Acrocomia aculeata oil: Beneficial effects on cyclophosphamide-induced reproductive toxicity in male rats.

This study aimed to evaluate the effects of the extracted oil of Acrocomia aculeata pulp in preventing or mitigating the reproductive toxicity induced by cyclophosphamide (CP) in male rats. Adult male rats were segregated into seven groups that received vehicle, 100 mg/kg/day of CP, or 10 mg/kg/day of ß-carotene or 3 or 30 mg/kg/day of A. aculeata oil co-administered with CP. A. aculeata oil exhibited a high content of ß-carotene. CP treatment induced reproductive toxicity in the animals, as it changed the reproductive organs weight, hormone levels, sperm counts and testicular histology. In contrast, co-administration of A. aculeata improved CP-induced alterations in these parameters. A. aculeata oil also increased the gene Ckit expression and normalised the antioxidant enzymes levels which were changed by CP. The A. aculeata oil is capable of protecting the male reproductive system from the adverse effects of CP, possibly by acting as an antioxidant and increasing the Ckit gene expression.

Anti-inflammatory effects of essential oils from Mangifera indica.

Mangifera indica is widely found in Brazil, and its leaves are used as an anti-inflammatory agent in folk medicine. The aim of this study is to perform composition analysis of essential oils from the M. indica varieties, espada (EOMIL1) and coração de boi (EOMIL2), and confirm their anti-inflammatory properties. Twenty-three volatile compounds were identified via gas chromatography-mass spectrometry (GC-MS) in two essential oils from the leaves. Paw edema and myeloperoxidase (MPO) activity were evaluated using the carrageenan-induced paw model, while leukocyte migration was analyzed using the pleurisy model. At oral doses of 100 and 300 mg/kg, the essential oils significantly reduced edema formation and the increase in MPO activity induced by carrageenan in rat paws. For a dose of 300 mg/kg EOMIL1, 62 ± 8% inhibition of edema was observed, while EOMIL2 led to 51 ± 7% inhibition of edema. At a dose of 100 mg/kg, the inhibition was 54 ± 9% for EOMIL1 and 37 ± 7% for EOMIL2. EOMIL1 and EOMIL2 significantly reduced MPO activity at doses of 100 mg/kg (47 ± 5 and 23 ± 8%, respectively) and 300 mg/kg (50 ± 9 and 31 ± 7%, respectively). In the pleurisy model, inhibitions were also observed for EOMIL1 and EOMIL2 in the leukocyte migration test. The results of the present study show that essential oils from M. indica differ in chemical composition and anti-inflammatory activity in rats.

Chemical signatures in the developmental stages of Protopolybia exigua.

The chemical signature of social insects is very important for communication, and specific signs of each colony and its individuals are acquired throughout their development. This chemical signature can also be related to the composition of the materials employed during nest construction. Furthermore, the venom also contains important chemicals required for the maintenance of wasp colonies. Therefore, the objective of this study was to evaluate the chemical composition of the wasp Protopolybia exigua along its different development stages, including the nest substrate and the venom of adult worker at different ages. To achieve this, gas chromatography coupled to mass spectrometry was used. The results show that the chemical cuticular compounds present in different stages of development, and in the nest of P. exigua, are qualitatively similar, but quantitatively different, demonstrating that these compounds can be used as signals for the identification of co-specific individuals within the colonies. However, there was no significant variation in these compounds between different colonies, which may be related to the parental level between the colonies and due to them sharing the same resources. The non-polar compounds of the venom vary significantly according to the age of the workers, probably due to their different roles within the colony, and there was a clear increase in the complexity of the compounds as the wasps aged. Older wasps perform higher-risk activities, such as foraging, and therefore need to make more use of venom.

Campomanesia adamantium extract induces DNA damage, apoptosis, and affects cyclophosphamide metabolism.

Campomanesia adamantium (Cambess.) O. Berg. is originally from Brazil. Its leaves and fruits have medicinal properties such as anti-inflammatory, antidiarrheal and antiseptic properties. However, the mutagenic potential of this species has been reported in few studies. This study describes the mutagenic/antimutagenic, splenic phagocytic, and apoptotic activities of C. adamantium hydroethanolic extract with or without cyclophosphamide in Swiss mice. The animals orally received the hydroethanolic extract at doses of 30, 100, or 300 mg/kg with or without 100 mg/kg cyclophosphamide. Mutagenesis was evaluated by performing the micronucleus assay after treatment for 24, 48, and 72 h, while splenic phagocytic and apoptotic effects were investigated after 72 h. Short-term exposure of 30 and 100 mg/kg extract induced mild clastogenic/aneugenic effects and increased splenic phagocytosis and apoptosis in the liver, spleen, and kidneys. When the extract was administered in combination with cyclophosphamide, micronucleus frequency and apoptosis reduced. Extract components might affect cyclophosphamide metabolism, which possibly leads to increased clearance of this chemotherapeutic agent. C. adamantium showed mutagenic activity and it may decrease the effectiveness of drugs with metabolic pathways similar to those associated with cyclophosphamide. Thus, caution should be exercised while consuming these extracts, especially when received in combination with other drugs.

Cuticular signature in the development of Polistes versicolor.

Wasps belong to societies that are highly complex and diverse, especially considering social organization and parental care. They use chemicals in their daily communication, and act incisively in recognition of mates and non-nest mates, and can even identify individuals of different castes. In this study, cuticle composition was examined during the development of Polistes versicolor from the egg stage to adulthood, with the aim to assess changes in the chemical signature of the species, using gas chromatography. Linear alkanes and branched alkanes were identified that were important to distinguish the various developmental stages of P. versicolor. The variation in the linear alkanes and branched alkanes was quantitative, since most of them are present in all stages, with the exception of some linear alkanes that uniquely characterized the egg stage. At the egg stage, differences were predominant for linear alkanes C8, C24, C27, and C29. For the larval instars, there was a predominance of linear alkanes C8, C22 and C24 to C30. In the pre-pupae, pupae and adult stages, C8, C22, C24, and C26 to C30 showed a higher abundance during later developmental stages. The addition to branched alkanes increased their abundance from the egg (13.52%) to adult (22.96%) stages.

Mosquitoes in Bromeliads at Ground Level of the Brazilian Atlantic Forest: the Relationship Between Mosquito Fauna, Water Volume, and Plant Type.

Water accumulating in the axils of bromeliads provides habitat for numerous invertebrates, frequently among them, immature mosquitoes. To evaluate mosquito richness in bromeliads and the relationship between mosquito presence and biotic and abiotic variables, we performed a study in the Parque Nacional do Itatiaia, Rio de Janeiro, Brazil. Mosquitoes of genus Culex were the most abundant and varied in species richness, among which nine belonged to subgenus Microculex, Culex (Microculex) neglectus Lutz and Culex ocellatus Theobald being the most frequent species. Sabethines of genera Wyeomyia and Runchomyia were found in low numbers. Wyeomyia (Spilonympha) airosai Lane and Cerqueira and Wyeomyia (Spilonympha) finlayi Lane and Cerqueira tend to proliferate in bromeliads of the genus Bilbergia which hold less than 50 ml of water and grow either alone or with Runchomyia frontosa (Theobald). The larger the volume of water, the greater the chance of finding Culex, Anopheles as well as Wyeomyia (Phoniomyia) species, which seems to be the more generalist as it is present in different bromeliad types with a large range of plant water holding capacities.

Fourier transform infrared photoacoustic spectroscopy as a potential tool in assessing the role of diet in cuticular chemical composition of Ectatomma brunneum.

The cuticular chemical composition plays a significant role in the recognition of nest mates in social insects, thus functioning as a chemical signature of the colony. The structure of cuticular chemicals is subject to interference from genetic and exogenous factors, including diet. In this study, various colonies of the Ectatomma brunneum ant were removed from their natural environment and housed in a laboratory to monitor the response of the cuticular chemical composition to dietary changes. Analyses were performed using gas chromatography and Fourier transform infrared photoacoustic spectroscopy, which has not been previously used for this type of analysis. The results indicate that this method is useful for analyzing biological and natural systems. We observed changes in the chemical signature with food traces in the first 30 days under feed control. Therefore, genetic information may not be the only criterion that can be used to describe the chemical signature of a species; environmental variations also influence recognition signals. Furthermore, these results reinforce the reliability of the Fourier transform infrared photoacoustic spectroscopy method.

Ocimum gratissimum essential oil in the transport water of Brycon hilarii: implications at water quality, blood parameters and residues in tissue and plasma.

Transporting live fish is a common practice in fish farming, and is certainly one of the main problems that affect fish homeostasis. In this scenario, the use of natural additives has shown promise in improving fish resistance to adverse situations. This study aimed to assess the impact of Ocimum gratissimum L. essential oil (OGEO) on water quality, hematological parameters, and residue levels in the plasma, fillet, and liver of juvenile piraputanga (Brycon hilarii) during a two-hour transportation period. The fish were divided into plastic bags (4 L) and exposed to three different OGEO concentrations (10, 20, and 30 mg L-1), while a control group received no OGEO (three repetitions each). After the two-hour transportation, blood samples were collected, as well as portions of the fillet and liver for quantifying essential oil compounds, which were also measured in the plasma. Oxygen levels remained high throughout the transportation period, in all groups, while the pH decreased. Hemoglobin, MCHC, and MCH increased in fish exposed to OGEO concentrations of 20 and 30 mg L-1, compared to the control group. However, lymphocyte counts and the concentrations of essential oil compounds in plasma, fillet, and liver increased with higher OGEO concentrations. The use of 10 mg L-1 OGEO in the two-hour transport water is promising to ensure the survival and well-being of Brycon hilarii juveniles (weighing 16 g), showing to be safe and effective. The residual concentration of eugenol the major compound of OGEO in the fillet remains below the maximum limit of the recommended daily intake.

Phytochemical analysis, toxicity and evaluation of antioxidant and antimicrobial activities of leaves of Dipteryx alata Vogel.

In this study, our objective was to conduct a comprehensive phytochemical analysis, determine toxicity levels, and assess the antioxidant and antimicrobial properties of extracts derived from the leaves of Dipteryx alata Vogel, a native species of the Brazilian cerrado flora. Three distinct extracts were prepared utilizing assisted ultrasound and the Soxhlet apparatus, namely, Ultrasound Crude Extract (UCE), Soxhlet Crude Extract (SCE), and the Soxhlet Ethanol Extract (SEE). The phytochemical analysis revealed the presence of flavonoids, tannins, phytosterols, and saponins in all extracts. Additionally, alkaloids were specifically identified in the SCE and SEE extracts. In the analysis using LC-DAD, the compounds gallic acid, rutin, quercetin, luteolin and kampefrol were determined in higher concentrations in the SCE, followed by the SEE and UCE, respectively. The GC-MS analysis revealed the presence of campesterol, stigmasterol and ß-sitosterol in all extracts, with UCE and SCE showing a higher concentration of ß-sitosterol. SCE showed the highest concentration of all identified compounds. In the analysis of antioxidant activity by DPPH• and ABTS•+, SEE showed greater efficiency (IC50 = 2.98 ± 2.92 and 6.57 ± 0.89 µg/mL, respectively). In the toxicity test with Allium cepa, all extracts stimulated root growth at 50 g/mL; UCE and SEE stimulated root growth at 250 g/mL; and SEE inhibited root growth at 750 g/mL. In the Artemia salina toxicity, all extracts were non-toxic. Antibacterial activity was identified in the microorganisms S. aureus and S. mutans; however, the extracts did not show antifungal action against the strain of C. albicans. The extracts of D. alata have therapeutic potential for applicability in dentistry.

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

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

Parkia from Cerrado: phytochemical bioprospection, toxicity and in vitro bioactivities of bark and flower extracts.

Parkia platycephala is the only species of the genus Parkia that is endemic to the brazilian Cerrado and the tree symbol of the state of Tocantins, but there are still few studies regarding its bioprospecting. In this study, we aimed to investigate the phytochemical composition, toxicity and bioactivities of the bark and flower of Parkia platycephala. Hot sequential extractions (Soxhlet) were performed using methanol and hydroethanolic solution (70%), after degreasing the sample (hexane). The presence of flavonoids, tannins, steroids and alkaloids was detected in the preliminary screening. Trilinolein, (Z)-9-octadecenamide, 3-O-methyl-d-glucose were detected by Gas Chromatography coupled to Mass Spectrometry (GC-MS). In the Liquid Chromatography with Diode Array Detector (LC-PDA) analysis, it was detected exclusively ferulic acid (bark) and ellagic acid (flower). The ethanolic extract of the bark (IC50=10.69 ± 0.35 µgmL-1) has an antioxidant potential (DPPH• radical) higher than that of the rutin standard (IC50=15.85 ± 0.08 µgmL-1). All extracts showed excellent anticholinesterase potential (Ellman), with emphasis on the ethanol extract of the flower (IC50 =5.34 ± 0.12 µgmL-1). Regarding toxicity (Artemia salina), the methanolic extract of the bark and the ethanolic extract of the flower presented high and moderate levels, respectively. Such results limit the concentrations of biological activities in this study, however, the antioxidant and anticholinesterase indices fall short of toxicity. The results demonstrated promising antioxidant and anticholinesterase activities of both the bark and the flower of Parkia platycephala.

Distinct linear hydrocarbon profiles and chemical strategy of facultative parasitism among Mischocyttarus wasps.

Insects use pheromones as a means of chemical communication. Pheromones act on individual receptors and produce specific behavioral or physiological responses that are fundamental to intra- and interspecific recognition. The objective of this study was to evaluate the interspecific differences among the linear hydrocarbon profiles of the cuticles of 3 wasp species of the genus Mischocyttarus. The chemical strategy that permits an interaction among 2 of these species was also examined about their hydrocarbon profiles. The cuticular hydrocarbons present on the abdomen of each individual were extracted with hexane in an ultrasonic bath and analyzed using gas chromatography with a flame ionization detector. The results suggested that the wasp species have distinct chemical signatures as the linear hydrocarbons of their cuticles. However, these signatures are more alike in those with similar morphological and behavioral aspects. These similarities facilitate an interaction of facultative parasitism observed among these species, in which the invaders acquire parts of the chemical signature of the host colony, possibly as a chemical strategy to increase the likelihood that an invader will be accepted by the females of a host colony. Both invader and host have their own chemical signature changed by the first contact between the species.

Cross-sectional hardness of enamel from human teeth at different posteruptive ages.

This study evaluated, by cross-sectional hardness (CSH), enamel maturation at different depths in sound human teeth at different posteruptive ages (12 per age group): unerupted (molars; control), 2-3 years (premolars), 4-10 years (premolars) and >10 years (molars). Cross-sectional Knoop hardness was measured at 10, 30, 50, 70, 90, 110, 220 and 330 mm depth from the surface. The data were subjected to two-way ANOVA followed by Bonferroni's pairwise tests (a = 0.05). The CSH significantly increased with posteruptive age, indicating that the time of exposure in the oral cavity might result in deep enamel maturation.

The quality of life of patients on the lung transplantation waiting list.

INTRODUCTION: Lung transplantation (LTx) candidates present incapacitating symptoms related to their mobility and activities of daily living, thereby affecting their work, social and emotional relations, and quality of life (QoL). OBJECTIVE: To study the QoL of LTx candidates, seeking to identify domains that suffer the greatest impact and verify if there are differences among these impairments according to the original lung disease. METHODS: We applied the Short Form-36 questionnaires and St George's Respiratory Questionnaire (SGRQ). All data were analyzed by one-way analysis of variance and the Kruskal Wallis test for the probability with significance at P < 0.05. RESULTS: Fifty patients were divided into groups of emphysema (n = 16), bronchiectasis (n = 12), idiopathic pulmonary fibrosis (n = 7), and cystic fibrosis (n = 15). The functional capacity, physical aspects, general status, and vitality domains showed average values below 50 points. The cystic fibrosis group showed higher functional capacity scores (46 +/- 23) than the emphysema (12 +/- 13) or idiopathic pulmonary fibrosis cohort (7 +/- 5). The limitation caused by pain affected the bronchiectasis more than the cystic fibrosis group (52 +/- 28 vs 81 +/- 25, respectively). The SGRQ scores showed impairment among all groups in all domains with average values over 50. The activities domain shows the highest score value; the emphysema (92 +/- 10) and idiopathic pulmonary fibrosis cohorts (91 +/- 9) were extremely affected compared with the cystic fibrosis (69 +/- 21) and bronchiectasis subjects (79 +/- 16). The impact domain show that subjects with cystic fibrosis were less emotionally affected by the disease. CONCLUSION: LTx candidates showed great impairment of their QoL due to their health problems, above all in the physical-functional aspects; the cystic fibrosis patients were the least affected by their health status.

Dental plaque disclosure as an auxiliary method for infants' oral hygiene.

AIM: This was to evaluate the effectiveness of plaque disclosure as an auxiliary method for early childhoods' oral hygiene. METHODS: The study was performed with 20 mothers and their children (aged 6-36 months), members of a preventive programme, which two groups used one of two approaches: conventional oral hygiene/group I (tooth brushing) and plaque disclosure with subsequent oral hygiene/group II (tooth brushing with prior plaque disclosure). Ten mothers started the study in group I and the other 10 in group II, after one month interval they changed to be in the alternate group. Each group consisted of baseline and three additional visits at weekly intervals. The effectiveness of oral hygiene was assessed in a blind fashion by plaque quantity estimation, using the Green Vermilion index for smooth surfaces and the plaque thickness index for occlusal surfaces. Statistical comparisons were performed using repeated measures ANOVA/Fisher's post hoc test and paired t-test (p < 0.05). RESULTS: For smooth and occlusal surfaces at first and second visits, group II recorded significantly lower plaque indices when compared with group I. Additionally, when considering the mean dental plaque index of all visits, group II also presented lower plaque scores than group I. CONCLUSION: Dental plaque disclosure before toothbrushing helps mothers to enhance the effectiveness of early childhood oral hygiene. REGISTRATION NUMBER AND NAME OF TRIAL REGISTRY: RBR-7fyc7g; Avaliação do Treinamento e de Métodos Auxiliares na Efetividade da Escovação Dentária Materna em Bebês. Where the full trial protocol can be accessed: http://www.ensaiosclinicos.gov .

Superconducting and magnetic behaviour of niobium doped RuSr(2)Gd(1.5)Ce(0.5)Cu(2)O(10-δ).

Polycrystalline samples of Ru(1-x)Nb(x)Sr(2)Gd(1.5)Ce(0.5)Cu(2)O(10-δ), 0≤x≤0.5, have been synthesized and structurally characterized by x-ray diffraction (XRD). Resistivity, magnetization and AC susceptibility measurements have been done and analysed considering a phase separation scenario. A strong suppression of the cluster glass (CG) transition associated with niobium doping was identified. In fact, the CG phase was not present in samples for x≥0.2, leading to changes in the magnetic hysteresis loops measured at low temperatures. These hysteresis loops can be explained as a result of the contribution of two distinct magnetic phases: the canted AFM phase and embedded Ru(4+)-rich clusters which order as a CG in low temperatures. Interestingly, the significant changes in the magnetic response of the material do affect the superconducting transition temperature T(c). It was found that both T(c) and the superconducting fraction are reduced in samples which present the spin glass phase. Therefore, our results point to some coupling between magnetism and superconductivity in this ruthenocuprate family, the presence of the magnetic moment being deleterious for the superconductivity.

Effect of xylitol varnishes on remineralization of artificial enamel caries lesions in situ.

OBJECTIVES: Analyze the effect of varnishes containing xylitol compared to commercial fluoridated varnishes on the remineralization of artificial enamel caries lesions in situ. METHODS: Twenty subjects took part in this crossover, double-blind study performed in four phases of 5days each. Each subject worn palatal appliances containing four predemineralized bovine enamel specimens. Artificial caries lesions were produced by immersion in 30ml of lactic acid buffer containing 3mM CaCl2·2H2O, 3mM KH2PO4, 6µM tetraetil metil diphosphanate (pH 5.0) for 6days. The specimens in each subject were treated once with the following varnishes: 20% xylitol (experimental); Duofluorid™ (6% NaF, 6% CaF2), Duraphat™ (5% NaF, positive control) and placebo (no-F/xylitol, negative control). The varnishes were applied in a thin layer and removed after 6h. Fifteen subjects were able to finish all phases. The enamel alterations were quantified by surface hardness and transversal microradiography. The percentage of surface hardness recovery (%SHR), the integrated mineral loss and lesion depth were statistically analyzed by Friedmann and Dunn's tests test (p<0.05). RESULTS: Enamel surface remineralization was significantly increased by Duraphat™, Duofluorid™ and 20% xylitol formulations. Significant subsurface mineral remineralization could also be seen for the experimental and commercial varnishes, except for Duraphat™, when the parameter "lesion depth" was considered. CONCLUSIONS: 20% xylitol varnish seem to be a promising alternative to increase surface and subsurface remineralization of artificial caries lesions in situ. CLINICAL SIGNIFICANCE: effective vehicles are desirable for caries control. Xylitol varnishes seem to be promising alternatives to increase enamel remineralization in situ, which should be confirmed by clinical studies.

Dentifrice pH but not consistency may affect fluoride uptake in plaque.

OBJECTIVE: Test the ability of acidic fluoridated solutions to enhance fluoride (F) bound on bacteria (1) and the effect of dentifrice consistency on plaque fluid F uptake (2). METHODS: (1) Streptococcus mutans isolates were grown in BHI medium (37°C/18h). Bacteria were washed either with EDTA or CaCl2 both at 1mM to remove or add calcium, respectively. Pellets were incubated with 12 mM NaF at pH 4.5 or 7 for 1 min and F was quantified in the lysates and supernatants with the electrode, after HMDS-facilitated diffusion. (2) A randomized, double-blind, crossover clinical trial was performed in three phases with nineteen adults (20-35 years) that used one of the dentifrices: commercial toothpaste (1100 ppm F, pH7.0 and conventional viscosity (Sorriso Fresh(®))); experimental liquid dentifrice (ELD) (1100 ppm F, pH7.0 and low viscosity [1.1% carboxymethylcellulose (CMC)]) and ELD (1100 ppm F and high viscosity pH7.0 (2.2% CMC)). F concentration in plaque fluid was analyzed using an inverted F electrode. RESULTS: (1) Significantly higher F amounts were detected in the lysates of bacteria incubated with NaF solution at pH4.5 compared to the supernatant, which was not seen at pH7.0, being this effect calcium-dependent. (2) Significantly higher F concentrations in plaque fluid were found 1h after toothbrushing compared to 12h, but no significant differences were seen among the toothpastes. CONCLUSIONS: F at low pH binds more efficiently to S. mutans than at neutral pH and dentifrice viscosity does not interfere in plaque fluid fluoride incorporation. CLINICAL SIGNIFICANCE: pH of the dentifrice but not consistency may be important to F uptake in plaque.

The role of matrix metalloproteinases and cysteine-cathepsins on the progression of dentine erosion.

OBJECTIVE: To evaluate in vitro the effect of the inhibition of endogenous dentinal enzymes (matrix metalloproteinases-MMPs and cysteine cathepsins-CCs) on dentine erosion. DESIGN: Dentine blocks (4mm×4mm×2mm) from sound human teeth were randomly divided into 7 groups (n=17) according to the treatment: MMP- and CC-inhibitor chlorhexidine digluconate (CHX, 10mM); MMP-inhibitor galardin (G, 0.2mM); specific cathepsin B inhibitor (CCB, 0.2mM); non-specific CC inhibitor (CCE-64, 0.5µM); fluoride (F, 1.23% NaF); placebo (P) and untreated (UT). Inhibitors were applied as gels once for 1min. Specimens were submitted to 5 days of pH cycling including the erosive challenge (Coke, pH 2.64, 90s/day) and remineralisation (artificial saliva). Demineralised organic surface loss was determined profilometrically. Demineralised organic matrix (DOM) was removed with collagenase and the profile was re-evaluated in the absence of collagen fibrils. The differences in profilometric results and DOM thickness among the groups were analysed with ANOVA and Tukey's test (p<0.05). RESULTS: Loss of demineralised tissue (µm, mean±SD) was: CHX 8.4±1.7 b, G 8.6±1.9 b, CCB 9.6±1.4 a, CCE-64 9.9±1.3 a, F 9.9±1.7 a, P 10.9±2.2 a, UT 11.0±1.5 a. Loss of mineralised tissue was: CHX 15.4±2.2 b, G 16.0±1.8 b, CCB 17.6±2.4 a, CCE-64 17.6±2.0 a, F 17.3±2.8 a, P 19.1±2.1 a, UT 18.9±2.4 a. MMP-inhibitors significantly reduced organic matrix and mineral loss in comparison to all the other groups (p<0.05). No statistically significant differences were found in the thickness of the remaining DOM (p=0.845). CONCLUSION: Dentine endogenous MMPs seem to be the main enzymes responsible for DOM loss and erosion.

In vitro biological screening of the anticholinesterase and antiproliferative activities of medicinal plants belonging to Annonaceae.

The aim of this research was to investigate the antiproliferative and anticholinesterase activities of 11 extracts from 5 Annonaceae species in vitro. Antiproliferative activity was assessed using 10 human cancer cell lines. Thin-layer chromatography and a microplate assay were used to screen the extracts for acetylcholinesterase (AchE) inhibitors using Ellman's reagent. The chemical compositions of the active extracts were investigated using high performance liquid chromatography. Eleven extracts obtained from five Annonaceae plant species were active and were particularly effective against the UA251, NCI-470 lung, HT-29, NCI/ADR, and K-562 cell lines with growth inhibition (GI50) values of 0.04-0.06, 0.02-0.50, 0.01-0.12, 0.10-0.27, and 0.02-0.04 µg/mL, respectively. In addition, the Annona crassiflora and A. coriacea seed extracts were the most active among the tested extracts and the most effective against the tumor cell lines, with GI50 values below 8.90 µg/mL. The A. cacans extract displayed the lowest activity. Based on the microplate assay, the percent AchE inhibition of the extracts ranged from 12 to 52%, and the A. coriacea seed extract resulted in the greatest inhibition (52%). Caffeic acid, sinapic acid, and rutin were present at higher concentrations in the A. crassiflora seed samples. The A. coriacea seeds contained ferulic and sinapic acid. Overall, the results indicated that A. crassiflora and A. coriacea extracts have antiproliferative and anticholinesterase properties, which opens up new possibilities for alternative pharmacotherapy drugs.