Urolithins, ellagic acid-derived metabolites produced by human colonic microflora, exhibit estrogenic and antiestrogenic activities

Urolithins A and B (hydroxy-6H-dibenzo[b,d]pyran-6-one derivatives) are colonic microflora metabolites recently proposed as biomarkers of human exposure to dietary ellagic acid derivatives. Molecular models suggest that urolithins could display estrogenic and/or antiestrogenic activity. To this purpose, both urolithins and other known phytoestrogens (genistein, daidzein, resveratrol, and enterolactone) were assayed to evaluate the capacity to induce cell proliferation on the estrogen-sensitive human breast cancer MCF-7 cells as well as the ability to bind to alpha- and beta-estrogen receptors. Both urolithins A and B showed estrogenic activity in a dose-dependent manner even at high concentrations (40 microM), without antiproliferative or toxic effects, whereas the other phytoestrogens inhibited cell proliferation at high concentrations. Overall, urolithins showed weaker estrogenic activity than the other phytoestrogens. However, both urolithins displayed slightly higher antiestrogenic activity (antagonized the growth promotion effect of 17-beta-estradiol in a dose-dependent manner) than the other phytoestrogens. The IC(50) values for the ERalpha and ERbeta binding assays were 0.4 and 0.75 microM for urolithin A; 20 and 11 microM for urolithin B; 3 and 0.02 for genistein; and 2.3 and 1 for daidzein, respectively; no binding was detected for resveratrol and enterolactone. Urolithins A and B entered into MCF-7 cells and were metabolized to yield mainly urolithin-sulfate derivatives. These results, together with previous studies regarding absorption and metabolism of dietary ellagitannins and ellagic acid in humans, suggest that the gut microflora metabolites urolithins are potential endocrine-disrupting molecules, which could resemble other described "enterophytoestrogens" (microflora-derived metabolites with estrogenic/antiestrogenic activity). Further research is warranted to evaluate the possible role of ellagitannins and ellagic acid as dietary "pro-phytoestrogens".     

Safety evaluation of an oak-flavored milk powder containing ellagitannins upon oral administration in the rat

Ellagitannins are phenolic compounds that occur in a number of dietary sources such as walnuts, raspberries, pomegranate, strawberries, and oak-aged wines. Ellagitannins have been acknowledged to have antioxidant and cancer chemopreventive activities. However, ellagitannins have also been considered to be "antinutrients" and have been reported to provoke adverse effects in animals. In this context, risk assessment studies are demanded to ensure that chemicals present in food do not compromise food safety. Oak-flavored milk can be obtained by thermal treatment of milk in the presence of oak shavings. The oak-flavored milk can be used to impart wood-like flavor to different recipes. The present study evaluated, under a subchronic approach, the safety of an oak-flavored milk powder containing ellagitannins in rats upon repeated oral administration for 96 days. The effect of 30% oak-flavored milk powder in the diet was evaluated. This amount involved a margin of safety equivalent to the daily intake of 134 yogurts by a 70 kg person for 96 days. Growth rate, food utility index, 9 hematological, and 13 serobiochemical parameters as well as the metabolism and tissue distribution of ellagitannins and derived metabolites were studied. Overall, no significant differences were found in oak-milk-fed rats in any blood parameter analyzed. No effect was found on growth rate and food utility index. Urolithin A was identified in feces of rats fed oak-flavored milk and the glucuronide derivative in urine. No metabolites were found in the liver, kidney, or uterus. Histopathological analysis of liver and kidney corroborated the absence of toxicity. Therefore, these results suggest that the intake of the oak-flavored milk powder assayed in this study is safe and does not involve potential hazard to human health.     

A comparative molecular field analysis study of obtusifoliol 14α-methyl demethylase inhibitors

This report describes the development of a Comparative Molecular Field Analysis (CoMFA) model from a set of obtusifoliol 14α-methyl demethylase (DM) inhibitors to aid in the design of herbicides targeting sterol biosynthesis. CoMFA is a three-dimensional (3-D) quantitative structure–activity relationship (QSAR) method that is useful in the probing of receptor binding sites when experimental structure data are unavailable. Conformational analysis and SAR of some rigid and active analogs were used to build the initial model using the active analog hypothesis. The model was subsequently used to design compounds that retain the active site shape requirements, but incorporate physical properties that favor soil-applied herbicidal action. In addition, a second-generation CoMFA model incorporating the newly designed inhibitors was developed and represents the current understanding of the DM binding site. This model was derived from a pharmacophore developed from two methods, the active analog approach as well as from the Catalyst program. The fact that two independent methods produced a similar pharmacophore strengthens the validity of the model. © 1999 Society of Chemical Industry     

Changes in amino acid levels in bacteria adhering to rumen epithelium in sheep after oral administration of very low doses of mercury

The effect of low concentrations of mercury taken in the diet (4 mg per a head and day) on the amino acid composition in the proteins of rumen bacteria adhering to the dorsal and ventral parts of the rumen was studied in six sheep. Though the mercury did not influence the amino acid concentration in hydrolyzates of epimural bacteria, low-mercury doses caused significant changes of some amino acids, as compared with the control group of animals. The levels of alanine, histidine, threonine, serine and glycine were significantly decreased and at the same time the levels of proline, valine, isoleucine, tyrosine, lysine and phenylalanine were significantly increased. It was stated in our experiment that the levels of histidine, alanine and threonine in hydrolyzates of epimural bacteria of the rumen were significantly decreased both in the sheep given the balanced feed ration with additions of mercury and nitrogen (according to the standard) and in the sheep given a low-nitrogen diet. Similarly, the levels of proline, tyrosine and phenylalanine in the hydrolizates of epimural bacteria in rumen were significantly increased both with the low-nitrogen diet and in the normal -nitrogen diet with mercury supplement. The results suggest that unexplained disorders frequently occurring in the microbial synthesis in the rumen might be caused by long-continued administration of feed contaminated with extremely low concentrations of heavy metals. The problem deserves detailed investigation in future studies, oriented to explaining the biosynthesis disorders in rumen, reducing the production ability of ruminants.     

A new species of Acanthochondria (Copepoda: Chondracanthidae) parasitizing the flounder Xystreurys rasile (Pleuronectiformes: Paralichthyidae) from Argentina

A new copepod species, Acanthochondria sagitta sp. n., is described based on specimens collected from the flounder Xystreurys rasile (Jordan) (Pleuronectiformes, Paralichthyidae), caught in the coastal waters off Necochea, Buenos Aires Province, Argentina. The new species differs from its congeners by the following combination of characters: presence of three pairs of cephalic outgrowths; the triangular shape of the trunk with diverging postero-lateral processes; leg 2 of an intermediate shape between Types C and D, which projects laterally from the trunk, and Type B-V antennule bearing two proximal processes (one ventral and one dorsal) on the swollen basal portion.     

Diagnosis of infectious pancreatic necrosis (IPN) of trout using immunofluorescence

Until recently, demonstration of infectious pancreatic necrosis was conditioned by the isolation of virus. By means of the immunofluorescence method it is possible to make a rapid and accurate diagnosis, as well as to determine the development of infection. In acute cases, the virus in fry was demonstrated mainly in the cytoplasm of epithelial cells of guts; in Pd1-2 trout, the virus was proved in the cytoplasm of epithelial cells of primary kidney tubules and in the cytoplasm of epithelial cells of pancreatic acini. In all cases the virus occurrence demonstrated by immunofluorescence was in correlation with the results achieved by cultivation methods.     

Amino acid production in the isolated and perfused rumen wall of sheep

The metabolism of amino acids and urea in an isolated perfused wall of sheep rumen was studied by the method of organ perfusion, using 3 kinds of perfusate. In experiments with semisynthetic perfusate containing urea (SPurea) the levels of lysine, valine, glutamic acid and alanine increased at a simultaneous decrease of levels of leucine, arginine, aspartic acid, threonine and methionine. In perfusions with the semisynthetic medium without urea (SP) the levels of lysine, valine, phenylalanine, isoleucine, glutamic acid, glycine and alanine increased, the level of threonine dropped. In perfusions with whole autologous blood (KP) the levels of valine, glutamic acid and alanine increased, the level of arginine decreased. The level of urea decreased sharply at using SPurea and KP, however, the equal cumulation of ammonia (200-400 mumol x 1(-1)) was found in all three types of perfusate after 2-hr perfusion. Moreover, in experiments with SP a low level of urea was found already after 5-min. of perfusion. The results of the experiments suggest that the rumen wall is capable of producing amino acids also without the contents of rumen, and that independently on the presence or absence of urea as a nitrogen source.     

TRACHEITIS IN SHEEP AFTER ORAL ADMINISTRATION OF A MINERAL SUPPLEMENT

Outbreaks of illness and deaths in sheep after the concurrent administration of a cobalt, copper and zinc mineral supplement and laevamisole are described. Clinical illness was characterised by profound dyspnoea and inability to exercise. The consistent postmortem finding was severe, necrotising tracheitis with partial occlusion of the airway. This lesion was reproduced experimentally by intratracheal administration of the mineral supplement dissolved in one particular formulation of laevamisole but not in water or another preparation of laevamisole. Further experiments revealed that combined zinc sulphate and copper sulphate dissolved in the particular formulation of laevamisole would produce the tracheal lesions.