In view of compliance with increasingly stringent environmental legislation imposed by regional, national, and supranational (e.g., European Union) authorities, innovative environmental technologies for the treatment of dye-contaminated effluents are necessary in the color industry. In this study, effluents of an industrial dye producer were subjected to distinct treatment trains following an initial qualitative characterization. The effectiveness of ozonation and a treatment using white rot fungi (WRF) and their enzymes were compared with respect to parameters such as residual color, toxicity on human cells, and genotoxicity. A combined ozonation/WRF process was also investigated. The effluent exhibited significant toxicity that was reduced by only 10% through ozonation, whereas the fungal treatment achieved a 35% reduction. A combined treatment (ozone/WRF) caused an abatement of the toxicity by more than 70%. In addition, the initial genotoxicity of the effluent was still present after the ozone treatment, while it was completely removed through the fungal treatment.

The recent regulations, which impose limits on single use plastics and packaging, are encouraging the development of bioplastics market. Some bioplastics are labelled as compostable with the organic waste according to a specific certification (EN 13432), however the conditions of industrial composting plants are generally less favourable than the standard test conditions. Aiming at studying the effective degradation of marketable bioplastic products under composting, the current research stresses novel elements which can strongly influence bioplastics degradation: the simulation of industrial composting conditions and the thickness of bioplastic products, ranging between 50 and 500 µm. The research approaches these critical aspects simulating a composting test of 20 days of thermophilic phase followed by 40 days of maturation phase, on starch-based polymer Mater-Bi® (MB), polybutylene adipate terephthalate (PBAT), polylactic acid (PLA) of different thickness. Conventional low density polyethylene (LDPE) was introduced as negative control. An overall study with Fourier Transform InfraRed (FTIR), ThermoGravimetric Analysis (TGA), Gel Permeation Chromatography (GPC), Scanning Electron Microscope (SEM) and visual inspections was applied. Results highlighted that MB film presented the highest degradation rate, 45 ± 4.7% in terms of weight loss. Both MB and PBAT were subjected to physico-chemical features change, while LDPE presented slight degradation signs. The most critical observations have been done for PLA, which is strongly influenced both by thickness and thermophilic phase duration, shorter than the EN 13432 conditions.

In this study, a selective substrate for cytochrome P450 2D6 was designed using a small molecule model developed by M. J. De Groot et al. [(1997) Chem. Res. Toxicol. 10, 41-48]. The substrate, 7-methoxy-4-(aminomethyl)coumarin (MAMC), and its putative O-demethylated metabolite 7-hydroxy-4-(aminomethyl)coumarin (HAMC) were synthesized, and their respective fluorescence properties were characterized. The selectivity of MAMC for P450 2D6 was characterized using microsomes containing single human P450 isoenzymes and human liver microsomes. Formation of the metabolic product HAMC was easily assessed in real time with fluorescence spectroscopy, since MAMC and HAMC excitation and emission wavelengths differed significantly. HPLC analysis confirmed that HAMC was the single metabolic product of MAMC and that HAMC formation accounts for the total increase in fluorescence. It was found that, in microsomes from yeast or lymphoblastoid cells selectively expressing P450 isoenzymes, MAMC was selective for P450 2D6 at a concentration of 25 microM with only P450 1A2 contributing significantly to the formation of HAMC. P450s 2A6, 2B6, 2C8, 2C9, 2C19, 2E1, 3A4, and 3A5 were shown not to metabolize MAMC at a concentration of 25 microM. K(m) and v(max) values of MAMC for P450 2D6 were found to be 26.2 +/- 2.8 microM and 2.9 +/- 0.07 min(-)(1), respectively. For P450 1A2, MAMC was found to have a K(m) value of 29.7 +/- 6.2 microM and a v(max) of 0.57 +/- 0.07 min(-)(1). Formation of HAMC in human liver microsomes could be completely inhibited by quinidine, at a concentration of 0.5 microM selective for P450 2D6, and furafylline, at a concentration of 30 microM selective for P450 1A2. In conclusion, O-demethylation of 7-methoxy-4-(aminomethyl)coumarin is a rapid and easily determined parameter for P450 2D6 activity and, due to the fluorescent properties of the metabolite formed, may be a valuable new tool for high-throughput screening purposes.

The recent development of several promising new thiourea-containing drugs has renewed interest in the thiourea functionality as a potential toxicophore. Most adverse reactions of thiourea-containing compounds are attributed to the thionocarbonyl moiety. Oxidation of these thionocarbonyl compounds by flavin-containing monooxygenases (FMO) and cytochrome P450 isoenzymes (P450) to reactive sulfenic, sulfinic, or sulfonic acids leads to alkylation of essential macromolecules. To more rationally design thiourea-containing drugs, structure-toxicity relationships (STRs) must be derived. Since for the development of STRs a large number of thiourea-containing compounds must be investigated, it is important to develop rapid in vitro assays for alkylating potential. In this study, the utility of activation of microsomal glutathione S-transferase (mGST) and inactivation of P450 1A1 as markers of the alkylating potential of metabolites of thiourea-containing compounds was investigated. It was found that metabolites of thiourea-containing compounds inactivate P450 1A1 in a time-dependent manner, as evidenced by a decrease in 7-ethoxyresorufin O-dealkylation (EROD) activity. An extent of inactivation of P450 1A1 by 100 microM N-phenylthiourea (PTU) of 64% was found after 10 min. This inactivation was dependent on the presence of NADPH and the presence of the thionosulfur, since the carbonyl analogue of PTU was not found to inactivate P450 1A1, and was partially prevented by heat treatment of the microsomes which is known to selectively inactivate FMO enzymes. Inactivation of P450 1A1 could be reversed by treatment with dithiothreitol, indicating the formation of disulfide bonds. However, thiourea-containing compounds also inhibited the EROD activity of P450 1A1 in a competitive manner. This property complicates the usefulness of the EROD activity of P450 1A1 as a marker for the alkylating potential of thiourea-containing compounds. It was found that metabolites of thiourea-containing compounds could transiently activate the mGST. A maximal level of activation by 100 microM PTU of 162+/-16% was found after 10 min. Activation of mGST by 100 microM PTU was dependent on the presence of NADPH and the presence of the thionosulfur, since the carbonyl analogue of PTU was not found to activate mGST. Activation was completely prevented by heat treatment of the microsomes, indicating involvement of FMO in the bioactivation process. Finally, a series of structurally diverse thiourea-containing compounds were tested for their ability to activate mGST. It appeared that their potency in alkylating mGST was inversely related to their Vmax/Km value for the FMO enzyme. From this study, it is concluded that, whereas activation of mGST in rat liver microsomes may be a useful system with which to investigate the relationship between structure and alkylating potential of thiourea-containing compounds in vitro, inactivation of P450 1A1 is not.