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Heavy metals present in water environment and hazardous sites as single compounds or mixture may drastically affect human health. In the present work, we investigated the risk assessment of wastewater effluents and leachate with a focus on three heavy metals-nickel (Ni), cadmium (Cd), and lead (Pb)-and their combined effect on mammalian cells, using Chinese hamster ovary cells transfected with the heat-shock protein (HSP) 47 promoter. The heavy metal mixture model was designed based on the concentrations of metals in wastewater effluents and leachate sampled in Tunisia. Using a ternary diagram, we investigated the stress response of the interaction model. This research indicated that the single heavy metals induced the stress response on HSP(+) cells even at concentrations lower than the local and international guidelines. Differences in water quality likely influenced the metal responses such that the organic composition of the leachate increased the stress response induced by the heavy metals exclusively, whereas the effluents included organic compounds that were able to mask the heavy metal effect. The mixture characterization discovered the key role played by the high levels of Ni or combination of Cd and Pb to induce the highest stress response following 3-h incubation. Heat-shock protein 47 has proven its effectiveness for assessing the heavy metal mixture effect even at low concentrations. Furthermore, the combination of a bioassay system with a statistical model proved extremely useful for better understanding the major contributors to the stress response of the mixture., (Copyright (c) 2010 SETAC.)

The physicochemical and biological properties of fulvic acid extracted and purified from excess sludge and solubilized excess sludge were studied. Solubilization was introduced to improve the recovery rate of fulvic acid from the sludge. The structural features of fulvic acid from excess sludge and solubilized excess sludge were characterized by using an elemental analysis, Fourier transform infrared spectroscopy and (1)H-nuclear magnetic resonance spectroscopy, and were compared with fulvic acid extracted from peat which had an inhibitory effect on the type I allergy in our previous study. The results show that they had a higher aliphatic characteristic with lower oxygen group content than fulvic acid from peat, and that the aliphatic characteristic was further strengthened by the use of solubilization. The biological properties of fulvic acid from excess sludge and solubilized excess sludge showed an inhibitory effect on beta-hexosaminidase release at the antigen-antibody binding stage and antigen-receptor binding stage by using rat basophilic leukemia cells.

Indoleacetic acid falcarindiol ester (compound 1) has previously been isolated and purified using an SiO2 column and ODS HPLC from an acetone extract of Japanese ivy (Hedera rhombea). Here we investigate the differentiation-inducing activity of compound 1 using the human promyelocytic leukemia HL-60 cell line. The effect of compound 1 on HL-60 cell viability and proliferation was determined at different treatment times using the 3-(4,5-dimethythiazol-2-yl)-2,5-diohenyl-2 H-tetrazolium bromide (MTT) assay and flow cytometry analysis. Also cell cycle kinetics were examined using propidium iodide staining of DNA. Cell differentiation was assessed by specific and non-specific esterase double staining assays, and by detection of the cell surface differentiation markers CD11b and CD14 using flow cytometry. The results showed HL-60 cell growth inhibition at 0.1 and 1.0 microg/mL compound 1, whereas 10 microg/mL was cytotoxic. The growth suppression induced by compound 1 was accompanied by G0/G1 phase arrest in the cell cycle at 1.0 microg/mL. Moreover, staining and immunochemical analysis indicated that compound 1 induced granulocytic differentiation in HL-60 cells. This is the first report describing granulocytic differentiation activity of a falcarindiol derived polyacetylenic compound on leukemia cells.