Effect of fluoridated dentifrice and acidulated phosphate fluoride application on early artificial carious lesions

Purpose : To evaluate the effect of a combination of fluoride dentifrice and acidulated phosphate fluoride (APF) on enamel with incipient caries lesions. Materials and Methods: Bovine dental enamel blocks with artificial caries lesions were divided into four treatment groups and submitted to a pH-cycling model: (1) Placebo non-fluoridated dentifrice (PD); (2) Fluoridated dentifrice (171)), 1,100 ppm F; (3) APF (12,300 ppm F, pH 3.5) + PD; 4) APF + FD. APF was applied to enamel blocks of groups APF+PD and APF+FD before the pH-cycling regimen and all of them were exposed to dentifrice during the cycling. Results: The results showed that 61-81% of the fluoride previously deposited in enamel by APF was released to the media during the pH-cycling. The final concentration of fluoride in enamel in the group APF+PD was lower than that before the cycling (P 0.05). FD treatment was significantly more efficient than PD in rehardening the enamel surface and increasing the hardness of the caries lesions. APF+PD treatment was not more efficient than PD in increasing enamel hardness and an additive effect of APF+FD was not observed.162919

Occlusion of dentin tubules by desensitizing agents.

Purpose: To evaluate the occluding effect of five desensitizing agents on human dentin tubules. Methods: 30 buccal and lingual surfaces were prepared from 15 extracted intact third molars. Each surface was polished with aluminum oxide abrasive papers to remove enamel and to expose the underlying dentin in cervical area. The flat dentin surfaces were treated with 0.5 M EDTA for 2 minutes to expose dentin tubule orifices. The samples were randomly divided into six groups: AS - immersed in artificial saliva at 37degreesC for 2 weeks (control); OX - Oxagel (monopotassium oxalate), DU - Duraphat (sodium fluoride), DE - Desensibilize (strontium chloride), OD - Odahcam (acidulated phosphate fluoride) and SE - Sensodyne (strontium chloride + calcium carbonate abrasive). Dentin desensitizers were applied during 2 weeks and after each application the samples were kept in artificial saliva at 37degreesC. The samples were prepared according to the scanning electron microscope procedures and were examined at x2000 magnification. Results: The results were expressed in percent (%) of tubule occlusion and analyzed by ANOVA and Duncan's multiple range test (P< 0.05): AS- 45.41 +/- 11.65(a); OX- 42.65 +/- 11.79(a); DU- 47.25 +/- 8.59(ab); DE- 49.36 +/- 18.27(ab); OD- 64.43 +/- 15.55(b) and SE- 65.44 +/- 10.93(b) . Results suggest that the dentin surfaces treated with OD and SE showed higher tubule occlusion when compared to AS and OX, but were not different compared to DU and DE treatments.17536837

Enzymatic activity and proteomic profile of class III peroxidases during sugarcane stem development

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Class III peroxidases are present as large multigene families in all land plants. This large number of genes together with the diversity of processes catalyzed by peroxidases suggests possible functional specialization of each isoform. However, assigning a precise role for each individual peroxidase gene has continued to be a major bottleneck. Here we investigated the enzyme activity and translational profile of class III peroxidases during stem development of sugarcane as a first step in the estimation of physiological functions of individual isoenzymes. Internodes at three different developmental stages (young, developing and mature) were divided into pith (inner tissue) and rind (outer tissue) fractions. The rind of mature internodes presented the highest enzymatic activity and thus could be considered the ideal tissue for the discovery of peroxidase gene function. In addition, activity staining of 2DE gels revealed different isoperoxidase profiles and protein expression regulation among different tissue fractions. In-gel tryptic digestion of excised spots followed by peptide sequencing by LC-MS/MS positively matched uncharacterized peroxidases in the sugarcane database SUCEST. Multiple spots matching the same peroxidase gene were found, which reflects the generation of more than one isoform from a particular gene by post-translational modifications. The identified sugarcane peroxidases appear to be monocot-specific sequences with no clear ortholog in dicot model plant Arabidopsis thaliana. (C) 2012 Elsevier Masson SAS. All rights reserved.556676Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)FAPESP [2008/58035-6

Suspension cell culture as a tool for the characterization of class III peroxidases in sugarcane

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Secreted class III peroxidases (EC 1.11.1.7) are implicated in a broad range of physiological processes throughout the plant life cycle. However, the unambiguous determination of the precise biological role of an individual class III peroxidase isoenzyme is still a difficult task due to genetic redundancy and broad substrate specificity in vitro. In addition, many difficulties are encountered during extraction and analysis of cell wall proteins. Since class III peroxidases are also secreted into the apoplast, the use of suspension cell cultures can facilitate isolation and functional characterization of individual isoforms. Here, we report on the characterization of class III peroxidases secreted in the spent medium of sugarcane suspension cell cultures. After treatment with specific inducers of cell wall lignification, peroxidases were isolated and activities assayed with guaiacol, syringaldazine and coniferyl alcohol. Enzymatic activity was not significantly different after treatments, regardless of the substrate, with the exception of methyl-jasmonate treatment, which led to a decreased guaiacol peroxidase activity. Remarkably, peroxidases isolated from the medium were capable of oxidizing syringaldazine, an analog to sinapyl alcohol, suggesting that sugarcane cultures can produce peroxidases putatively correlated to lignification. A proteomic approach using activity staining of 2-DE gels revealed a complex isoperoxidase profile, composed predominantly of cationic isoforms. Individual spots were excised and analyzed by LC-ESI-Q-TOF and homology-based search against the Sugarcane EST Database resulted in the identification of several proteins. Spatio-temporal expression pattern of selected genes was determined for validation of identified class III peroxidases that were preferentially expressed during sugarcane stem development. (C) 2012 Elsevier Masson SAS. All rights reserved.62110Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)FAPESP [BIOEN 08/58035-6

The Efficacy of Laser Fluorescence to Detect in Vitro Demineralization and Remineralization of Smooth Enamel Surfaces

Objective: The purpose of this study was to evaluate the efficacy of the laser fluorescence (LF) device in detecting in vitro demineralization and remineralization of smooth surface caries-like lesions. Background Data: The early detection of smooth surface caries-like lesions is important to provide proper management of carious lesions, and allows monitoring of them over time. Also, some authors suggest that LF could be useful in monitoring the caries pathological process. Materials and Methods: Seventy-eight blocks of bovine teeth were obtained, and then submitted to artificial caries lesion induction and to a pH-cycling process. Superficial microhardness (SMH) and laser fluorescence analysis were performed at baseline, after induction of caries-like lesions, and after the pH-cycling regimen to promote remineralization. Results: Friedman's and multiple comparison tests were performed for all variables. SMH analysis showed significant differences (p < 0.05) between baseline (286.77 +/- 1.49 Vickers hardness number [VHN] units), before (38.48 +/- 0.85 VHN), and after remineralization (131.93 +/- 2.63 VHN). Baseline values for LF were extremely low (2.71 +/- 0.05), and a statistically significant difference was observed only after remineralization (3.61 +/- 0.08), as demonstrated by the increase in LF values. Conclusion: The LF device did not show efficacy for monitoring in vitro demineralization and remineralization of smooth enamel surfaces.o TEXTO COMPLETO DESTE ARTIGO, ESTARÁ DISPONÍVEL À PARTIR DE AGOSTO DE 2015.271576

Effect of luting cement on dental biofilm composition and secondary caries around metallic restorations in situ

Since the importance of luting cement on secondary caries in enamel and dentin is unknown, an in situ crossover study was conducted in three phases over 21 days using a fluoride-containing toothpaste. One hundred and twenty-six metallic restorations were cemented into the dentinoenamel junction of slabs of human teeth with zinc phosphate (ZP), resin-modified glass ionomer (GI) or resinous cement (RC). The slabs were inserted onto flanges of the removable partial acrylic dentures of 14 volunteers and covered with gauze to enhance dental plaque accumulation. The volunteers used fluoride toothpaste (1.100 mug F/g, w/w). After 21 days, the biofilm that formed on the slabs was collected for biochemical and microbiological analyses, and the demineralization in enamel-dentin around the restorations was evaluated. The fluoride concentration of biofilm in the GI group was higher (p<0.05) than the ZP and RC groups. Also, the concentration of Zinc in biofilm formed on the slabs cemented with ZP was higher (p<0.05) than the other groups. However, the effect of the luting material on enamel or dentin demineralization was not statistically significant (p&GT;0.05). The data suggest that when fluoride toothpaste is used, the anti-cariogenic property of the luting cement may not be relevant to the reduction of secondary caries.29550951

In situ effect of frequent sucrose exposure on enamel demineralization and on plaque composition after APF application and F dentifrice use

Since the effect of the combination of methods of fluoride use on enamel demineralization and on plaque composition is not clearly established, this study examined the effect of the combination of acidulated phosphate fluoride (APF) application and F dentifrice on enamel demineralization and on plaque composition. In this crossover study, 16 volunteers, wearing a palatal appliance containing bovine enamel blocks, were subjected to 4 treatment groups: non-fluoridated dentifrice (PD), FD, APF+PD, and APF+FD. The APF was applied to the enamel before the 14-day experimental period. During the experimental period, test dentifrices were applied 3x/day, and a 20% sucrose solution was applied 4x and 8x/day by being dripped on the blocks. Although APF application was able either to increase F concentration in plaque or to reduce the % of mutans streptococci, its combination with F dentifrice use neither reduced enamel mineral loss nor changed any other measured plaque variable with respect to the FD group alone.831717

Characterization of hNek6 Interactome Reveals an Important Role for Its Short N-Terminal Domain and Colocalization with Proteins at the Centrosome

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Physical protein protein interactions are fundamental to all biological processes and are organized in complex networks One branch of the kinome network is the evolutionarily conserved NIMA-related serine/threonine kinases (Neks) Most of the 11 mammalian Neks studied so far are related to cell cycle regulation and due to association with diverse human pathologies, Neks are promising chemotherapeutic targets Human Nek6 was associated to carcinogenesis but its interacting partners and signaling pathways remain elusive Here we introduce hNek6 as a highly connected member in the human kinase interactome In a more global context, we performed a broad data bank comparison based on degree distribution analysis and found that the human kinome is enriched in hubs Our networks include a broad set of novel hNek6 interactors as identified by our yeast two hybrid screens classified into 18 functional categories All of the tested interactions were confirmed and the majority of tested substrates were phosphorylated in vitro by hNek6 Notably, we found that hNek6 N-terminal is important to mediate the interactions with its partners Some novel interactors also colocalized with hNek6 and gamma-tubulin in human cells, pointing to a possible centrosomal interaction The interacting proteins link hNek6 to novel pathways, for example, Notch signaling and actin cytoskeleton regulation, or give new insights on how hNek6 may regulate previously proposed pathways such as cell cycle regulation, DNA repair response, and NF-kappa B signaling Our findings open new perspectives in the study of hNek6 role in cancer by analyzing its novel interactions in specific pathways in tumor cells which may provide important implications for drug design and cancer therapy91262986316Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Associacao Brasileira de Tecnologia de Luz Sincrotron (ABTLuS)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Temporal relationship between sucrose-associated changes in dental biofilm composition and enamel demineralization

The aim of this study was to investigate the temporal relationship between changes in biofilm composition and enamel demineralization following exposure to sucrose. A crossover blind study was conducted in situ in three phases, during which 12 volunteers, divided into three groups, subjected enamel slabs 8 times/day to water ( negative control), 10% glucose + 10% fructose ( active control) or 20% sucrose solution. Biofilms accumulated for 3, 7 and 14 days were collected and analyzed biochemically and microbiologically, and mineral loss from enamel (Delta Z) was evaluated. Significantly higher (Delta Z) was found in the sucrose group after 7 days. However, on the 3rd day, lactobacilli, insoluble extracellular polysaccharide (EPS) and intracellular polysaccharide were significantly higher, and the calcium, inorganic phosphorus and fluoride concentrations in the biofilm were significantly lower in the sucrose group than in the negative controls. The only significant difference compared to glucose + fructose treatment was a higher insoluble EPS concentration. The data suggest that, although sucrose induces significant enamel demineralization only after 7 days of biofilm accumulation, changes in the biofilm composition are observed earlier. Copyright (c) 2007 S. Karger AG, Basel41540641

Effect of sucrose containing iron (II) on dental biofilm and enamel demineralization in situ

Since the effect of iron (Fe) on the cariogenicity of sucrose in humans is unexplored, this study assessed in situ the effect of Fe co-crystallized with sucrose (Fe-sucrose) topically applied in vitro on the acidogenicity, biochemical and microbiological composition of the dental biofilm formed in vivo and on the demineralization of the enamel. During two phases of 14 days each, 16 volunteers wore palatal appliances containing blocks of human enamel, which were submitted to four groups of separate treatments: (1) water; (2) 20% sucrose; (3) 20% (w/v) sucrose plus 18 mug Fe/ml, and (4) 20% (w/v) sucrose plus 70 mug Fe/ml. The solutions were dripped onto the blocks 8 times per day. The biofilms formed on the blocks were analyzed with respect to acidogenicity, biochemical and microbiological composition. Mineral loss was determined on enamel by surface and cross-sectional microhardness. Lower demineralization was found in the blocks subjected to Fe-sucrose (70 mug Fe/ml) than in those treated with sucrose (p < 0.05). This concentration of Fe also reduced significantly the populations of mutans streptococci in the biofilm formed on the blocks. In conclusion, our data suggest that Fe may reduce in situ the cariogenic potential of sucrose and the effect seems to be related to the reduction in the populations of mutans streptococci in the dental biofilm formed. Copyright (C) 2005 S. Karger AG, Basel.39212312