Proliferative effect of whey from cows' milk varying in phyto-oestrogens in human breast and prostate cancer cells.

Intake of dietary phyto-oestrogens has received a great deal of attention owing to their potential influence on hormone-sensitive cancers such as breast and prostate cancer. Cows' milk contains phyto-oestrogens and the content varies according to the composition of the feed and the type and amount of legumes used. In this study we evaluated the proliferative effect of milk (whey) with different phyto-oestrogen content in human breast (MCF-7) and prostate cancer cells (PC-3). Milk was obtained from cows fed either a birdsfoot trefoil-timothy silage based ration (B1) or two different red clover silage based diets (R1 and R2) resulting in total phyto-oestrogen contents of 403, 1659 and 1434 ng/ml for the B1, R1 and R2 diets, respectively. Whey was produced from the milk and added to cell culture medium in concentrations up to 10% for MCF-7 cells and 5% for PC-3 cells. Cell proliferation was measured fluorometrically after 7 d for MCF-7 cells and 5 d for PC-3 cells. There was no significant difference in the proliferative effect of whey from the different dietary treatments at any of the whey concentrations tested. An anti-proliferative effect (P<0·01) of 5 and 10% whey was seen when tested in the presence of 10 pM oestradiol in the medium. This effect was independent of dietary treatment of cows. Whey induced a significant (P<0·01) proliferative response in PC-3 cells independent of dietary treatment. Purified equol in concentrations similar to equol concentrations in milk decreased PC-3 cell proliferation, and therefore the stimulatory effect of whey in PC-3 cells is believed to be mediated by other bioactives than equol. In conclusion, our results suggest that using whey in these proliferation assays, it was not possible to discriminate between milk with high or low levels of phyto-oestrogens.

Effect of feeding fermentable fibre-rich feedstuffs on meat quality with emphasis on chemical and sensory boar taint in entire male and female pigs.

Skatole, androstenone and other compounds such as indole cause boar taint in entire male pork. However, female pigs also produce skatole and indole. The purpose of this experiment was to minimise boar taint and increase overall impression of sensory quality by feeding entire male and female pigs with fibre-rich feedstuffs. The pigs have been fed three organic diets for either 1 or 2 weeks prior to slaughter of which two diets contained different fermentable fibre-rich feedstuffs - 10-13.3% dried chicory roots or 25% blue lupines. These two treatments were compared with pigs fed with an organic control diet for either 1 or 2 weeks prior to slaughter. Lupines significantly reduced skatole in blood and backfat for both genders after 1 week. Moreover, lupines showed negative impact on growth rate and feed conversion whilst chicory showed no significant differences in this respect. However, the indole concentration was significantly lower in chicory than lupine fed pigs. From a sensory perspective, chicory and lupine feeding reduced boar taint since odour and flavour of manure related to skatole and urine associated to androstenone were minimised. The level of boar taint in the entire male pigs was most effectively reduced after 14 days by both fibre-rich feeds while lupine had the largest influence on "boar" taint reduction in female pigs.

Bone biochemical markers for assessment of bone responses to differentiated phosphorus supply in growing-finishing pigs.

Phosphorus (P) is essential for building and maintaining a healthy and strong skeleton. Moreover, dietary P supply may play a role for bone turnover, and the excretion of bone turnover metabolites may be useful as markers for sufficient dietary P supply. The objective was to study the long-term effects of low, medium, and high dietary P supply on bone metabolism in terms of serum concentration and urinary excretion of bone turnover components and metabolites in healthy growing-finishing pigs compared with bone mineral content (BMC) and bone mineral density (BMD) of humerus and femur. Pigs were fed diets containing low [LP; 4.1 g/kg dry matter (DM)], medium (MP; 6.2 g/kg DM), or high dietary P (HP; 8.9 g/kg DM) from 39.7 kg body weight (BW) until slaughter at 110 kg BW. Urine and blood were collected at 40, 70, and 110 kg BW while bones were collected at slaughter. Serum was analyzed for osteocalcin (OC), bone alkaline phosphatase (BAP), and C-terminal telopeptides of type I collagen (CTX-I), whereas urine was analyzed for pyridinoline (PYD), deoxypyridinoline (DPD), CTX-I, hydroxylysine (HYL), galactosyl-hydroxylysine (GAL-HYL), glycosyl-galactosyl-hydroxylysine (GLC-GAL-HYL), and hydroxyproline (HYP). Humerus and femur were analyzed for BMC and BMD. The LP diet caused reduced OC and increased BAP and CTX-I concentrations in serum. Furthermore, BAP was increased in response to the HP diet. Urine metabolites of bone resorption were all increased in pigs fed the LP diet, but only a few responses were obtained in response to the HP diet. Furthermore, age-related decreases were identified for BAP, HYL, GAL-HYL, and GLC-GAL-HYL. Bone mineral content and BMD were markedly lowered in pigs fed the LP diet but were not affected in pigs fed the HP diet. In conclusion, OC, BAP, and CTX-I in serum have proved useful for P adequacy in growing-finishing pigs. In addition, urine bone resorption metabolites have also proved useful for P adequacy and analysis of PYD, DPD, and CTX-I was considered to be the most relevant markers due to their specificity for bone and their negative correlation with BMD, BMC, ash, calcium (Ca), and P contents. Finally, DPD may be the preferred marker in long-term P feeding assessments.

Plagiotropism and auxin in Abies nordmanniana.

Main branches of Abies nordmanniana Spach. were examined through their first growth season from subapical buds around the leader bud to fully expanded shoots. Plagiotropism was evident in branch orientation, which was almost horizontal, as well as in the orientation of buds developing on the branches. Auxin transport capacity was predominantly basipetal (> 90%) and consistently higher in the middle part of the branch than in the distal and proximal ends. Auxin transport capacity was higher on the dorsal side of the branch during the short initial hyponastic growth phase, but the difference disappeared when the branch became horizontal. No dorsal-ventral differences could be detected in young horizontal branches in concentrations of indole acetic acid, cytokinins, gibberellins or abscisic acid. Branch orientation was unaffected by decapitation of the leader apex or by decapitation and replacement with exogenous auxin. However, decapitation resulted in a less plagiotropic bud arrangement on the branches, and auxin application to the leader bud scar counteracted this effect. Thus, a signal originating in the stem seems to be involved in regulating branch bud positioning, whereas the horizontal branch orientation must rely on a different mechanism, presumably autonomic within the branch.

Phytoestrogens and their metabolites in bulk-tank milk: effects of farm management and season.

Phytoestrogens have structures similar to endogenous steroids and may induce or inhibit the response of hormone receptors. The objectives of the present study were to compare the effects of long-term vs. short-term grassland management in organic and conventional dairy production systems, compare organic and conventional production systems and assess seasonal variation on phytoestrogen concentrations in bulk-tank milk. The concentrations of phytoestrogens were analyzed in bulk-tank milk sampled three times in two subsequent years from 28 dairy farms: Fourteen organic (ORG) dairy farms with either short-term or long-term grassland management were paired with 14 conventional (CON) farms with respect to grassland management. Grassland management varied in terms of time since establishment. Short-term grassland management (SG) was defined as establishment or reseeding every fourth year or more often, and long-term grassland management (LG) was defined as less frequent establishment or reseeding. The proportion of red clover (Trifolium pretense L.) in the herbage was positively correlated with milk concentrations of the mammalian isoflavone equol. Therefore, organically produced bulk-tank milk contained more equol than conventionally produced milk, and milk from ORG-SG farms had more equol than milk from ORG-LG farms. Milk produced during the indoor-feeding periods had more equol than milk produced during the outdoor feeding period, because pastures contained less red clover than fields intended for silage production. Organically produced milk had also higher concentrations of the mammalian lignan enterolactone, but in contrast to equol, concentrations increased in the outdoor-feeding periods compared to the indoor-feeding periods. There were no indications of fertility problems on ORG-SG farms who had the highest red clover proportions in the herbage. This study shows that production system, grassland management, and season affect milk concentrations of phytoestrogens. However, compared to soy products, milk concentrations of phytoestrogens are low and future studies are required to investigate if the intake of phytoestrogens from dairy products has physiological effects in humans.

Effect of flavonoids on stress responses in myotube cultures.

Effects of flavonoids on stress response of myotube cultures was studied by monitoring the release of [14C] taurine, leukotriene production, and 2',7'-dichlorodihydroflourescein (DCFH2) oxidation. Stress was induced by hypotonic shock, which was accompanied by cell swelling leading to increased leukotriene production and a concomitant increase in reactive oxygen species and osmolyte release. In this model system, addition of the flavonoids catechin and quercetin decreased leukotriene production, DCFH2 oxidation, and taurine efflux, indicating a reduction of cellular stress. High concentrations of epigallocatechin gallate (EGCG) and tea extract increased leukotriene production and initial DCFH2 oxidation, indicating an increased cellular stress (possibly toxicity). However, taurine efflux was reduced, and also longer exposure time as well as lower concentrations of EGCG and tea reduced DCFH2 oxidation. Trolox and alpha-tocopherol did not significantly affect taurine efflux or leukotriene production, and it was therefore concluded that suppression of these responses was not confined to redox activity in a myotube culture.

Validation of a simple liquid chromatographic method for determination and quantitation of residual ivermectin and doramectin in pig liver.

A rapid and quantitative method for the extraction, derivatization, and liquid chromatography with fluorescence detection of ivermectin (IVM) and doramectin (DOM) residues in porcine liver was developed and validated. IVM and DOM were extracted from the liver samples with acetonitrile, the supernatant was evaporated to dryness at 37 degrees C under nitrogen, and the residue was reconstituted in 1-methylimidazole solution. After 2 min at room temperature, IVM and DOM were converted to a fluorescent derivative and then separated on a Hypersil ODS column. The derivatives of IVM and DOM were detected and quantitated with high specificity by fluorescence (excitation: 365 nm, emission: 475 nm). Abamectin was used as an internal standard. The mean extraction efficiencies from fortified samples (15 ng/g) were 75% for IVM and 70% for DOM. The limit of detection was 0.8 ng/g for both IVM and DOM.