No abstract

Many health benefits of pomegranate products have been attributed to the potent antioxidant action of their tannin components, mainly punicalagins and ellagic acid. While moving through the intestines, ellagitannins are metabolized by gut bacteria into urolithins that readily enter systemic circulation. In this study, the antioxidant properties of seven urolithin derivatives were evaluated in a cell-based assay. This method is biologically more relevant because it reflects bioavailability of the test compound to the cells, and the antioxidant action is determined in the cellular environment. Our results showed that the antioxidant activity of urolithins was correlated with the number of hydroxy groups as well as the lipophilicity of the molecule. The most potent antioxidants are urolithins C and D with IC(50) values of 0.16 and 0.33 microM, respectively, when compared to IC(50) values of 1.1 and 1.4 microM of the parent ellagic acid and punicalagins, respectively. The dihydroxylated urolithin A showed weaker antioxidant activity, with an IC(50) value 13.6 microM, however, the potency was within the range of urolithin A plasma concentrations. Therefore, products of the intestinal microbial transformation of pomegranate ellagitannins may account for systemic antioxidant effects.

Pomegranate juice derived ellagitannins and their intestinal bacterial metabolites, urolithins, inhibited TCDD-induced CYP1-mediated EROD activity in vitro with IC(50) values ranging from 56.7 microM for urolithin A to 74.8 microM for urolithin C. These compounds exhibited dose- and time-dependent decreases in cell proliferation and clonogenic efficiency of HT-29 cells. Inhibition of cell proliferation was mediated through cell cycle arrest in the G(0)/G(1) and G(2)/M stages of the cell cycle followed by induction of apoptosis. These results indicate that the ellagitannins and urolithins released in the colon upon consumption of pomegranate juice in considerable amounts could potentially curtail the risk of colon cancer development, by inhibiting cell proliferation and inducing apoptosis.

The consumption of pomegranate products leads to a significant accumulation of ellagitannins in the large intestines, where they interact with complex gut microflora. This study investigated the effect of pomegranate tannin constituents on the growth of various species of human gut bacteria. Our results showed that pomegranate byproducts and punicalagins inhibited the growth of pathogenic clostridia and Staphyloccocus aureus. Probiotic lactobacilli and bifidobacteria were generally not affected by ellagitannins, while relatively small growth inhibition by ellagic acid likely resulted from decreasing media quality due to the formation of tannin-protein complexes. The effect of pomegranate ellagitannins on bifidobacteria was species- and tannin-dependent. The growth of Bifidobacterium animalis ssp. lactis was slightly inhibited by punicalagins, punicalins, and ellagic acid. POMx supplementation significantly enhanced the growth of Bifidobacterium breve and Bifidobacterium infantis.

Aplysinopsins are tryptophan-derived marine natural products isolated from numerous genera of sponges and scleractinian corals, as well as from one sea anemone and one nudibranch. Aplysinopsins are widely distributed in the Pacific, Indonesia, Caribbean, and Mediterranean regions. Up to date, around 30 analogues occurring in Nature have been reported. Natural aplysinopsins differ in the bromination pattern of the indole ring, variation in the structure of the C ring, including the number and position of N-methylation, the presence and configuration of the C-8-C-1′ double bond, and the oxidation state of the 2-aminoimidazoline fragment. Aplysinopsins can also occur in the form of dimers. This review summarizes 30 years’ research on aplysinopsins. The origin, isolation sources, chemistry, bioactivity, and ecological functions of aplysinopsins are comprehensively reviewed.

The cytochrome P450 enzyme, CYP1B1, is an established target in prostate cancer chemoprevention. Compounds inhibiting CYP1B1 activity are contemplated to exert beneficial effects at three stages of prostate cancer development, that is, initiation, progression, and development of drug resistance. Pomegranate ellagitannins/microbial metabolites were examined for their CYP1B1 inhibitory activity in a recombinant CYP1B1-mediated ethoxyresorufin-O-deethylase (EROD) assay. Urolithin A, a microbial metabolite, was the most potent uncompetitive inhibitor of CYP1B1-mediated EROD activity, exhibiting 2-fold selectivity over CYP1A1, while urolithin B was a noncompetitive inhibitor with 3-fold selectivity. The punicalins and punicalagins exhibited potent CYP1A1 inhibition with 5-10-fold selectivity over CYP1B1. Urolithins, punicalins, and punicalagins were tested for their 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced CYP1 inhibitory activity in the 22Rv1 prostate cancer cell line. Urolithins A and B showed a decrease in their CYP1-mediated EROD inhibitory IC50 values upon increasing their treatment times from 30 min to 24 h. Urolithin C, 8-O-methylurolithin A, and 8,9-di-O-methylurolithin C caused a potent CYP1-mediated EROD inhibition in 22Rv1 cells upon 24 h of incubation. Neutral red uptake assay results indicated that urolithin C, 8-O-methylurolithin A, and 8,9-di-O-methylurolithin C induced profound cytotoxicity in the proximity of their CYP1 inhibitory IC50 values. Urolithins A and B were studied for their cellular uptake and inhibition of TCDD-induced CYP1B1 expression. Cellular uptake experiments demonstrated a 5-fold increase in urolithin uptake by 22Rv1 cells. Western blots of the CYP1B1 protein indicated that the urolithins interfered with the expression of CYP1B1 protein. Thus, urolithins were found to display a dual mode mechanism by decreasing CYP1B1 activity and expression.

Lichens produce a great number of secondary metabolites that participate in ecological interactions and respond to environmental changes. We examined the influence of heavy metal accumulations on lichen secondary metabolism. Thalli of Hypogymnia physodes were transplanted for 6 months to the Cracow-Silesia industrial region. Based on heavy metal accumulations in lichen, two of the investigated sites were classified as highly polluted. The highest concentrations of Cd, Pb, and Zn were found in lichens transplanted in the vicinity of a Zn-Pb smelter. Significant accumulations of Cr and Ni were detected in Hypogymnia transplanted near a chemical industry. Physodic, physodalic, hydroxyphysodic acids, and atranorin were identified and analyzed in extracts obtained from specimen samples. The most detrimental changes were observed in lichen transplanted into the vicinity of a chemical industry producing chromium, phosphor, and sulfur compounds that contained 340-fold higher Cr levels than control thalli. Decreases in the levels of physodic acid, hydroxyphysodic acid, and atranorin were detected, and one additional polar compound (probably product of degradation of lichen acids) appeared in the extract. The content of physodalic acid increased in every thalli sample transplanted, suggesting a possible role of this compound in defense against stress caused by accumulated pollutants. The levels of physodic acid decreased in thalli from both of the most polluted sites compared to those of the controls--but were not changed in thalli transplanted to less polluted sites. Our results illustrate that lichen compounds are sensitive to heavy metal accumulation and could be used as biomarkers in environmental studies.

Vaccinium myrtillus L. is a common shrub in the herbaceous layer of pine forests. In connection with resistance to pollution it has been widely used in the environmental monitoring based on heavy metal accumulations in the foliage. The present study investigates leaves of bilberry growing under influence of emissions from a Zn-Pb smelter. Determination of heavy metal levels by AAS revealed enhanced accumulations of Zn, Pb, Cd, and Fe, and the level of Zn fell into the toxic range compiled for plants of average sensitivity. To verify the significance of metal accumulations occurring in the studied plant, the concentrations of phenolics were quantified by measuring UV absorption of foliage extracts. Phenolics can be used as a potential biomarker of pollution because they participate in plant's response to accumulation of heavy metals, acting as antioxidants able to scavenge free radicals produced by metal ions. Leaves collected near the smelter contained significantly higher amounts of phenolics. Since these compounds are also responsible for ecological interactions, changes in their concentrations resulting from heavy metal accumulations might also influence other organisms in the ecosystem. To study the role of phenolics in heavy metal accumulations at the cellular level, a special fixation was applied for TEM, involving glutaraldehyde with caffeine, to precipitate phenolics in the cell. A histochemical reaction showed significantly higher electron-opacity, most probably deposits of phenolics, in cells of the leaves collected near the smelter. The subsequent degenerative changes in mesophyll of these leaves were characterized by increased contents of heterogeneous multivesicular structures, degeneration of chloroplasts, and disorganisation of membranes leading to death of the cell and necrosis.