In vitro evaluation of the antimicrobial activity of a castor oil-based irrigant

The antimicrobial activity of irrigating solutions - Endoquil (castor oil detergent), 2% chlorhexidine gluconate solution, and 0.5% NaOCI solution - was evaluated against Gram-positive cocci (Micrococcus luteus, Staphylococcus aureus, Enterococcus faecalis, Staphylococcus epidermidis, Streptococcus mutans, and Streptococcus sobrinus), Gramnegative rods (Escherichia coli and Pseudomonas aeruginosa), and the yeast Candida albicans. Activity was evaluated using the two-layer agar diffusion technique. The base layer was obtained by pouring 10.0 ml of Muller Hinton Medium or 10.0 ml of Brain Heart Infusion agar in a Petri dish. After solidification a 5.0 ml seed layer of Muller Hinton Medium or Brain Heart Infusion agar with inoculum (106/ml) was added. Absorbent paper disks (6.0 mm in diameter) immersed in the solutions were placed at equidistant points. Plates were maintained at room temperature for 2 h for prediffusion of the solutions and incubated at 37°C for 24 h. The candle jar system was used for the Brain Heart Infusion agar plates. All tests were performed in duplicate. After incubation the medium was optimized with 0.05 g% triphenyltetrazolium chlorate gel and inhibition halos were measured. All bacterial strains were inhibited by 2.0% chlorhexidine gluconate. Endoquil was effective against Grampositive microorganisms, and 0.5% NaOCI was effective only against S. aureus. Copyright © 2001 by The American Association of Endodontists.

IL-12 and neutralization of endogenous IL-10 revert the in vitro antigen-specific cellular immunosuppression of paracoccidioidomycosis patients

Treatment of patients with paracoccidioidomycosis is still a challenge. Patients present defective lymphoproliferation and IFN-γ responses to the main Paracoccidioides brasiliensis antigen (gp43), which correlates with disease severity. Here, we demonstrated that the patients show also a defective synthesis of interleukin (IL)-12. Therefore, we attempted to revert this immune disfunction by adding IL-12 and neutralizing anti-IL-10 antibody to gp-43-stimulated peripheral blood mononuclear cell cultures. Both treatments increased IFN-γ secretion to levels observed with healthy sensitized individuals, but affected proliferation only modestly. When combined, the treatments further increased IFN-γ synthesis and cell proliferation. The addition of suboptimal concentrations of IL-2 also further increased the IL-12-mediated secretion of IFN-γ. Interestingly, the immune modulation was mostly antigen-specific, since the responses to Candida albicans' antigen were not affected. These results suggest that appropriate immune intervention with cytokines and/or anti-cytokines may help in the treatment of PCM. © 2002 Elsevier Science Ltd. All rights reserved.

Effectiveness of Microwave Sterilization on Three Hard Chairside Reline Resins

Purpose: The aim of this study was to evaluate the effectiveness of microwave irradiation sterilization on hard chairside reline resins. Materials and Methods: Specimens of three reline resins (Kooliner, Tokuso Rebase, and Ufi Gel Hard) were fabricated and subjected to ethylene oxide sterilization. The specimens were then individually inoculated (107 cfu/mL) with Tryptic Soy Broth media containing one of the tested microorganisms (C albicans, S aureus, B subtilis, and P aeruginosa). After 48 hours at 37°C, the samples were vortexed for 1 minute and allowed to stand for 9 minutes, followed by a short vortex to resuspend any organisms present. After inoculation, 40 specimens of each material were immersed in 200 mL of water and subjected to microwave irradiation at 650 W for 6 minutes. Forty non-irradiated specimens were used as positive controls. Replicate specimens (25 μL) of suspension were plated at dilutions of 10-3 to 10-6 on plates of selective media appropriate for each organism. All plates were incubated at 37°C for 48 hours. After incubation, colonies were counted, and the data were statistically analyzed by the Kruskal-Wallis test. Twelve specimens of each material were prepared for SEM. Results: All immersed specimens showed consistent sterilization of all the individual organisms after microwave irradiation. SEM examination indicated an alteration in cell morphology after microwave irradiation. Conclusion: Microwave sterilization for 6 minutes at 650 W proved to be effective for the sterilization of hard chairside reline resins.

Epidemiologia de candidíase hospitalar: Importância da identificação específica

Nosocomial infections with Candida species are recognized as a significant cause of morbidity and mortality in both seriously ill immunocompetent and immunocompromised patients. Infections with Candida albicans and non-albicans Candida species have become a significant cause of infection in humans. Several of the more commonly Candida spp isolates are less susceptible to the antifungal drugs currentlly applied in clinical treatment, a factor that means significant difficulties for effective treatment. The modern mycology laboratory has an important role to play in several aspects relating to these organisms, including therapy, detection, identification and epidemiological analysis. In this study, we have provided an initial comparison of differences in species distribution among Candida isolates from four general hospitals of São Paulo,SP. Overall, 40 isolates of C. albicans, C. parapsilosis and C. tropicalis were isolated respectively in 35%, 50% and 15%, revealed a tendency of higher frequency of non-albicans species. The species distribution in patients with candidemia showed that the most commonly species were C. parapsilosis (45,4%), followed by C. albicans (36,4%) and C. tropicalis (18,2%); thus, we have an increase of non-albicans species. The three different species were include in 6, 3, and 4 different biotypes, respectively C. albicans, C. parapsilosis e C. tropicalis. This study emphasizes the importance of periodic evaluation of Candida species distribution especially in centers caring for patients at risk.

In vitro antifungal susceptibility of Candida spp. isolates from patients with chronic periodontitis and from control patients

Superinfection by Candida can be refractory to conventional periodontal treatments in specific situations, such as in immunocompromised patients. In these cases, the systemic therapy with antifungal drugs could be indicated. The aim of this study was to analyse antifungal susceptibility of Candida spp. strains isolated from chronic periodontitis patients and from control individuals. A total of 39 C. albicans isolates, 9 C. tropicalis, 2 C. glabrata and 5 Candida spp. from control individuals and 30 C. albicans, 3 C. tropicalis and 2 C. glabrata from periodontitis patients were tested. In the control group, 1 isolate of C. glabrata was resistant to ketoconazole and 1 Candida spp. was resistant to amphotericin B, ketoconazole and miconazole. Among the isolates of periodontitis group, 1 (3.33%) C. albicans isolate was resistant to flucytosine and ketoconazole. According to the obtained results, it could be concluded that fluconazole was the most effective drug against the several Candida species studied. There were not expressive differences in the susceptibility of isolates from periodontitis patients or from control individuals.

rDNA-RFLP identification of Candida species in immunocompromised and seriously diseased patients

PCR was used to amplify a targeted region of the ribosomal DNA of 76 Candida spp. isolates from immunocompromised and seriously diseased patients. Thirty-seven strains isolated from different anatomical sites of 11 patients infected with HIV (Vitória, ES, Brazil), 26 isolates from patients under treatment at Odilon Behrens Hospital and 13 isolates from skin and urine samples from São Marcos Clinical Analysis Laboratory (Belo Horizonte, Brazil) were scored. Fragments of rDNA were amplified using primer pairs ITS1-ITS4, for the amplification of ITS1 and ITS2 regions, including the gene for the 5.8s subunit. Amplification resulted in fragments ranging in size from 350 to 950 bp. Amplicons were digested with eight restriction enzymes. A pattern of species-specificity among the different medically important Candida species could be identified following restriction digestion of the PCR products. Candida albicans was the species most frequently observed, except for the group of newborns under treatment at the Odilon Behrens Hospital and for the isolates from the clinical analysis laboratory. C. parapsilosis was the species most frequently observed in these two groups.

Prevalence of yeasts in the oral cavity of children treated with inhaled corticosteroids

The aim of this study was to observe the prevalence of Candida spp. in the oral cavity of children undergoing treatment with inhaled corticosteroids. Thirty children treated with inhaled corticosteroids and thirty control children were studied. Saliva samples were collected through oral rinses with phosphate buffered saline (PBS). The samples were plated on Sabouraud's dextrose agar and incubated at 37 degrees C for 48 h. After this period, the number of colony-forming units per ml (cfu/ml) of saliva was calculated. The isolates were identified by phenotypic characterization. Candida spp. was isolated from 43.33% of the samples of children treated with corticosteroids, with a mean of 780 cfu/ml of saliva, and from 30% of the samples of the control group, with a mean of 560 cfu/ml of saliva. No significant statistical difference was observed between the groups. C. albicans was the prevalent species in both groups, followed by C. guilliermondii, C. parapsilosis and C. stellatoidea. Furthermore, Rhodotorula rubra and C. lusitaniae were also isolated from the treated group. We concluded that there was no significant increase in the prevalence and number of Candida spp. in the oral cavity of children treated with inhaled corticosteroids.

Communiols E-H: New polyketide metabolites from the coprophilous fungus Podospora communis

Communiols E-H (1-4), four new polyketide-derived natural products containing furanocyclopentane, furanocyclopentene, cyclopentene, or γ-lactone moieties, have been isolated from two geographically distinct isolates of the coprophilous fungus Podospora communis. The structures of these compounds were determined by analysis of NMR and MS data. © 2005 American Chemical Society and American Society of Pharmacognosy.

Oral manifestations during chemotherapy for acute lymphoblastic leukemia: A case report

A 14-year-old, male patient was referred for the treatment of mucositis, idiopathic facial asymmetry, and candidiasis. The patient had been undergoing chemotherapy for 5 years for acute lymphoblastic leukemia. He presented with a swollen face, fever, and generalized symptomatology in the mouth with burning. On physical examination, general signs of poor health, paleness, malnutrition, and jaundice were observed. The extraoral clinical examination showed edema on the right side of the face and cutaneous erythema. On intraoral clinical examination, generalized ulcers with extensive necrosis on the hard palate mucosa were observed, extending to the posterior region. Both free and attached gingivae were ulcerated and edematous with exudation and spontaneous bleeding, mainly in the superior and inferior anterior teeth region. The tongue had no papillae and was coated, due to poor oral hygiene. The patient also presented with carious white lesions and enamel hypoplasia, mouth opening limitation, and foul odor. After exfoliative cytology of the affected areas, the diagnosis was mixed infection by Candida albicans and bacteria. Recommended treatment was antibiotics and antifungal administration, periodontal prophylaxis, topical application of fluor 1.23%, and orientation on and control of proper oral hygiene and diet during the remission phase of the disease.

2,3,4,6-Tetra-O-(3-nitropropanoyl)-O-β-D-glucopyranoside, a new antimicrobial from the roots of Heteropteris aphrodisiaca

A new 2,3,4,6-tetra-O-(3-nitropropanoyl)-O-β-D-glucopyranoside anomer was isolated from the roots of Heteropteris aphrodisiaca and characterized by spectroscopic methods. Activity against Staphylococcus aureus, Bacillus subtilis, Candida albicans, C. parapsilosis, C. krusei, and C. tropicalis was demonstrated.

Antibiótico antifungico produzido por um estreptomiceto da região de Araraquara

With the significant increase in the incidence of invasive fungal infections during the last decade, mainly in patients with cancer, AIDS and other hospitalized patients who stay for long periods in intensive care units, there is an urgent need to screen for new antifungal agents possessing some advantages over known ones. This article reports a search in the field for a microorganism producing antibacterial and antifungal substances. Strains from soil samples collected in the region of Araraquara, Brazil, were isolated and analyzed for their antimicrobial potential against standard microorganisms (fungi Candida albicans and Aspergillus oryzae and bacteria Staphylococcus aureus and Escherichia coli). Out of the 64 strains isolated, 34 produced detectable antimicrobial activity. The streptomycete strain Ar4014 was chosen for further study, owing to its good antimicrobial activity against Candida albicans. Two of the fermentation media tested, 608-K and 602-B, were found to be best for the production and extraction of the antibiotic from Ar4014. After chromatographic separation of the crude extract on a silica column, the active fractions obtained showed UV-VIS absorption peaks characteristic of normal pentaenic antibiotics. The antibiotic was provisionally designated Ara 4014-75.

Effectiveness of microwave irradiation on the disinfection of complete dentures

Purpose: The purpose of this study was to evaluate the effectiveness of microwave irradiation on the disinfection of simulated complete dentures. Materials and Methods: Eighty dentures were fabricated in a standardized procedure and subjected to ethylene oxide sterilization. The dentures were individually inoculated (10 7 cfu/mL) with tryptic soy broth (TSB) media containing one of the tested microorganisms (Candida albicans, Streptoccus aureus, Bacillus subtilis, and Pseudomonas aeruginosa). After 48 hours of incubation at 37°C, 40 dentures were individually immersed in 200 mL of water and submitted to microwave irradiation at 650 W for 6 minutes. Forty nonirradiated dentures were used as positive controls. Replicate aliquots (25 μL) of suspensions were plated at dilutions of 10 -3 to 10 -6 on plates of selective media appropriate for each organism. All plates were incubated at 37°C for 48 hours. TSB beakers with the microwaved dentures were incubated at 37°C for 7 more days. After incubation, the number of colony-forming units was counted and the data were statistically analyzed by Kruskal-Wallis test (α = .05). Results: No evidence of growth was observed at 48 hours for S aureus, B subtilis, and C albicans. Dentures contaminated with P aeruginosa showed small growth on 2 plates. After 7 days incubation at 37°C, no growth was visible in the TSB beakers of S aureus and C albicans. Turbidity was observed in 3 broth beakers, 2 from P aeruginosa and 1 from B subtilis. Conclusion: Microwave irradiation for 6 minutes at 650 W produced sterilization of complete dentures contaminated with S aureus and C albicans and disinfection of those contaminated with P aeruginosa and B subtilis.

Effect of the association of nystatin with a tissue conditioner on its ultimate tensile strength

Purpose: This study evaluated the ultimate tensile strength of a tissue conditioner without nystatin incorporation (GI - control group) and the same tissue conditioner modified by the addition of nystatin in two concentrations: GII - 500,000 International Units (U) and GIII - 1,000,000 U, in which each milligram of the medicament corresponded to 6079 U. Materials and Methods: Dumbbell-shaped specimens (N = 7) with a central cross-sectional area of 33 × 6 × 3 mm were produced for the three experimental groups. After polymerization following manufacturer's instructions, specimens were immersed in distilled water at 37°C for either 24 hours or 7 days and then tested in tension in the MTS 810 at 40 mm/minute. Data were analyzed by two-way ANOVA followed by Tukey's test, at 95% level of confidence. Results: The means (force-grams (gf) ± standard deviation) of the ultimate tensile strength were: GI - 634.29 ± 122.80; GII - 561.92 ± 133.56; and GIII - 547.30 ± 73.47 for 24-hour storage, and GI - 536.68 ± 54.71; GII - 467.50 ± 143.51; and GIII - 500.62 ± 159.76 for 7-day storage. There were no statistically significant differences among the three experimental groups (p > 0.05). The ultimate tensile strength means of all experimental groups after 7 days were significantly lower than those observed after 24 hours (p = 0.04). Conclusions: The results of this study suggest that the addition of nystatin into the tissue conditioner investigated in concentrations below 1,000,000 U did not affect its ultimate tensile strength. Copyright © 2006 by The American College of Prosthodontists.

Antimicrobial activity of Uncaria tomentosa against oral human pathogens

Uncaria tomentosa is considered a medicinal plant used over centuries by the peruvian population as an alternative treatment for several diseases. Many microorganisms usually inhabit the human oral cavity and under certain conditions can become etiologic agents of diseases. The aim of the present study was to evaluate the antimicrobial activity of different concentrations of Uncaria tomentosa on different strains of microorganisms isolated from the human oral cavity. Micropulverized Uncaria tomentosa was tested in vitro to determine the minimum inhibitory concentration (MIC) on selected microbial strains. The tested strains were oral clinical isolates of Streptococcus mutans, Staphylococcus spp., Candida albicans, Enterobacteriaceae and Pseudomonas aeruginosa. The tested concentrations of Uncaria tomentosa ranged from 0.25-5% in Müeller-Hinton agat. Three percent Uncaria tomentosa inhibited 8% of Enterobacteriaceae isolates, 52% of S. mutans and 96% of Staphylococcus spp. The tested concentrations did not present inhibitory effect on P. aeruginosa and C. albicans. It could be concluded that micropulverized Uncaria tomentosa presented antimicrobial activity on Enterobacteriaceae, S. mutans and Staphylococcus spp. isolates.

In vitro antimicrobial activity of endodontic sealers, MTA-based cements and Portland cement.

The aim of this study was to evaluate the antimicrobial activity of different root-end filling materials - Sealer 26, Sealapex with zinc oxide, zinc oxide and eugenol, white and gray Portland cement, white and gray MTA-Angelus, and gray Pro Root MTA - against six different microorganism strains. The agar diffusion method was used. A base layer was made using Müller-Hinton agar (MH) and wells were formed by removing the agar. The materials were placed in the wells immediately after manipulation. The microorganisms used were: Micrococcus luteus (ATCC9341), Staphylococcus aureus (ATCC6538), Escherichia coli (ATCC10538), Pseudomonas aeruginosa (ATCC27853), Candida albicans (ATCC 10231), and Enterococcus faecalis (ATCC 10541). The plates were kept at room temperature for 2 h for prediffusion and then incubated at 37 degrees C for 24 h. Triphenyltetrazolium chloride 0.05% gel was added for optimization, and the zones of inhibition were measured. Data were subjected to the Kruskal-Wallis and Dunn tests at a 5% significance level. The results showed that all materials had antimicrobial activity against all the tested strains. Analysis of the efficacy of the materials against the microbial strains showed that Sealapex with zinc oxide, zinc oxide and eugenol and Sealer 26 created larger inhibition halos than the MTA-based and Portland cements (P < 0.05). On the basis of the methodology used, it may be concluded that all endodontic sealers, MTA-based and Portland cements evaluated in this study possess antimicrobial activity, particularly the endodontic sealers.