Effect of the inclusion of artichoke silage in the ration of lactating ewes on the properties of milk and cheese characteristics during ripening.

The effect of including artichoke silage in the rations of dairy ewes on milk characteristics and biochemical changes of ripened cheeses was evaluated. Four groups of lactating ewes were fed rations containing 0, 10, 20, or 30% artichoke silage on a dry matter basis. Bulk milk samples were collected 3 times during the feeding period, and semi-hard cheeses were manufactured and sampled during ripening. Milk composition and cheese yield were not affected by diet. Inclusion of 20 and 30% artichoke silage reduced the firmness of the curds at a level only detected by the Gelograph (Gelograph-NT, Gel-Instrumente, Thalwil, Switzerland) probe. Inclusion of artichoke silage in ewes' diet decreased fat and total free fatty acids content of these cheeses and increased total free amino acids content. Despite the effect of diet on cheese ripening characteristics, the overall sensory scores for cheeses corresponding to artichoke silage diets were statistically higher than those for the control cheeses.

Effect of the inclusion of whole citrus in the ration of lactating ewes on the properties of milk and cheese characteristics during ripening.

The effect of including citrus fruits (CF) in the rations of dairy ewes on the milk characteristics and biochemical changes of cheeses during ripening was evaluated. For this purpose, 48 lactating ewes (Guirra breed) were divided into 4 homogeneous groups and fed with isoenergetic and isoprotein rations containing CF at 0, 10, 20, and 30% on a dry matter basis in substitution of dry barley and pelleted beet pulp. During the experimental period, 3 batches of bulk milk were collected from each group and semi-hard cheeses were manufactured. Cheeses were sampled at 15, 30, and 60 d of ripening. Milk coagulation parameters and cheese yield were not negatively affected by the inclusion of CF in the ration. Physicochemical composition of cheeses at 60 d showed statistical differences for lower total solids and fat content of 30% CF cheeses. Proteolysis of cheeses measured by water-soluble nitrogen and total free amino acids content was not influenced by the ration. Differences between rations with respect to free fatty acids were significant for medium- and long-chain free fatty acids, and therefore for total content, but differences did not show a trend related to the increase of CF in the diet. The inclusion of CF in the ration of lactating ewes up to levels of 30% did not negatively affect the properties of milk and the biochemical and sensory characteristics of cheeses.

Effects of high pressure on proteolytic enzymes in cheese: relationship with the proteolysis of ewe milk cheese.

Ewe milk cheeses were submitted to 200, 300, 400, and 500 MPa (2P to 5P) at 2 stages of ripening (after 1 and 15 d of manufacturing; P1 and P15). The high-pressure-treated cheeses showed a more important hydrolysis of beta-casein than control and 2P1 cheeses. Degradation of alpha(s1)-casein was more important in 3P1, 4P1, and P15 cheeses than control and 2P1 cheeses. The 5P1 cheeses exhibited the lowest degradation of alpha(s)-caseins, probably as a consequence of the inactivation of residual chymosin. Treatment at 300 MPa applied on the first day of ripening increased the peptidolytic activity, accelerating the secondary proteolysis of cheeses. The 3P1 cheeses had extensive peptide degradation and the highest content of free amino acids. Treatments at 500 MPa, however, decelerated the proteolysis of cheeses due to a reduction of microbial population and inactivation of enzymes.

Interactions between calvatic acid and related compounds with rat liver microsomes.

In this study we examined the interactions of liver microsomes with the antibiotic calvatic acid and with structural analogues, some of which had shown antimicrotubular properties. These drugs decreased cytochrome P-450 content differently according to the substitutions on the azoxy function and the ethoxycarbonyl derivatives were found to be the most effective ones. The decrease in cytochrome P-450 could be prevented by addition of cysteine or GSH, suggesting an involvement of sulphydryl groups. Furthermore, chromatographic analyses showed that ethoxycarbonyl derivatives were completely metabolized, and this would explain the different behaviour of these compounds towards microtubular protein when they were incubated with purified bovine brain protein or with liver or hepatoma extracts.

Inhibition of human placenta glutathione transferase P1-1 by calvatic acid.

The inhibition mechanism of the dimeric human placenta glutathione transferase (GST P1-1) by the antibiotic p-carboxyphenylazoxycyanide (calvatic acid) has been investigated at pH 7.0 and 30.0 degrees C. Experiments performed at different calvatic acid/GST P1-1 molar ratios indicate that one mole of calvatic acid inactivates one mole of the homodimeric enzyme molecule, containing two catalytically equivalent active sites. The apparent second order rate constant for GST P1-1 inactivation is 2.4 +/- 0.3 M-1 s-1. The recovery of all the 5,5'-dithio-bis(2-nitro-benzoic acid)-titratable thiol groups as well as the original catalytic activity of GST P1-1 after treatment of the inhibited enzyme with dithiothreitol indicates that two disulfide bridges per dimer, likely between Cys47 and Cys101, have been formed during the reaction with calvatic acid. To the best of the authors knowledge, calvatic acid represents a unique case of enzyme inhibitor acting also throughout its reaction product(s).

The in vitro metabolic inversion of R(-) to S(+) indoprofen.

The paper reports a study on the metabolic inversion of indoprofen (2-[4-(2-isoindolinyl-1-one)-phenyl]-propionic acid) following incubation of the drug with liver microsomes from non-induced and phenobarbital-induced rats. The enantiomeric composition of the drug was determined after different incubation times of the racemate and the individual isomers. The S(+)/R(-) ratio was evaluated by densitometry following HPTLC separation of the R(+)-1-phenylethylamides. After incubation of the racemate and the individual isomers, no detectable amounts of indoprofen catabolites were extracted from the acidified incubation mixture. An appreciable enrichment in the S(+) enantiomer was observed after incubation of both racemate and R(-)-indoprofen; the S(+)/R(-) ratio reached a maximum after 1 h. Values were higher in the case of induction. After incubation of S(+)-indoprofen, a small but statistically significant decrease of the S(+)/R(-) ratio was observed. The increase of the S(+)-isomer concentration observed following incubation of R(-)-indoprofen can be ascribed to metabolic inversion by phenobarbital-inducible liver enzymes.

Changes in the three-dimensional organization of LoVo cells associated with resistance to doxorubicin.

LoVo cells and a derived subline resistant to doxorubicin were compared in the spheroids system. The resistant line, unlike the parent one, was unable to grow as spheroids, but formed irregular loose aggregates. Moreover treatment of the resistant cells with membrane-active agents able to reverse pleiotropic drug resistance had no effect on the capability of these cells to grow as spheroids. The results indicate that the inability of resistant cells to form spheroids is not related to the resistance mechanism.