Detection of Bisphenol A in dental wastewater after grinding of dental resin composites.

OBJECTIVES: This study evaluated the release of bisphenol A (BPA) in wastewater after grinding of resin composites and tested three filtration materials. METHODS: Three resin composites (Ceram X, Filtek Supreme XTE and Core-X flow) were used. Samples (5mm×2mm, n=10) were prepared using a metal mold and were polymerized for 20s according to manufacturers' instructions. A dental unit was disconnected from wastewater circulation and composite samples were ground under standardized procedures (200,000rpm; 90s). Wastewater was collected in glass bottles. Water samples were collected as control by performing the same procedure without grinding resin composite. All samples were stored at 7°C for 6 months to simulate storage. Then they were analyzed by HPLC-FLD. Three filtration materials (Zeosorb, Katalox Light and Catalytic Carbon) were used for water treatment to remove BPA. BPA-water solutions were prepared; corresponding to the highest amount released by the resin composites. These solutions were analyzed before and after filtration by HPLC-FLD and their efficacy (%) was calculated. RESULTS: BPA was detected in all composite solutions: Ceram X and Filtek Supreme XTE showed similar findings (p>0.05) which were significantly higher than the control (p<0.001) and Core-X flow (p=0.001). The efficacy of the filtration materials was: Katalox Light (5.09%)

Random mutagenesis of the multidrug transporter AcrB from Escherichia coli for identification of putative target residues of efflux pump inhibitors.

Efflux is an important mechanism of bacterial multidrug resistance (MDR), and the inhibition of MDR pumps by efflux pump inhibitors (EPIs) could be a promising strategy to overcome MDR. 1-(1-Naphthylmethyl)-piperazine (NMP) and phenylalanine-arginine-ß-naphthylamide (PAßN) are model EPIs with activity in various Gram-negative bacteria expressing AcrB, the major efflux pump of Escherichia coli, or similar homologous pumps of the resistance-nodulation-cell division class. The aim of the present study was to generate E. coli AcrB mutants resistant to the inhibitory action of the two model EPIs and to identify putative EPI target residues in order to better understand mechanisms of pump inhibition. Using an in vitro random mutagenesis approach focusing on the periplasmic domain of AcrB, we identified the double mutation G141D N282Y, which substantially compromised the synergistic activity of NMP with linezolid, was associated with similar intracellular linezolid concentrations in the presence and absence of NMP, and did not impair the intrinsic MICs of various pump substrates and dye accumulation. We propose that these mutations near the outer face of the distal substrate binding pocket reduce NMP trapping. Other residues found to be relevant for efflux inhibition by NMP were G288 and A279, but mutations at these sites also changed the susceptibility to several pump substrates. Unlike with NMP, we were unable to generate AcrB periplasmic domain mutants with resistance or partial resistance to the EPI activity of PAßN, which is consistent with the modes of action of PAßN differing from those of NMP.

Human gingival keratinocyte response to substances eluted from silorane composite material reveal impact on cell behavior reflected by RNA levels and induction of apoptosis.

OBJECTIVES: The aim of this study was the characterization of siloran-derived composite eluates in conjunction with their putative impact on human gingival keratinocytes (HGK), i.e. levels of total RNA and induction of apoptosis compared to a methacrylate-based material. METHODS: Standardized Filtek™ Silorane specimens (n = 20) were subjected to scanning ion monitoring to detect monomer masses between 100 and 1000, after storage in human saliva, and 75% ethanol for up to 28 days. In order to evaluate the effect on cells, HGK were exposed to eluates from Filtek™ Silorane, Filtek™ Supreme XT and control medium for 1 and 4 days, prior to isolation of total RNA, and Annexin-5 fluorescence labeling indicating induction of apoptosis. RESULTS: Irrespective of the mode and storage time, SIM identified discrete peaks, corresponding to masses of "393" and "337". In response to both composite eluates, an effect on HGK was reflected by drastically reduced levels of isolated total RNA at each time period (after 1 day: control: 302 ng/µl; Filtek™ Silorane: 128 ng/µl, Filtek™ Supreme XT: 129 ng/µl and after 4 days: control: 528 ng/µl; Filtek™ Silorane: 162 ng/µl, Filtek™ Supreme XT: 166 ng/µl). Exposure to eluates from both composite materials yielded apoptosis induction in HGK, as demonstrated by a significant increase of cells exhibiting Annnexin-5 fluorescence. SIGNIFICANCE: Two distinct peaks were identified, which indicated the presence of corresponding substances. The composite-derived effects on HGK strongly suggest a negative impact on cells, as revealed by a clear reduction of total RNA levels, and significant increase in induction of apoptosis.

Release of monomers from four different composite materials after halogen and LED curing.

PURPOSE: To evaluate the release of monomers from four different composite materials (Ceram X, Filtek Supreme XT, Tetric Flow, Tetric EvoCeram), polymerized using either halogen or LED unit. METHODS: Ten specimens were made for each material/unit combination. Each specimen was stored in 1 ml 75% ethanol. The storage medium was renewed after 1, 7 and 28 days. Aliquots of this medium were analyzed by LC-MS/MS. RESULTS: The effect of the curing unit on monomers' release differed significantly among the materials (P < 0.0001). The amount of BisGMA and TEGDMA released from Ceram X was not influenced by the unit used (P > 0.05). Curing with LED reduced the amount of Bisphenol A released from Ceram X compared to halogen. For Filtek Supreme XT, the type of unit exerted a significant effect on the elution of BisGMA (P < or = 0.05). LED curing resulted in a higher release of TEGDMA and UDMA compared to halogen (P < or = 0.05). For Tetric Flow, LED curing resulted in lower monomer release (P < 0.0001). For Tetric EvoCeram, the amounts of BisGMA, UDMA and Bisphenol A were higher when polymerizing with LED compared to halogen. The release of substances was more material dependent and less influenced by the curing unit used.

Therapeutic drug monitoring of posaconazole in hematology patients: experience with a new high-performance liquid chromatography-based method.

Parallel administration of the proton pump inhibitor (PPI) esomeprazole has been shown to decrease oral bioavailability of posaconazole in healthy volunteers. We prospectively analyzed serum samples (n = 59) obtained from hematology patients (n = 27) under posaconazole prophylaxis. Patients treated concomitantly with pantoprazole had significantly lower posaconazole levels than patients without PPI treatment (median levels of 630 microg/liter versus 1,125 microg/liter, respectively). These results suggest that drug monitoring is relevant when posaconazole and pantoprazole are administered concomitantly.

Determination of the antifungal agent posaconazole in human serum by HPLC with parallel column-switching technique.

Posaconazole is a new broad-spectrum antifungal agent that is currently only available as an oral suspension and shows high intra- and inter-individual differences in oral bioavailibility. Pre-existing methods for the determination of the substance involve the use of internal standards or require a quite complicated and time-consuming sample pre-treatment. Our HPLC method is fast and fully-automated and there is no need for any manual sample pre-treatment. On-line transfer of posaconazole from the extraction column was followed by chromatographic separation on a C18 column and fluorescence detection (lambda(ex): 261 nm, lambda(em): 357 nm). Retention time of posaconazole was about 11.7 min, the lower limit of quantification was found to be 0.1 mg/l. A linear calibration curve was obtained over the concentration range of 0.1-5 mg/l using a 50 microl sample (r(2)=0.999). The relative standard deviations of intra-day variations ranged from 2.3% to 9.4%, intra-day accuracy from 88.8% to 114.8%.

Release of monomers from different core build-up materials.

OBJECTIVE: The aim of the present study was to evaluate and compare the elution of monomers from three different core build-up composite materials and correlate it with the degree of conversion. METHODS: Three different core build-up composite materials (a chemically cured, a photo-cured, and a dual-cured) were tested. Ten samples (diameter: 4.5 mm and thickness: 2 mm) of each material were fabricated to evaluate the release of monomers. The photo-cured samples were polymerized for 40s and the dual-cured samples for 20 s. The samples remained undisturbed for 10 min and then were stored in 1 ml of 75% ethanol at room temperature, and the storage medium was renewed after 24 h, 7 and 28 days. From the storage medium that was removed, samples were prepared and analyzed by LC-MS/MS. Additionally, four samples of each material were tested for the degree of conversion by using a FT-IR spectrometer. RESULTS: The three composite materials differed significantly concerning the elution of monomers (BisGMA: p<0.0001; TEGDMA: p<0.0001; and Bisphenol A: p<0.0001). A significantly higher amount of BisGMA and TEGDMA was released from the chemically cured composite compared to the other two materials. Between the photo-cured and the dual-cured material the latter eluted significantly higher amounts of BisGMA and TEGDMA. During the storage of the samples, the amounts of the eluted substances decreased. The degree of conversion of the chemically cured composite was significantly lower compared to the other two materials. SIGNIFICANCE: Using the present parameters, the photo-cured material released less monomer and therefore they might be less dangerous with respect to toxicological effects.

Photo-degradation of the antimicrobial ciprofloxacin at high pH: identification and biodegradability assessment of the primary by-products.

Photo-treatment for the removal of pharmaceuticals in effluents is a topic currently under discussion. In some countries effluents from hospitals are directly emitted into open ditches without any further treatment and with very little dilution. Under such circumstances photo-degradation in the environment can occur. However, photo-degradation does not necessarily end up with the complete mineralization of a chemical. Therefore, photo-product biodegradability and toxicity against environmental bacteria is of interest. Hospital effluents have often a pH around 9. Therefore, photo-oxidation (150W medium-pressure Hg-lamp, batch reactor) of ciprofloxacin (CIP) was studied at pH 9. The primary elimination of CIP was monitored and structures of photo-products were assessed by liquid chromatography ion trap mass spectrometry (LC-MS/MS). Five compounds were identified as probable products of photo-defluorination, -decarboxylation and loss of the piperazine moiety. These photo-products were not biodegradable in the Closed Bottle test - OECD 301D. They did not affect Vibrio fisheri in the applied concentrations.

Urethane dimethacrylate: a molecule that may cause confusion in dental research.

In recent years, the elution of monomers from dental materials has been a cause for public concern. Urethane dimethacrylate, commonly abbreviated to UDMA, is one of the monomers that are most often tested with regard to elution from and cytotoxicity of resin-based materials. Although each chemical name represents the chemical type, chemical structure, and molecular weight of a molecule, it does not seem to be the same with UDMA. In the present paper, the different forms of UDMA are presented. These include those used by dental manufacturers to produce composite materials and the different types of urethane dimethacrylate used in studies concerning the elution of monomers from composite materials. High-performance liquid chromatography (HPLC) is usually used to detect the eluted monomers, but it does not appear to be adequate in determining the different forms of UDMA. The combination of HPLC with mass spectrometry is shown to be able to specifically identify the compounds eluted in addition to those compounds used as standards in the various studies. The fact that the same name is given to different molecules causes confusion about the results of studies testing the elusion of monomers from composite materials and their possible toxicity.

Long-term release of monomers from modern dental-composite materials.

The elution of monomers from composite materials influences the biocompatibility of dental restorations. The purpose of the present study was to investigate the elution of monomers [bisphenol A glycidyl methacrylate (BisGMA), triethylene glycol dimethacrylate (TEGDMA), urethane dimethacrylate (UDMA), and bisphenol A (BPA)] from two light-cured materials (nanohybrid and ormocer) and from a chemically cured composite material, after different curing times (0, 20, 40 and 80 s) and different storage periods (24 h, 7 d, 28 d, and 1 yr after curing). Each specimen was stored in 1 ml of 75% ethanol. This medium was renewed after 24 h, 7 d, 28 d, and 1 yr. The ethanol samples were analyzed using liquid chromatography tandem mass spectrometry (LC-MS/MS). The amount of monomers released from the nanohybrid and the chemically cured composite was significantly higher than released from the ormocer. The curing time exerted a significant effect on the release of monomers. For the nanohybrid, less monomer was released after increasing the curing time. For the ormocer, 80 s of curing resulted in a higher degree of monomer release. The effect of storage differed between the monomers. Although the elution of TEGDMA was significantly decreased after storage for 28 d and 1 yr, a similar amount of BisGMA was released at each storage time-point analyzed, even after 1 yr. The present study showed that ormocer released a very small amount of monomers compared with the other materials.

Deactivation and transformation products in biodegradability testing of beta-lactams amoxicillin and piperacillin.

Antibiotics have increasingly been detected in effluents and the environment. However, information on the degree of deactivation and mineralization, and the nature of possible formed dead-end transformation products is scarce but desirable for proper risk assessment. An important group of antibiotics is the beta-lactams. We studied the transformation of the closely structurally related beta-lactams piperacillin and amoxicillin in two OECD biodegradability batch tests. None of the antibiotics was biodegraded in the closed bottle test (CBT). However, primary abiotic elimination as monitored by HPLC-UV-VIS was 20% and 100% in the CBT within 14 days, respectively. With HPLC-UV-VIS and ion trap LC-MS/MS primary elimination was shown to be more than 94% for both antibiotics within seven days in the Zahn-Wellens test (ZWT). Both compounds were deactivated by hydrolysis. For piperacillin, a dead-end transformation product resulted after hydrolysis of the beta-lactam ring. For amoxicillin full mineralization of the transformation products was observed.

Effect of bleaching on the elution of monomers from modern dental composite materials.

OBJECTIVE: The aim of this study was to evaluate the effect of bleaching on the elution of monomers from two modern composite materials. METHODS: Two different resin composites (a nanohybrid the Filtek Supreme XT and an ormocer the Ceram X) were bleached with two products (hydrogen peroxide 38% for 45 min and carbamide peroxide 15% for 56 h). Four groups (n=10, diameter: 4.5 mm, thickness: 2 mm) of each material were fabricated, two for each bleaching product. One group was used as unbleached control and the other one was bleached. Then the samples were stored in 1 ml of 75 vol% ethanol at room temperature, and the storage medium was renewed after 24 h, 7 days, and 28 days. From the storage medium that was removed samples were prepared and analysed with LC-MS/MS. RESULTS: None of the bleaching products had an effect on the amount of Bis-GMA and TEGDMA released from Ceram X. The amount of Bisphenol A released from the bleached samples of Ceram X was significantly lower compared to the control samples. Bleaching reduced significantly the amount of Bis-GMA released from Filtek Supreme XT. The amount of TEGDMA released from Filtek Supreme XT was not affected by bleaching. SIGNIFICANCE: The bleaching agents tested in the present study reduced the amount of some of the monomers released from the two composite materials. Bleaching of modern composite materials does not increase the release of monomers.

Fast, fully automated analysis of voriconazole from serum by LC-LC-ESI-MS-MS with parallel column-switching technique.

Voriconazole is a novel broad-spectrum antifungal agent. We developed an on-line LC-LC-MS-MS method for fully automated and direct analysis of voriconazole in raw human serum. After injection of human serum size-selective sample fractionation and analyte extraction was achieved using an extraction column (25 mm x 4 mm) packed with a restricted access material (RAM, LiChrospher ADS C(8), 25 microm). On-line transfer of voriconazole from the extraction column was followed by chromatography separation on a C(18) column. Detection was done by ESI-MS-MS. The total analysis time was 13 min, managed by parallel extraction and chromatographic separation. This LC-MS assay was fully validated. The lower limit of quantification was 0.05 microg/ml. The automated inline extraction of voriconazole described here eliminates the need for difficult and time-consuming sample pre-treatment. Other advantages of the new method are that only a small quantity (5 microl) of serum is needed and that the method is very specific.

Long term azithromycin therapy in cystic fibrosis patients: a study on drug levels and sputum properties.

BACKGROUND: Following reports on the treatment of diffuse panbronchiolitis (DPB), recent studies demonstrate that long term therapy with azithromycin (AZM) is effective in cystic fibrosis (CF) patients. However, the underlying mechanisms remain uncertain. Some macrolides, including AZM, display inhibition of virulence factors and other antipseudomonal effects at subinhibitory levels in vitro. OBJECTIVES: Drug doses used for CF and DPB therapy were investigated to determine whether they achieve corresponding sputum drug levels in CF patients in vivo. METHODS: In an open, prospective study, 14 CF patients with chronic Pseudomonas aeruginosa airway infection received 250 mg AZM either daily ('high dose') or twice weekly ('low dose') for 12 weeks. Viscoelasticity of sputum was assessed by magnetic microrheology. RESULTS: AZM accumulated in sputum by two orders of magnitude over a period of four weeks. In the following steady state, median AZM concentrations in sputum were 9.5 microg/mL (0.6 to 79.3 microg/mL, interquartiles 1.4 to 33.4 microg/mL) and 0.5 microg/mL (range less than 0.1 [below detection level] to 5.2 microg/mL, interquartiles 0.2 to 1.4 microg/mL) in the high and low dose groups, respectively. Viscoelasticity improved in all patients but one. CONCLUSIONS: The findings suggest that antipseudomonal activity has to be considered among the potential mechanisms of macrolide therapy. Further, viscoelasticity may be a valuable parameter in future clinical trials.