Antimicrobial and mechanical properties of acrylic resins with incorporated silver-zinc zeolite - part I

Objective: The purpose of this in vitro study was to evaluate the antimicrobial activity of acrylic resins containing different percentages of silver and zinc zeolite, and to assess whether the addition of zeolite alters the flexural and impact strength of the resins. Background: The characteristics of acrylic resins support microorganism development that can threaten the health of the dentures user. Material and methods: A microwave-polymerised (Onda-Cryl) and two heat-polymerised (QC20 and Lucitone 550) acrylic resins were used. The materials were handled according to the manufacturers` instructions. Fifty rectangular-shaped specimens (8 x 10 x 4mm) were fabricated from each resin and assigned to 5 groups (n = 10) according to their percentage of Irgaguard B5000 silver-zinc zeolite (0%- control, 2.5%, 5.0%, 7.5% and 10%). Flexural strength and Izod impact strength were evaluated. The antimicrobial activity against two strains of Candida albicans and two strains of Streptococcus mutans was assessed by agar diffusion method. Data were analysed statistically by one-way ANOVA and Tukey`s test at 5% significance level. Results: The addition of 2.5% of Irgaguard B5000 to the materials resulted in antimicrobial activity against all strains. Flexural strength decreased significantly with the addition of 2.5% (QC20 and Lucitone 550) and 5.0% (Onda-Cryl) of Irgaguard B5000. The impact strength decreased significantly with the addition of 2.5% (Lucitone 550) and 5.0% (QC20 and Onda-Cryl) of zeolite. Conclusion: The addition of silver-zinc zeolite to acrylic resins yields antimicrobial activity, but may affect negatively the mechanical properties, depending on the percentage of zeolite.

Microbial culture and checkerboard DNA-DNA hybridization assessment of bacteria in root canals of primary teeth pre- and post-endodontic therapy with a calcium hydroxide/chlorhexidine paste

Aim. To investigate the root canal microbiota of primary teeth with apical periodontitis and the in vivo antimicrobial effects of a calcium hydroxide/chlorhexidine paste used as root canal dressing. Design. Baseline samples were collected from 30 root canals of primary teeth with apical periodontitis. Then, the root canals were filled with a calcium hydroxide paste containing 1% chlorhexidine for 14 days and the second bacteriologic samples were taken prior to root canal filling. Samples were submitted to microbiologic culture procedure to detect root canal bacteria and processed for checkerboard DNA-DNA hybridization. Results. Baseline microbial culture revealed high prevalence and cfu number of anaerobic, black-pigmented bacteroides, Streptococcus, and aerobic microorganisms. Following root canal dressing, the overall number of cfu was dramatically diminished compared to initial contamination (P < 0.05), although prevalence did not change (P > 0.05). Of 35 probes used for checkerboard DNA-DNA hybridization, 31 (88.57%) were present at baseline, and following root canal dressing, the number of positive probes reduced to 13 (37.14%). Similarly, the number of bacterial cells diminished folowing application of calcium hydroxide/chlorhexidine root canal dressing (P = 0.006). Conclusion. Apical periodontitis is caused by a polymicrobial infection, and a calcium hydroxide/chlorhexidine paste is effective in reducing the number of bacteria inside root canals when applied as a root canal dressing.

Use of the checkerboard DNA-DNA hybridisation technique for in vivo detection of cariogenic microorganisms on metallic brackets, with or without use of an antimicrobial agent

Objective: Using checkerboard DNA-DNA hybridisation (CDDH) assay, this randomised clinical study evaluated the contamination of metallic brackets by four cariogenic bacterial strains (Streptococcus mutans, Streptococcus sobrinus, Lactobacillus casei and Lactobacillus acidophilus) and the efficacy of 0.12% chlorhexidine gluconate (CHX) mouthwashes in reducing bacterial contamination. Methods: Thirty-nine 11-33-year-old patients under treatment with fixed orthodontic appliances were enrolled in the study and had 2 new metallic brackets bonded to premolars. Nineteen patients used a 0.12% CHX mouthwash (Periogard (R)) and 20 patients used a placebo mouthwash (control) twice a week. After 30 days, the brackets were removed and samples were obtained for analysis by CDDH. Data were analysed statistically by the Kruskal-Wallis test (alpha = 0.05) using the SAS software. Results: S. mutans, S. sobrinus, L. casei and L. acidophilus were detected in 100% of the samples from both groups. However, brackets of the control group were more heavily contaminated by S. mutans and S. sobrinus (P < 0.01). In the experimental group, although all counts decreased after rinsing with the chlorhexidine solution, there was significant difference only for S. mutans (P = 0.03). Conclusions: The use of 0.12% chlorhexidine gluconate mouthwashes can be useful in clinical practice to reduce the levels of cariogenic microorganisms in patients under treatment with fixed orthodontic appliances. (C) 2011 Elsevier Ltd. All rights reserved.

Efficacy of Microwaves and Chlorhexidine on the Disinfection of Pacifiers and Toothbrushes: An In Vitro Study

Purpose: The purpose of this study was to evaluate, in vitro, the contamination of toothbrushes and pacifiers by Streptococcus mutans, and the efficacy of microwave and chlorhexidine for their disinfection. Methods: Sixty pacifiers and 60 toothbrushes were contaminated with S mutans and then divided into groups according to the disinfection protocol: Group 1-chlorhexidine solution; Group 2-microwave sterilization; and Group 3-sterile tap water. The devices were evaluated microbiologically as to the formation of S mutans colonies/biofilms and were examined by scanning electron microscopy. The results were submitted for statistical analysis by Friedman`s test at a 5% significance level. Results: The results of both types of evaluation showed a large number of S mutans colonies/biofilms after spraying with sterile tap water, and chlorhexidine spraying and microwaving were effective in eliminate colonies/biofilms. Groups 1 and 2 were statistically similar to each other (P>.05) and differed significantly from Group 3 (P<.05). Conclusions: The 0.12% chlorhexidine solution spray and 7 minutes of microwave irradiation were effective for disinfection of pacifiers and toothbrushes. (Pediatr Dent 2011;33:10-3) Received July 29, 2009 I Last Revision January 26, 2010 I Accepted March 10, 2010

Morphological differentiation between S. mutans and S. sobrinus on modified SB-20 culture medium

Due to the major role of Streptococcus mutans and Streptococcus sobrinus in the etiology of dental caries, it is important to use culture media that allow for differentiating these bacterial species. The aim of this study was to evaluate the suitability of a modified SB-20 culture medium (SB-20M) for the isolation and morphological differentiation of S. mutans and S. sobrinus, compared to biochemical identification (biotyping). Saliva samples were collected using the spatula method from 145 children, seeded on plates containing the SB-20M, in which sucrose was replaced by coarse granular cane sugar, and incubated in microaerophilia at 37 degrees C during 72 h. Identification of the microorganisms was performed under stereomicroscopy based on colony morphology of 4904 colonies. The morphological identification was examined by biochemical tests of 94 randomly selected colonies with the macroscopic characteristic of S. mutans and S. sobrinus using sugar fermentation, resistance to bacitracin and production of hydrogen peroxide. There was no statistically significant difference (p> 0.05) between morphological identification in the SB-20M medium and biochemical identification (biotyping). Biotyping confirmed that S. mutans and S. sobrinus colonies were correctly characterized in the SB-20M in 95.8% and 95.5% of the cases, respectively. Of the mutans streptococci detected in the children 98% were S. mutans and 2% S. sobrinus. The SB-20M medium is reliable for detection and direct morphological identification of S. mutans and S. sobrinus. (C) 2010 Elsevier GmbH. All rights reserved.

Effect of Three Methods for Cleaning Dentures on Biofilms Formed In Vitro on Acrylic Resin

Purpose: The aim of this study was to evaluate the effect of three denture hygiene methods against different microbial biofilms formed on acrylic resin specimens. Materials and methods: The set (sterile stainless steel basket and specimens) was contaminated (37 degrees C for 48 hours) by a microbial inoculum with 106 colony-forming units (CFU)/ml (standard strains: Staphylococcus aureus, Streptococcus mutans, Escherichia coli, Candida albicans, Pseudomonas aeruginosa, and Enterococcus faecalis; field strains: S. mutans, C. albicans, C. glabrata, and C. tropicalis). After inoculation, specimens were cleansed by the following methods: (1) chemical: immersion in an alkaline peroxide solution (Bonyplus tablets) for 5 minutes; (2) mechanical: brushing with a dentifrice for removable prostheses (Dentu Creme) for 20 seconds; and (3) a combination of chemical and mechanical methods. Specimens were applied onto a Petri plate with appropriate culture medium for 10 minutes. Afterward, the specimens were removed and the plates incubated at 37 degrees C for 48 hours. Results: Chemical, mechanical, and combination methods showed no significant difference in the reduction of CFU for S. aureus, S. mutans (ATCC and field strain), and P. aeruginosa. Mechanical and combination methods were similar and more effective than the chemical method for E. faecalis, C. albicans (ATCC and field strain), and C. glabrata. The combination method was better than the chemical method for E. coli and C. tropicalis, and the mechanical method showed intermediate results. Conclusion: The three denture hygiene methods showed different effects depending on the type of microbial biofilms formed on acrylic base resin specimens.

Radiographic Findings Among Children Hospitalized With Severe Community-Acquired Pneumonia

Background: Community-acquired pneumonia (CAP) is a leading cause of childhood death. There are few published reports of radiographic findings among children with severe CAP. Objective: To describe chest X-ray (CXR) findings and assess association between these radiographic findings and pneumococcal isolation in children with severe CAP. Methods: A prospective, multicenter, observational study was conducted in 12 centers in Argentina, Brazil, and the Dominican Republic. Children aged 3-59 months, hospitalized with severe pneumonia, were included. On admission, blood and pleural effusion cultures were performed. Streptococcus pneumoniae was identified according to standard procedures in the respective national reference laboratory. Chest X-rays were taken on admission and read before the culture results were reported. Results: Out of 2,536 enrolled patients, 283 (11.2%) had S. pneumoniae isolated, in 181 cases (7.1%) from blood. The follow radiographic patterns were observed: alveolar infiltrate (75.2%), pleural effusion (15.6%), and interstitial infiltrate (9.2%). Overall, pleural effusion was associated with pneumococcal isolation and pneumococcal bacteremia (P < 0.001). Infiltrates were unilateral (78.7%) or bilateral (21.3%), right-sided (76%) or left-sided (24%), in the lower lobe (53.6%) or the upper lobe (46.4%). Multivariate analysis including patients with affection of only one lobe showed that upper lobe affection and pleural effusion were associated with pneumococcal isolation (OR 1.8, 95% CI, 1.3-2.7; OR 11.0, 95% CI, 4.6-26.8, respectively) and with pneumococcal bacteremia (OR 1.7, 95% CI, 1.2-2.6; OR 3.1, 95% CI, 1.2-8.0, respectively). Conclusions: Three-quarters of the patients studied had alveolar infiltrates. Upper lobe compromising and pleural effusion were associated with pneumococcal invasive disease. Pediatr Pulmonol. 2010; 45:1009-1013. (C) 2010 Wiley-Liss, Inc.

The role of respiratory viral infections among children hospitalized for community-acquired pneumonia in a developing country

We report an investigation for 16 bacteria and viruses among 184 children hospitalized with pneumonia in Salvador, Brazil. Etiology was established in 144 (78%) cases. Viral, bacterial, and mixed infections were found in 110 (60%), 77 (42%), and 52 (28%) patients, respectively. Rhinovirus (21%) and Streptococcus pneumoniae (21%) were the most common pathogens. Our results demonstrate the importance of viral and pneumococcal infections among those patients.

A new type of Brazilian propolis: Prenylated benzophenones in propolis from Amazon and effects against cariogenic bacteria

Ethanol extracts of four propolis samples (E1-E4) from Manaus (Brazilian Amazon) were analysed by HPLC/DAD/ESI-MS/MS and GC/EIMS. The major constituents of E2 and E4 were analysed by NMR ((1)H and (13)C) and ESI/MS/MS. The main constituents of E2 and E4 are polyprenylated benzophenones: 7-epi-nemorosone, 7-epi-clusianone (major E4 constituents), xanthochymol and gambogenone (major E2 constituents), making up a chemical profile so far unreported for Brazilian propolis. Aristhophenone, methyl insigninone, 18-ethyloxy-17-hydroxy-17,18-dihydroscrobiculatone B, and derivatives of dimethyl weddellianone A and B, propolones, and a scrobiculatone derivative, were detected as minor constituents. Triterpenoids (beta-amyrins, beta-amyrenone, lupeol and lupenone) were ubiquitous and predominant in El and E3. The extracts E2 and E4 were highly active against the cariogenic bacteria Streptococcus mitis, Streptococcus mutans and Streptococcus salivarius. E2 was more active than E4, probably due to a higher content of 2-epi-nemorosone, while the latter was richer in di-hydroxylated compounds. (C) 2010 Elsevier Ltd. All rights reserved.

Ohr (Organic Hydroperoxide Resistance Protein) Possesses a Previously Undescribed Activity, Lipoyl-dependent Peroxidase

The Ohr (organic hydroperoxide resistance) family of 15-kDa Cys-based, thiol-dependent peroxidases is central to the bacterial response to stress induced by organic hydroperoxides but not by hydrogen peroxide. Ohr has a unique three-dimensional structure and requires dithiols, but not monothiols, to support its activity. However, the physiological reducing system of Ohr has not yet been identified. Here we show that lipoylated enzymes present in the bacterial extracts of Xylella fastidiosa interacted physically and functionally with this Cys-based peroxidase, whereas thioredoxin and glutathione systems failed to support Ohr peroxidase activity. Furthermore, we could reconstitute in vitro three lipoyl-dependent systems as the Ohr physiological reducing systems. We also showed that OsmC from Escherichia coli, an orthologue of Ohr from Xylella fastidiosa, is specifically reduced by lipoyl-dependent systems. These results represent the first description of a Cys-based peroxidase that is directly reduced by lipoylated enzymes.

Transfer of antibodies across the placenta and in breast milk from mothers on intravenous immunoglobulin

We studied the levels of immunoglobulins in colostrum, milk and sera from two common variable immunodeficiency (CVID) mothers (M1 and M2), and in sera from their newborn infants. During pregnancy they continued intravenous immunoglobulin therapy (IVIG). Antibody levels from maternal and cord blood collected at delivery and colostrum and milk, collected on the 3rd and 7th post-partum days, respectively, were analyzed. Although cord/maternal blood ratios of total immunoglobulins and subclasses, as well as specific antibodies differed between M1 and M2, both showed good placental transfer of anti-protein and anti-polysaccharide antibodies, despite lower cord/maternal blood ratios in M2. Anti-Streptococcus pneumoniae antibody avidity indexes were similar between paired maternal and cord serum. Both mothers` colostrum and milk samples showed only traces of IgA, and IgM and IgG levels in colostrum were within normal range in M1, whereas M2 presented elevated IgG and low IgM levels, when compared with healthy mothers. The study of colostrum and milk activity showed that they strongly inhibited enteropathogenic Escherichia coli adhesion in vitro. CVID patients must be informed about the relevance of regular IVIG administration during pregnancy, not only for their own health but also for their immune immature offspring. Breast-feeding should be encouraged as colostra from these CVID patients strongly inhibited E. coli adhesion to human epithelial cells thus providing immunological protection plus nutritional and psychological benefits for the infant.

Microbiological diversity of generalized aggressive periodontitis by 16S rRNA clonal analysis

Background/aim: The purpose of this study was to determine the bacterial diversity in the subgingival plaque of subjects with generalized aggressive periodontitis by using culture-independent molecular methods based on 16S ribosomal DNA cloning. Methods: Samples from 10 subjects with generalized aggressive periodontitis were selected. DNA was extracted and the 16S rRNA gene was amplified with the universal primer pairs 9F and 1525R. Amplified genes were cloned, sequenced, and identified by comparison with known 16S rRNA sequences. Results: One hundred and ten species were identified from 10 subjects and 1007 clones were sequenced. Of these, 70 species were most prevalent. Fifty-seven percent of the clone (40 taxa) sequences represented phylotypes for which no cultivated isolates have been reported. Several species of Selenomonas and Streptococcus were found at high prevalence and proportion in all subjects. Overall, 50% of the clone libraries were formed by these two genera. Selenomonas sputigena, the species most commonly detected, was found in nine of 10 subjects. Other species of Selenomonas were often present at high levels, including S. noxia, Selenomonas sp. EW084, Selenomonas sp. EW076, Selenomonas FT050, Selenomonas sp. P2PA_80, and Selenomonas sp. strain GAA14. The classical putative periodontal pathogens, such as, Aggregatibacter actinomycetemcomitans, was below the limit of detection and was not detected. Conclusion: These data suggest that other species, notably species of Selenomonas, may be associated with disease in generalized aggressive periodontitis subjects.

Some factors associated with dental caries in the primary dentition of children with Down syndrome

Aim It is well reported in the scientific literature that there is a high level of periodontal disease and lower caries prevalence in Down Syndrome (DS) individuals, when compared with age-matched non DS individuals. This study was conducted to investigate the process of dental caries in DS children. Materials and methods In this study the following parameters were considered: oral hygiene habits, levels of Streptococcus mutans (SM) and Lactobacillus spp. (LB), Modified Gingival Index (MGI), and Simplified Oral Hygiene Index (OHI-S). A case group with DS children (n=69) and a control group of non DS children (n=69) were formed to perform this study Dental caries severity was determined using the DMFT index. Samples of non-stimulated saliva were collected to determine the Lactobacillus spp levels. For SM levels, MSB agar plates were used. Results The findings revealed that the case group attended, dental check-ups more frequently brushed their teeth more times per day, flossed less, and also more frequently had SM levels classified as ""high count"". The MGI was higher and the OHI-S was lower than the control group (p<0.001). Conclusion No significant differences were found between the DMFT indexes of children from the two groups (p=0.345). The logistic regression analysis showed that in the case group, age, MGI, and SM count were positively related to dental caries (p<0.05).

The Challenges of Antimicrobial Resistance in Brazil

Brazil is a country with continental proportions with high geographic and economic diversity. Despite its medical centers of excellence, antimicrobial resistance poses a major therapeutic challenge. Rates of methicillin-resistant Staphylococcus aureus are up to 60% and are related to an endemic Brazilian clone. Local resistance to vancomycin in Enterococci was first related to Enterococcus faecalis, which differs from European and American epidemiology. Also, local Klebsiella pneumoniae and Escherichia coli isolates producing extended-spectrum beta-lactamases have a much higher prevalence (40%-50% and 10%-18%, respectively). Carbapenem resistance among the enterobacteriaceae group is becoming a major problem, and K. pneumoniae carbapenemase isolates have been reported in different states. Among nonfermenters, carbapenem resistance is strongly related to SPM-1 (Pseudomonasaeruginosa) and OXA-23 (Acinetobacter baumannii complex) enzymes, and a colistin-only susceptible phenotype has also emerged in these isolates, which is worrisome. Local actions without loosing the global resistance perspective will demand multidisciplinary actions, new policies, and political engagement.

Susceptibility of Staphylococcus aureus to porphyrin-mediated photodynamic antimicrobial chemotherapy: an in vitro study

The ability of Staphylococcus aureus to develop multidrug resistance is well documented, and the antibiotic resistance showed by an increasing number of bacteria has shown the need for alternative therapies to treat infections, photodynamic therapy (PDT) being a potential candidate. The aim of this study was to determine the effect of photodynamic therapy as a light-based bactericidal modality to eliminate Staphylococcus aureus. The study investigated a technique based on a combination of light and a photosensitizer that is capable of producing oxidative species to induce a cytotoxic effect. A Staphylococcus aureus suspension was exposed to a light emitting diode (LED) emitting at 628 nm, 14.6 mW/cm(2), and energy density of 20J/cm(2), 40J/cm(2), or 60 J/cm(2) in the presence of different porphyrin concentrations (PhotogemA (R)). Three drug concentrations were employed: 12 mu l/ml, 25 mu l/ml, and 50 mu l/ml. The treatment response was evaluated by the number of bacterial colony forming units (CFU) after light exposure. The results indicated that exposure to 60 J/cm(2) eliminated 100% (10 log(10) scales) of bacteria, on average. The best PDT response rate to eliminate Staphylococcus aureus was achieved with exposure to LED light in combination with the photosensitizer at concentrations ranging from 25 mu l/ml to 50 mu l/ml. These data suggest that PDT has the potential to eliminate Staphylococcus aureus in suspension and indicates the necessary drug concentration and light fluency.