A rapid and sensitive cytotoxicity screening assay for trichothecenes in cereal samples. 2003

J Widestrand, and T Lundh, and H Pettersson, and J E Lindberg
Department of Animal Nutrition and Management, Swedish University of Agricultural Sciences, S-75007 Uppsala, Sweden.

The application of a bioassay for toxicity screening of cereal samples has been studied. The sensitive Swiss mouse 3T3 fibroblasts (3T3 cells) were used to determine the cytotoxicity of Fusarium trichothecenes T-2 toxin (T-2), HT-2 toxin (HT-2), deoxynivalenol (DON) and nivalenol (NIV) in cereal samples. Clean-up of the sample extracts was performed using the MycoSep #225 column and the cytotoxic effects of the samples were determined using the 5-bromo-2'-deoxyuridine (BrdU) bioassay assessing DNA-synthesis. Trichothecene-free samples of wheat, wheat bran and barley had no toxic effect on the 3T3 cells at sample concentrations up to 400 mg/ml cell culture medium while a few oat samples had an inhibiting effect on the DNA-synthesis. IC(50) values (50% response compared with untreated cells) of trichothecene-free wheat and oats spiked with T-2, HT-2, DON and NIV were similar to IC(50) values of pure toxins. A cytotoxicity screening of 33 different cereal samples showed that all samples contaminated with a trichothecene concentration higher than the IC(50) values of T-2, HT-2, DON and NIV had a clear inhibiting effect on DNA-synthesis of the 3T3 cells. The culture of 3T3 cells on microtiter plates in combination with the simple and rapid sample clean-up using the MycoSep #225 column provides a sensitive and easy method for the screening of trichothecenes in cereal samples.

UI MeSH Term Description Entries
D007202 Indicators and Reagents Substances used for the detection, identification, analysis, etc. of chemical, biological, or pathologic processes or conditions. Indicators are substances that change in physical appearance, e.g., color, at or approaching the endpoint of a chemical titration, e.g., on the passage between acidity and alkalinity. Reagents are substances used for the detection or determination of another substance by chemical or microscopical means, especially analysis. Types of reagents are precipitants, solvents, oxidizers, reducers, fluxes, and colorimetric reagents. (From Grant & Hackh's Chemical Dictionary, 5th ed, p301, p499) Indicator,Reagent,Reagents,Indicators,Reagents and Indicators
D009183 Mycotoxins Toxic compounds produced by FUNGI. Fungal Toxins,Mycotoxin,Toxins, Fungal
D011500 Protein Synthesis Inhibitors Compounds which inhibit the synthesis of proteins. They are usually ANTI-BACTERIAL AGENTS or toxins. Mechanism of the action of inhibition includes the interruption of peptide-chain elongation, the blocking the A site of ribosomes, the misreading of the genetic code or the prevention of the attachment of oligosaccharide side chains to glycoproteins. Protein Synthesis Antagonist,Protein Synthesis Antagonists,Protein Synthesis Inhibitor,Antagonist, Protein Synthesis,Antagonists, Protein Synthesis,Inhibitor, Protein Synthesis,Inhibitors, Protein Synthesis,Synthesis Antagonist, Protein,Synthesis Inhibitor, Protein
D001973 Bromodeoxyuridine A nucleoside that substitutes for thymidine in DNA and thus acts as an antimetabolite. It causes breaks in chromosomes and has been proposed as an antiviral and antineoplastic agent. It has been given orphan drug status for use in the treatment of primary brain tumors. BUdR,BrdU,Bromouracil Deoxyriboside,Broxuridine,5-Bromo-2'-deoxyuridine,5-Bromodeoxyuridine,NSC-38297,5 Bromo 2' deoxyuridine,5 Bromodeoxyuridine,Deoxyriboside, Bromouracil
D002470 Cell Survival The span of viability of a cell characterized by the capacity to perform certain functions such as metabolism, growth, reproduction, some form of responsiveness, and adaptability. Cell Viability,Cell Viabilities,Survival, Cell,Viabilities, Cell,Viability, Cell
D002523 Edible Grain SEEDS used as a major nutritional source, most often the grain from the POACEAE family. Cereals,Cereal Grain,Cereal,Cereal Grains,Edible Grains,Grain, Cereal,Grain, Edible,Grains, Cereal,Grains, Edible
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D000963 Antimetabolites Drugs that are chemically similar to naturally occurring metabolites, but differ enough to interfere with normal metabolic pathways. (From AMA Drug Evaluations Annual, 1994, p2033) Antimetabolite
D001467 Hordeum A plant genus of the family POACEAE. The EDIBLE GRAIN, barley, is widely used as food. Barley,Hordeum vulgare
D001681 Biological Assay A method of measuring the effects of a biologically active substance using an intermediate in vivo or in vitro tissue or cell model under controlled conditions. It includes virulence studies in animal fetuses in utero, mouse convulsion bioassay of insulin, quantitation of tumor-initiator systems in mouse skin, calculation of potentiating effects of a hormonal factor in an isolated strip of contracting stomach muscle, etc. Bioassay,Assay, Biological,Assays, Biological,Biologic Assay,Biologic Assays,Assay, Biologic,Assays, Biologic,Bioassays,Biological Assays

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