E. Fusi

Effects of retinoids on proliferation and plasminogen activator expression in a bovine mammary epithelial cell line

Effects of two natural (retinol and retinoic acid, RA) and one synthetic N-(4-hydroxyphenyl) retinamide (4-HPR) retinoids on proliferation and expression of urokinase-plasminogen activator (u-PA) by bovine mammary epithelial cells were examined. The BME-UV1 established bovine mammary epithelial cell line was used as a model system. All retinoids tested (retinol, RA and 4-HPR) were effective inhibitors of cell proliferation. When cells were cultured in the absence of fetal bovine calf serum (FBCS), inhibition occurred at concentrations as low as 1 nM for all retinoids tested. The effect of retinoids on cell proliferation was not dose-related when cells were cultured in the absence of FBCS. All retinoids (retinol, RA, 4-HPR), when used in the range 1 nM-10 microM (noncytotoxic concentrations), were equally effective and had identical inhibition patterns. Inhibition of cell proliferation by RA was apparent by 6 h and was higher after 24 h in culture. In contrast, when cells were cultured in the presence of FBCS, the effect of RA and retinol on cell proliferation was dose-related. RA and retinol inhibited cell proliferation (P<0.01) when added to the culture medium in concentrations as low as 10 nM and 100 nM, respectively. 4-HPR was inhibitory (P<0.01) in concentrations as low as 1 nM. Higher concentrations of 4-HPR in the range 1 nM-1 microM had no further effect on cell proliferation. None of the retinoids tested, when added to cultures in the presence or absence of FBCS, could completely arrest cell proliferation at noncytotoxic concentrations. RA at 1 microM inhibited (P<0.05) insulin or IGF-I-induced cell proliferation but had no effect (P>0.05) on u-PA mRNA levels or u-PA activity. Furthermore, RA inhibited cell proliferation in the presence of FBCS but had no effect (P>0.05) on u-PA mRNA levels. Thus, retinoids are effective inhibitors of bovine mammary epithelial cell proliferation and this growth inhibition does not seem to correlate with any changes in u-PA mRNA or u-PA activity.

Sampling feed for mycotoxins: acquiring knowledge from food

Abstract The occurrence and control of mycotoxins in feed and food are items of great interest to researchers, producers, manufacturers and regulatory agencies. In order to implement knowledge of control measures for mycotoxins in the entire food production chain, coordinated inspection programmes aimed to check the presence and concentration of mycotoxins in feedingstuffs are recommended by the Commission of the European Communities. Reliability of measured levels of mycotoxins in feed and food is greatly affected by the collection of representative samples. Because of the heterogeneous distribution of mycotoxins, the variability associated with a mycotoxin test procedure usually depends heavily on the sampling plan. European legislation dealing with sampling plans for mycotoxins in foodstuffs has been recently revised. The aim of the following overview is to discuss the role of sampling in mycotoxin-contaminated feed by considering the evolution of legislation dealing with sampling plans for food. A sampling procedure is a multistage process and consists of three distinct phases: sampling, sample preparation and analysis. The variability associated with each step of a sampling procedure and the aspects related to feedstuffs, matrix/mycotoxin combination and level of contamination are discussed.

Alpha-Tocopherol Counteracts the Cytotoxicity Induced by Ochratoxin A in Primary Porcine Fibroblasts

The aims of the current study were to determine the half-lethal concentration of ochratoxin A (OTA) as well as the levels of lactate dehydrogenase release and DNA fragmentation induced by OTA in primary porcine fibroblasts, and to examine the role of α-tocopherol in counteracting its toxicity. Cells showed a dose-, time- and origin-dependent (ear vs. embryo) sensitivity to ochratoxin A. Pre-incubation for 3 h with 1 nM α-tocopherol significantly (P < 0.01) reduced OTA cytotoxicity, lactate dehydrogenase release and DNA damage in both fibroblast cultures. These findings indicate that α-tocopherol supplementation may counteract short-term OTA toxicity, supporting its defensive role in the cell membrane.

Heterologous Expression of Biologically Active Porcine Lactoferrin in Pichia Pastoris Yeast

INTRODUCTIONLactoferrin is a glycoprotein of about 80kDa, belonging to the transferrin family. It is foundin colostrum, milk and other biological fluids. This protein has antibacterial, antiviral andanticarcinogenicactivity;itisinvolvedinironabsorptionandimmunomodulationanditalsopromotes the growth of some strains of lactic acid bacteria (Brock, 2002; Kim

Rumen-protected choline and vitamin E supplementation in periparturient dairy goats: effects on milk production and folate, vitamin B 12 and vitamin E status

We investigated the effects of rumen-protected choline (RPC) and vitamin E (VITE) administration on milk production and status of folate, vitamin B12 and vitamin E during the periparturient period of dairy goats. Forty-eight Saanen multiparous goats were selected for the 72-day experiment, being moved to a maternity pen 30 days before expected parturition and assigned to one of the four experimental groups: control (CTR), no choline or vitamin E supplementation; choline (RPC), supplemented with 4 g/day choline chloride in rumen-protected form; vitamin E (VITE), supplemented with 200 IU/day vitamin E in rumen-protected form; and choline and vitamin E (RPCE), supplemented with 4 g/day RPC chloride and 200 IU/day vitamin E. Supplements were administered individually before the morning feed to ensure complete consumption, starting 30 days before kidding and continuing for 35 days after. During the experiment, milk yield and 4% fat-corrected milk (FCM) yield were, respectively, 210 and 350 g/day higher in RPC-supplemented goats than in non-supplemented goats. Milk fat concentration and fat yield were also increased by RPC treatment. Milk yield and composition were unaffected by vitamin E supplementation. There were no significant interactions between RPC and VITE for any of the variables measured. Plasma metabolites did not differ between treatments before and after kidding except that plasma folate at parturition was higher in RPC-supplemented goats. Neither choline nor vitamin E affected vitamin B12 plasma concentrations, while a time effect was evident after the second week of lactation, when B12 levels in each treatment group started to increase. Vitamin E administration resulted in plasma α-tocopherol levels that were 2 to 2.5 times higher than in non-supplemented goats. Overall, these results suggest that greater choline availability can improve milk production and methyl group metabolism in transition dairy goats.

Evaluation of the damage induced by ochratoxin A and the protective role of α-tocopherol in cultured bovine mammary epithelial cells

Evaluation of the damage induced by ochratoxin A and the protective role of α-tocopherol in cultured bovine mammary epithelial cells E. Fusi & R. Rebucci & C. Pecorini & L. Rossi & F. D’Ambrosio & A. Baldi Published online: 7 August 2008 # Springer Science + Business Media B.V. 2008

Effects of putrescine, cadaverine, spermine, spermidine and β-phenylethylamine on cultured bovine mammary epithelial cells

Abstract A bovine mammary epithelial cell line (BME-UV1) and three-dimensional collagen primary bovine organoids were used to evaluate the effects of cadaverine, putrescine, spermine, spermidine and β-phenylethyla-mine on mammary epithelial cells. Each biogenic amine was diluted in several concentrations (0-50 mM in BME-UV1 and 0-4 mM in primary bovine organoids) in the appropriate saline solution for the cell culture considered. In order to determine the activity of each compound tritiated thymidine incorporation was used. At low concentrations, all amines induced cell proliferation in both cultures. In BME-UV1, spermine significantly inhibited cell proliferation (P<0.001), while the other amines inhibited at higher concentrations (50mM). In primary bovine organoids, β−phenylethylamine significantly (P<0.001) inhibited cell proliferation at 4 mM. Organoids cultured in the presence of all amines, except β-phenylethylamine, had stellate projections indicating intense cell proliferation. Proliferation of mammary epithelial cells was stimulated at low concentrations, while at high concentrations it was inhibited. Our results suggested that the effects of each compound on mammary epithelial cells could be related to the compound itself and not to mediating by the bovine amino oxidase, responsible of the formation of toxic metabolites.

Cell-based models for mycotoxin screening and toxicity evaluation: an update

This review presents the applications of cell-based models in mycotoxin research, with a focus on models for mycotoxin screening and cytotoxicity evaluation. Various cell-based models, cell and cell culture condition related factors, toxicity endpoints and culture systems as well as predictive value of cell-based bioassays are reviewed. Advantages, drawbacks and technical problems regarding set up and validation of consistent, robust, reproducible and high-throughput cell-based models are discussed. Various cell-based models have been developed and used as screening tests for mycotoxins but the data obtained are difficult to compare. However, the results highlight the potential of cell-based models as promising in vitro platforms for the initial screening and cytotoxicity evaluation of mycotoxins and as a significant analytical approach in mycotoxin research before any animal or human clinical studies. To develop cell-based models as powerful high-throughput laboratory platforms for the analysis of large ...

Effects of Rumen-protected Choline Supplementation in Periparturient Dairy Goats

D’Ambrosio, F., Campagnoli, A., Susca, F., Fusi, E., Rebucci, R., Agazzi, A, Pinotti, L. and Baldi, A., 2007. Effects of rumen-protected choline supplementation in periparturient dairy goats. Veterinary Research Communications, 31(Suppl. 1), 393–396

Milk Choline, α-Tocopherol and Neutrophil Chemotaxis in the Periparturient Dairy Cow

L. Pinotti*, R. Rebucci, E. Fusi, L. Rossi and A. Baldi Department of Veterinary Sciences and T echnology for Food Safety, Faculty of Veterinary Medicine, University of Milan, Milan, Italy *Correspondence: Dipartimento di Scienze e T ecnologie Veterinarie per la Sicurezza Alimentare, Facolta di Medicina Veterinaria, Universita degli Studi di Milano, V ia T rentacoste 2, 20134 Milano, Italy E-mail: luciano.pinotti@unimi.it