citrinin and thiazolyl-blue

Food safety organizations indicate the likelihood of constant human and animal exposure to mycotoxin mixtures as a possible negative public health impact. Risk assessment demonstrates that certain mycotoxins of Aspergillus and Penicillium spp. are toxic and hold a significant genotoxic efficacy at nanomolar concentrations. The aim of the current study was to investigate the potential cytogenetic effects of sterigmatocystin (STER), ochratoxin A (OTA) and citrinin (CTN) alone or in combination, at pM to μΜ concentrations, on the human hepatocellular cancer cell line Hep3B. MTT reduction, mitotic divisions, cell cycle delays and sister chromatid exchange rates (SCE) were determined as endpoints of metabolic activity, cytotoxicity, cytostaticity, and genotoxicity, respectively. All mycotoxin treatments induce SCE rates from 10-12 M, while their cytotoxic and cytostatic potential varies. In PRI and MI assays, but not at MTT, STER alone or in combination with OTA + CTN appeared cytostatic and cytotoxic, even at 10-12 M, while CTN alone and all other combinations displayed substantial cellular survival inhibition in doses ≥ 10-8 M. Co-administration of STER + OTA or STER + CTN in concentrations ≤ 10-1 M, increased the MI and MTT activity, while it did not affect the PRI. Mycotoxin co-treatments revealed in general similar-to-additive or antagonistic genotoxic and cytotoxic effects. Our results for the first time describe that STER alone or in combination with OTA and/or CTN share a cytotoxic and cytogenetic potential even at picoMolar concentrations on human hepatoma cells in vitro.

Topics: Cell Cycle; Cell Line, Tumor; Citrinin; Humans; Mitotic Index; Mutagenicity Tests; Ochratoxins; Sister Chromatid Exchange; Sterigmatocystin; Tetrazolium Salts; Thiazoles

Topics: Antibiotics, Antineoplastic; Cell Line, Tumor; Chromatography, High Pressure Liquid; Citrinin; Culture Media; Dimerization; Fermentation; Humans; Magnetic Resonance Spectroscopy; Penicillium chrysogenum; Salts; Spectrophotometry, Ultraviolet; Tetrazolium Salts; Thiazoles

Hundreds of mycotoxins are known to date and many of them are of great interest with regard to human and animal health since they are detected frequently in plant-derived products. Various mycotoxins may occur simultaneously, depending on the environmental and substrate conditions. Considering this coincident production, it is very likely, that humans and animals are always exposed to mixtures rather than to individual compounds. Therefore, future risk assessments should consider mixture toxicity data. This is particularly true for ochratoxin A (OTA), ochratoxin B (OTB), citrinin (CIT) and occasionally for patulin (PAT) as they are all produced by a number of Penicillium and Aspergillus species. Therefore, these four toxins were chosen to study the interactive effects in vitro, using the well-established porcine renal cell line LLC-PK1 and the MTT reduction test as a cytotoxicity endpoint. By application of a step-wise approach to test combination toxicity, using various full factorial as well as a central composite experimental designs, the interactive (synergistic) cytotoxic effects of the these four toxins were assessed. The results obtained in this study confirm a potential for interactive (synergistic) effects of CIT and OTA and possibly other mycotoxins in cells of renal origin.

Topics: Algorithms; Animals; Citrinin; Drug Synergism; Indicators and Reagents; Kidney; LLC-PK1 Cells; Models, Biological; Mycotoxins; Ochratoxins; Patulin; Swine; Tetrazolium Salts; Thiazoles

1. The cytotoxicities of the nephrotoxic mycotoxins, citrinin and ochratoxin A were assayed on HeLa, C3H/10T1/2, NIH/3T3, MDCK (canine kidney), and HeLa P3 cell lines, using the MTT colorimetric assay. 2. Citrinin was less toxic than ochratoxin A in all of the cell lines examined. 3. The MDCK cells were more susceptible to both citrinin and ochratoxin A, in comparison with other cell lines. 4. Dose-responses, as measured by activities of leucine aminopeptidase and alkaline phosphatase of MDCK cells, were less sensitive than MTT colorimetric assay, indicating that these enzymes were not specifically inhibited in MDCK cells. 5. The LD50 of both toxins, calculated at 72 hr incubation, was in the same order as those reported from animal experiments using rats and mice.

Topics: Alkaline Phosphatase; Animals; Cell Line; Cell Survival; Citrinin; Colorimetry; Dogs; Humans; Lethal Dose 50; Leucyl Aminopeptidase; Mice; Mycotoxins; Ochratoxins; Succinate Dehydrogenase; Tetrazolium Salts; Thiazoles