Non-mutagenic or genotoxic effects of medicinal aqueous extracts from the Agaricus blazei mushroom in V79 cells

Agaricus blazei Murill is a mushroom largely consumed due to its medicinal properties. Effects of aqueous extract from its lineage AB97/11 in 2 fruiting body development stages (closed and opened pileus) were evaluated on chinese hamster V79 cells using cytokinesis blocking micronucleus (CBMN) and comet assays. The cells were treated at 0.15% concentration of aqueous extract prepared at different temperatures: ice-cold (4°C), room temperature (21°C) and warm (60°C). The extracts were applied in mutagenicity and antimutagenicity protocols (simultaneous, pre-incubation and continuous). The results showed that the aqueous extracts of Agaricus blazei lineage AB97/11 obtained at the 3 temperatures and both development stages did not present mutagenic or antimutagenic effect in V79 cells either in CBMN or comet assay.

Genotoxic and antigenotoxic effects of organic extracts of mushroom Agaricus blazei Murrill on V79 cells

Agaricus blazei Murrill, popularly known as the sun mushroom, is a native mushroom in SP, Brazil, that has been widely used in the treatment of cancer and many other pathologies in different parts of the world. A water-soluble protein-polysaccharide complex (1 → 6)β-D-glucan has been isolated from its fruiting body that showed immune-modulation activity. From organic extracts, linoleic acid has been isolated and determined to be the main substance with antimutagenic activity. Using both the micronucleus (MN) and comet (single cell microgel electrophoresis) assays, this study determined the genotoxic and antigenotoxic potential of A. blazei (AB) obtained from commercial sources or the following strains: a) strains AB 97/29 (young and sporulated phases); b) a mixture taken from AB 96/07, AB 96/09 and AB 97/ 11 strains; and c) commercial mushrooms from Londrina, PR and Piedade, SP, designated as AB PR and AB SP, respectively. The extracts from these mushrooms were isolated in chloroform:methanol (3:1) and used in vitro at three different concentrations. V79 cells (Chinese hamster lung cells) were exposed to the extracts under pre-, simultaneous and post-treatment conditions, combined with methyl methanesulfonate (MMS). Under the circumstances of this study, these organic extracts did not show any genotoxic or mutagenic effects, but did protect cells against the induction of micronuclei by MMS. Copyright by the Brazilian Society of Genetics.

Fluorescence spectroscopy for tongue carcinoma detection: Study in an animal model

Fluorescence diagnosis of malignant lesions has been showed as an attractive optical technique due especially to its real-time response and a more objective and quantitative evaluation. Even though the oral cavity allows a direct examination many lesions are diagnosed when it is already in advanced stage, compromising the patient prognosis. In this study, the fluorescence spectroscopy was used to the detection of chemically induced carcinoma at the lateral border of the tongue in a hamster model. Two excitations wavelengths in visible region were applied: 442 and 532 nm. All the spectra results were analyzed comparing with the histopathological diagnosis. The better results were achieved with the 442 nm laser excitation. The spectra from carcinoma showed new emission bands and these were used to determined different ratios for a quantitative analysis. Using the 625-645 nm fluorescence range under 442 nm excitation (A3 coefficient) the percentage of false negative was of 9.1%, however the false positive percentage was of 18.5%. The 532 nm excitation provided a better normal tissue detection compared to 442 nm excitation. The ideal clinical condition is probably the use of multiple wavelengths excitation for a broader tissue fluorescence investigation.

Benzopyrans from Curvularia sp., an endophytic fungus associated with Ocotea corymbosa (Lauraceae)

An isolate of Curvularia sp. was obtained from the leaves of Ocotea corymbosa, a native plant of the Brazilian Cerrado. The ethyl acetate extract from culture of this fungus afforded two benzopyran derivatives: (2′S)-2-(propan-2′-ol)-5-hydroxy-benzopyran-4-one (2) and 2,3-dihydro-2-methyl-benzopyran-4,5-diol (4); and two known benzopyrans: 2-methyl-5-methoxy-benzopyran-4-one (1) and (2R)-2,3-dihydro-2-methyl-5-methoxy- benzopyran-4-one (3). The structures of 2 and 4 were established on the basis of comprehensive spectroscopic analysis, mainly using 1D and 2D NMR experiments. The benzopyrans 1 and 2 showed weak in vitro antifungal activity against Cladosporium sphaerospermum and C. cladosporioides. Analyses of the biological activities were also carried out on HeLa (human cervix tumor) and CHO (Chinese hamster ovary) cells, aiming to evaluate their potential effects on mammalian cell line proliferation. Results from both cell lines indicated that compound 2 was able to induce cell proliferation: 70% on HeLa cells and 25% on CHO cells. © 2005 Elsevier Ltd. All rights reserved.

Ex vivo biocompatibility tests of regular and white forms of mineral trioxide aggregate

Aim: To examine the genotoxicity and cytotoxicity of regular and white mineral trioxide aggregate (MTA) ex vivo by the single-cell gel (comet) assay and trypan blue exclusion test, respectively. Methodology: Aliquots of 1 × 10 4 Chinese hamster ovary cells were incubated at 37°C for 3 h with grey and white forms of MTA at final concentrations ranging from 1 to 1000 μg mL -1. The negative control group was treated with vehicle control phosphate buffer solution for 3 h at 37°C and the positive control group was treated with methyl metasulfonate (at 1 μg mL -1) for 1 h at 37°C. After incubation, the cells were centrifuged at 180 g for 5 min and washed twice with fresh medium and resuspended with fresh medium. Each individual treatment was repeated three times consecutively to ensure reproducibility. Parameters from single-cell gel (comet) and cytotoxicity assays were assessed by the Kruskal-Wallis nonparametric test. Results: Neither compounds produced genotoxic effects with respect to the single-cell gel (comet) assay in all concentrations evaluated. In the same way, the dose-response relationships of all compounds tested at concentrations ranging from 1 to 1000 μg mL -1 on cell viability assessed by the trypan blue assay displayed no statistically significant differences (P > 0.05) for either endodontic material. Conclusions: Regular (grey) and white MTA are not genotoxins and do not induce cellular death. © 2006 International Endodontic Journal.

Blefarospasmo essencial e espasmo hemifacial: Características dos pacientes, tratamento com toxina botulínica A e revisão da literatura

Purpose: To evaluate the characteristics of the essential blepharospasm and hemifacial spasm patients and the feasible treatment with botulinum toxin. A. Methods: Thirty-four essential blepharospasm or hemifacial spasm patients were evaluated according to gender, ocular complaint, time of disease, treatment outcome and complications. Results: Age median was 63 years and the mean was 61 years, with no difference regarding sex; 66.66% of the patients had hemifacial spasm and 33.33%, essential blepharospasm. Many patients complained of dry eye associated with involuntary spasm. Botulinum toxin A showed a positive outcome in 91.30% of the treated patients and complications observed after treatment were eyelid ptosis (8.33%) and buccal angle deviation (8.33%). Conclusion: Essential blepharospasm and hemifacial spasm occurred in the elderly, of both sexes. Treatment with botulinum toxin A was useful, with very low complication rates.

Interdisciplinary approach to treat dyskeratosis congenita associated with severe aplastic anemia: A case report

This paper reports on a 4-year-old male who had dyskeratosis congenita and who acquired severe aplastic anemia. The patient developed hyperpigmentation of the face, neck and chest region, arms, shoulders and legs. In addition, he had dry skin, deformed fingernails and toenails, sparse hair and eyebrows and hyperkeratosis of the dorsum of the hands and feet. Laboratory and histological analysis revealed severe pancytopenia and dyserythropoiesis of red blood cells, hypocellularity of white blood cells and decreased megakaryocytes with dysplasia. The intraoral examination identified bleeding gums; petechiae of the palate, tongue and cheek mucosa; and an atrophic, smooth and shining dorsal surface of the tongue. There were deep carious lesions in the deciduous mandibular molars and maxillary anterior teeth; as well as mobility of mandibular left canine, which had bone loss. The treatment for oral lesions included diet changes, improved oral hygiene, and extraction of the deciduous teeth destroyed by caries.

Esthetic influence of negative space in the buccal corridor during smiling

The purpose of this study was to measure and verify the esthetic influence of the bilateral spaces between maxillary teeth and lip corners, called negative space (NS), during smile. The sample was comprised of 60 smile photographs obtained from 60 individuals (30 men and 30 women) aged 18 to 25 years old. Two orthodontists and two lay people evaluated these pictures regarding esthetics by a visual analogue scale. In each picture, the right and left NS were measured in millimeters and in proportion to the smile width (SW). Data were analyzed for statistical significance (P = .05). The mean NS of the sample was 6.68 ± 1.99 mm, and the NS proportion in relation to the SW was 9.6 ± 2.56%, for both sides of the arch. No significant asymmetries were observed between the right and left sides. The NS was significantly larger in men than in women when measured in millimeters (P = .028) (7.08 ± 2.24 mm in men vs 6.28 ± 1.62 mm in women), but the NS proportion to the SW was similar (9.94 ± 2.24% in men vs 9.26 ± 1.61% in women). When the 12 individuals with the smallest NS in proportion to SW were compared with the 12 individuals with the largest NS in proportion to SW, there was no statistical difference regarding the esthetic evaluation (P = .11). It was concluded that the NS did not influence the esthetic evaluation of smile photographs in the sample in this study, for both orthodontists and lay people. © 2006 by The EH Angle Education and Research Foundation, Inc.

Study of DNA damage induced by dental bleaching agents in vitro

Dental bleaching is a simple and conservative procedure for aesthetic restoration of vital and non-vital discolored teeth. Nevertheless, a number of studies have demonstrated the risk of tissue damage from the contact of these agents with the oral mucosa. In the current study, the genotoxic potential associated with exposure to dental bleaching agents was assessed by the single cell gel (comet) assay in vitro. Chinese hamster ovary (CHO) cells in vitro were exposed to six commercial dental bleaching agents (Clarigel Gold - Dentsply; Whitespeed - Discus Dental; Nite White - Discus Dental; Magic Bleaching - Vigodent; Whiteness HP - FGM and Lase Peroxide - DMC). The results pointed out that all dental bleaching agents tested contributed to DNA damage as depicted by the mean tail moment, being the strongest effect observed with the highest dose of hydrogen peroxide (Whiteness HP and Lase peroxide, at a 35% concentration). On the other hand, Magic Bleaching (Vigodent) induced the lowest level of DNA breakage. Negative and positive controls displayed absence and presence of DNA-damaging, respectively. Taken together, these results suggest that dental bleaching agents may be a factor that increases the level of DNA damage. A higher concentration of hydrogen peroxide produced higher noxious activities in the genome as detected by single cell gel (comet) assay.

Cytologic analysis of alterations induced by smoking and by alcohol consumption

Objective: To analyze cytologically the buccal mucosa of smoking and nonsmoking volunteers to determine what cellular changes are induced by cigarettes and alcohol consumption. Study Design: In order to evaluate cellular changes induced by smoking and alcohol consumption, exfoliative cytology was used for the analysis of mucosal smears obtained from the buccal mucosa of 25 smokers and 25 nonsmokers. The number of cigarettes consumed, duration of smoking, presence or absence of alcohol ingestion, ingested alcohol dose and frequency of consumption, and most frequently used type of alcoholic beverage were determined using a questionnaire. Three smears from each individual stained by the Papanicolaou method were analyzed quantitatively and qualitatively under a light microscope by 2 experienced examiners in terms of inflammatory and dysplastic alterations and of the degree of epithelial maturation. Results: Although numerous alterations were observed in smokers they corresponded up to only Papanicolaou class II and were not significantly different from nonsmokers (Mann-Whitney and χ 2 tests, p < 0.05). A higher proportion of inflammatory cells (polymorphonuclear and mononuclear cells) were obtained from smokers as compared to nonsmokers, while the proportion of bacteria was similar in the 2 groups. Conclusion: The findings indicate that even after a short period of cigarette use and alcohol consumption, inflammatory alterations were detectable on exfoliative cytology of the buccal mucosa in a young group, demonstrating the usefulness of cytology for early detection in smokers. © The International Academy of Cytology.

Antimicrobial endodontic compounds do not modulate alkylation-induced genotoxicity and oxidative stress in vitro

Objective: To investigate if formocresol, paramonochlorophenol, or calcium hydroxide modulate the genotoxic effects induced by the oxidatively damaging agent hydrogen peroxide (H 2O 2) or the alkylating agent methyl methanesulfonate (MMS) in vitro by using single cell gel (comet) assay. Study design: Chinese hamster ovary (CHO) cells in culture were exposed directly to formocresol, paramonochlorophenol, or calcium hydroxide (adjusted to 100 μg/mL) for 1 hour at 37°C. Subsequently the cultures were incubated with increasing concentrations (0-10 μmol/L) of MMS in phosphate-buffered solution (PBS) for 15 minutes at 37°C or of H 2O 2 at increasing concentrations (0-100 μmol/L) in distilled water for 5 minutes on ice. The negative control cells were treated with PBS for 1 hour at 37°C. The parameter from the comet assay (tail moment) was assessed by the Kruskal-Wallis nonparametric test followed by a post hoc analysis (Dunn test). Results: Clear concentration-related effects were observed for the genotoxin-exposed CHO cells. Increase of MMS-induced DNA damage was not significantly altered by the presence of the compounds tested. Similarly, no significant changes were observed when hydrogen peroxide was used with the endodontic compounds evaluated. Conclusion: Formocresol, paramonochlorophenol, and calcium hydroxide are not able to modulate alkylation-induced genotoxicity or oxidative DNA damage as depicted by the single cell gel (comet) assay. © 2006 Mosby, Inc. All rights reserved.

In vitro metabolism effect on genotoxicity and antigenotoxicity of Agaricus blazei organics and aqueous extracts by the comet assay

There is high interest in the natural products properties due to their use in popular medicine. Agaricus blazei Murrill ss. Heinem. (Ab) is native to Brazil and has been widely disseminated because its medicinal properties. In the present study, the genotoxic and antigenotoxic potential of Ab extracts were investigated using the comet assay. The cells utilized were the non drug-metabolizing line CHO-k1 (Chinese hamster ovary) and the drug-metabolizing line HTC (rat hepatoma). Cells were treated for 3 h in the absence of fetal bovain serum (FBS) with methanolic, hexanic and n-butanolic extracts at 50 μg/ml and 0.75% aqueous extract to test for genotoxicity. Antigenotoxic effects of extracts were determined in cells exposed to the DNA damage inducing agent ethyl methanesulfonate under simultaneous or simultaneous with 1 h pre-incubation conditions. The extracts did not show genotoxicity in HTC, while they were genotoxic in CHO-k1. No antigenotoxic effect was observed with any extract under any condition. These results demonstrate that the metabolism in presence or in absence has a direct influence on the genotoxicity of these extracts. © 2006 The Japan Mendel Society.

External location of the buccinator muscle to facilitate electromyographic analysis

Electromyography is frequently used to measure the activity of masticatory muscles. It requires the precise setting of the electrodes, which demands the accurate location of the muscle to be evaluated. The purpose of this study was to investigate the accuracy of an external method to locate the buccinator muscle. Fifteen human cadavers were evaluated and planes were etermined on the face using anatomic landmarks. An angle (α) was obtained at the intersection of these planes on the central point of buccinator muscle and measured with a protractor. The value of the angle allows locating the central point of buccinator muscle based on anatomic landmarks on the face. Statistical analysis of the collected data indicated an angle of 90° with 95% reliability, thus proving the efficacy of the proposed method.

Effects of mandibular fixed implant-supported prostheses on masticatory and swallowing functions in completely edentulous elderly individuals

Purpose: To evaluate the effect of oral rehabilitation with immediately loaded fixed implant-supported mandibular prostheses on chewing and swallowing in elderly individuals. Materials and Methods: Fifteen completely edentulous patients aged more than 60 years (10 women and rive men), wearing removable dentures in both arches, had a mandibular denture replaced by an implant-supported prosthesis. All individuals were evaluated before surgery and again 3, 6, and 18 months later with regard to mastication and swallowing conditions. Examinations entailed an interview, evaluation of tactile sensitivity of the face, and observation of food intake, masticatory type, formations of bolus, and pain during mastication. The swallowing evaluation comprised observation of clinical signs related to the oral and pharyngeal stages of swallowing, as well as the presence of oral residue. The findings of different evaluations before and 3, 6, and 18 months after the surgical-prosthetic procedure were statistically compared by analysis of variance for repeated measurements at a significance level of 5%. Results: The questionnaire revealed a reduction in complaints of masticatory and swallowing disturbances, a decreased need for liquid ingestion, and reduced choking and coughing. Clinical evaluations showed improved oral function and bolus propulsion for both solid and paste-consistency foods; pain during mastication was also resolved. Conclusion: Treatment with mandibular implant-supported dentures had positive effects on the clinical aspects of mastication and swallowing in elderly individuals.

Topical Calendula officinalis L. successfully treated exfoliative cheilitis: A case report

Authors describe a case of recurrent exfoliative cheilitis that responded to treatment with a standardized topical preparation of Calendula officinalis L. An eighteen-year-old man was referred to UNESP - São Paulo State University, Department of Biosciences and Oral Diagnosis, São José dos Campos Dental School to investigate a chronic dry scaling lesion on his lips. The patient's main chief was aesthetic compromising. Corticoid therapy was suspended and Calendula officinalis ointment 10% for ad libitum use has been prescribed. The results presented allow the authors to consider Calendula officinalis L. as a potential therapy in cases of cheilitis exfoliative. © 2009 Roveroni-Favaretto et al; licensee BioMed Central Ltd.