Protein-polyphenol reactions: 1. Nutritional and metabolic consequences of the reaction between oxidized caffeic acid and the lysine residues of casein

1. Studies were made on the lysine content of casein reacted with caffeic acid oxidized aerobically under alkaline conditions or enzymically with tyrosinase (EC 1.14.18.1). 2. Loss of fluorodinitrobenzene (FDNB)-reactive lysine was rapid at pH 10 and increased with time and the temperature of the reaction, with concentration of caffeic acid and with the oxygenation of the mixture. In presence of the enzyme mushroom tyrosinase, maximum reduction of reactive lysine occurred at pH 7 and was dependent on the reaction time and on the concentration of caffeic acid. 3. Reaction of α-formyl-L-[U- 14C]lysine with caffeic acid at pH 10 showed the rapid formation of five reaction products which appeared to polymerize gradually as the reaction progressed. 4. The nutritionally available lysine content of the casein-caffeic acid mixtures, as assayed with rats, was reduced after both alkaline and enzymic reactions, as were faecal digestibility, net protein ratio and net protein utilization. Biological value however was not reduced. 5. In metabolic studies using goat milk casein labelled with L-[3H]lysine and reacted with caffeic acid in the same way, the lysine-caffeoquinone reaction products were not absorbed by the rat but were excreted directly in the faeces. 6. The importance of the reaction of proteins with caffeoquinone and chlorogenoquinone (formed by the oxidation of caffeic and chlorogenic acids respectively) is discussed in relation to the production of sunflower protein, leaf protein and other vegetable-protein concentrate

Fluoropyrimidine sensitivity of human MCF-7 breast cancer cells stably transfected with human uridinehosphorylase

The relationship between uridine phosphorylase (UP) expression level in cancer cells and the tumour sensitivity to fluoropyrimidines is unclear. In this study, we found that UP overexpression by gene transfer, and the subsequent efficient metabolic activation of 5-fluorouracil (5-FU) by the ribonucleotide pathway, does not increase the fluoropyrimidine sensitivity of MCF-7 human cancer cells. © 2001 Cancer Research Campaign http://www.bjcancer.co

Diagnose différentielle de l'espèce sur les carcasses et les abats des moutons et des chèvres de l'Afrique tropicale de l'Ouest

Les auteurs étudient région par région, les caractères immédiats et médiats de la diagnose d'espèce des carcasses et des abats des petits ruminants de l'Afrique de l'Ouest. Ils montrent que la diagnose est facile lorsque la tête et la queue restent sur la carcasse ou la demi-carcasse et qu'elle est difficile lorsqu'elles en sont séparées. Ils distinguent trois types de caractères différentiels: ceux qui sont communs aux deux groupes régionaux, ceux qui sont particuliers aux espèces des régions tempérées et ceux qui sont propres aux ovins et aux caprins de l'Afrique de l'Oues

Laser Shock Adhesion Test (LASAT) of electron beam physical vapor deposited thermal barrier coatings (EB-PVD TBCs)

International audienceDamage prediction, adhesion strength and remaining lifetime of TBC are highly important data for understanding and preventing TBC spallation on blades. LAser Shock Adhesion Test (LASAT) is a powerful method to measure adhesion of coating due to its rapidity, simplicity and capabilities to distinguish different strength levels and the easy damage observation in case of TBCs. A new protocol of LASAT has been introduced in order to measure the adhesion level of the ceramic coating from the exploitation of the two-dimensional effects that promotes a shock wave pressure-dependent size of the damage. Finite element modeling, taking into account the TBCs dimensions, showed the edges effect on interfacial stress applied by laser shock

Etude par choc laser de l'adhérence de barrières thermiques aéronautiques

National audienceL'estimation de la durée de vie des barrières thermiques aéronautiques (BT) déposées sur les aubes de turbines haute pression nécessite de comprendre les mécanismes induisant l'endommagement entre la sous-couche et la zircone. Le LASAT, LAser Shock Adhesion Test ou essai d'adhérence par choc laser, est une technique exploitant tout son potentiel dans le cas des BTs, en particulier par sa rapidité, simplicité et au comportement optique de la couche de zircone. Ce dernier avantage permet de dimensionner l'endommagement généré sans réaliser de coupes métallographiques. Un nouveau protocole de l'essai LASAT a été mis en place utilisant les ondes de choc bidimensionnelles et exploitant la dépendance entre l'intensité du choc appliqué, le niveau d'adhérence du dépôt, et la dimension de la fissure générée

Utilization of galactomannan from Gleditsia triacanthos in polysaccharide-based films : effects of interactions between film constituents on film properties

The objective of this work was to evaluate the effect of the concentrations of Gleditsia triacanthos galactomannan and glycerol and the presence of corn oil in the physical properties of edible films. The influence of interactions between those constituents on films' permeability to gases (water vapour, CO2 and O2), solubility in water, mechanical properties and colour was evaluated. The effects of those variables were analysed according to a 23 factorial design; regression coefficients were used to understand the influence of each variable (factor) on the studied properties, and a multifactor model was developed. Results show that galactomannan concentration is the most significant factor affecting the studied properties; moreover, the increase of plasticizer concentration and the presence of oil showed to be the most influent in the particular cases of solubility and transport properties (water vapour permeability and O2 permeability), respectively. These results show that galactomannan films' properties can be tailored to allow their use as alternative to non-biodegradable, non-edible packaging materials.The author M. A. Cerqueira is recipient of a fellowship from Fundacao para a Ciencia e Tecnologia (FCT, SFRH/BPD/72753/2010) and B. W. S. Souza is a recipient of a fellowship from the Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior, Brazil (Capes, Brazil)

Combined effects of GSTP1 and MRP1 in melanoma drug resistance

Glutathione-S-transferase Pi1 (GSTP1) and multidrug resistance protein 1 (MRP1) are overexpressed in melanoma, a skin cancer notoriously resistant to all current modalities of cancer therapy. To investigate the involvement of these detoxifying enzymes in the drug resistance of melanoma, an inducible (Tet-On™ system) antisense (AS) RNA strategy was used to specifically inhibit GSTP1 expression in A375 cells, a human melanoma cell line expressing high levels of GSTP1 and MRP1. Stable transfectant clones were established and analysed for GSTP1 inhibition by AS RNA. The clone A375-ASPi1, presenting a specific 40% inhibition of GSTP1 expression in the presence of doxycycline, was selected. Lowering the GSTP1 level significantly increased (about 3.3-fold) the sensitivity of A375-ASPi1 cells to etoposide. Inhibitors of glutathione synthesis (BSO), GSTs (curcumin, ethacrynic acid), and also of MRPs (MK571, sulphinpyrazone) improved the sensitising effect of GSTP1 AS RNA. All these inhibitors had stronger sensitising effects in control cells expressing high GSTP1 level (A375-ASPi1 cells in the absence of doxycycline). In conclusion, GSTP1 can act in a combined fashion with MRP1 to protect melanoma cells from toxic effects of etoposide

Role of platelet-derived endothelial cell growth factor/thymidine phosphorylase in fluoropyrimidine sensitivity

Platelet-derived endothelial cell growth factor (PD-ECGF)/thymidine phosphorylase (TP) catalyses the reversible phosphorolysis of thymidine to thymine and 2-deoxyribose-1-phosphate and is involved in the metabolism of fluoropyrimidines. It can also activate 5'-deoxyfluorouridine (5'DFUR) and possibly 5-fluorouracil (5FU) and Ftorafur (Ft), but inactivates trifluorothymidine (TFT). We studied the contribution of TP activity to the sensitivity for these fluoropyrimidines by modulating its activity and/or expression level in colon and lung cancer cells using a specific inhibitor of TIP (TPI) or by overproduction of TIP via stable transfection of human TP. Expression was analysed using competitive template-RT-PCR (CT-RT-PCR), Western blot and an activity assay. TP activity ranged from nondetectable to 70678 pmol h(-1) 10(-6) cells, in Colo320 and a TP overexpressing clone Colo320TPI, respectively. We found a good correlation between TIP activity and mRNA expression (r = 0.964, P <0.01) in our cell panel. To determine the role of TIP in the sensitivity to 5FU, 5'DFUR, Ft and TFT, cells were cultured with the various fluoropyrimidines with or without TPI and differences in IC50's were established. TPI modified 5'DFUR, increasing the IC50's 2.5- to 1396-fold in WiDR and Colo320TPI, respectively. 5-Fluorouracil could be modified by inhibiting TP but to a lesser extent than 5'DFUR: IC50's increased 1.9- to 14.7-fold for WiDR and Colo320TPI, respectively. There was no effect on TFT or Ft. There appears to be a threshold level of TP activity to influence the 5'DFUR and 5FU sensitivity, which is higher for 5FU. Even high levels of TP overexpression only had a moderate effect on 5FU sensitivity. (C) 2003 Cancer Research UK