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Raffaele Piccolomini

Publications and source records attributed to Raffaele Piccolomini.

8 recordsLinked to original sources

Biofilm formation by the emerging fungal pathogen Trichosporon asahii: development, architecture, and antifungal resistance.

Trichosporon asahii is the most common cause of fatal disseminated trichosporonosis, frequently associated with indwelling medical devices. Despite the use of antifungal drugs to treat trichosporonosis, infection is often persistent and is associated with high mortality. This drove our interest in evaluating the capability of T. asahii to form a biofilm on biomaterial-representative polystyrene surfaces through the development and optimization of a reproducible T. asahii-associated biofilm model. Time course analyses of viable counts and a formazan salt reduction assay, as well as microscopy studies, revealed that biofilm formation by T. asahii occurred in an organized fashion through four distinct developmental phases: initial adherence of yeast cells (0 to 2 h), germination and microcolony formation (2 to 4 h), filamentation (4 to 6 h), and proliferation and maturation (24 to 72 h). Scanning electron microscopy and confocal scanning laser microscopy revealed that mature T. asahii biofilms (72-h) displayed a complex, heterogeneous three-dimensional structure, consisting of a dense network of metabolically active yeast cells and hyphal elements completely embedded within exopolymeric material. Antifungal susceptibility testing demonstrated a remarkable rise in the MICs of sessile T. asahii cells against clinically used amphotericin B, caspofungin, voriconazole, and fluconazole compared to their planktonic counterparts. In particular, T. asahii biofilms were up to 16,000 times more resistant to voriconazole, the most active agent against planktonic cells (MIC, 0.06 microg/ml). Our results suggest that the ability of T. asahii to form a biofilm may be a major factor in determining persistence of the infection in spite of in vitro susceptibility of clinical isolates.

Antifungal Agents↗

Effectiveness of ultrasonic instruments in the therapy of severe periodontitis: a comparative clinical-microbiological assessment with curettes.

Patients with deep periodontal pockets were treated with either Vector System (TG) or manual instruments (CG). Clinical assessments by supragingival plaque (PL+), gingival index (GI), bleeding on probing (BOP), probing depth (PD), clinical attachment level (CAL) and subgingival plaque collection for microbiological analysis were made prior to and after treatment. Multiplex Polymerase Chain Reaction was used to determine the presence of Actinobacillus actinomycetemcomitans, Campylobacter rectus, Eikenella corrodens, Fusobacterium nucleatum, Prevotella intermedia, Porphyromonas gingivalis, Tannerella forsythensis and Treponema denticola. GI, PD, CAL and the number of BOP+ sites underwent a significant reduction over time in both groups. When compared to baseline, the pair-wise analyses showed significantly lower PD and CAL at 6 months in the CG and significant reductions in the GI, PD, CAL and a number of BOP+ sites at 3 and 6 months in the TG. For microbiological results, significant reductions were seen for C. rectus and R. gingivalis in the CG and for T. forsythensis, E. corrodens and T. denticola in the TG. The total bacterial count underwent a reduction in both groups. Both ultrasonic and manual debridement are equally effective in non-surgical periodontal therapy of severe periodontitis in terms of clinical and microbiological effects.

Adult↗

Clinical and microbiological effects of different restorative materials on the periodontal tissues adjacent to subgingival class V restorations.

OBJECTIVES: The relationship between subgingival dental restorations and periodontal health has been thoroughly investigated for many years. However, longitudinal data on the subgingival microflora features after the placement of well-finished subgingival restorations are still lacking. Therefore, this study compares the short-term clinical and microbiological features occurring in the gingiva after the completion of different subgingival restorations. MATERIAL AND METHODS: Sixteen systemically healthy subjects, 10 males and six females (ages: 31.7-45.8 years; mean age 39.3+/-5.1 years), who were non-smokers and were positive for the presence of three cervical abrasion/erosion defects to be restored in three different adjacent teeth were enrolled in this study. The cervical abrasion/erosion defects were each restored by using one of three different materials: amalgam, glass ionomer cement, or composite resin. Immediately before class V cavity preparations and restorations (baseline), clinical monitoring and subgingival plaque sampling were performed in the mid-buccal aspect of each experimental restored tooth and in one adjacent sound, non-treated, control tooth. These procedures were repeated every 4 months over the following 1 year. RESULTS: Throughout the study, the clinical parameters recorded did not change significantly in any of the experimental groups, and no differences were detected among them at each clinical session. Over this time, no significant changes in the composition of the subgingival microflora were observed in amalgam, glass ionomer cement, and control groups. Conversely, in the composite resin group, there was a significant increase in the total bacterial counts, and a significant (p<0.05) decrease in Gram-positive, aerobic bacteria, which was associated with a significant (p<0.05) increase in the Gram-negative, anaerobic microbiota. CONCLUSIONS: Over a 1-year observation period, amalgam, glass ionomer cement, and composite resin subgingival restorations do not significantly affect the clinical parameters recorded. However, composite resin restorations may have some negative effects on the quantity and quality of subgingival plaque.

Adult↗

Clinical and microbiological effects of subgingival administration of two active gels on persistent pockets of chronic periodontitis patients.

OBJECTIVES: The present controlled, single-blind study was performed to assess and compare the clinical healing and the microbiological findings following repeated intrasulcular applications of 1% metronidazole or 1% chlorhexidine gels in persistent periodontal pockets previously treated by scaling and root planing (SRP). MATERIAL AND METHODS: Sixty-three systemically healthy subjects, 25 males and 38 females (mean age 48.4+/-7.2 years), diagnosed for chronic periodontitis were enrolled in this study. They underwent SRP and received oral hygiene instructions (OHI). Three months later, at baseline, a single persistent pocket with a probing depth (PD) of 5-9 mm was chosen as the experimental site in each patient; the subjects were stratified into three matched experimental groups on the basis of the treatment to be performed, which consisted of the subgingival administration of 1% metronidazole gel (MG, n=19), 1% chlorhexidine gel (CG, n=20) or placebo gel (PG, n=24). The treatments consisted of four repeated administrations of subgingival gels, each separated by 7 days, starting at the baseline. Clinical assessment was performed at the baseline and at the 180-day follow-up, after the end of treatment. For microbiological evaluations, subgingival plaque was sampled from the experimental sites at baseline, prior to the first subgingival gel administration, and at 7, 15, 30 and 90 days after the end of the treatment (days 28, 36, 51 and 111 from baseline). RESULTS: Plaque accumulation did not change significantly in all three groups. Bleeding on probing and clinical attachment levels reduced in the MGs and CGs only. PD was significantly reduced by the same amount in all experimental groups. In the MGs and CGs a remarkable reduction in the frequencies of detection of several periodontopathic micoorganisms was recorded after the treatment. The same was not seen for the PGs. CONCLUSIONS: Subgingival administration of MG or CG, both at 1%, may have a role in the management of persistent pockets during chronic periodontitis.

Adult↗

Biofilm formation by Stenotrophomonas maltophilia: modulation by quinolones, trimethoprim-sulfamethoxazole, and ceftazidime.

We investigated the in vitro effects of seven fluoroquinolones (ciprofloxacin, grepafloxacin, levofloxacin, moxifloxacin, norfloxacin, ofloxacin, and rufloxacin), compared to those of trimethoprim-sulfamethoxazole (SXT) and ceftazidime on total biomass and cell viability of Stenotrophomonas maltophilia biofilm. S. maltophilia attached rapidly to polystyrene, within 2 h of incubation, and then biofilm formation increased over time, reaching maximum growth at 24 h. In the presence of fluoroquinolones at one-half and one-fourth the MIC, biofilm biomass was significantly (P < 0.01) reduced to 55 to 70% and 66 to 76% of original mass, respectively. Ceftazidime and SXT did not exert any activity. Biofilm bacterial viability was significantly reduced by all antibiotics tested at one-half the MIC. At one-fourth the MIC all antibiotics, except levofloxacin, significantly reduced viability. Treatment of preformed biofilms with bactericidal concentrations (500, 100, and 50 micro g/ml) of all fluoroquinolones caused, except for norfloxacin, significant reduction of biofilm biomass to 29.5 to 78.8, 64.1 to 83.6, and 70.5 to 82.8% of original mass, respectively. SXT exerted significant activity at 500 micro g/ml only. Ceftazidime was completely inactive. Rufloxacin exhibited the highest activity on preformed biofilm viability, significantly decreasing viable counts by 0.6, 5.4, and 17.1% at 500, 100, and 50 micro g/ml, respectively. Our results show that (i) subinhibitory (one-half and one-fourth the MIC) concentrations of fluoroquinolones inhibit adherence of S. maltophilia to polystyrene and (ii) clinically achievable concentrations (50 and 100 micro g/ml) of rufloxacin are able to eradicate preformed S. maltophilia biofilm.

Anti-Infective Agents↗

In vitro pharmacodynamic characteristics of amphotericin B, caspofungin, fluconazole, and voriconazole against bloodstream isolates of infrequent Candida species from patients with hematologic malignancies.

Time-kill and postantifungal effect (PAFE) of amphotericin B, caspofungin, fluconazole, and voriconazole were determined against clinical isolates of Candida guilliermondii, Candida kefyr, and Candida lusitaniae. Azoles displayed fungistatic activity and no measurable PAFE, regardless of the concentration tested. Amphotericin B and caspofungin demonstrated concentration-dependent fungicidal activity, although amphotericin B only produced a significant dose-dependent PAFE against all isolates tested.

Amphotericin B↗

Comparison of Etest, agar dilution, broth microdilution and disk diffusion methods for testing in vitro activity of levofloxacin against Staphylococcus spp. isolated from neutropenic cancer patients.

The susceptibility to levofloxacin of 194 consecutive staphylococcal (45 Staphylococcus aureus and 149 coagulase-negative staphylococci) isolates from neutropenic patients was determined by Etest and the results compared with those obtained using NCCLS-methods (broth microdilution, agar dilution and disk diffusion). Overall agreement at +/- 1log(2) dilution for Etest compared with broth microdilution and agar dilution was 99.0 and 83.5%, respectively. The Etest category agreement with broth microdilution and disk diffusion was 95.9 and 89.7%, respectively. Comparison of categories with Etest and agar dilution method gave only 67.0% absolute categorical agreement, with 29.9% minor errors and 10.7% major errors. No very major errors occurred by the four methods tested. Our results show that Etest is a valid alternative to the reference NCCLS-methods for monitoring the clinical usefulness of levofloxacin against staphylococci isolates from neutropenic patients.

Agar↗

Bacteriologic evaluation of the effect of Nd:YAG laser irradiation in experimental infected root canals.

The aim of this study was to evaluate the efficacy of the Pumped Diodium-Nd:YAG laser in sterilizing contaminated root canals. After hand instrumentation, 30 teeth were inoculated with Actinomyces naeslundii CH-12 and 30 teeth with Pseudomonas aeruginosa ATCC 27853 and incubated for 24 h. The teeth were divided into three subgroups: subgroup A received no treatment; subgroup B was irradiated with laser (5 Hz for 15 s or 10 Hz for 15 s); and subgroup C was irrigated with 5.25% NaOCl. The number of viable bacteria in each group was evaluated by using the surface-spread plate technique. The results indicated an average of 34.0% decrease in colony-forming units for A. naeslundii CH-12 and 15.7% for P. aeruginosa ATCC 27853 with the 5 Hz/15 s laser treatment, and for the 10 Hz laser frequency, a decrease of the 77.4% for A. naeslundii CH-12 and 85.8% for P. aeruginosa ATCC 27853. No bacteria were detected in the canals treated with 5.25% NaOCl. The results show an antibacterial effect of the Pumped Diodium Nd:YAG laser, depending on the radiation frequency. However, 5.25% NaOCl was more effective than either laser application.

Actinomyces↗