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See detailFood interactions : effects on health, consumer perception and impact on agro-food industries “FOODINTER”.
Scippo, Marie-Louise ULg; Blust, Ronny; Boniver, Delphine et al

Report (2011)

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See detailPotential of an in vitro toolbox combined with exposure data as a first step for the risk assessment of dietary chemical contaminants
Ribonnet, Laurence; Van Der Heiden, Edwige ULg; Nobels, Ingrid et al

in Food Additives & Contaminants (2011), 28(9), 1136-1158

In vitro risk assessment of dietary contaminants has become a priority in human food safety. This paper proposes an in vitro approach associating different complementary tools in an original toolbox and ... [more ▼]

In vitro risk assessment of dietary contaminants has become a priority in human food safety. This paper proposes an in vitro approach associating different complementary tools in an original toolbox and aims to improve the assessment of the toxicological impact of dietary contaminants at realistic human exposure levels, with a special focus on the intestinal compartment. The system is based on the use of four complementary cellular tools, namely stress gene induction in transgenic strains of Escherichia coli, modulation of the activity of key biotransformation enzymes (cytochrome P-450 (CYP) 1A1 and 3A4) in a human intestinal cell line, and activation of aryl hydrocarbon receptor (AhR) and oestrogenic receptor (ER)-dependent genes in agonistic and antagonistic assays with luciferase reporter cells. It was applied to four chosen model molecules: ochratoxin A (OTA) and deoxynivalenol (DON), two common food-borne mycotoxins, and imazalil (IMA) and benomyl (BEN), two fungicides widely occurring in foodstuffs. All these assays were performed at or around a realistic intestinal concentration, determined through a deterministic approach based on the calculation of a theoretical maximum daily intake (TMDI). Using the four model molecules, it is clearly highlighted that induction of CYP1A1 activity and inhibition of CYP3A4 activity occurred in Caco-2 cells at a realistic intestinal concentration of IMA. Furthermore, some bacterial stress genes were induced in a range of realistic concentrations, following exposure to DON and IMA. In addition, BEN clearly provoked an ER agonistic activity in a human oestrogen sensitive reporter cell line. All these results are in accordance with the literature, suggesting that the in vitro toolbox constitutes an interesting approach in order to obtain a first 'fingerprint' of dietary contaminants at realistic human exposure for further risk assessment. [less ▲]

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See detailModulation of CYP1A1 activity by a Ginkgo biloba extract in the human intestinal Caco-2 cells
Ribonnet, Laurence; Callebaut, Alfons; Nobels, Ingrid et al

in Toxicology Letters (2011), 202(3), 193-202

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See detailRelationships between vitamin A and PCBs in grey seal mothers and pups during lactation
Vanden Berghe, Marie; Mat, Audrey; Arriola, Aline et al

in Environmental Pollution (2010), 158(5), 1570-1575

A previous study has shown a simultaneous increase of vitamin A and PCBs in grey seal (Halichoerus grypus) milk at late lactation (Debier et al., 2004). Here we sought to understand this unexpected ... [more ▼]

A previous study has shown a simultaneous increase of vitamin A and PCBs in grey seal (Halichoerus grypus) milk at late lactation (Debier et al., 2004). Here we sought to understand this unexpected relationship by comparing the dynamics of vitamin A and PCBs in the different tissue compartments of transfer. Lactating grey seals and their pups were sampled longitudinally in Scotland during the 2006 breeding season. As blubber reserves decreased, concentrations of vitamin A and PCBs increased during lactation in the inner layer of maternal blubber. A concomitant rise was observed in milk and consequently in the serum of suckling pups. The similar dynamics of vitamin A and PCBs in milk and inner blubber suggest a common mechanism of mobilisation from maternal body stores and transfer into the milk. A panel data analysis highlighted a negative impact of PCBs in milk and pup serum on vitamin A status in pup serum. © 2009 Elsevier Ltd. All rights reserved. [less ▲]

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See detailCYP1A1 and CYP3A4 modulation by dietary flavonoids in human intestinal Caco-2 cells.
Sergent, Thérèse; Dupont, Isabelle; Van Der Heiden, Edwige ULg et al

in Toxicology Letters (2009), 191

Flavonoids have been proposed to exert beneficial effects in a multitude of disease states. However, evidence of potential toxic actions has also emerged. Since large doses of flavonoids can be ... [more ▼]

Flavonoids have been proposed to exert beneficial effects in a multitude of disease states. However, evidence of potential toxic actions has also emerged. Since large doses of flavonoids can be encountered in the intestine simultaneously with ingested drugs and pollutants, this study aimed at investigating nine individual flavonoid compounds and their interactions with the major intestinal isoforms of cytochrome P450, i.e. CYPs 1A1 and 3A4, using human intestinal Caco-2 cells cultivated in a serum-free medium. Genistein, quercetin and chrysin provoked a dose-dependent inducing effect on the CYP1A1 activity, measured with the EROD assay. However, they did not affect the CYP1A1 mRNA expression, suggesting they are not aryl hydrocarbon receptor-ligands in intestinal cells and act at a post-transcriptional level. Chrysin, at 50muM, was detected as a potent inhibitor of the TCDD-induced CYP1A1 activity, leading the activity to ca. 10% of the TCDD-control value (n=3), this effect involving, at least partly, direct interactions at the enzyme level. Quercetin was also shown to significantly inhibit the constitutive CYP3A4 activity, measured by the 6beta-(OH)-testosterone assay, and to impair its induction by 1,25-vitamin D(3). Chrysin, quercetin and genistein, were detected as significant inhibitors of the 1,25-vitamin D(3)-induced CYP3A4 activity. In vivo, these effects could result in reduced activation of procarcinogens and/or in drug bioavailability limitation. They underline the importance of intestinal studies to assess food safety and health risks linked to the ingestion of flavonoid-enriched supplements or functional foods. [less ▲]

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See detailCYP1A1 induction and CYP3A4 inhibition by the fungicide imazalil in the human intestinal Caco-2 cells-Comparison with other conazole pesticides
Sergent, Thérèse; Dupont, Isabelle; Jassogne, Coralie et al

in Toxicology Letters (2009), 184(3), 159-168

Imazalil (IMA) is a widely used imidazole-antifungal pesticide and. therefore. a food contaminant. This compound is also used as a drug (enilconazole). As intestine is the first site of exposure to ... [more ▼]

Imazalil (IMA) is a widely used imidazole-antifungal pesticide and. therefore. a food contaminant. This compound is also used as a drug (enilconazole). As intestine is the first site of exposure to ingested drugs and pollutants, we have investigated the effects of IMA, at realistic intestinal concentrations, on xenobiotic-metabolizing enzymes and efflux pumps by using Caco-2 cells, as a validated in vitro model of the human intestinal absorptive epithelium. For comparison, other conazole fungicides, i.e. ketoconazole, propiconazole and tebuconazole. were also studied. IMA induced cytochrome P450 (CYP) 1A1 activity to the same extent as benzo(a)pyrene (B(a)P) or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), in a dose-and time-dependent manner. Cell-free aryl hydrocarbon receptor (AhR) binding assay and reporter gene assay suggested that IMA is not an AhR-ligand, implying that IMA-mediated induction should involve an AhR-independent pathway. Moreover, IMA strongly inhibited the CYP3A4 activity in 1,25-vitamin D-3-induced Caco-2 cells. The other fungicides had weak or nil effects on CYP activities. Study of the apical efflux pump activities revealed that ketoconazole inhibited both P-glycoprotein (Pgp) and multidrug resistance-associated protein 2 (MRP-2) or breast cancer resistance protein (BCRP), whereas IMA and other fungicides did not. Our results imply that coingestion of IMA-contaminated food and CYP3A4- or CYP1A1-metabolizable drugs or chemicals could lead to drug bioavailability modulation or toxicological interactions, with possible adverse effects for human health. (C) 2008 Elsevier Ireland Ltd. All rights reserved. [less ▲]

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See detailLC-MS/MS multi-analyte method for mycotoxin determination in food supplements
Di Mavungu, Jose Diana; Monbaliu, Sofie; Scippo, Marie-Louise ULg et al

in Food Additives & Contaminants (2009), 26(6), 885-895

A multi-analyte method for the liquid chromatography-tandem mass spectrometric determination of mycotoxins in food supplements is presented. The analytes included A and B trichothecenes (nivalenol ... [more ▼]

A multi-analyte method for the liquid chromatography-tandem mass spectrometric determination of mycotoxins in food supplements is presented. The analytes included A and B trichothecenes (nivalenol, deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxyvalenol, neosolaniol, fusarenon-X, diacetoxyscirpenol, HT-2 toxin and T-2 toxin), aflatoxins (aflatoxin-B-1, aflaxoin-G(1) and aflatoxin-G(2)). Alternaria toxins (alternariol, alternariol methyl ether and altenuene), fumonisins (fumonisin-B-1, fumonisin-B-2 and fumonisin-B-3), ochratoxin A, zearalenone, beauvericin and sterigmatocystin. Optimization of the stimulataneous extraction of these toxins and the sample pretreatment procedure, as well as method validation were performed on maca (Lepidium meyenii) food supplements. The results indicated that the solvent mixture ethyl acetate/formic acid (95:5, v/v) n-hexane was applied as partial clean-up step to remove excess of co-extracted non-polar components. Further clean-up was performed on Oasis HLB(TM) cartidges. Samples were analysed using an Acquity UPLC system coupled to a Micromass Quattro Micro triple quadrupole mass spectrometer equipped with an electrospray interface operated in the positive-ion mode. Limits of detection and quantification were in the range of 0.3-30 ng g(-1) and 1-100 ng g(-1), respectively. Recovery yields were above 60% for most of the analytes, except for different food supplements such as soy (Glycine max) isoflavones, St John's wort (Hypericum perforatum), garlic (Allium sativum), Ginkgo biloba, and black radish (Raphanus niger) demonstrated the general applicability of the method. Due to different matrix effects observed in different food supplement samples, the standard addition approach was applied to perform correct quantitative analysis. In 56 out of 62 samples analysed, none of the 23 mycotoxins investigated was detected. Positive samples contained at least one of the toxins fumonisin-B-1, fumonisin-B-2, fumonisin-B-3 and ochratoxin A. [less ▲]

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See detailAnalysis of EU priority polycyclic aromatic hydrocarbons in food supplements using high performance liquid chromatography coupled to an ultraviolet, diode array or fluorescence detector
Danyi, Sophie ULg; Brose, François ULg; Brasseur, Catherine ULg et al

in Analytica Chimica Acta (2009), 633

High performance liquid chromatography coupled to an ultraviolet, diode array or fluorescence detector (HPLC/UV-FLD) has been used to set up a method to detect the 15(+1) EU priority polycyclic aromatic ... [more ▼]

High performance liquid chromatography coupled to an ultraviolet, diode array or fluorescence detector (HPLC/UV-FLD) has been used to set up a method to detect the 15(+1) EU priority polycyclic aromatic hydrocarbons (PAHs) in food supplements covering the categories of dried plants and plant extracts excluding oily products. A mini validation was performed and the following parameters have been determined: limit of detection, limit of quantification, precision, recovery and linearity. They were in close agreement with quality criteria described in the Commission Regulation (EC) No 333/2007 concerning the PAH benzo[a]pyrene in foodstuffs, except the not fluorescent cyclopenta[c,d]pyrene for which the UV detection leads to a higher limit of detection. Analysis of twenty commercial food supplements covering mainly the class of dried plants was performed to evaluate their PAHs contamination levels and to test the applicability of the method to various plant matrices. Fifty percent of analyzed samples showed concentration exceeding 2 gkg−1 for one or more PAHs. [less ▲]

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See detailFood flavonoid aryl hydrocarbon receptor-mediated agonistic/antagonistic/synergic activities in human and rat reporter gene assays
Van Der Heiden, Edwige ULg; Bechoux, Nathalie ULg; Muller, Marc ULg et al

in Analytica Chimica Acta (2009), 637

Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor mediating the adverse effects of dioxins and polycyclic aromatic hydrocarbons (PAHs). In this study, we investigated the genetic ... [more ▼]

Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor mediating the adverse effects of dioxins and polycyclic aromatic hydrocarbons (PAHs). In this study, we investigated the genetic-, time-, dose-, species- and tissue-dependent AhR-mediated agonistic/ antagonistic activities of three food flavonoids: quercetin, chrysin and genistein. To that end, four stably transfected cell lines were used in cell-based luciferase reporter gene assays: three lines were transformed with the ptKLuc vector harbouring four dioxinresponsive elements (DREs) upstream of the thymidine kinase promoter and the luciferase gene (HepG2-Luc, T-47D-Luc and H4IIE-ULg). The fourth is a patented cell line transformed with a different construct: H4IIE DR-CALUX®. Both H4IIE cells were compared for their genetic construction. Human hepatoma (HepG2-Luc) and human breast tumour (T-47D-Luc) cells were compared for tissue-dependent effects. Rat hepatoma (H4IIE-ULg) and human hepatoma (HepG2-Luc) cellswere compared for species-dependent activities.We concluded that quercetin, chrysin and genistein act in a time-, dose-, species- and tissue-specific way. For example, genistein displayed agonistic activities when exposed to rat hepatoma cells during 6h but not after 24 h. Flavonoids displayed agonistic/antagonistic activities in human breast tumour cells, depending on the exposure time, while in human hepatoma cells, only antagonistic activities of flavonoids were measured. In addition, we report, in all the cells, a synergy between an isoflavone and two food contaminants; the 2,3,7,8-tetrachlorodibenzop- dioxin and 3-methylcholanthrene, a PAH. In rat cells, this synergy occurred when cells were exposed to flavonoids and contaminant for 6h, while it was observed in human cells only after 24 h. [less ▲]

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See detailModulation of cytochrome P450 1A1 expression and activity in intestinal Caco-2 cells by components of Ginkgo biloba-based dietary supplements
Ribonnet, Laurence; Callebaut, Alfons; Scippo, Marie-Louise ULg et al

in Toxicology Letters (2009), 189

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See detailEstablishing a Belgian Nutrition Society (BNS): Filling The Void
Cani, Patrice; Clarys, Peter; Clinquart, Antoine ULg et al

in Archives of Public Health (2009), 67

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See detailImazalil modulates CYPs 1A1 and 3A4 activities in the human Caco-2 cells as an intestinal model to assess food safety
Sergent, Thérèse; Ribonnet, Laurence; Jassogne, C. et al

in Toxicology Letters (2007), 172

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See detailMaintenance threonine requirement and efficiency of its use for accretion of whole-body threonine and protein in Atlantic salmon (Salmo salar L.) fry.
Rollin, Xavier; Wauters, Jean*-Baptiste; Bodin, Noe Lie et al

in British Journal of Nutrition (2006), 95(2), 234-45

Eighteen groups of seventy Atlantic salmon (Salmo salar L.) fry (initial mean body weight 0.8 (sd 0.01) g) were fed on semi-purified diets containing graded levels of l-threonine (Thr) in 15 litres ... [more ▼]

Eighteen groups of seventy Atlantic salmon (Salmo salar L.) fry (initial mean body weight 0.8 (sd 0.01) g) were fed on semi-purified diets containing graded levels of l-threonine (Thr) in 15 litres aquaria at a temperature of 14.5+/-1 degrees C. Doses of Thr represented 1, 31, 41, 51, 62, 72, 83 and 93 % of its ideal level for optimum protein deposition. Indispensable amino acids other than Thr were included in the same proportion (on a g/16 g N basis) as in the Atlantic salmon fry whole-body carcass. Following 36 d of feeding and a 36 h fast, fry were killed for whole-body protein and amino acid analysis. Weight gain (r2 0.98), protein accretion (r2 0.97), and Thr accretion (r2 0.97) were linear (P<0.01) functions of Thr intake. Slope of the Thr accretion regression line showed that the efficiency of Thr utilisation above maintenance was 76 %. At zero Thr intake, fry lost 5.4 mg Thr/kg body weight0.75 per d. The Thr maintenance requirement was 7.2 mg/kg body weight0.75 per d and the Thr requirement for growth was 66 mg for 1 g protein deposition. Increasing doses of Thr resulted in increased (P<0.05) concentrations of histidine and lysine, and decreased concentrations of isoleucine in whole-body protein. The maintenance need for Thr represented 13.4 % of the total need for Thr. The data suggest that efficiency of Thr utilisation above maintenance is constant at all levels of Thr intake between 1 and 93 % of the level required for optimum protein deposition. [less ▲]

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See detailDynamics of PCB transfer from mother to pup during lactation in UK grey seals Halichoerus grypus: differences in PCB profile between compartments of transfer and changes during the lactation period
Debier, Cathy; Pomeroy, Paddy P.; Dupont, Cédric et al

in Marine Ecology-Progress Series (2003), 247

The composition of the PCB mixture which is transferred from mother to pup in grey seals Halichoerus grypus throughout lactation was measured in the successive compartments of transfer: maternal blubber ... [more ▼]

The composition of the PCB mixture which is transferred from mother to pup in grey seals Halichoerus grypus throughout lactation was measured in the successive compartments of transfer: maternal blubber, maternal serum, milk and pup serum. Mother-pup pairs were captured regularly between birth and weaning on the Isle of May, Scotland. The PCB profiles of the different compartments of transfer consisted mainly of congeners 153, 138, 180, 187, 170 and 101, which made up 85% of the total PCB burden. Outer blubber contained higher relative concentrations (R-153) Of hepta-, octa-, and nona-chlorinated congeners (higher chlorinated congeners) than inner blubber, at both early (less than or equal to 5 d) and late (greater than or equal to 11 d) lactation. There was no change in the composition of the outer blubber layer between early and late lactation, while in the inner blubber, there was a slight increase in the R153 of higher chlorinated congeners at late lactation. In maternal serum, profiles changed towards having a higher proportion of higher chlorinated congeners as lactation progressed, but in milk, the PCB pattern stayed constant throughout lactation. Surprisingly, in pup serum profiles also changed towards higher R153 of higher chlorinated congeners at late lactation compared to early lactation, which was different to the pattern observed in milk. In all body compartments, higher proportions of higher chlorinated congeners were associated with higher concentrations of total PCBs. Maternal blubber and pup serum contained a higher R153 of higher chlorinated congeners than milk and, to a lesser extent, maternal serum. Thus biomagnification of the higher chlorinated congeners was recorded in pup serum. [less ▲]

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See detailQuantitative dynamics of PCB transfer from mother to pup during lactation in UK grey seals Halichoerus grypus
Debier, Cathy; Pomeroy, Paddy P.; Dupont, Cédric et al

in Marine Ecology-Progress Series (2003), 247

PCB contamination was measured in the milk and serum of grey seal Halichoerus grypus mothers and in the serum of their pups sampled from 2 to 5 times between parturition and weaning on the Isle of May ... [more ▼]

PCB contamination was measured in the milk and serum of grey seal Halichoerus grypus mothers and in the serum of their pups sampled from 2 to 5 times between parturition and weaning on the Isle of May, Scotland, in 1998 and 2000. Blubber biopsies were also taken from the lactating females at early and late lactation in 2000. Concentrations of PCBs in milk stayed constant during the first part of lactation (0.31 +/- 0.17 mug g milk(-1)) and then increased at late lactation (0.67 +/- 0.42 mug g milk(-1)). Curiously, it did not follow the changes of milk lipid content, which increased at early lactation and then stayed constant until the end of the nursing period. As a result, even when expressed per unit of milk lipids, PCBs underwent a rise at the end of lactation. The changes in milk PCBs were accompanied by similar dynamics in maternal serum as well as in pup serum. Increased concentrations of PCBs at late lactation in serum and milk may be explained in part by the changes observed in maternal blubber. PCB levels increased significantly between early and late lactation in inner blubber, suggesting that PCBs are less easily mobilised from blubber than lipids. At late lactation, the retention capacity of the reduced blubber layer for PCBs might have reached its maximum. The mobilization of less polar lipids from blubber might also occur at this stage. In both cases, this could result in a higher mobilization of PCBs at this time. While inner blubber was significantly less concentrated than outer blubber at early lactation (1.26 +/- 0.72 mug g lipids(-1) in inner blubber vs 3.16 +/- 1.34 mug g lipids(-1) in outer blubber), these variations disappeared at late lactation (3.24 +/- 2.60 mug g lipids(-1) in inner blubber vs 3.59 +/- 1.46 mug g(-1) lipids in outer blubber). Newborn pups already had significantly higher serum levels of PCBs than their mothers, revealing an important placental transfer (11.9 +/- 7.0 ng ml serum(-1) in pups vs 6.7 +/- 3.5 ng ml serum(-1) in mothers). These differences were even greater in late lactation, due to the ingestion of milk (27.9 +/- 18.1 ng ml serum(-1) in pups vs 12.2 +/- 7.2 ng ml serum(-1) in mothers). As lactation progressed, PCB levels in pup serum increased exponentially as compared to the levels in the serum of their mothers. [less ▲]

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