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.

Faecal contamination (viral and bacteria) detection in groundwater used for drinking purposes in São Paulo, Brazil

This project has been developed to evaluate the possible relationship between the cesspit (pit latrine) in as far as it degrades the quality of underground water. Its importance is due to the fact that in the rural communities in the State of São Paulo (Brazil) this type of cesspit is very common as a means of sewage disposal and these communities use underground water for their supply of drinking water. Rural properties distributed over the rural area in the municipality of São José do Rio Preto were selected. A preliminary study was then set up to determine the social situation and health of the households as well as qualitative evaluations on the type of water supply and sewage disposal of these communities. Campaigns of water sampling then followed and laboratory analyses of water taken from wells were carried out. Parameters were set up to evaluate the potability according to Brazilian legislation (2004) paying attention to microbiologic (coliforms, Crytosporidium sp., and adenovirus). The analyses showed evidence of possible interaction between the wells and the sewage effluents and drainage in these communities. A PCR reaction to detect adenovirus showed a presence in 53.3% of the samples. The tests for the detection of Cryotosporidium sp all showed a negative result.

Use of skin graft punch graft type for the healing of leg ulcers in treated Hansen's disease patients

Hansen's disease is an infectious illness caused by Mycobacterium leprae. It affects preferentially the skin and the peripheral nervous system leading to incapacities, such as leg ulcers, which happens due to the direct action of the bacillus on the organs or its indirect action on the peripheral nervous system. Leg ulcers can occur by two physiopathologic processes. There are many treatments for general leg ulcers, which include the ones caused by Hansen's disease sequels. Among them, surgical treatment shows to be effective when using skin graft, which can be performed by several techniques. Considering the low number of techniques known for treating leg ulcers in Hansen's disease sequels, the aims of this work were to standardize alternative techniques, to detect the main bacteria found in ulcer secretion cultures, to analyze the patients profile and the ulcers, to describe the histophatologies found, and to correlate these data with those of literature from all over the world. Skin graft punch type was carried out and analyzed; males had a mean age of 59.4 years old and females, 54.2 years old. Patients were 73.6% male and 26.3% female. Lepromatous type was present in 89.4% patients and tuberculoid type was seen in 10.5% of them. Associated systemic diseases were observed in 26.3% patients. Mean time of ulcers evolution was 11.6 years in male and 12.8 years in women. The average diameter of ulcers in the pre-treatment period was 8.5 X 9.5 cm in male and 10.2 X 6.8 cm in women. After the graft, their average diameters were 3.2 X 2.7 cm in male and 5.1 X 5.6 cm in women. Statistical analysis showed that there was no significant correlation between the ulcer diameter and its reduction or not in the post-surgery period (p=0.269732). The mean age of patients whose ulcers diameter did not change or reduced by only 20% was 63.5 years. Using the Spearman's coefficient, it was possible to observe that there was no significant correlation between the patients' age and the ulcers diameter reduction after the skin graft (p=0.222531). Evolution time of ulcers that did not present any satisfactory result in the post-surgery period was 12.1 years. The Spearman's coefficient showed that there was no significant correlation between the ulcers evolution time and the ulcers diameter reduction in the post-surgery period (p=0.191655). Cultures presented 50% of cases with Pseudomonas aeruginosa. Statistical analysis showed there is no correlation between the bacterial types found and the ulcer evolution in the post-surgery period (p=0.697531). The average of the ulcers diameter reduction was 42.4%, and in 26.3% of the patients the lesions disappeared after the skin graft.

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.

Heating time required for an MgO-graphite refractory during a hot modulus of rupture test

In the work described in the present paper, an analytical solution of the general heat conduction equation was employed to assay the temperature profile inside a solid slab which is initially at room temperature and is suddenly plunged into a fluid maintained at a high temperature. The results were then extrapolated to a simulation of a hot modulus of rupture test of typical MgO-graphite refractory samples containing different amounts of graphite in order to evaluate how fast the temperature equilibrates inside the test specimens. Calculations indicated that, depending on the graphite content, the time to full temperature homogenization was in the range of 80 to 200 s. These findings are relevant to the high temperature testing of such refractories in oxidizing conditions in view of the graphite oxidation risks in the proper evaluation of the hot mechanical properties.

Production of monoclonal antibodies against Streptococcus mutans antigens

Several studies have been conducted in the last decades aiming to obtain an anti-canies vaccine, however some studies have demonstrated cross reactivity between Streptococcus mutans surface antigens and the human cardiac tissue. In this work, the reactivity of five anti-Streptococcus mutans monoclonal antibodies (MoAb) (24A, 56G, G8, E8 and F6) was tested against oral streptococci, cardiac antigens and skeletal and cardiac myosins, aiming to evaluate the specificity of these MoAb. The hybrid producers of immunoglobulins of the IgG 2b class were cloned by limit dilution and expanded in vivo. MoAb were tested by ELISA. The hybrid 24A reacted with S. mutans CCT 1910, S. salivarius CCT 0365 and S. pyogenes T23. No reactivity difference was observed among the tested species. Cross reactivity with heart and cardiac myosin was not confirmed and only reaction with myosin of skeletal muscle was observed (p = 0.0381). The hybrid 56G reacted with all the tested microorganisms and there was statistically significant difference between S. mutans and S. pyogenes T23 (p < 0.001). This hybrid also reacted with myosin of skeletal muscle (p = 0.0095). C8, E8 and F6 presented low reactivity against oral streptococci strains and no reactivity against cardiac antigens. The data of this study showed that the 24A and 56G anti-S. mutans MoAb presented reactivity with S. pyogenes and S. salivarius, reinforcing the occurrence of common antigens between these species. The tested MoAb presented low cross-reactivity with myosin of skeletal muscle, but anti-heart activity could not be confirmed.

Effect of 10% carbamide peroxide dental bleaching on microhardness of filled and unfilled sealant materials

Purpose: The purpose of this study was to quantitatively evaluate the effect of 10% carbamide peroxide on the microhardness of pit and fissure sealant materials. Methods: Fluroshield, Vitroseal Alfa, and one unfilled (Clinpro) sealants were placed in Teflon matrices (4 mm in diameter by 2 mm in height) and polymerized for 40 seconds. A total of 20 specimens were prepared for each material, in which half were assigned as the control group (stored in artificial saliva and no bleaching treatment). For the remaining half, Clarigel Gold bleaching agent (10% carbamide peroxide) was placed over the specimen surface for 4 hours/day during 4 weeks. When specimens were not under bleaching treatment, they were kept in artificial saliva. Afterwards, specimens were subjected to Knoop microhardness testing using a 25-g load for 5 seconds. Five measurements were made on the sealants' surfaces and then calculated in Knoop hardness values. The data were statistically analyzed by two-way analysis of variance and Tukey's tests with a 5% confidence level. Results: The results of this in vitro study showed that the application of a carbamide peroxide-based bleaching material significantly affected the microhardness values of filled sealant materials. The bleaching agent did not affect the microhardness of the unfilled sealant. CLINICAL SIGNIFICANCE: The results of this in vitro study suggest that the bleaching agents altered the surface hardness of filled sealant restorative materials. This could possibly lead to increased wear and surface roughness. © 2006, Copyright the authors.

Importância de Yersinia enterocolitica em microbiologia médica

Y. enterocolitica is a human invasive enteropathogen which causes a number of intestinal and extraintestinal clinical symptoms of various degrees of severity, ranging from mild gastroenteritis to mesenteric lymphadenitis, which mimics appendicitis and in rare cases can evolve to septicemia. Infection by Y. enterocolitica can also lead to post-infection immunological sequelae including arthritis, erythema nodosum and glomerulonephritis. Pathogenic Y. enterocolitica strains have traditionally been linked to specific biotypes and serogroups and associated to a variety of phenotypic characteristics related to virulence. Molecular genetics studies have pointed to the importance of the pYV virulence plasmid, which encodes various virulence genes, as well that of specific chromosomal virulence genes, in determining the pathogenesis of this bacterium. Intestinal infections by Y. enterocolitica are mostly self-limiting and usually do not need an antibiotic treatment. The occurrence of this microorganism is not as frequently described in Brazil as it is in other countries, such as Japan, USA and many European countries. This review focuses on the general characteristics, pathogenesis, clinical symptoms, virulence characteristics, treatment and antibiotic susceptibility of Yersinia enterocolitica strains isolated in Brazil and around the world.

Rapid molecular diagnosis of tuberculosis and other mycobacteriosis in smear-negative clinical specimens

Delay in diagnosis of pulmonary and other forms of tuberculosis (TB) can be fatal, particularly in HIV-infected patients. Hence, techniques based on nucleic acid amplification, which are both rapid and of high specificity and sensitivity, are now widely used and recommended for laboratories that diagnose TB. In the present study, diagnostic methods based on mycobacterial DNA amplification were evaluated in comparative trials alongside tradicional bacterial methods, using negative smear samples from patients with clinically-suspected TB (sputum samples from 25 patients with suspected pulmonary TB, urine samples from two patients with suspected renal TB and cerebrospinal fluid samples from one patient with suspected meningeal TB). A specificity of 100% was achieved with DNA amplification methods and tradicional culture/identification methods, in relation to clinical findings and treatment results. For the smear-negative sputa, conventional PCR for M. tuberculosis was positive in 62% of suspected lung TB case, showing the same sensitivity as bacterial identification. Both techniques failed in the detection of extra-pulmonary samples. Nested PCR showed, after species-specific amplification, a sensitivity of 100% for M. avium and 85% for M. tuberculosis. For extra-pulmonary smear-negative samples, only Nested PCR detected M. tuberculosis and all cases were confirmed clinically. Nested PCR, in which two-step amplification reactions are performed, can identify the two most important mycobacteria in human pathology quickly and directly from clinical spicimens.