Abstract Objective To evaluate the protective effect of an experimental solution containing TiF4/NaF on the development of radiation-induced dentin caries lesions. Methodology bovine root samples were irradiated (70Gy) and distributed as following (n=12/group): Commercial Saliva (BioXtra), NaF (500 ppm F-), TiF4 (500 ppm F), TiF4/NaF (TiF4: 300 ppm F-, NaF: 190 ppm F-), and Phosphate buffer solution (PBS, negative control). Biofilm was produced using biofilm from irradiated patients and McBain saliva (0.2% of sucrose, at 37oC and 5% CO2) for five days. The treatments were applied 1x/day. Colony-forming units (CFU) were counted and demineralization was quantified by transversal microradiography. The ANOVA/Tukey test was applied for all parameters. Results All treatments reduced CFU for total microorganisms. TiF4 reduced Lactobacillus sp. (7.04±0.26 log10 CFU/mL) and mutans streptococci (7.18±0.28) CFU the most, when compared to PBS (7.58±0.21 and 7.75±0.17) and followed by NaF (7.12±0.31 and 7.34±0.22) and TiF4/NaF (7.16±0.35 and 7.29± 0.29). TiF4 and Commercial saliva showed the lowest integrated mineral loss (ΔZ-vol%.mm) (1977±150 and 2062±243, respectively) when compared to PBS (4540±335), followed by NaF (2403±235) and TiF4/NaF (2340±200). Commercial saliva was the only to significantly reduce mineral loss (LD-µm) (111±25) compared to PBS (153±24).Mean mineral loss (R-vol%) decreased by 35.2% for TiF4 (18.2±3.3) when compared to PBS (28.1±2.9) Conclusion: TiF4/NaF has a comparable anti-cariogenic effect to TiF4 and Commercial saliva under the model in this study. TiF4NaF TiFNaF TiF radiationinduced radiation induced lesions 70Gy Gy (70Gy n=12/group n12group ngroup n 12 group (n=12/group) BioXtra, BioXtra , (BioXtra) 500 (50 F, F F-) F) (TiF4 30 F- 19 PBS, (PBS control. control . control) 0.2% 02 0 2 (0.2 sucrose oC 5 CO2 CO days 1xday xday 1x day x 1x/day Colonyforming Colony forming (CFU microradiography ANOVATukey ANOVA Tukey parameters microorganisms sp 7.04±0.26 704026 7 04 26 (7.04±0.2 log log1 CFU/mL CFUmL mL 7.18±0.28 718028 18 28 (7.18±0.28 most 7.58±0.21 758021 58 21 (7.58±0.2 7.75±0.17 775017 75 17 7.12±0.31 712031 31 (7.12±0.3 7.34±0.22 734022 34 22 7.16±0.35 716035 16 35 (7.16±0.3 729 29 7.29 0.29. 029 0.29 0.29) ΔZvol%.mm ΔZvolmm ΔZ vol%.mm vol mm (ΔZ-vol%.mm 1977±150 1977150 1977 150 (1977±15 2062243 2062 243 2062±243 respectively 4540±335, 4540335 4540±335 4540 335 (4540±335) 2403±235 2403235 2403 235 (2403±235 2340±200. 2340200 2340±200 2340 200 (2340±200) LDµm LD µm (LD-µm 111±25 11125 111 25 (111±25 153±24.Mean 15324Mean Mean 153±24 .Mean 153 24 Rvol% Rvol R vol% (R-vol% 352 35.2 18.2±3.3 18233 3 (18.2±3.3 28.1±2.9 28129 1 9 (28.1±2.9 Conclusion anticariogenic anti cariogenic study (n=12/group (BioXtra 50 (5 (TiF 0.2 (0. 7.04±0.2 70402 (7.04±0. 7.18±0.2 71802 (7.18±0.2 7.58±0.2 75802 (7.58±0. 7.75±0.1 77501 7.12±0.3 71203 (7.12±0. 7.34±0.2 73402 7.16±0.3 71603 (7.16±0. 72 7.2 ΔZvol volmm 1977±15 197715 197 15 (1977±1 206224 206 2062±24 454033 4540±33 454 33 (4540±335 2403±23 240323 240 23 (2403±23 234020 2340±20 234 20 (2340±200 111±2 1112 11 (111±2 15324 153±2 (R-vol 35. 18.2±3. 1823 (18.2±3. 28.1±2. 2812 (28.1±2. ( 0. (0 7.04±0. 7040 (7.04±0 7.18±0. 7180 (7.18±0. 7.58±0. 7580 (7.58±0 7.75±0. 7750 7.12±0. 7120 (7.12±0 7.34±0. 7340 7.16±0. 7160 (7.16±0 7. 1977±1 19771 (1977± 20622 2062±2 45403 4540±3 45 (4540±33 2403±2 24032 (2403±2 23402 2340±2 (2340±20 111± (111± 1532 153± 18.2±3 182 (18.2±3 28.1±2 281 (28.1±2 7.04±0 704 (7.04± 7.18±0 718 (7.18±0 7.58±0 758 (7.58± 7.75±0 775 7.12±0 712 (7.12± 7.34±0 734 7.16±0 716 (7.16± 1977± (1977 2062± 4540± 4 (4540±3 2403± (2403± 2340± (2340±2 (111 18.2± (18.2± 28.1± (28.1± 7.04± 70 (7.04 7.18± 71 (7.18± 7.58± (7.58 7.75± 77 7.12± (7.12 7.34± 73 7.16± (7.16 (197 (4540± (2403 (2340± (11 18.2 (18.2 28.1 (28.1 7.04 (7.0 7.18 (7.18 7.58 (7.5 7.75 7.12 (7.1 7.34 7.16 (19 (4540 (240 (2340 (1 18. (18. 28. (28. 7.0 (7. 7.1 7.5 7.7 7.3 (454 (24 (234 (18 (28 (7 (45 (2 (23 (4

Abstract There are many suitable strategies for addressing caries, which is an ongoing worldwide problem. Although white spot lesions (WSLs) can be either remineralized naturally or treated with non- or micro-invasive strategies, their whitish and opaque appearance may persist. Objective To evaluate the effects of tooth bleaching as a complement to fluoride-enhanced remineralization or resin infiltration in masking WSLs, as well as in enamel surface roughness relative to that of the adjacent enamel. Methodology Flattened rectangular bovine enamel fragments (6×3×~2.9 mm length, width and thickness) were divided into six groups (L/N, F/N, F.BL/BL, I/N, I.BL/BL, N/N; n=15). Treatments applied to the 3×3 mm left half included: L (Lesion) - WSL simulation with 50 mM acetate buffer, 96 hours, 37ºC; F (Fluoride) - WSL treatment with 2% NaF neutral gel, 1x/week, 8 weeks; I (Infiltration) - WSL treatment with H3PO4 37%/10 s; Icon®-Dry/30 s; Icon®-Infiltrant/3 min+1 min; N (Nothing) - sound enamel/control. Treatments applied to both halves after F and I included: BL (Bleaching) - Opalescence Boost 40%, 3×/20 min each; N (Nothing) - control. The differences in color (ΔE00, ΔL, Δa, Δb) and surface roughness (ΔRa) between the left and right halves were measured. Kruskal-Wallis/post-hoc tests were applied to ΔE00, ΔL, Δa and ΔRa, and 1-way ANOVA/Tukey tests to Δb (α=0.05). Results The factor under study significantly influenced ΔE00 (p=0.0001), ΔL (p=0.0024), Δb (p=0.0015), and ΔRa (p<0.001), but not Δa (p=0.1592). Both fluoride-enhanced remineralization and resin infiltration were able to mask WSL, regardless of subsequent bleaching. However, when bleaching was performed, ΔE00 median values did not exceed the acceptability threshold for color difference. Only resin infiltration reduced ΔRa between WSL and the adjacent enamel. Conclusions Both remineralization and infiltration, particularly if complemented by bleaching, fostered satisfactory esthetic results. Only infiltration without bleaching led to really good results in surface roughness. caries problem WSLs (WSLs non microinvasive micro invasive persist fluorideenhanced fluoride enhanced 6×3×~2.9 6329 6 3 2 9 (6×3×~2. length thickness L/N, LN (L/N FN F/N FBLBL F.BL/BL IN I/N IBLBL I.BL/BL N/N NN n=15. n15 n n=15 . 15 n=15) 33 3× included Lesion (Lesion 5 buffer hours 37ºC ºC Fluoride (Fluoride gel 1xweek xweek 1x week x 1x/week weeks Infiltration (Infiltration HPO H PO H3PO 3710 37 10 37%/1 s Icon®Dry/30 IconDry30 IconDry Icon® Dry/30 Icon Dry 30 Icon®-Dry/3 Icon®Infiltrant/3 IconInfiltrant3 IconInfiltrant Infiltrant/3 Infiltrant Icon®-Infiltrant/ min1 1 min+ Nothing (Nothing enamelcontrol control enamel/control Bleaching (Bleaching 40 40% 320 20 3×/2 each ΔE (ΔE00 (ΔRa measured KruskalWallis/posthoc KruskalWallisposthoc Kruskal Wallis/post hoc Wallis post 1way way ANOVATukey ANOVA Tukey α=0.05. α005 α α=0.05 0 05 (α=0.05) ΔE0 p=0.0001, p00001 p p=0.0001 , 0001 (p=0.0001) p=0.0024, p00024 p=0.0024 0024 (p=0.0024) p=0.0015, p00015 p=0.0015 0015 (p=0.0015) p<0.001, p0001 p<0.001 001 (p<0.001) p=0.1592. p01592 p=0.1592 1592 (p=0.1592) However performed difference 6×3×~2. 632 (6×3×~2 L/N n1 n=1 371 37%/ Icon®Dry/3 IconDry3 Dry30 Dry/3 Icon®-Dry/ Icon®Infiltrant/ Infiltrant3 Infiltrant/ Icon®-Infiltrant 4 32 3×/ (ΔE0 KruskalWallis posthoc Wallispost α00 α=0.0 (α=0.05 p0000 p=0.000 000 (p=0.0001 p0002 p=0.002 002 (p=0.0024 p=0.001 (p=0.0015 p000 p<0.00 00 (p<0.001 p0159 p=0.159 159 (p=0.1592 6×3×~2 63 (6×3×~ n= 37% Icon®Dry/ Dry3 Dry/ Icon®-Dry Icon®Infiltrant (ΔE α0 α=0. (α=0.0 p=0.00 (p=0.000 (p=0.002 (p=0.001 p00 p<0.0 (p<0.00 p015 p=0.15 (p=0.159 6×3×~ (6×3× Icon®Dry α=0 (α=0. p=0.0 (p=0.00 p0 p<0. (p<0.0 p01 p=0.1 (p=0.15 6×3× (6×3 α= (α=0 p=0. (p=0.0 p<0 (p<0. (p=0.1 6×3 (6× (α= p=0 (p=0. p< (p<0 6× (6 (α p= (p=0 (p< ( (p= (p

Abstract Microcosm biofilms can reproduce the complexity of a dental biofilm. However, different forms of cultivation have been used. The impact of the culture atmosphere on the development of microcosm biofilms and their potential to cause tooth demineralization has not yet been deeply studied. Objective This study analyzes the effects of three experimental cultivation models (microaerophile vs. anaerobiosis vs. experimental mixed) on the colony-forming units (CFU) of the cariogenic microorganisms and tooth demineralization. Methodology 90 bovine enamel and 90 dentin specimens were distributed into different atmospheres: 1) microaerophilia (5 days, 5% CO2); 2) anaerobiosis (5 days, jar); 3) mixed (2 days microaerophilia and 3 days anaerobiosis), which were treated with 0.12% chlorhexidine (positive control – CHX) or Phosphate-Buffered Saline (negative control – PBS) (n=15). Human saliva and McBain’s saliva containing 0.2% sucrose were used for microcosm biofilm formation, for 5 days. From the second day to the end of the experiment, the specimens were treated with CHX or PBS (1x1 min/day). Colony-forming units (CFU) were counted, and tooth demineralization was analyzed using transverse microradiography (TMR). Data were subjected to two-way ANOVA and Tukey’s or Sidak’s test (p<0.05). Results CHX was able to reduce total microorganism’s CFU compared to PBS (differences of 0.3–1.48 log10 CFU/mL), except for anaerobiosis and microaerophilia in enamel and dentin biofilm, respectively. In the case of dentin, no effect of CHX on Lactobacillus spp. was observed. CHX significantly reduced enamel demineralization compared to PBS (78% and 22% reductions for enamel and dentin, respectively). Enamel mineral loss did not differ when compared with the other atmospheres; however, the enamel lesion depth was greater under anaerobiosis. Dentin mineral loss was lower under anaerobiosis when compared with the other atmospheres. Conclusion The type of atmosphere has, in general, little influence on the cariogenic ability of the microcosm biofilm. However studied microaerophile vs colonyforming colony forming (CFU 9 atmospheres 1 ( CO2 CO CO2) 2 jar jar) anaerobiosis, , anaerobiosis) 012 0 12 0.12 positive PhosphateBuffered Phosphate Buffered negative n=15. n15 n n=15 . 15 (n=15) McBains McBain s 02 0.2 formation experiment 1x1 x (1x min/day. minday min/day min min/day) Colonyforming Colony counted TMR. TMR (TMR) twoway two way Tukeys Tukey Sidaks Sidak p<0.05. p005 p p<0.05 05 (p<0.05) microorganism differences 03148 48 0.3–1.4 log log1 CFU/mL, CFUmL CFU/mL mL CFU/mL) respectively spp observed 78% 78 (78 22 respectively) however general 01 0.1 n1 n=1 (n=15 0. 1x (TMR p00 p<0.0 (p<0.05 0314 4 0.3–1. 7 (7 n= (n=1 p0 p<0. (p<0.0 031 0.3–1 (n= p<0 (p<0. 03 0.3– (n p< (p<0 0.3 (p< (p

Abstract Fluoride (F) has been widely used to control dental caries, and studies suggest beneficial effects against diabetes when a low dose of F is added to the drinking water (10 mgF/L). Objectives This study evaluated metabolic changes in pancreatic islets of NOD mice exposed to low doses of F and the main pathways altered by the treatment. Methodology In total, 42 female NOD mice were randomly divided into two groups, considering the concentration of F administered in the drinking water for 14 weeks: 0 or 10 mgF/L. After the experimental period, the pancreas was collected for morphological and immunohistochemical analysis, and the islets for proteomic analysis. Results In the morphological and immunohistochemical analysis, no significant differences were found in the percentage of cells labelled for insulin, glucagon, and acetylated histone H3, although the treated group had higher percentages than the control group. Moreover, no significant differences were found for the mean percentages of pancreatic areas occupied by islets and for the pancreatic inflammatory infiltrate between the control and treated groups. Proteomic analysis showed large increases in histones H3 and, to a lesser extent, in histone acetyltransferases, concomitant with a decrease in enzymes involved in the formation of acetyl-CoA, besides many changes in proteins involved in several metabolic pathways, especially energy metabolism. The conjunction analysis of these data showed an attempt by the organism to maintain protein synthesis in the islets, even with the dramatic changes in energy metabolism. Conclusion Our data suggests epigenetic alterations in the islets of NOD mice exposed to F levels comparable to those found in public supply water consumed by humans. (F caries (1 mgFL mgF/L . mgF L mgF/L) treatment total 4 groups 1 weeks period insulin glucagon H Moreover extent acetyltransferases acetylCoA, acetylCoA acetyl CoA, CoA acetyl-CoA metabolism humans (

Abstract The objective of this study was to compare the protein profile of the acquired enamel pellicle (AEP) formed in vivo in patients with or without gastroesophageal reflux disease (GERD), and with or without erosive tooth wear (ETW). Twenty-four volunteers were divided into 3 groups: 1) GERD and ETW; 2) GERD without ETW; and 3) control (without GERD). The AEP formed 120 min after prophylaxis was collected from the lingual/palatal surfaces. The samples were subjected to mass spectrometry (nLC-ESI-MS/MS) and label-free quantification by Protein Lynx Global Service software. A total of 213 proteins were identified, or 119, 92 and 106 from each group, respectively. Group 2 showed a high number of phosphorylated and calcium-binding proteins. Twenty-three proteins were found in all the groups, including 14-3-3 protein zeta/delta and 1-phosphatidylinositol. Several intracellular proteins that join saliva after the exfoliation of oral mucosa cells might have the potential to bind hydroxyapatite, or participate in forming supramolecular aggregates that bind to precursor proteins in the AEP. Proteins might play a central role in protecting the dental surface against acid dissolution. (AEP GERD, , (GERD) ETW. ETW . (ETW) Twentyfour Twenty four groups 1 GERD. GERD) 12 lingualpalatal lingual palatal surfaces nLCESIMS/MS nLCESIMSMS nLC ESI MS/MS MS (nLC-ESI-MS/MS labelfree label free software 21 identified 119 9 10 group respectively calciumbinding calcium binding Twentythree three 1433 14 14-3- zetadelta zeta delta 1phosphatidylinositol. 1phosphatidylinositol phosphatidylinositol phosphatidylinositol. 1-phosphatidylinositol hydroxyapatite dissolution (GERD (ETW nLCESIMS MSMS 11 143 14-3 14-

Abstract Quantification of collagen degradation is an important parameter to evaluate dentin caries for preventive aid. Objectives: Evaluate preventive methods against root collagen degradation by the hydroxyproline assay (HYP) and microradiography technique (MRT). Methodology: Five bovine root dentin blocks were obtained and subjected to an artificial demineralization process by acetate buffer (pH 5) to induce carious lesion formation. Samples were subjected to the following therapeutic treatments: 1) 0.12% chlorhexidine for 1 min, 2) 2% fluoride for 1 min, 3) Nd:YAG Laser (400 μm diameter optical fiber, 10 Hz frequency, 60 mJ/pulse energy, 48 J/cm2 energy density, in noncontact mode for 10 s), 4) deionized water (control) for 1 min, 5) MRT control group (without treatment and removal of collagen). Samples were exposed to degradation by a collagenase enzyme for five days. The enzyme solution was collected, by colorimetry in a spectrophotometer, from the collagen matrix for the hydroxyproline release analysis. The same samples were subjected to an additional two days of demineralization to induce the progression of mineral loss. Samples were analyzed by MRT for the visualization of their degraded areas (estimation of lesion depth and mineral loss). ANOVA was applied to compare hydroxyproline release rates. MRT data were subjected to the Kruskal-Wallis test, followed by the Dunn’s test. Comparisons between the initial five-day and the subsequent two-day demineralization processes were performed by repeated t-test or Wilcoxon (p<0.05) measurements. Results: The amount of HYP released from the dentin samples failed to show significant differences among the groups (p=0.09). Fluoride and chlorhexidine were able to interact with the samples, reducing the progression of dentin caries after removal of the demineralized organic matrix. CHX was the only treatment able to show significant lower lesion depth than the negative control. Conclusion: Chlorhexidine and fluoride were effective in reducing root caries progression.

Abstract Objectives To analyze the effect of 5 toothpastes containing different percentages of S-PRG fillers compared to NaF toothpaste and NaF varnish on the dentin hydraulic conductance (Lp). Methodology Dentin disks (1.0±0.2 mm thickness) were cut from third molars, and their Lp values were evaluated using Flodec. The specimens were allocated into 7 groups (n=8). The minimum (smear layer) and the maximum (after acid etching) Lp values were recorded. Lp was also assessed after treatment with either a 0wt.%, 1wt.%, 5wt.%, 20wt.%, or 30wt.% S-PRG toothpaste, a NaF toothpaste, or a NaF varnish. Toothpastes were applied by brushing for 15 s, allowing it to settle for 1 min, and rinsing with deionized water. The NaF varnish was applied for 4 min and was removed with a probe. Specimens were exposed to citric acid (6%, pH 2.1, 1 min) and their final Lp was recorded. The pH of all products was recorded (n=3) and specimens from each group were analyzed by Laser Scanning Confocal Microscopy (LSCM). Data were subjected to 2-way repeated measures ANOVA and post-hoc Bonferroni (a=0.05). Results The highest Lp reduction was noticed for the 5wt.% S-PRG toothpaste, NaF toothpaste, and NaF varnish. However, the toothpastes containing 5wt.%, 20wt.%, and 30wt.% of S-PRG were similar to all toothpastes but differed from the NaF varnish. After erosion, all groups retrieved their maximum Lp values, except for the NaF varnish. The LSCM evidenced deposits on the surface of specimens treated with 5%, 20%, and 30% S-PRG-based toothpastes and NaF toothpaste. Even more deposits were observed for the NaF varnish. After the erosive challenge, the deposits were diminished in all groups. Conclusion Toothpastes containing 5wt.%, 20wt.%, and 30wt.% of S-PRG fillers behaved similarly to a conventional NaF toothpaste, even after an erosive challenge. The NaF varnish promoted better reduction of the Lp, but its effect was also diminished after erosion.

Abstract The initial characteristics of white spot lesion (WSLs), such as the degree of integrated mineral loss (ΔZ), depth and pattern of mineral distribution, have an impact on further demineralization and remineralization. However, these lesion parameters have not been evaluated in WSLs produced from microcosm biofilms. Objective: This study characterized artificial white spot lesions produced on human enamel under microcosm biofilm for different experimental periods. Methodology: In total, 100 human enamel specimens (4x4mm) were assigned to 5 distinct groups (n=20/group) differing according to the period of biofilm formation (2, 4, 6, 8 or 10 days). Microcosm biofilm was produced on the specimens from a mixture of human and McBain saliva at the first 8h. Enamel samples were then exposed to McBain saliva containing 0.2% sucrose. WSLs formed were characterized by quantitative light-induced fluorescence (QLF) and transverse microradiography (TMR). Data were analyzed by ANOVA/Tukey or Kruskal-Wallis/Dunn tests (p<0.05). Results: A clear time-response pattern was observed for both analyses, but TMR was able to better discriminate among the lesions. Regarding QLF analysis, median (95%CI; %) changes in fluorescence ∆Z were -7.74(-7.74:-6.45)a, -8.52(-8.75:-8.00)ab, -9.17(-10.00:-8.71)bc, -9.58(-10.53:-8.99)bc and -10.01(-11.44:-9.72)c for 2, 4, 6, 8, and 10 days, respectively. For TMR, median (95%CI; vol%.µm) ∆Z were 1410(1299-1479)a, 2420(2327-2604)ab, 2775(2573-2899)bc, 3305(3192-3406)cd and 4330(3972-4465)d, whereas mean (SD; µm) lesion depth were 53.7(12.3)a, 71.4(12.0)a, 103.8(24.8)b, 130.5(27.2)bc, 167.2(39.3)c for 2, 4, 6, 8 and 10 days, respectively. Conclusion: The progression of WSLs formed on human enamel under microcosm biofilm can be characterized over 2-10 days, both by QLF and TMR analyses, although the latter provides better discrimination among the lesions.

Abstract: This randomized three-armed controlled clinical trial compared the effect of titanium tetrafluoride (TiF4) and sodium fluoride (NaF) varnishes on caries control in smooth surfaces of permanent dentition and children's acceptability. Sixty children (6-8 y/o) were randomly divided into TiF4 (2.45% F–), NaF (2.26% F–) or placebo (control) groups. Varnishes were applied on permanent teeth once a week for the first 4 weeks and after the 6th and 12th months of the study. The variables were as follows: International Caries Detection and Assessment System (ICDAS) scores, quantitative fluorescence changes, visual plaque index (VPI) and degree of acceptability. Two-way RM-ANOVA, ANOVA/Tukey and χ2 tests were performed (p < 0.05). No differences were found between the treatments with respect to ICDAS scores (p = 0.32). Only TiF4 reduced the mean fluorescence loss significantly at 18 months compared to the baseline (p = 0.003). TiF4 showed a lower percentage of new caries lesions by tooth surface than the placebo, while NaF did not induce such a change (p < 0.014). Regardless of the treatment, more than 95% of the participants reported being satisfied. For all groups, the VPI decreased significantly at 3 months compared to the baseline value (p < 0.001), with no differences between the treatments (p = 0.17). TiF4 had a similar ability to control caries lesions as NaF; however, only TiF4 differed from the placebo (p = 0.004). The acceptability of TiF4 varnish was similar to that of NaF varnish.

Abstract Background: Almost all Tityus characterized toxins are from subgenera Atreus and Tityus, there are only a few data about toxins produced by Archaeotityus, an ancient group in Tityus genus. Methods: Tityus (Archaeotityus) mattogrossensis crude venom was fractionated by high performance liquid chromatography, the major fractions were tested in a frog sciatic nerve single sucrose-gap technique. Two fractions (Tm1 and Tm2) were isolated, partially sequenced by MALDI-TOF/MS and electrophysiological assayed on HEK293 Nav 1.3, HEK293 Nav 1.6, DUM and DRG cells. Results: The sucrose-gap technique showed neurotoxicity in four fractions. One fraction caused a delay of action potential repolarization and other three caused a reduction in amplitude. An electrophysiological assay showed that Tm1 is active on HEK293 Nav 1.3, HEK293 Nav 1.6, DUM and DRG cells, and Tm2 on HEK293 Nav 1.3 and DRG cells, but not in HEK293 Nav 1.6. In addition, Tm1 and Tm2 did promote a shift to more negative potentials strongly suggesting that both are α-NaScTx. Conclusion: Although Tityus (Archaeotityus) mattogrossensis is considered an ancient group in Tityus genus, the primary structure of Tm1 and Tm2 is more related to Tityus subgenus. The patch clamp electrophysiological tests suggest that Tm1 and Tm2 are NaScTx, and also promoted no shift to more negative potentials, strongly suggesting that both are α-NaScTx. This paper aimed to explore and characterize for the first time toxins from the ancient scorpion Tityus (Archaeotityus) mattogrossensis.

Abstract The Theraphosidae family includes the largest number of species of the Mygalomorphae infraorder, with hundreds of species currently catalogued. However, there is a huge lack on physiologic and even ecologic information available, especially in Brazil, which is the most biodiverse country in the world. Over the years, spiders have been presented as a source of multiple biologically active compounds with basic roles, such as primary defense against pathogenic microorganisms or modulation of metabolic pathways and as specialized hunters. Spider venoms also evolved in order to enable the capture of prey by interaction with a diversity of molecular targets of interest, raising their pharmaceutical potential for the development of new drugs. Among the activities found in compounds isolated from venoms and hemocytes of Brazilian Theraphosidae there are antimicrobial, antifungal, antiparasitic and antitumoral, as well as properties related to proteinase action and neuromuscular blockage modulated by ionic voltage-gated channel interaction. These characteristics are present in different species from multiple genera, which is strong evidence of the important role in spider survival. The present review aims to compile the main results of studies from the last decades on Brazilian Theraphosidae with special focus on results obtained with the crude venom or compounds isolated from both venom and hemocytes, and their physiological and chemical characterization.

Abstract Objective This in vitro study evaluated the effect of commercial whitening dentifrices on erosive tooth wear (ETW) of bovine enamel samples, in comparison with commercial regular dentifrices. Methodology Sixty bovine crowns were embedded in acrylic resin, polished and then had their baseline profile determined. They were randomly assigned to 5 groups (n=12/group), according to the type of commercial dentifrice to be tested: GI – Crest Anti-cavity Regular; GII – Crest 3D White; GIII – Colgate Total 12 Clean Mint; GIV – Colgate Optic White; GV – Placebo (negative control, fluoride-free dentifrice). The samples were submitted to daily erosive and abrasive challenges for 3 days. The erosive challenges were performed 3 times a day by immersing the specimens in 0.1% citric acid solution (pH 2.5) for 90 s. Each day after the first and last erosive challenges, the specimens were subjected to the abrasive challenge for 15 s, using a toothbrushing machine (Biopdi, São Carlos, SP, Brazil), soft toothbrushes and slurry (1:3 g/ml) of the tested toothpastes (1.5 N). The specimens were kept in artificial saliva between the challenges. The final profile was obtained and the ETW (µm) was calculated. Data were analyzed by Kruskal-Wallis and Dunn’s tests (p<0.05). Results All dentifrices tested significantly reduced the enamel wear in comparison with the Placebo, except GIII. The median (95% CI) ETW was 1.35 (1.25-1.46)bc for GI, 1.17 (1.01-1.34)cd for GII, 1.36 (1.28-1.45)ab for GIII, 1.08 (1.04-1.14)d for GIV and 2.28 (2.18-2.39)a for GV. Conclusion When dentifrices from the same manufacturer were compared, the whitening dentifrices led to similar or less wear than the regular ones.

Abstract The increased consumption of citrus sweets can contribute to the development of erosive tooth wear (ETW). Objective This in vitro study evaluated the erosive potential of citrus sweets on bovine enamel samples regarding the quantification of wear. Methodology Ninety bovine crowns were prepared and samples were randomly distributed into 6 groups (n=15): 0.1% citric acid solution (pH 2.5); Coca-Cola ® Soft Drink (pH 2.6); Fini ® Diet (lactic and citric acid, pH 3.3); Fini ® Jelly Kisses (lactic and citric acid, pH 3.5); Fini ® Fruit Salad Bubblegum (maleic acid, pH 2.6); Fini ® Regaliz Acid Tubes (maleic and citric acid, pH 3.1). Sweets were dissolved in the proportion of 40 g/250 mL of deionized water. Enamel samples were submitted to erosive challenges for 7 days (4 daily acid immersion cycles for 90 s each). Enamel wear was measured using contact profilometry (μm), and data (median values [interquartile range]) were submitted to Kruskal-Wallis/Dunn’s test (p<0.0001). Results All citrus sweets tested present a high erosive potential, Fini Diet ® (2.4 [1.2]) and Fini Regaliz Tubs ® (2.2 [0.5]) show the highest erosive potential, similar to 0.1% citric acid (2.3 [0.7]); Fini Regaliz Tubs ® is more erosive than Coca-Cola ® (1.4 [0.9]). Conclusion The evaluated citrus sweets have great erosive potential and play a key role in the development of ETW.

Abstract Objectives: The aim of this study was to assess the effect of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves hydroalcoholic extracts on viability and metabolism of a microcosm biofilm and on enamel demineralization prevention. Methodology: Microcosm biofilm was produced on bovine enamel using inoculum from pooled human saliva mixed with McBain saliva, under 0.2% sucrose exposure, for 14 days. The biofilm was daily-treated with the extracts for 1 min. At the end, it was analyzed with respect to viability by fluorescence, CFU counting and extracellular polysaccharides (phenol-sulphuric acid colorimetric assay) and lactic acid (enzymatic assay) production. The demineralization was measured by TMR. The data were compared using ANOVA or Kruskal-Wallis (p<0.05). Results: M. urundeuva All. at 100, 10 and 0.1 μg/mL and Q. grandiflora Mart. at 100 and 0.1 μg/mL reduced biofilm viability similarly to positive control (chlorhexidine) and significantly more than the negative-vehicle control (35% ethanol). M. urundeuva at 1000, 100 and 0.1 μg/mL were able to reduce both lactobacilli and mutans streptococci CFU counting, while Q. grandiflora (1000 and 1.0 μg/mL) significantly reduced mutans streptococci CFU counting. On the other hand, the natural extracts were unable to significantly reduce extracellular polysaccharides and lactic acid productions neither the development of enamel carious lesions. Conclusions: The extracts showed antimicrobial properties on microcosm biofilm, however, they had no effect on biofilm metabolism and caries protection.

Abstract Microcosm biofilm has been applied to induce carious lesions in dentin. However, no study has been done to compare the impact of the type of model for providing nutrients to microcosm biofilm formation on dentin. Objective This study compared the performance of two kinds of models (static and semi-dynamic) on the biofilm formation and the development of dentin carious lesions. Material and Methods In both models, biofilm was produced using inoculum from pooled human saliva mixed with McBain saliva for the first 8 h (5% CO2 and 37°C). Afterwards, for the static model, the samples were placed in 24-wells microplate containing McBain saliva with 0.2% sucrose, which was replaced at 24 h. In the semi-dynamic model, the samples were submitted to artificial mouth system with continuous flow of McBain saliva with 0.2% sucrose (0.15 ml/min, 37°C) for 10 h a day (for the other 14 h, no flow was applied, similarly to the static model). After 5 days, biofilm viability was measured by fluorescence and dentin demineralization by transverse microradiography. Results Biofilm viability was significantly lower for the static compared with semi-dynamic model, while dentin demineralization was significantly higher for the first one (p<0.05). The static model was able to produce a higher number of typical subsurface lesions compared with the semi-dynamic model (p<0.05). Conclusions The type of model (static and semi-dynamic) applied in the microcosm biofilm may have influence on it's viability and the severity/profile of dentin carious lesions.