María José Ruiz

Effects of four carbamate compounds on antioxidant parameters.

The effect of four carbamates, aldicarb and its metabolites (aldicarb sulfone and aldicarb sulfoxide) and propoxur on glutathione content and the activity of the enzymes involved in the sulfur-redox cycle in the mammalian cellular model CHO-K1 cells after 24-h exposure were determined. Carbamate exposure resulted in a depletion of intracellular reduced glutathione (GSH) content, no change was observed in oxidized glutathione (GSSG) and a decrease in GSH/GSSG ratio was detected. After carbamates exposition a GSH/GSSG decreases in ranged from 12.44% to 21.35% of control was observed. Depletion of GSH levels was accompanied by the induction of glutathione reductase (GR) after 24h exposure with each of the four carbamates to CHO-K1 cells. After aldicarb sulfone, aldicarb sulfoxide, and propoxur exposure, glutathione peroxidase (GPx) activity increased in CHO-K1 cells by 198%, 32%, and 228% of control, respectively. After aldicarb sulfone and propoxur exposure, glutathione transferase (GST) activities increased by 49% and 230% of control, respectively. Due to the role played by GSH in preventing cytotoxicity via free-radical scavenging, results obtained suggest that high concentrations of aldicarb sulfone and propoxur closely resembling oxidative stress in CHO-K1 cells.

Exposure estimates to Fusarium mycotoxins through cereals intake

Mycotoxins are harmful substances produced by fungi in several commodities with a widespread presence in foodstuffs. Human exposure to mycotoxins occurs mainly by contaminated food. The quantitation of mycotoxins in cereal-based food, highly consumed by different age population, is of concern. In this survey, 159 cereal-based samples classified as wheat, maize and rice-based, have been evaluated for the occurrence of patulin, deoxynivalenol, 3-acetyl-deoxynivalenol, fusarenon-X, diacetoxyscirpenol, nivalenol, neosolaniol, HT-2, T-2 and zearalenone by gas chromatography-tandem mass spectrometry. Intakes were calculated for average consumers among adults, children and infants and compared with the tolerable daily intakes (TDI). Data obtained were used to estimate the potential exposure levels. 65.4% of the samples were contaminated with at least one mycotoxin and 15.7% of the analyzed samples showed co-occurrence of mycotoxin. The dietary exposure to HT-2 and T-2 toxins was estimated as 0.010 and 0.086 μg kg(-1) bw d(-1), amounting to 10% and 86% of the TDI, for adults and infants respectively. These results back up the necessity to take a vigilant attitude in order to minimize human intake of mycotoxins.

Application of capillary electrophoresis-mass spectrometry for determining organic food contaminants and residues.

Food contamination continues to be a serious problem around the world. Surveillance of chemical contaminants in foods is important not only for public health but also because of the negative economic impact of contamination. From the analytical perspective, analysis of contaminants in food is an extremely challenging area. There is a wide variety of questions, ranging from the quantification of extremely low levels of individual components to the detailed assessment and evaluation of the analytical technique possibilities. This review considers the applications of CE coupled to MS detection (CE‐MS) for the analysis of organic contaminants in food. Analytical information on sample concentration techniques, as well as on the CE separation conditions and recoveries obtained from water and food are provided. Different sections include several fields of application, such as pesticides, drug residues, or toxic formed during food processing in different matrices. A number of tables report a comprehensive listing of CE‐MS applications. As a result, this work presents an update overview on the principal application of CE‐MS together with a discussion of their main advantages and drawbacks, and an outline of future trends on analysis of organic contaminants.

Comparison of basal cytotoxicity of seven carbamates in CHO-K1 cells

The cytotoxic effects of seven carbamate pesticides, aldicarb, aldicarb sulfone, aldicarb sulfoxide, benfuracarb, pirimicarb, propoxur and thiobencarb, were compared in Chinese hamster ovary (CHO-K1) cell line of Circetulus griseus. The endpoints evaluated were lysosomal function by neutral red assay and mitochondrial integrity by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT assay). The carbamates tested were evaluated in both serum-free medium and in serum-containing medium. Results demonstrate that CHO-K1 lysosomes appeared more susceptible to propoxur, aldicarb and its metabolites than mitochondria. Aldicarb was the most toxic carbamate pesticide tested on CHO-K1 cells and aldicarb sulfoxide the least. The presence of foetal calf serum (FCS) diminished significantly the cytotoxicity of tested carbamates.

Formation of Fumonisin B1−Glucose Reaction Product, in Vitro Cytotoxicity, and Lipid Peroxidation on Kidney Cells

Fumonisin B(1) (FB(1)) content in corn products decreases during the heating process in foods containing reducing sugars, mainly because of the formation of N-(carboxymethyl)fumonisin B(1). In this study, a rapid method has been developed for the determination of both compounds in corn products using a high-speed blender, Ultra-Turrax, for solvent extraction and liquid chromatography-tandem mass spectrometry. The kinetics of FB(1) degradation and the formation of the Maillard adduct were studied in a model system constituted by corn bread spiked with FB(1) and heated at 160, 180, and 200 degrees C for 3, 6, 10, 15, and 20 min. FB(1) decreased from 0.96 to 0.3 mg/kg and N-(carboxymethyl)fumonisin B(1) increased to 0.1 mg/kg. Cytotoxicity and lipid peroxidation were studied in monkey kidney cells (Vero cells). After 24 h exposure, FB(1) revealed an IC(50) (median inhibitory concentration) of 55 +/- 7 microM with neutral red uptake, but no IC(50) was obtained after N-(carboxymethyl)fumonisin B(1) exposure at the studied concentrations. Lipid peroxidation was assessed using the thiobarbituric acid reactive substance (TBARS) method for 90 min and 24 and 48 h. FB(1) significantly increased the production of malondialdehyde in Vero cells exposed to 1 microM FB(1) after 24 h, while malondialdehyde increased after 5 microM N-(carboxymethyl)fumonisin B(1) exposure. These findings showed that the transformation products exhibit lower cytotoxicity than fumonisin B(1) and lipid peroxidation may be involved in the cytotoxicity induced by both toxins.

Antibacterial activity of the enniatin B, produced by Fusarium tricinctum in liquid culture, and cytotoxic effects on Caco-2 cells.

The enniatins (ENs) are bioactive compounds of hexadepsipeptidic structure produced by several strains of Fusarium sp. The EN B was purified from extracts of Fusarium tricinctum growth on liquid culture of potato dextrose broth (PDB), using a semipreparative liquid chromatography (LC) followed by an analytical LC. The purity and the structure of the isolated compound were confirmed by the determination of the extinction coefficient and with electrospray ionization–mass spectrometry (ESI-MS) study. The pure fraction of EN B was utilized to determine the antibiotic effects on several bacterial strains that are considered normally pathogens of the intestinal tract: Escherichia coli, Enterococcus faecium, Salmonella enterica, Shigella dysenteriae, Listeria monocytogenes, Yersinia enterocolitica, Clostridium perfringens, Pseudomonas aeruginosa, and Staphylococcus aureus, and to study the cytotoxic effects on Caco-2 differentiated and undifferentiated cells. The results obtained demonstrated that in several antibiograms, EN B induced the inhibition of the grown microorganisms tested and no significant differences over control were detected when Caco-2 cells were exposed to EN B, at any of the concentrations used.

A Review of the Mycotoxin Enniatin B

Mycotoxin enniatin B (ENN B) is a secondary metabolism product by Fusarium fungi. It is a well-known antibacterial, antihelmintic, antifungal, herbicidal, and insecticidal compound. It has been found as a contaminant in several food commodities, particularly in cereal grains, co-occurring also with other mycotoxins. The primary mechanism of action of ENN B is mainly due to its ionophoric characteristics, but the exact mechanism is still unclear. In the last two decades, it has been a topic of great interest since its potent mammalian cytotoxic activity was demonstrated in several mammalian cell lines. Moreover, the co-exposure in vitro with other mycotoxins enhances its toxic potential through synergic effects, depending on the concentrations tested. Despite its clear cytotoxic effect, European Food Safety Authority stated that acute exposure to ENNs, such as ENN B, does not indicate concern for human health, but a concern might be the chronic exposure. However, given the lack of relevant toxicity data, no firm conclusion could be drawn and a risk assessment was not possible. In fact, very few studies have been carried out in vivo and, in these studies, no adverse effects were observed. So, research on toxicological effects induced by ENN B is still on-going. Recently, some studies are dealing with new advances regarding ENN B. This review summarizes the information on biochemical and biological activity of ENN B, focusing on toxicological aspects and on the latest advances in research on ENN B.

Production, purification, and mass spectrometry characterization of the cyclohexadepsipeptide enniatin J3 and study of the cytoxicity on differentiated and undifferentiated Caco-2 cells

The enniatins (EN) are bioactive compounds of hexadepsipeptidic structure produced by several strains of Fusarium spp. The enniatin J3 (EN J3) was purified from extracts of Fusarium solani growth on solid medium of kamut, using semipreparative liquid chromatography (LC) followed by an analytical LC. The purity and the structure of the isolated compound were confirmed by electrospray ionization-mass spectrometry study-linear ion trap (ESI-MS-LIT). The pure fraction of EN J3 was utilized to study the cytotoxicity on differentiated and undifferentiated Caco-2 cells. The use of both chromatographic techniques allowed us to produce and purify 50 mg of EN J3 completely characterized with the technique of the ESI-MS-LIT. No statistically significant differences over control were detected on Caco-2 cells exposed to EN J3 at any of the concentrations used.

Antioxidant capacity of trans-resveratrol dietary supplements alone or combined with the mycotoxin beauvericin

Trans-resveratrol (trans-RSV) is a polyphenol with multiples biological properties, such as anti-inflammatory, antioxidant, anti-aging, anti-diabetic, and antiplatelet. It occurs naturally in grapes and derivate, peanuts and berries. Beauvericin (BEA) is a mycotoxin present in cereals that produces cytotoxicity, intracellular reactive oxygen species and lipid peroxidation. The general objective of this research was to evaluate whether trans-RSV could be used as a good polyphenol against damages produced by BEA. Because trans-RSV can be ingested through dietary supplements, to reach this goal, the following specific objectives were proposed: to determine a) the trans-RSV content in different polyphenol dietary supplements by capillary electrophoresis, b) the antioxidant capacity of the trans-RSV in polyphenol supplements, and c) the influence of BEA in the antioxidant capacity of trans-RSV when they are in combination by photochemioluminiscence assay. The results obtained in this study showed that all polyphenol dietary supplements present higher RSV content that the content of the label. The polyphenol supplements present antioxidant capacity. And the combination of trans-RSV and BEA did not affect the antioxidant capacity of trans-RSV. Thus, RSV could contribute to decrease oxidant effects produced by BEA.

In vitro mechanisms of Beauvericin toxicity: A review

Beauvericin (BEA) is a mycotoxin produced by many species of fungus Fusarium and by Beauveria bassiana; BEA is a natural contaminant of cereals and cereals based products and possesses a wide variety of biological properties. The mechanism of action seems to be related to its ionophoric activity, that increases ion permeability in biological membranes. As a consequence, BEA causes cytotoxicity in several cell lines and is capable to produce oxidative stress at molecular level. Moreover, BEA is genotoxic (produces DNA fragmentation, chromosomal aberrations and micronucleus) and causes apoptosis with the involvement of mitochondrial pathway. However, several antioxidant mechanisms protect cells against oxidative stress produced by BEA. Despite its strong cytotoxicity, no risk assessment have been still carried out by authorities due to a lack of toxicity data, so research on BEA toxicological impact is still going on. This review reports information available regarding BEA mechanistic toxicology with the aim of updating information regarding last researches on this mycotoxin.