Effects of different primers and colouring solutions on orthodontic bonding: shear bond strength and colour change.

OBJECTIVES: This in vitro study evaluated the effect of different colouring solutions and primer systems used in the bonding of brackets on enamel colour change and bond strength. MATERIALS AND METHODS: 120 premolar teeth were divided into four main groups; brackets were bonded with 37% orthophosphoric acid + Transbond XT Primer in Group 1, 3 M Single Bond Universal in Group 2, Transbond Plus SEP in Group 3, and G-Premio Bond in Group 4. Each group was divided into three subgroups, and the teeth were placed in a cup containing coffee and tea mixture, in a cup containing cola and in distilled water. A bond strength test was applied to all teeth. Colour measurements of all teeth were performed at 2 different times: before bonding and after the bond strength test. RESULTS: The average bond strength of the 37% orthophosphoric acid group was higher than that of the other groups. The effect of primer and solution groups on colour change was statistically significant (p = 0.001 and p = 0.023, respectively). CONCLUSIONS: In this study, the bond strength was clinically sufficient in all primer groups. The highest colour change was observed when the tea-coffee solution and Transbond Plus SEP primer were used. CLINICAL RELEVANCE: This study has identified enamel discoloration and bond strength from different colouring solutions and primer systems used for bonding braces, which can be used to inform clinicians and patients to achieve better treatment results.

High-Resolution Characterization of Enamel Remineralization Using Time-of-Flight Secondary Ion Mass Spectrometry and Electron Microscopy.

INTRODUCTION: The aim of this in vitro study was to assess the suitability of high-resolution time-of-flight secondary ion mass spectrometry (ToF-SIMS) for visualizing cross-sectional changes in human enamel microstructure and chemical composition during treatment and remineralization cycling of artificially generated caries lesions underneath an artificial plaque. METHODS: Treatments consisted of exposure to twice daily toothpaste/water slurries prepared from 0, 1,100, and 5,000 µg/g fluoride (F) NaF/silica toothpastes. In addition, treatments with slurries prepared from 1,100 µg/g F SnF2/silica toothpastes were done using 44Ca in the remineralization solution to allow for differentiation of newly formed mineral and exploration of incorporated metal dopants using ToF-SIMS. Complementary microhardness, scanning electron microscopy, and high-resolution transmission electron microscopy (HR-TEM) investigations were performed on enamel cross sections. RESULTS: HR-TEM was used for the first time to determine the change in crystallinity during remineralization revealing distinct microstructural zones within one lesion. Chemical mapping using ToF-SIMS demonstrated that the distribution of F, while observed primarily in the new mineral phase, was widespread throughout the lesion with 44Ca substantially limited to the remineralizing mineral. Both penetrated the inter-rod spaces of the sound enamel illustrating how acid damage propagates into the native mineral as the caries lesion deepens. HR-TEM examination revealed different regions within the lesion characterized by distinct micro- and ultrastructures. Importantly, HR-TEM revealed a return of crystallinity following remineralization. F dose-response observations verified the ability of these high-resolution techniques to differentiate remineralization efficacy. CONCLUSION: The collective results provided new insights such as the visualization of F or calcium penetration pathways, as well as new tools to study the caries process.

EFFECTS OF PROFESSIONAL ORAL HYGIENE AND TEETH WHITENING ON THE MICROELEMENT COMPOSITION OF ENAMEL.

OBJECTIVE: Aim: The objective of this study is to investigate the impact of professional teeth cleaning and the substances used in modern dentistry for whitening on the microelement composition of tooth enamel. PATIENTS AND METHODS: Materials and Methods: To study the morphology and microelement composition of the enamel, scanning electron microscopy was performed using the MiraLM microscope equipped with a Schottky field emission electron gun from Tescan. RESULTS: Results: A comparative analysis between the areas subjected to mechanical cleaning and those where it was not applied revealed a significant difference in the research results, particularly in carbon, which changed from 25.16±1.04 to 32.02±1.8. An analysis of the enamel's chemical composition before and after whitening revealed a decrease in carbon from 45.91±1.20 to 42.46±1.74. The change in phosphorus content was determined to be from 9.77±0.39 to 9.56±0.75. A decrease in calcium from 15.96±0.64 to 15.21±1.22 and magnesium from 0.07±0.01 to 0.01±0.01 was also observed. CONCLUSION: Conclusions: Professional dental hygiene does not have a direct impact on the microelement composition of enamel, such as the levels of calcium, phosphorus, fluoride, and other microelements. However, it can have an indirect and temporary influence due to the use of abrasive materials that affect dental deposits, pellicle, and the surface layer of enamel. Teeth whitening can affect the microelement composition of enamel, but these changes are mostly temporary and associated with processes of demineralization/ remineralization and oxygenation.

Bioinspired caries preventive strategy via customizable pellicles of saliva-derived protein/peptide constructs.

Dental caries has been the most widespread chronic disease globally associated with significant health and financial burdens. Caries typically starts in the enamel, which is a unique tissue that cannot be healed or regrown; nonetheless, new preventive approaches have limitations and no effective care has developed yet. Since enamel is a non-renewable tissue, we believe that the intimate overlaying layer, the acquired enamel pellicle (AEP), plays a crucial lifetime protective role and could be employed to control bacterial adhesion and dental plaque succession. Based on our identified AEP whole proteome/peptidome, we investigated the bioinhibitory capacities of the native abundant proteins/peptides adsorbed in pellicle-mimicking conditions. Further, we designed novel hybrid constructs comprising antifouling and antimicrobial functional domains derived from statherin and histatin families, respectively, to attain synergistic preventive effects. Three novel constructs demonstrated significant multifaceted bio-inhibition compared to either the whole saliva and/or its native proteins/peptides via reducing biomass fouling and inducing biofilm dispersion beside triggering bacterial cell death. These data are valuable to bioengineer precision-guided enamel pellicles as an efficient and versatile prevention remedy. In conclusion, integrating complementary acting functional domains of salivary proteins/peptides is a novel translational approach to design multifunctional customizable enamel pellicles for caries prevention.

Human mobility in a Bronze Age Vatya 'urnfield' and the life history of a high-status woman.

In this study, we present osteological and strontium isotope data of 29 individuals (26 cremations and 3 inhumations) from Szigetszentmiklós-Ürgehegy, one of the largest Middle Bronze Age cemeteries in Hungary. The site is located in the northern part of the Csepel Island (a few kilometres south of Budapest) and was in use between c. 2150 and 1500 BC, a period that saw the rise, the apogee, and, ultimately, the collapse of the Vatya culture in the plains of Central Hungary. The main aim of our study was to identify variation in mobility patterns among individuals of different sex/age/social status and among individuals treated with different burial rites using strontium isotope analysis. Changes in funerary rituals in Hungary have traditionally been associated with the crises of the tell cultures and the introgression of newcomers from the area of the Tumulus Culture in Central Europe around 1500 BC. Our results show only slight discrepancies between inhumations and cremations, as well as differences between adult males and females. The case of the richly furnished grave n. 241 is of particular interest. The urn contains the cremated bones of an adult woman and two 7 to 8-month-old foetuses, as well as remarkably prestigious goods. Using 87Sr/86Sr analysis of different dental and skeletal remains, which form in different life stages, we were able to reconstruct the potential movements of this high-status woman over almost her entire lifetime, from birth to her final days. Our study confirms the informative potential of strontium isotopes analyses performed on different cremated tissues. From a more general, historical perspective, our results reinforce the idea that exogamic practices were common in Bronze Age Central Europe and that kinship ties among high-rank individuals were probably functional in establishing or strengthening interconnections, alliances, and economic partnerships.

Bronze Age innovations and impact on human diet: A multi-isotopic and multi-proxy study of western Switzerland.

The archaeological Bronze Age record in Europe reveals unprecedented changes in subsistence strategies due to innovative farming techniques and new crop cultivation. Increasing cultural exchanges affected the economic system. The inhabitants of Switzerland played a pivotal role in this European context through relationships with the Mediterranean, the High and Middle Danube regions and the Alps thanks to the area's central position. This research aims to reconstruct, for the first time in Switzerland, human socio-economic systems through the study of human diet, herding and farming practices and their changes throughout the Bronze Age (2200-800 BCE) by means of biochemical markers. The study includes 41 human, 22 terrestrial and aquatic animal specimens and 30 charred seeds and chaff samples from sites in western Switzerland. Stable isotope analyses were performed on cereal and legume seeds (δ13C, δ15N), animal bone collagen (δ13Ccoll, δ15N, δ34S), human bone and tooth dentine collagen (δ13Ccoll, δ15N,) and human tooth enamel (δ13Cenamel). The isotopic data suggest a) an intensification of soil fertilization and no hydric stress throughout the Bronze Age, b) a human diet mainly composed of terrestrial resources despite the proximity of Lake Geneva and the Rhone river, c) a diet based on C3 plants during the Early and Middle Bronze Age as opposed to the significant consumption of 13C-enriched resources (probably millet) by individuals from the Final Bronze Age, d) no important changes in dietary patterns throughout an individual's lifespan but a more varied diet in childhood compared to adulthood, e) no differences in diet according to biological criteria (age, sex) or funerary behavior (burial architecture, grave goods).

Desperately seeking stress: A pilot study of cortisol in archaeological tooth structures.

OBJECTIVES: Cortisol is a glucocorticoid hormone produced through activation of the hypothalamic pituitary adrenal axis. It is known as the "stress hormone" for its primary role in the body's stress response and has been the focus of much modern clinical research. Within archaeology, only a few studies have analyzed cortisol in human remains and these have been restricted to hair (Webb et al., 2010; Webb, White, van Uum, & Longstaffe, 2015a; Webb, White, van Uum, & Longstaffe, 2015b). This study examines the utility of dentine and enamel, which survive well archaeologically, as possible reservoirs for detectable levels of cortisol. MATERIALS AND METHODS: Then, 69 teeth from 65 individuals from five Roman and Post-Roman sites in France were tested via competitive enzyme-linked immunosorbent assay (ELISA) to assess and quantify the cortisol concentrations present within tooth dentine and enamel. RESULTS: In both tooth dentine and enamel, detectable concentrations of cortisol were identified in multiple teeth. However, concentrations were low and not all teeth yielded results that were measurable through cortisol ELISA. Differences in cortisol values between dentine and enamel could suggest different uptake mechanisms or timing. DISCUSSION: These results suggest that cortisol is incorporated within tooth structures and merits further investigation in both modern and archaeological contexts. Analysis of the results through liquid chromatographic-mass spectrometry would verify current results and might yield values that could be better integrated with published cortisol studies. Future studies of cortisol in tooth structures would greatly expand the research potential of cortisol in the past and could have implications for studies of human stress across deep time.

Qualitative and quantitative assessment of the potential effect of cigarette smoking on enamel of human smokers' teeth.

OBJECTIVE: This study aimed to examine the potential changes in enamel surface of human smokers' teeth. MATERIALS AND METHODS: Forty extracted permanent, human, noncarious anterior teeth were used in this study. Half of these teeth were obtained from heavy smokers, while the other half of teeth were collected from nonsmokers (control teeth). The teeth were then subjected for scanning electron microscopic examination together with energy dispersive X ray and micro-hardness analysis to evaluate the qualitative and quantitative effect of smoking respectively. RESULTS: SEM of smokers' teeth showed variable degrees of destruction from small areas of demineralization as holes and pits to destruction and deterioration of the organizational pattern of the rod substance. Moreover, areas of defective remineralization were detected. The microhardness, calcium and phosphorus weight % significantly decreased whereas the Ca/P ratio was significantly increased. CONCLUSION: Cigarette smoking adversely affected the ultrastructure and mechanical properties of enamel and even hindered the normal remineralization process thus cigarette smoking cessation should be promoted in the dental office daily practices.

New evidence on the earliest domesticated animals and possible small-scale husbandry in Atlantic NW Europe.

The distribution of the first domesticated animals and crops along the coastal area of Atlantic NW Europe, which triggered the transition from a hunter-gatherer-fisher to a farmer-herder economy, has been debated for many decades among archaeologists. While some advocate a gradual transition in which indigenous hunter-gatherers from the very beginning of the 5th millennium cal BC progressively adopted Neolithic commodities, others are more in favor of a rapid transition near the end of the 5th millennium caused by a further northwest migration of farmers-herders colonizing the lowlands. Here, radiocarbon dated bones from sheep/goat and possibly also cattle are presented which provide the first hard evidence of an early introduction of domesticated animals within a hunter-gatherer context in NW Belgium, situated ca. 80 km north of the agro-pastoral frontier. Based on their isotope signal it is suggested that these first domesticates were probably not merely obtained through exchange with contemporaneous farmers but were kept locally, providing evidence of small-scale local stockbreeding in the lowlands maybe as early as ca. 4800/4600 cal BC. If confirmed by future in-depth isotope analyses, the latter testifies of intense contact and transmission of knowledge in this early contact period, which is also visible in the material culture, such as the lithic and pottery technology. It also implies direct and prolonged involvement of farmer-herders, either through visiting specialists or intermarriage, which follows recent genetic evidence demonstrating much more hunter-gatherer ancestry in early farmer's genes in western Europe compared to central and SE Europe.

Specialized cattle farming in the Neolithic Rhine-Meuse Delta: Results from zooarchaeological and stable isotope (δ18O, δ13C, δ15N) analyses.

Schipluiden (3630-3380 cal BC), the earliest known year-round settlement in the Rhine-Meuse Delta in the Netherlands, is a key site for addressing the nature of Neolithic subsistence in the wetlands of northwestern Europe. A preliminary zooarchaeological study suggested that cattle husbandry was a major activity at Schipluiden. In contrast, stable carbon and nitrogen isotope analyses of human remains from the site indicated a marine-oriented diet, implying that the Mesolithic-Neolithic dietary transition continued well into the mid-4th Millennium BC in this region. Here, we re-investigate the role and nature of cattle husbandry at Neolithic Schipluiden using mortality profiles and stable isotope analysis (δ18O, δ13C, δ15N) of animal bone collagen and tooth enamel. The age-at-death analysis suggests that cattle were managed for both meat and milk production. The δ18O and δ13C analysis of tooth enamel provide evidence that calving spread over five-and-a-half-months, which would have led to a longer availability of milk throughout the year. Cattle were grazing in open, marshy environments near the site and winter foddering was practiced occasionally. The faunal isotopic data also reveal that the high 15N in human bone collagen is more likely to signal the consumption of products from cattle that grazed on 15N-enriched salt marsh plants around the site, rather than a marine-oriented diet. This undermines the previous interpretation of the dietary practices at Schipluiden by showing that human diet in mid-4th millennium BC Rhine-Meuse area was fully "Neolithic", based primarily on products from domesticates, especially cattle, with some input from wild terrestrial and aquatic resources available in their surroundings, contrary to what has been proposed before. Collating these results demonstrates a high level of investment in cattle husbandry, highlighting the social and economic importance of cattle at the lower Rhine-Meuse Delta during the 4th millennium BC.

Influence of salivary conditioning and sucrose concentration on biofilm-mediated enamel demineralization.

INTRODUCTION: The acquired pellicle formation is the first step in dental biofilm formation. It distinguishes dental biofilms from other biofilm types. OBJECTIVE: To explore the influence of salivary pellicle formation before biofilm formation on enamel demineralization. METHODOLOGY: Saliva collection was approved by Indiana University IRB. Three donors provided wax-stimulated saliva as the microcosm bacterial inoculum source. Acquired pellicle was formed on bovine enamel samples. Two groups (0.5% and 1% sucrose-supplemented growth media) with three subgroups (surface conditioning using filtered/pasteurized saliva; filtered saliva; and deionized water (DIW)) were included (n=9/subgroup). Biofilm was then allowed to grow for 48 h using Brain Heart Infusion media supplemented with 5 g/l yeast extract, 1 mM CaCl2.2H2O, 5% vitamin K and hemin (v/v), and sucrose. Enamel samples were analyzed for Vickers surface microhardness change (VHNchange), and transverse microradiography measuring lesion depth (L) and mineral loss (∆Z). Data were analyzed using two-way ANOVA. RESULTS: The two-way interaction of sucrose concentration × surface conditioning was not significant for VHNchange (p=0.872), ∆Z (p=0.662) or L (p=0.436). Surface conditioning affected VHNchange (p=0.0079), while sucrose concentration impacted ∆Z (p<0.0001) and L (p<0.0001). Surface conditioning with filtered/pasteurized saliva resulted in the lowest VHNchange values for both sucrose concentrations. The differences between filtered/pasteurized subgroups and the two other surface conditionings were significant (filtered saliva p=0.006; DIW p=0.0075). Growing the biofilm in 1% sucrose resulted in lesions with higher ∆Z and L values when compared with 0.5% sucrose. The differences in ∆Z and L between sucrose concentration subgroups was significant, regardless of surface conditioning (both p<0.0001). CONCLUSION: Within the study limitations, surface conditioning using human saliva does not influence biofilm-mediated enamel caries lesion formation as measured by transverse microradiography, while differences were observed using surface microhardness, indicating a complex interaction between pellicle proteins and biofilm-mediated demineralization of the enamel surface.

High-Frequency Raman Analysis in Biological Tissues Using Dual-Wavelength Excitation Raman Spectroscopy.

A dual-wavelength excitation Raman probe with laser inputs at 866 nm or 1064 nm is customized and integrated into a compact Raman spectrometer that is based on an InGaAs detector. Under 1064 nm illumination, the spectrometer detects fingerprint Raman signals below 2000 cm-1. While under 866 nm illumination, the spectral range is extended to cover high-frequency region (2400-4000 cm-1) that includes major C-H and O-H Raman vibrations. We demonstrate that the dual excitation InGaAs Raman is beneficial in detecting high-frequency Raman signals, especially water contents in high-fluorescent biological samples such as human dental tissues, grape skin, and plum skin due to the suppressed fluorescence interference.

Influence of salivary conditioning and sucrose concentration on biofilm-mediated enamel demineralization

Abstract The acquired pellicle formation is the first step in dental biofilm formation. It distinguishes dental biofilms from other biofilm types. Objective To explore the influence of salivary pellicle formation before biofilm formation on enamel demineralization. Methodology Saliva collection was approved by Indiana University IRB. Three donors provided wax-stimulated saliva as the microcosm bacterial inoculum source. Acquired pellicle was formed on bovine enamel samples. Two groups (0.5% and 1% sucrose-supplemented growth media) with three subgroups (surface conditioning using filtered/pasteurized saliva; filtered saliva; and deionized water (DIW)) were included (n=9/subgroup). Biofilm was then allowed to grow for 48 h using Brain Heart Infusion media supplemented with 5 g/l yeast extract, 1 mM CaCl2.2H2O, 5% vitamin K and hemin (v/v), and sucrose. Enamel samples were analyzed for Vickers surface microhardness change (VHNchange), and transverse microradiography measuring lesion depth (L) and mineral loss (∆Z). Data were analyzed using two-way ANOVA. Results The two-way interaction of sucrose concentration × surface conditioning was not significant for VHNchange (p=0.872), ∆Z (p=0.662) or L (p=0.436). Surface conditioning affected VHNchange (p=0.0079), while sucrose concentration impacted ∆Z (p<0.0001) and L (p<0.0001). Surface conditioning with filtered/pasteurized saliva resulted in the lowest VHNchange values for both sucrose concentrations. The differences between filtered/pasteurized subgroups and the two other surface conditionings were significant (filtered saliva p=0.006; DIW p=0.0075). Growing the biofilm in 1% sucrose resulted in lesions with higher ∆Z and L values when compared with 0.5% sucrose. The differences in ∆Z and L between sucrose concentration subgroups was significant, regardless of surface conditioning (both p<0.0001). Conclusion Within the study limitations, surface conditioning using human saliva does not influence biofilm-mediated enamel caries lesion formation as measured by transverse microradiography, while differences were observed using surface microhardness, indicating a complex interaction between pellicle proteins and biofilm-mediated demineralization of the enamel surface.

Effect of grape seed extract-containing phosphoric acid formulations on bonding to enamel and dentin.

The aim was to evaluate the effect of 2% grape seed extract (GSE) containing phosphoric acid (PhA) on the bond strength to enamel and dentin. The control group was 37% PhA. The following three PhA formulations with 2% GSE and 20% ethanol were obtained: GSE5 = 5% PhA; GSE10 = 10% PhA; and GSE20 = 20% PhA. The enamel and dentin surfaces of molars were etched with the acid solutions, followed by Scotchbond Multi-Purpose adhesive and composite resin application. The tensile bond strength (TBS) test evaluated the bond to enamel after 24 h, and the microtensile bond strength (µTBS) test evaluated the bond to dentin after 24 h and 12-month water storage. Etched enamel and dentin were observed by scanning electron microscopy (SEM) and atomic force microscopy (AFM), respectively. The TBS data were submitted to one-way ANOVA, while µTBS data were submitted to two-way ANOVA and Tukey's test (α = 0.05). The TBS (MPa) to enamel did not significantly differ among the control (48.1 ± 15.7), GSE5 (46.1 ± 9.6), GSE10 (49.8 ± 13.6) and GSE20 (44.1 ± 11.9) groups (p = 0.537). The µTBS (MPa) to dentin of the control (28.4 ± 14.4) and GSE20 (24.1 ± 8.1) groups were significantly higher than those of the GSE5 (16.8 ± 7.4) and GSE10 (17.5 ± 6.6) groups at 24 h (p < 0.006). After 12-month storage, only GSE5 (21.0 ± 7.8) and GSE10 (17.6 ± 8.0) did not show significantly decreased µTBS (p > 0.145). SEM micrographs showed a shallower enamel etching pattern for GSE5. AFM images showed the formation of collagenous globular structures for GSE5 and GSE10. The different acid solutions did not influence the TBS to enamel, and the µTBS to dentin was stable over time when dentin was etched with GSE5 and GSE10.

Role of Candida albicans on enamel demineralization and on acidogenic potential of Streptococcus mutans in vitro biofilms.

There is growing evidence that C. albicans is associated with dental caries, but its role on caries development needs to be better clarified. Label="OBJECTIVE">To evaluate at the hard tissue level the effect of C. albicans on the cariogenic potential of S. mutans biofilms focusing on the mineral profile of induced carious lesions. This study also aimed to evaluate the effect of C. albicans on the acidogenic potential of S. mutans biofilms. METHODOLOGY Dual-species (CA+SM) and single-species biofilms (CA or SM) were grown on the surface of enamel slabs in the presence of glucose/sucrose supplemented culture medium for 24, 48 and 72 hours. Demineralization was evaluated through percentage of surface microhardness change (%SMC) and transversal microradiography analysis (ILM and LD) and pH of the spent medium was recorded daily. Data were analyzed by two-way ANOVA followed by Bonferroni correction. RESULTS%SMC was statistically different among the biofilms at each time point being the highest for SM biofilms and the lowest for CA biofilms which also differed from CA+SM biofilms [SM (24 h: 47.0±7.3; 48 h: 66.3±8.3; 72 h: 75.4±3.9); CA (24 h: 7.3±3.3; 48 h: 7.1±6.4; 72 h: 6.6±3.6); CA+SM (24 h: 35.9±7.39.1; 48 h: 47.2±9.5; 72 h: 47.6±9.5)]. pH of spent medium was statistically lower for SM biofilms compared to the other biofilms at each time point and remained constant over time while pH values increased from 24 to 72 h for both CA and CA+SM biofilms [SM (24 h: 4.4±0.1; 48 h: 4.4±0.1; 72 h: 4.5±0.1); CA (24 h: 6.9±0.3; 48 h: 7.2±0.2; 72 h: 7.5±0.2); CA+MS (24 h: 4.7±0.2; 48 h: 5.1±0.1; 72 h: 6.1±0.6)]. IML and LD for SM biofilms increased over time while no difference was observed from 24 to 72 h for the other biofilms. CONCLUSIONS The present data suggest that C. albicans has low enamel demineralization potential and the presence of C. albicans can reduce both the cariogenic and acidogenic potentials of S. mutans biofilms.

The effect of cerebral palsy on neonatal line thickness and enamel components.

OBJECTIVE: The aim of the study was to compare the neonatal line width as well as the composition of the pre-natal and post-natal enamel in deciduous teeth of children with cerebral palsy (CP) to deciduous teeth of healthy children. DESIGN: 58 extracted or normally exfoliated deciduous teeth were collected for the study, 29 teeth from children with cerebral palsy and 29 pair matched teeth from healthy children who served as controls. The teeth were cut along the bucco-lingual/palatal axis and polished up to a thickness of 50-100 microns. The sections were examined using a light microscope and scanning electron microscope (SEM). The neonatal line was identified in 25 teeth and measured in width and the enamel concentrations of calcium, phosphorus, silicon, magnesium, sodium, oxygen, and carbon were measured in the pre-natal and post-natal enamel using ESD program. RESULTS: The neonatal line was significantly narrower in the deciduous teeth collected from children with cerebral palsy. In both groups, the concentration of magnesium and sodium in the pre-natal enamel was significantly higher than in the post-natal enamel. Magnesium and sodium levels were significantly higher in pre-natal enamel in the CP group compared to the pre-natal enamel of the control group. CONCLUSIONS: The neonatal line was narrower in the deciduous teeth collected from children with cerebral palsy than in those collected from healthy children. Children with cerebral palsy have a higher concentration of magnesium and sodium in the enamel that developed before birth.

Comparing the accuracy of PCR-capillary electrophoresis and cuticle microhistological analysis for assessing diet composition in ungulates: A case study with Pyrenean chamois.

The study of diet composition is required to understand the interactions between animal and plant ecosystems. Different non-invasive techniques applied on faecal samples have commonly been used for such purposes, with cuticle microhistological analysis (CMA) and emerging DNA-based methods being the most relevant. In this work, we refined and optimized a qualitative DNA-based approach combining PCR amplification of long trnL(UAA) and ITS2 fragments and capillary electrophoresis (PCR-CE), instead of short trnL(UAA) fragments and massive sequencing technologies commonly reported. To do so, we developed a controlled diet assay using a stabled Pyrenean chamois specimen (Rupicapra pyrenaica pyrenaica), which included representative herbaceous and shrubby plant species. We also assessed the impact of sample freshness on the diet determination of this mountain caprinae by exposing faecal samples to the outdoor environment for three weeks. Faecal samples from both experiments were analysed by qualitative PCR-CE and semi-quantitative CMA in order to compare the pros and cons of both approaches. Our results show that all of the offered plant species were detected by both methodologies although CMA over-detected shrubs compared to herbaceous species. At the same time, sample degradation due to sustained climate exposure is a limiting factor for molecular analysis, but not for CMA. Taken all together, our results suggest that the qualitative information obtained by CMA and PCR-CE can be interchangeable when faecal samples are fresh (less than one week after deposition) but, afterwards, molecular analysis underestimates diet composition probably due to DNA degradation. CMA, however, can accurately be used at least three weeks after defecation. Moreover, by combining the results of simultaneous PCR amplification of two complementary genes, this optimized PCR-CE methodology provides a reliable, feasible and more affordable alternative for multiple and routine analyses of complex samples. Neither CMA nor PCR-CE seems to solve comprehensively the quatification of herbivore diets and thus further research needs to be done.

Inhibition Effects of Different Toothpastes on Demineralisation of Incipient Enamel Lesions.

PURPOSE: To evaluate the inhibitory effects of different toothpastes on demineralisation of incipient enamel lesions using a toothbrush simulator. MATERIALS AND METHODS: Fifty enamel specimens were prepared from extracted human molars. The specimens were randomly assigned to the following groups (n = 10/group): 1. no treatment (control); 2. toothpaste containing arginine (ProRelief, Colgate;); 3. fluoride toothpaste (Pronamel, Sensodyne GlaxoSmithKlein); 4. tooth mousse containing casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) (Recaldent, GC); 5. toothpaste (Restore, Dr. Collins) containing bioactive glass (NovaMin, GlaxoSmithKlein). All specimens were exposed to pH cycling. The remineralising agents were applied to the samples with a toothbrush simulator for 2 min twice a day for five days. The weight percentage of mineral changes for the elements calcium (Ca), phosphorus (P), sodium (Na) and silica (Si) were measured by SEM energy-dispersive x-ray spectroscopy (SEM-EDX). SEM revealed properties of treated enamel surfaces. The data were analysed using one-way ANOVA. RESULTS: Statistically significantly higher levels of Ca and P were found in all groups compared to the control (p < 0.05). CONCLUSIONS: The toothpastes' efficacy of inhibiting demineralisation depended on the active ingredients in the respective toothpaste. The demineralisation inhibition efficacy of the tested toothpastes depended on the active ingredients in the toothpaste.

A new method for estimating time since death by analysis of substances deposited on the surface of dental enamel in a body immersed in seawater.

The present investigation was performed with the objective of developing a method to estimate how long a corpse had been immersed in water after death (the time since death). Accurate determination of the time elapsed since death may lead to identification of the place of drowning, and therefore, serves not only as a piece of information useful for determination of the cause of death but also leads to prompt identification of the body. The results showed that diatoms attached to the surface of dental enamel increased with prolongation of immersion time in water. Further, as the immersion time increased, the quantity of O, Si, Mg, K, Al, and S detected on the surface of dental enamel increased, while the quantity of the main dental components (Ca and P) that were detected gradually decreased. Based on these results, we calculated a regression formula to estimate the immersion time. Our method is considered to be a breakthrough technique for evaluating the time since death more objectively, compared to the conventional method of determination based on the degree of decomposition of the corpse.