Cytotoxicity and estrogenicity in simulated dental wastewater after grinding of resin-based materials.

OBJECTIVE: This study evaluated the cytotoxic and estrogenic effects of dust and eluates released into simulated wastewater after grinding of dental resin-based materials. METHODS: Four materials were used: ceram.x® universal, Filtek™ Supreme XTE, Lava™ Ultimate and Core-X™ flow. From each composite material, samples (5 × 2 mm, n = 50) were prepared according to the manufacturers' instructions. Lava™ Ultimate was used as blocks. All samples were ground to dust with a diamond bur (106 µm) and suspended in distilled water at 60 mg/mL. After storage for 72 h, the suspensions were separated into a soluble (eluate) and a particulate (dust) fraction. Eluates and dusts were evaluated for inhibition of Vibrio fischeri bioluminescence and cytotoxicity on human A549 lung cells (WST-1-Assay). The estrogenic activity was assessed by YES-Assay using Saccharomyces cerevisiae. Additionally, dental monomers (BisGMA, BisEMA, UDMA, TEGDMA, HEMA) and Bisphenol A were investigated. RESULTS: All eluates showed inhibition of V. fischeri bioluminescence at concentrations above 1.1 mg/mL (p < 0.05). The activity of the eluates of ceram.x® universal and Filtek™ Supreme XTE was significantly higher than Lava™ Ultimate and Core-X™ flow (p < 0.05). In the WST-1-Assay, all materials induced cytotoxic effects at concentrations of 0.1 mg/mL (p < 0.05), while no significant differences were detected among them. The tested materials revealed no estrogenic activity. All dental monomers and Bisphenol A showed concentration dependent cytotoxic effects (p < 0.05), whereas only Bisphenol A induced an estrogenic effect (p < 0.01). SIGNIFICANCE: Dust and eluates of resin-based dental materials released into wastewater exert bactericidal and cytotoxic effects in vitro. However, they reveal no estrogenic effect.

Higher blood concentrations of synthetic musks in women above fifty years than in younger women.

Synthetic musk compounds are widely used as fragrance ingredients in many consumer products. Little is known about their accumulation in humans and especially in older persons. In this study, we determined concentrations of 11 synthetic musks in women above fifty years and compared the results with earlier results from samples of young females. Blood was taken from 53 women above 50 years of age, visiting outpatients of the Department of Angiology at the Hanusch-Krankenhaus in Vienna, Austria. The used analytical methods consist of an extraction and clean-up step and a chromatographic analysis by GC/MS. Tonalide-D3 was used as recovery standard in all samples. Hexachlorobenzene (13)C(6) was used as internal standard. Study participants also completed a questionnaire on the use of cosmetics, about nutrition and other life-style aspects. The two substances which could be detected in higher percentages of the blood plasma samples were galaxolide (89 percent, maximum concentration 6900 ng/L) and musk xylene (62 percent, maximum concentration 190 ng/L). Regression analysis revealed a significant association of galaxolide concentration with frequent use of perfumes, deodorants and shampoos. Frequent use of soaps and fabric softener was associated with higher plasma concentrations of musk xylene. Nutrition habits, skin type, body mass index or surface area were not related to plasma concentration of these musk compounds. From the study group investigated older persons showed higher plasma concentrations. These findings could be due to the higher use of lotions and crèmes on face and hands and a more frequent use of skin care products because older persons reported more frequently dry skin. In addition, physiological aging related changes might be responsible for higher dermal absorption of synthetic musks. These results indicate that more focus on aging tissues is needed.

12-O-retinoylphorbol-13-acetate (RPA) behaves like a retinoid-derivative in binding to retinol-binding-protein: a correlation with its specific action as an incomplete tumor promoter?

12-O-retinoylphorbol-13-acetate (RPA), an incomplete tumor promoter of the phorbol ester type and protein kinase C (PKC) activator, consists of two characteristic structural elements: the phorbol body and the retinoyl ester chain. Therefore, possible binding of the incomplete tumor promoter RPA to the human transport protein retinol-binding-protein (RBP) has been examined by molecular modeling methods and experimental binding studies. The calculated prediction of binding properties was primarily based on a comparative geometrical approach. It was shown that the beta-ionone-ring of RPA was not altered within the binding pocket of RBP (molecular modeling) compared to retinoic acid (X-ray crystallographic data). The torsion angle C5'-C6'-C7'-C8', determining the conformation of the RBP-beta-ionone-ring relative to the isoprene tail, is rotated by 42 degrees for RPA compared to retinol and to retinoic acid, respectively. Combining all the results from force field calculations, MD simulations and geometrical comparisons, the conclusion could be drawn that RPA should be able to bind to RBP. This interaction should be less strong than that with its natural ligand retinol or with retinoic acid. This prediction was proven experimentally. RPA was able to compete with retinoic acid for binding at RBP in human plasma. The binding properties were investigated using 3H-labeled retinoic acid in homologous and heterologous competition studies in a one-dimensional native polyacrylamide gel electrophoresis system. An approximately 2000-fold weaker binding of RPA to RBP as compared to retinoic acid was determined experimentally, confirming the prediction of the molecular modeling approach. The characteristic behaviour of RPA as an incomplete promoter, due to possible binding to PKC and RBP, is discussed.

Protective effects of Brussels sprouts towards B[a]P-induced DNA damage: a model study with the single-cell gel electrophoresis (SCGE)/Hep G2 assay.

The aim of this study was to investigate the chemoprotective effects of Brussels sprouts juice towards benzo[a]pyrene (B(a)P)-induced DNA damage in the single-cell gel electrophoresis (SCGE)/Hep G2 test system. This assay combines the advantages of the SCGE assay with that of the use of human-derived cells possessing inducible phase I and phase II enzymes. Co-treatment of Hep G2 cells with small amounts of Brussels sprouts juice (0.25-2.0 microl/ml) and B(a)P reduced the genotoxic effect of the latter in a dose-dependent manner. Contrary to the results with the crude juice, unexpected synergistic effects were observed with allyl isothiocyanate (AITC, 1.0-6.0 microM), a breakdown product of sinigrin, which is the most abundant glucosinolate in Brussels sprouts. Although these concentrations of AITC did not cause DNA damage per se, at higher concentrations (> or =25 microM), the compound caused a pronounced dose-dependent DNA damage by itself. Mechanistic studies showed that Brussels sprouts juice causes induction of activities of ethoxyresorufin O-deethylase (EROD) and glutathione S-transferase (GST) at dose levels which were protective towards B(a)P. In combined treatment experiments with (+/-)-anti-benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide (BPDE, 5.0 microM), the main genotoxic metabolite of B(a)P, and Brussels sprouts juice, only weak protection was found indicating that the mechanism of chemoprotection of Brussels sprouts is not mediated through inactivation of this metabolite. In conclusion, our findings show that Brussels sprouts are highly protective against B(a)P-induced DNA damage in human-derived cells.

Studies on the in vivo and in vitro mutagenicity and the lipid peroxidation of chlorinated surface (drinking) water in rats and metabolically competent human cells.

In the present study, DNA damaging and mutagenic effects of chlorinated drinking water (CDW) extracts obtained from polluted raw water resources were examined in metabolically competent human Hep G2 hepatoma cells using the in vitro micronucleus assay and the single cell gel electrophoresis (SCGE, comet assay). Additionally, the in vivo induction of micronuclei (MN) was studied in polychromatic erythrocytes (PCEs) derived from bone marrow of CDW-treated Wistar rats. Furthermore, we examined the influence of CDW on the lipid peroxidation (LpO) in blood, liver, kidney and testicle of rats. The results demonstrated significant increases of micronucleated PCEs in the bone marrow of rats fed with relatively low CDW doses (33.3ml/kg body weight per day). Similar effects, i.e. increases of MN frequencies, were found in Hep G2 hepatoma cells after CDW treatment (41 MN/1000 binucleated cells (BNCs) for 167ml CDW) in comparison to the vehicle control (24 MN/1000 BNC). Additionally, DNA damages caused by CDW were observed in the comet assay. As a product of LpO, the levels of malondialdehyde (MDA) were significantly enhanced almost in all animals and organs tested after CDW treatment. In livers and serum of rats dose-dependent increases of MDA were observed. The data indicated that extracts from CDW obtained from polluted raw water were able to cause oxidative damages and to induce various biological effects in mammalian cells in vivo and in vitro, i.e. clastogenicity and/or aneugenicity, DNA strand breaks and/or alkali-labile damages. The consistency of the results among the various biological systems and endpoints led to the conclusion that the consumption of chlorinated drinking water obtained from polluted raw water may enhance the body burden with mutagenic and/or carcinogenic substances and therefore, means a potential genetic hazard for human health.

Evaluation of health risks caused by musk ketone.

Among the nitro musks, musk ketone (MK) as a synthetic compound with a typical musk odor is widely used in cosmetics. In the European Community the total amount used in fragrances has been reported to be 110 tons/a. Additionally, relevant amounts of MK are used in Indian joss sticks. As a result of its inherently low biodegradability MK has been detected in the aquatic environment (surface water, sediments, edible fish). Moreover, it has been shown that MK concentrates in human fatty tissue and breast milk, indicating that humans are constantly exposed. Several studies provided convincing evidence of lack of a genotoxic potential for MK. However, MK was identified as a strong inducer of phase I enzymes in rodents and a cogenotoxicant in vitro in human derived cells in rather low doses, suggesting that exposure to MK might increase the susceptibility to health hazards caused by carcinogens in humans.

Dietary cancer risk from conditional cancerogens in produce of livestock fed on species of spurge (Euphorbiaceae). III. Milk of lactating goats fed on the skin irritant herb Euphorbia peplus is polluted by tumor promoters of the ingenane diterpene ester type.

Special procedures were developed to investigate poisonous milk of lactating goats fed experimentally on aerial parts of the herb Euphorbia peplus L. In extracts of the milk, weakly irritant in the mouse-ear assay, three diterpene ester toxins were detected by techniques of high-performance liquid chromatography. They are of the ingenane structural type: Euphorbia factor Pel (ingenol 20-acetate 3-angelate), Euphorbia factor Pe2 (20-deoxyingenol 3-angelate) and Euphorbia factor Pe4 (20-deoxyingenol-6alpha,7alpha-epoxide 3-angelate). From goats milk collected 15 days after cessation of the experimental feeding period, extracts were completely free of diterpene ester toxins. The toxins polluting the milk are identical to diterpene ester entities occurring in the aerial parts of E. peplus. Of these, Euphorbia factors Pel and Pe2 are known as promoters of tumors of mouse skin. Apart from the toxic Euphorbia factors, the non-toxic parent alcohol ingenol was also detected in the milk. It is absent in the plant, and may have been generated metabolically from a certain portion of the toxic diterpene esters picked up by the goats from their fodder. The results presented here provide, for the first time, data for a novel interpretation of the locally high incidence of esophageal cancer observed in certain areas in the Caspian littoral of Iran, associated with a greater consumption of goat's (and sheep's) milk.