Rich in Phenolics-Strong Antioxidant Fruit? Comparative Study of 25 Strawberry Cultivars.

Phenolic compounds of 25 newly introduced strawberry cultivars were profiled using spectrophotometry, electron paramagnetic resonance (EPR) spectroscopy, and high-performance liquid chromatography-mass spectrometry. Total phenolic and anthocyanin content (TPC and TACY, respectively), as well as vitamin C, and concentrations of individual phenolic compounds in fruits were evaluated to identify the most promising cultivars according to their phenolic profile. The highest values of TPC, TACY, and vitamin C were recorded in 'Premy' (1.53 mg eq GA g-1 FW), 'Sandra' (30.60 mg eq Pg-3-g 100 g-1 FW), and 'Laetitia' (56.32 mg 100 g-1 FW), respectively. The DPPH and •OH radicals scavenging activity of fruit methanolic extracts was estimated using EPR spectroscopy. All cultivars are almost uniformly effective in the scavenging of •OH radical, while 'Tea', 'Premy', and 'Joly' were marked as highly potent cultivars (over 70%) in terms of DPPH-antiradical activity. Specific peroxidase activities were the highest in 'Garda', 'Federica', and 'Rumba' (0.11, 0.08, and 0.06 U mg-1 prot, respectively). 'Laetitia', 'Joly', 'Arianna', 'Tea', and 'Mila' cultivars were distinguished from others as the richest concerning almost all flavonoids and phenolic acids, including some other parameters of bioactivity. These cultivars could be recommended to consumers as functional fruit foods.

Phenolic Profiling of 12 Strawberry Cultivars Using Different Spectroscopic Methods.

Phenolic compounds of 12 strawberry cultivars were profiled using spectrophotometry, spectrofluorometry, and high-performance liquid chromatography-mass spectrometry, coupled with multivariate regression analysis. Total phenolic content, total anthocyanin content, and total antioxidant capacity (TPC, TACY, and TAC, respectively) and concentrations of individual phenolics were evaluated, and the multivariate statistic was employed to identify the most promising cultivars based on their phenolic content. According to the principal component analysis, TAC was strongly correlated with the TPC (0.81), pointing out its importance in overall antioxidant activity. 'Joly', 'Laetitia', and 'Asia' cultivars were distinguished from others as the richest in concerning identified anthocyanins, almost all flavonoids and phenolic acids. Multivariate curve resolution with alternating least squares indicated the presence of two main types of fluorophores assigned to anthocyanins and phenolics, in which emission spectral ratios also showed the highest values in the referred cultivars. These cultivars could be recommended to consumers in terms of the health functionality of fruit.

Biochemical and histological characterization of succulent plant Tacitus bellus response to Fusarium verticillioides infection in vitro.

We present changes in Tacitus bellus antioxidative system that specifically correspond to subsequent phases of hemibiotroph Fusarium verticillioides infection revealed by histological analysis. T. bellus response to spore germination 6 h post inoculation (hpi), manifested as first oxidative burst, was characterized by transient decrease in malondialdehyde (MDA) content, transient increase in catalase (CAT), low level of superoxide dismutase (SOD) and peroxidase (POD) activity, as well as with transient decrease in total antioxidant capacity (TAC), total phenol content (TPC) and phenylalanine ammonium lyase activity (PAL), and no changes in polyphenol oxidase (PPO) activity, or phenolic profile. During the biotrophic phase of F. verticillioides infection, characterized by hyphae spread intercellularly in epidermal and mesophyll tissue, the host antioxidative system was suppressed. The transition to necrotrophic phase of F. verticillioides infection (inter- and intracellular colonization and sporulation), occurred 3-4 days post inoculation (dpi). During the necrotrophic phase, 5-7 dpi, slowed progression of colonization of T. bellus mesophyll cells occurred and it coincided with sharp increase in MDA content and CAT, SOD and POD activities, but the drop in TAC, TPC content, and PPO activity, as well as the production of phytotoxin fusaric acid. Presented results add to the knowledge of events and mechanisms related to the transition from biotrophy to necrotrophy in F. verticillioides.

Anticancer properties of Ganoderma lucidum methanol extracts in vitro and in vivo.

Anticancer activities of various extracts of the medicinal mushroom, Ganoderma lucidum, have been widely demonstrated and are mainly associated with the presence of different bioactive polysaccharides and triterpenoids. We have evaluated and compared in vitro and in vivo the antitumor effects of two preparations from Ganoderma lucidum: a methanol extract containing total terpenoids (GLme) and a purified methanol extract containing mainly acidic terpenoids (GLpme). Both extracts inhibited tumor growth of B16 mouse melanoma cells inoculated subcutaneously into syngeneic C57BL/6 mice and reduced viability of B16 cells in vitro, whereby GLme exhibited stronger effect. Furthermore, anticancer activity of GLme was demonstrated for the first time against two other rodent tumor cell lines, L929-mouse fibrosarcoma and C6-rat astrocytoma. The mechanism of antitumor activity of GLme comprised inhibition of cell proliferation and induction of caspase-dependent apoptotic cell death mediated by upregulated p53 and inhibited Bcl-2 expression. Moreover, the antitumor effect of the GLme was associated with intensified production of reactive oxygen species, whereas their neutralization by the antioxidant, N-acetyl cysteine, resulted in partial recovery of cell viability. Thus, our results suggest that GLme might be a good candidate for treatment of diverse forms of cancers.

Potential of Teucrium chamaedrys L. to modulate apoptosis and biotransformation in colorectal carcinoma cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Teucrum chamaedrys L. is one of the known medicinal plants, useful for treatment of various health problems, especially digestive. In this study, we investigated methanol, ethyl-acetate and acetone extracts of T. chamaedrys in respect to their anticancer properties in SW480 colorectal cancer cells. MATERIALS AND METHODS: Cytotoxicity and proapoptotic potential were assessed by MTT cell viability assay and AO/EB double staining. Molecular mechanisms of induced apoptosis were determined by monitoring Fas receptor protein expression through immunofluorescence, Caspase 8 and 9 activity, as well as concentrations of O2.- spectrophotometrically. Additionally, mRNA expression of biotransformation enzymes (CYP1A1, CYP1B1, GSTP1) and membrane transporters (MRP1 and MRP2) involved in drug resistance were investigated by qPCR method. Qualitative analysis of individual phenolic compounds was performed by reversed phase HPLC-MS analysis. RESULTS: Methanol extract shows the best cytotoxicity and selectivity compared to ethyl-acetate and acetone extracts, mainly causing apoptosis of SW480 cells, without affecting normal HaCaT keratinocytes. The increased expression of Fas receptor protein and caspase 8 activity indicate that the death receptor-mediated pathway plays a crucial role in the observed apoptosis. The increased caspase 9 activity and O2.- concentration suggest that mitochondria are also involved in the apoptosis. T. chamaedrys methanol extract inhibits mRNA expression of CYP1A1, CYP1B1, GSTP1, MRP1 and MRP2 in SW480 cells. CONCLUSIONS: Induction of apoptosis and inhibition of CYP1A1, CYP1B1, GSTP1, MRP1 and MRP2 mRNA expression implies that T. chamaedrys can serve as a valuable source of bioactive compounds as dietary supplements or selective anticancer agents, with the ability to induce apoptosis and modulate drug resistance in colorectal cancer cells.

Filter strip as a method of choice for apoplastic fluid extraction from maize roots.

Apoplastic fluid was extracted from maize (Zea mays L.) roots using two procedures: collection from the surface of intact plant roots by filter paper strips (AF) or vacuum infiltration and/or centrifugation from excised root segments (AWF). The content of cytoplasmic marker (glucose-6-phosphate, G-6-P) and antioxidative components (enzymes, organic acids, phenolics, sugars, ROS) were compared in the extracts. The results obtained demonstrate that AF was completely free of G-6-P, as opposed to AWF where the cytoplasmic constituent was detected even at mildest centrifugation (200×g). Isoelectric focusing of POD and SOD shows the presence of cytoplasmic isoforms in AWF, and HPLC of sugars and phenolics a much more complex composition of AWF, due to cytoplasmic contamination. Organic acid composition differed in the two extracts, much higher concentrations of malic acid being registered in AF, while oxalic acid due to intracellular contamination being present only in AWF. EPR spectroscopy of DEPMPO spin trap in the extracts showed persistent generation of hydroxyl radical adduct in AF. The results obtained argue in favor of the filter strip method for the root apoplastic fluid extraction, avoiding the problems of cytoplasmic contamination and dilution and enabling concentration measurements in minute regions of the root.