1 (1% CaGP/5.63% NaF/CaF2); experimental varnish 2 (5% CaGP/5.63% NaF/CaF2); and no varnish were set up. In stage 1, 60 acrylic blocks were randomly distributed into 6 groups (n = 10). Then 300 µg of each varnish was applied to each block. The blocks were immersed in deionized water, which was changed after 1, 8, 12, 24, 48 and 72 hours. Fluoride concentration in the water was analyzed using a fluoride electrode. In stage 2, 60 bovine enamel samples were distributed into 6 groups (n = 10), and treated with 300 µg of the respective varnish. After 6 h the varnish was removed and the samples were subjected to a 7-day in vitro pH cycle (6 h demineralization/18 h remineralization per day). The demineralization was measured using surface hardness. The results showed that both experimental varnishes released more fluoride than Duofluorid® and Duraphat® (p < 0.05), but Duraphat® showed the best preventive effect by decreasing enamel hardness loss (p < 0.05). Therefore, we conclude that even though (1) the experimental varnishes containing CaGP released greater amounts of F, (2) they did not increase in the preventive effect against enamel demineralization.

Objective: This study assessed the effect of fluoride varnishes on the progression of tooth erosion in vitro., Material and Methods: Forty-eight enamel and 60 root dentin samples were previously demineralized (0.1% citric acid, pH 2.5, 30 min), leading to a baseline and erosive wear of 12.9 and 11.4 µm, respectively. The samples were randomly treated (6 h) with a 4% TiF4 varnish (2.45%F-, pH 1.0), a 5.42% NaF varnish (2.45%F-, pH 5.0), a placebo varnish and no varnish (control). The samples were then subjected to erosive pH cycles (4x90 s/day in 0.1% citric acid, intercalated with artificial saliva) for 5 days. The increment of the erosive tooth wear was calculated. In the case of dentin, this final measurement was done with and without the demineralized organic matrix (DOM). Enamel and dentin data were analyzed using ANOVA/Tukey's and Kruskal-Wallis/Dunn tests, respectively (p<0.05)., Results: The TiF4 (mean±s.d: 1.5±1.1 µm) and NaF (2.1±1.7 µm) varnishes significantly reduced enamel wear progression compared to the placebo varnish (3.9±1.1 µm) and control (4.5±0.9 µm). The same differences were found for dentin in the presence and absence of the DOM, respectively: TiF4 (average: 0.97/1.87 µm), NaF (1.03/2.13 µm), placebo varnish (3.53/4.47 µm) and control (3.53/4.36 µm)., Conclusion: The TiF4 and NaF varnishes were equally effective in reducing the progression of tooth erosion in vitro.

Dentin organic matrix, with type I collagen as the main component, is exposed after demineralization in dentinal caries, erosion or acidic conditioning during adhesive composite restorative treatment. This exposed matrix is prone to slow hydrolytic degradation by host collagenolytic enzymes, matrix metalloproteinases (MMPs) and cysteine cathepsins. Here we review the recent findings demonstrating that inhibition of salivary or dentin endogenous collagenolytic enzymes may provide preventive means against progression of caries or erosion, just as they have been shown to retain the integrity and improve the longevity of resin composite filling bonding to dentin. This paper also presents the case that the organic matrix in caries-affected dentin may not be preserved as intact as previously considered. In partially demineralized dentin, MMPs and cysteine cathepsins with the ability to cleave off the terminal non-helical ends of collagen molecules (telopeptides) may lead to the gradual loss of intramolecular gap areas. This would seriously compromise the matrix ability for intrafibrillar remineralization, which is considered essential in restoring the dentin's mechanical properties. More detailed data of the enzymes responsible and their detailed function in dentin-destructive conditions may not only help to find new and better preventive means, but better preservation of demineralized dentin collagenous matrix may also facilitate true biological remineralization for the better restoration of tooth structural and mechanical integrity and mechanical properties.

This study evaluated the effect of an experimental paste containing hydroxyapatite in nanoparticles (nano-HA)/fluoride on dental de-remineralisation in situ. Thirteen subjects took part in this crossover/randomised/double-blind study performed in 4 phases (14 days each). Four sound and 4 pre-demineralised specimens were worn intraorally at each phase corresponding to the following treatments: Nanop Plus (10% HA, 0.2% NaF, nano-HA/fluoride), MI Paste Plus (casein phosphopeptide-amorphous calcium phosphate, 0.2% NaF), F (0.2% NaF) and placebo. Two-hundred and forty enamel and 240 dentine specimens were selected by using surface microhardness; half of them were subjected to pre-demineralisation and the other half remained sound. Sound specimens were further exposed to severe cariogenic challenge (20% sucrose in biofilm) in situ, while pre-demineralised specimens were not. All specimens were exposed to fluoride dentifrice slurry 2×1 min/day. Thereafter, the treatments were done for 4 min. The de-remineralisation was quantified by transversal microradiography. The data were statistically analysed by repeated-measures ANOVA/Tukey's tests (p<0.05). Generally, no huge differences were found among the treatments. However, Nanop Plus was the only treatment able to significantly reduce dentine demineralisation (x0394;Z, integrated mineral loss) and to improve enamel remineralisation (x0394;x0394;Z, integrated mineral uptake) compared to placebo. No treatments were able to reduce enamel demineralisation, while for dentine remineralisation all treatments were similarly effective in improving x0394;x0394;Z compared to placebo. Nanop Plus seems to have a positive influence on dental de-remineralisation, which should be further confirmed., (© 2015 S. Karger AG, Basel.)

Demineralization in dentinal caries and erosion exposes dentine organic matrix. This exposed matrix, containing type I collagen and non-collagenous proteins, is then degraded by host collagenolytic enzymes, matrix metalloproteinases (MMPs) and cysteine cathepsins. The knowledge of the identities and function of these enzymes in dentine has accumulated only within the last 15 years, but has already formed a field of research called 'dentine degradomics'. This research has demonstrated the role of endogenous collagenolytic enzymes in caries and erosion development. In demineralized dentine, the enzymes degrade triple-helical collagen molecules, leading to the gradual loss of collagen matrix. Even before that, they can cleave off the terminal non-helical ends of collagen molecules called telopeptides, leading to the structural changes at the intramolecular gap areas, which may affect or even prevent intrafibrillar remineralization, which is considered essential in restoring the dentine's mechanical properties. They may also cause the loss of non-collagenous proteins that could serve as nucleation sites for remineralization. Here we review the findings demonstrating that inhibition of salivary or dentine endogenous MMPs and cysteine cathepsins may provide preventive means against the progression of caries or erosion. Furthermore, we also suggest the future directions for the new experimental preventive research to gain more knowledge of the enzymes and their function during and after dentine demineralization, and the pathways to find the clinically acceptable means to prevent the functional activity of these enzymes., (2015 S. Karger AG, Basel)

Background: Dental erosion is caused by frequent exposure to acids without the involvement of microorganism. This study analyzed the effect of biguanides (polyhexamethylene biguanide - PHMB and chlorhexidine - CHX) on dentin erosion due to their possible influence on the enzymatic degradation of the demineralized organic matrix., Method: Sixty bovine dentin specimens were prepared. On both sides of their surface, nail varnish was applied to maintain the reference surfaces for the determination of dentin loss. Samples were cyclically de- and remineralized for 6 days. Demineralization was performed with a 0.87 M citric acid solution (6×5 min daily). Thereafter, samples were treated with distilled water (negative control), 0.12% CHX (positive control), 0.07% PHMB, Sanifill Perio Premium™ (0.07% PHMB plus 0.05% NaF), or F solution (0.05% NaF) for 1 min and then subjected to enzymatic challenge for 10 min using a bacterial collagenase (Clostridium hystoliticum, 100 μg/ml). Dentin loss was assessed using profilometry (μm) daily. Data were analyzed using 2-way repeated measures-ANOVA and Bonferroni's test (p < 0.05)., Results: Dentin loss progressed significantly for all groups during the 6 days. After the 3rd day, Sanifill Premium™, CHX, and PHMB significantly reduced dentin erosion compared to control. On the 6th day, the lowest mean (±SD) dentin loss was observed for Sanifill Perio Premium™ (94.4 ± 3.9 μm). PHMB and CHX led to intermediate dentin loss (129.9 ± 41.2 and 135.3 ± 33.5 μm, respectively) that was significantly lower than those found for negative control (168.2 ± 6.2 μm). F (157.4 ± 6.1 μm) did not significantly differ from negative control., Conclusions: Sanifill Perio Premium™ mouthwash has a good potential to reduce dentin loss, which might be associated with the presence of PHMB.

Administration of high doses of fluoride (F) can alter glucose homeostasis and lead to insulin resistance (IR). This study determined the profile of protein expression in the gastrocnemius muscle of rats with streptozotocin-induced diabetes that were chronically exposed to F. Male Wistar rats (60 days old) were randomly distributed into two groups of 18 animals. In one group, diabetes was induced through the administration of streptozotocin. Each group (D-diabetic and ND-non-diabetic) was further divided into 3 subgroups each of which was exposed to a different F concentration via drinking water (0 ppm, 10 ppm or 50 ppm F, as NaF). After 22 days of treatment, the gastrocnemius muscle was collected and submitted to proteomic analysis (2D-PAGE followed by LC-MS/MS). Protein functions were classified by the GO biological process (ClueGO v2.0.7+Clupedia v1.0.8) and protein-protein interaction networks were constructed (PSICQUIC, Cytoscape). Quantitative intensity analysis of the proteomic data revealed differential expression of 75 spots for ND0 vs. D0, 76 for ND10 vs.D10, 58 spots for ND50 vs. D50, 52 spots for D0 vs. D10 and 38 spots for D0 vs. D50. The GO annotations with the most significant terms in the comparisons of ND0 vs. D0, ND10 vs. D10, ND50 vs. D50, D0 vs. D10 and D0 vs. D50, were muscle contraction, carbohydrate catabolic processes, generation of precursor metabolites and energy, NAD metabolic processes and gluconeogenesis, respectively. Analysis of subnetworks revealed that, in all comparisons, proteins with fold changes interacted with GLUT4. GLUT4 interacting proteins, such as MDH and the stress proteins HSPB8 and GRP78, exhibited decreased expression when D animals were exposed to F. The presence of the two stress proteins indicates an increase in IR, which might worsen diabetes. Future studies should evaluate whether diabetic animals treated with F have increased IR, as well as which molecular mechanisms are involved.

Objective: This study aimed to assess the impact of in vitro erosion provoked by different cola-based drinks (Coke types), associated or not with toothbrushing, to bonding to enamel., Material and Methods: Fifty-six [Corrected] bovine enamel specimens were prepared and randomly assigned into seven groups (N=8): C- Control (neither eroded nor abraded), ERO-RC: 3x/1-minute immersion in Regular Coke (RC), ERO-LC: 3x/1-minute immersion in Light Coke (LC), ERO-ZC: 3x/1-minute immersion in Zero Coke (ZC) and three other eroded groups, subsequently abraded for 1-minute toothbrushing (EROAB-RC, EROAB-LC and EROAB-ZC, respectively). After challenges, they were stored overnight in artificial saliva for a total of 24 hours and restored with Adper Single Bond 2/Filtek Z350. Buildup coronal surfaces were cut in 1 mm2 -specimens and subjected to a microtensile test. Data were statistically analyzed by two-way ANOVA/Bonferroni tests (α=0.05). Failure modes were assessed by optical microscopy (X40). The Interface of the restorations were observed using Confocal Laser Scanning Microscopy (CLSM)., Results: All tested cola-based drinks significantly reduced the bond strength, which was also observed in the analyses of interfaces. Toothbrushing did not have any impact on the bond strength. CLSM showed that except for Zero Coke, all eroded specimens resulted in irregular hybrid layer formation., Conclusions: All cola-based drinks reduced the bond strength. Different patterns of hybrid layers were obtained revealing their impact, except for ZC.

Introduction: Methicillin-resistant Staphylococcus aureus (MRSA) strains have been responsible for many nosocomial outbreaks. Within hospitals, colonized employees often act as reservoirs for the spread of this organism. This study collected clinical samples of 91 patients admitted to the intensive care unit (ICU), hemodialysis/nephrology service and surgical clinic, and biological samples from the nasal cavities of 120 professionals working in those environments, of a University Hospital in Recife, in the State of Pernambuco, Brazil. The main objective of this study was to determine the occurrence and dissemination of methicillin- and vancomycin-resistant Staphylococcus spp., Methods: The isolates obtained were tested for susceptibility to oxacillin and vancomycin and detection of the mecA gene. In addition, the isolates were evaluated for the presence of clones by ribotyping-polymerase chain reaction (PCR)., Results: MRSA occurrence, as detected by the presence of the mecA gene, was more prevalent among nursing technicians; 48.1% (13/27) and 40.7% (11/27) of the isolates were from health professionals of the surgical clinic. In patients, the most frequent occurrence of mecA-positive isolates was among the samples from catheter tips (33.3%; 3/9), obtained mostly from the hemodialysis/nephrology service. Eight vancomycin-resistant strains were found among the MRSA isolates through vancomycin screening. Based on the amplification patterns, 17 ribotypes were identified, with some distributed between patients and professionals., Conclusions: Despite the great diversity of clones, which makes it difficult to trace the source of the infection, knowledge of the molecular and phenotypic profiles of Staphylococcus samples can contribute towards guiding therapeutic approaches in the treatment and control of nosocomial infections.

Background: The distribution of fluoride and calcium in plaque after the use of fluoride dentifrices has not yet been determined., Aim: To evaluate fluoride and calcium distribution in sections of biofilms generated in situ after the use of conventional and low-fluoride dentifrices., Design: Children (n = 11, 8–10 years old) brushed with placebo (fluoride-free), low-fluoride (513 mgF/kg), and conventional (1072 mgF/kg) dentifrices twice daily for 1 week, following a double-blind, cross-over protocol. Biofilms were generated using Leeds in situ devices, which were collected 1 and 12 h after brushing, and sectioned through their depth. Sections were grouped (10 x 5 μm) for fluoride and calcium analysis. Sections 4 lm thick were used for image analysis and determination of biomass fraction. Results were analysed by ANOVA, Tukey’s test, and linear regression analysis (P < 0.05)., Results: Fluoride and calcium were mostly located at the outer sections of biofilms for all dentifrices tested, and these ions were directly correlated throughout most of biofilm’s sections. Results for conventional dentifrice were significantly higher than for the placebo, but did not differ from those for the low-fluoride dentifrice., Conclusions: The use of a low-fluoride dentifrice did not promote a higher fluoride uptake in inner biofilms’ sections, as hypothesized. As plaque fluoride was significantly elevated only after the use of the conventional dentifrice, the recommendation of low-fluoride formulations should be done with caution, considering both risks and benefits.

Objective: This study aimed to assess the overall apatite crystals profile in the enamel matrix of mice susceptible (A/J strain) or resistant (129P3/J strain) to dental fluorosis through analyses by atomic force microscopy (AFM)., Material and Methods: Samples from the enamel matrix in the early stages of secretion and maturation were obtained from the incisors of mice from both strains. All detectable traces of matrix protein were removed from the samples by a sequential extraction procedure. The purified crystals (n=13 per strain) were analyzed qualitatively in the AFM. Surface roughness profile (Ra) was measured., Results: The mean (±SD) Ra of the crystals of A/J strain (0.58±0.15 nm) was lower than the one found for the 129P3/J strain (0.66±0.21 nm) but the difference did not reach statistical significance (t=1.187, p=0.247). Crystals of the 129P3/J strain (70.42±6.79 nm) were found to be significantly narrower (t=4.013, p=0.0013) than the same parameter measured for the A/J strain (90.42±15.86 nm)., Conclusion: enamel crystals of the 129P3/J strain are narrower, which is indicative of slower crystal growth and could interfere in the occurrence of dental fluorosis.

Objective: This randomized clinical trial evaluated the effect of pH and fluoride concentration of liquid dentifrices (LD) on caries progression and fluoride uptake in the toenails of 2-4-year-old children., Materials and Methods: Schoolchildren living in a fluoridated area, with (A) or without (I) active caries lesions, were randomly allocated into three groups according to the LD they would use over 12 months: group 1 (n = 48-A/56-I): 550 μg fluoride (F)/g, pH 4.5; group 2 (n = 56-A/48-I) 1,100 μg F/g, pH 7.0; and group 3 (n = 52-A/55-I): 550 μg F/g, pH 7.0. The number of lesions becoming active/cavities or inactive was clinically evaluated by determining progression or regression. Additionally, white spot lesions were evaluated by the quantitative light-induced fluorescence (QLF) method in 75 children. Toenail fluoride concentration was also evaluated. Data were analyzed by ANOVA and followed by Dunn's test or ANCOVA followed by Tukey's test (p < 0.05)., Results: Caries net increment followed a decreasing pattern according to the dentifrice used (G1 < G2 < G3), regardless caries activity, but significant differences were detected only for caries progression and net increment (G1 < G3) for the caries-active group when evaluated through visual inspection. For the regression, the values found for the three groups were more similar, without significant differences. QLF analysis detected no significant difference between groups 1 and 2, but they performed significantly better than group 3. A significantly lower toenail fluoride concentration was observed when children used the low-fluoride dentifrices., Conclusion: The low-fluoride acidic LD has good potential to prevent caries and reduce fluoride intake., Clinical Relevance: The low-F acidic dentifrice combines the desirable characteristics of caries prevention and reduced fluoride intake-for a product to be used by children, regardless their caries status.

Unlabelled: Fluoride varnishes play an important role in the prevention of dental caries, promoting the inhibition of demineralization and the increase of remineralization., Objective: This study aimed to analyze the amount of fluoride released into water and artificial saliva from experimental TiF4 and NaF varnishes, with different concentrations, for 12 h., Material and Methods: Fluoride varnishes were applied on acrylic blocks and then immersed in 10 ml of deionized water and artificial saliva in polystyrene bottles. The acrylic blocks were divided in seven groups (n=10): 1.55% TiF4 varnish (0.95% F, pH 1.0); 3.10% TiF4 varnish (1.90% F, pH 1.0); 3.10% and 4% TiF4 varnish (2.45% F, pH 1.0); 2.10% NaF varnish (0.95% F, pH 5.0); 4.20% NaF varnish (1.90% F, pH 5.0); 5.42% NaF varnish (2.45% F, pH 5.0) and control (no treatment, n=5). The fluoride release was analyzed after 1/2, 1, 3, 6, 9 and 12 h of exposure. The analysis was performed using an ion-specific electrode coupled to a potentiometer. Two-way ANOVA and Bonferroni's test were applied for the statistical analysis (p<0.05)., Results: TiF4 varnishes released larger amounts of fluoride than NaF varnishes during the first 1/2 h, regardless of their concentration; 4% TiF4 varnish released more fluoride than NaF varnishes for the first 6 h. The peak of fluoride release occurred at 3 h. There was a better dose-response relationship among the varnishes exposed to water than to artificial saliva., Conclusions: The 3.10% and 4% TiF4 -based varnishes have greater ability to release fluoride into water and artificial saliva compared to NaF varnish; however, more studies must be conducted to elucidate the mechanism of action of TiF4 varnish on tooth surface.

In recent years, different agents have been discussed as potential alternatives to fluoride in the prevention of dental erosion. These agents are intended to form acid-resistant layers on the surface, to induce repair of eroded lesions by mineral precipitation or to prevent the enzymatic degradation of demineralised collagen. The application of adhesives and/or fissure sealants is considered to be an effective alternative to fluoride, but requires professional application and, depending on the product used, a re-sealing of the surface every several months. Studies testing film-forming products, such as polymers, have suggested the potential effectiveness of some of these approaches, such as chitosan, although further studies are needed to confirm the effectiveness of this approach. Other studies have demonstrated that products designed to deliver calcium and/or phosphate have not been successful at providing a significant anti-erosive effect. In advanced erosive lesions, the demineralised collagenous dentine matrix can be degraded by host enzymes such as matrix metalloproteinases (MMPs). As well as fluorides, epigallocatechin gallate and chlorhexidine have been identified as effective MMP inhibitors, with the potential to reduce the progression of dentine erosion. While fluoride compounds have been shown to have an anti-erosive potential, particularly those containing tin, alternative approaches that provide even greater protective capacity still need to be developed and proven to be effective., (© 2014 S. Karger AG, Basel.)

The objective of this study was to evaluate comparatively the effect of fluoride in the expression of the receptor activator of nuclear factor kappa B ligand (RANKL), osteoprotegerin (OPG) and tartrate-resistant acid phosphatase (TRAP) in alveolar bone repair in rats. We used 3 groups of male Wistar rats (n = 5/group), which received drinking water containing different doses of F (NaF): 0, 5 and 50 ppm, for 60 days before the incisors extraction. The upper incisors were extracted and the animals were killed 7, 14, 21 and 30 days after extraction. The hemi-maxillae were collected for microscopic examination (histomorphometric and immunostaining for RANKL, OPG and TRAP). Histomorphometric analysis confirmed an increase in the volume density of neoformed bone between 7 and 30 days for groups control, 5 and 50 ppm of F, with a concomitant decrease in the volume density of connective tissue and blood clot. Higher blood clot for groups 5 and 50 ppm of F at 30 days was observed. The RANKL and OPG expressions were not changed by chronic exposure to fluoride in the drinking water during the studied periods; on the other hand, TRAP expression was changed (at 7 days) by chronic exposure to fluoride (p < 0.05). It was concluded that F in high concentrations can slow the blood clot remission and bone repair, and alter the TRAP expression in the beginning of the bone tissue repair. However, a better understanding about this blood clot remission phenomenon is required.

Dental erosion is the loss of dental hard tissues caused by non-bacterial acids. Due to acid contact, the tooth surface becomes softened and more prone to abrasion from toothbrushing. Dentifrices containing different active agents may be helpful in allowing rehardening or in increasing surface resistance to further acidic or mechanical impacts. However, dentifrices are applied together with brushing and, depending on how and when toothbrushing is performed, as well as the type of dentifrice and toothbrush used, may increase wear. This review focuses on the potential harmful and helpful effects associated with the use of dentifrices with regard to erosive wear. While active ingredients like fluorides or agents with special anti-erosive properties were shown to offer some degree of protection against erosion and combined erosion/abrasion, the abrasive effects of dentifrices may increase the surface loss of eroded teeth. However, most evidence to date comes from in vitro and in situ studies, so clinical trials are necessary for a better understanding of the complex interaction of active ingredients and abrasives and their effects on erosive tooth wear.

Purpose: This double-blind, crossover study evaluated whole plaque fluoride concentration (F), as well as whole plaque calcium concentrations (Ca) after brushing with a placebo (PD - fluoride free), low-fluoride (LFD, 513 microg F/g) and conventional (CD, 1,072 microg F/g) dentifrices., Methods: Children (n=20) were randomly assigned to brush twice daily with one of the dentifrices, during 7 days. On the 7th day, samples were collected at 1 and 12 hours after brushing. F and Ca were analyzed with an ion-selective electrode and with the Arsenazo III method, respectively. Data were analyzed by ANOVA, Tukey's test and by Pearson correlation coefficient (P< 0.05)., Results: The use of the fluoridated dentifrices significantly increased plaque [F]s 1 hour after brushing when compared to PD, returning to baseline levels 12 hours after. Positive and significant correlations were found between plaque [F] and (Ca) under most of the conditions evaluated. The mean increase in plaque [F] observed 1 hour after brushing with the CD were only about 47% higher than those obtained for the LFD. The use of a LFD promotes proportionally higher increases in plaque [F] when compared to a CD. Plaque F concentrations were also shown to be dependent on plaque Ca concentrations.