Effects of Cd and Al stress on secondary metabolites, antioxidant and antibacterial activity of Hypoxis hemerocallidea Fisch. & C.A. Mey.

This study investigated the effects of cadmium (Cd) and aluminium (Al) on the accumulation of phenolics, flavonoids and the bioactive compound hypoxoside in Hypoxis hemerocallidea. In addition, antioxidant scavenging and antibacterial activity were assessed to evaluate if Cd and Al stress affect the accumulation of bioactive compounds in H. hemerocallidea. In vitro grown plantlets of H. hemerocallidea were acclimatized for seven months in a greenhouse. Thereafter plants were exposed to various concentrations of Cd and Al both singularly and in combination in the form of Cd(NO3)2 (2, 5, 10 mg Cd/L); Al3(NO3)3 (500, 1000, 1500 mg Al/L) and combinations of Cd and Al (Cd 2:Al 500, Cd 5:Al 1000 and Cd 10:Al 1500 mg/L) for a further six weeks. The highest amounts of Cd and Al translocated to the shoot were 34 and 1608 mg/L respectively. Phytochemical screening showed significantly high amounts of total phenolics and flavonoids at the moderate Cd treatment (5 mg/L) compared to the controls. Exposure to Cd and Al significantly decreased the accumulation of hypoxoside. There was a significant increase in diphenylpicrylhydrazyl (DPPH) antioxidant scavenging activity in most of the metal-treated plants compared to the positive control ascorbic acid. Extracts from Cd 2 mg/L treatment exhibited moderate antibacterial activity against Staphylococcus aureus compared to the control. The results of the present study revealed that cultivating H. hemerocallidea on metal contaminated soils affects the accumulation of the bioactive compound, hypoxoside.

The design of dextran-based hypoxia-inducible hydrogels via in situ oxygen-consuming reaction.

Hypoxia plays a critical role in the development and wound healing process, as well as a number of pathological conditions. Here, dextran-based hypoxia-inducible (Dex-HI) hydrogels formed with in situ oxygen consumption via a laccase-medicated reaction are reported. Oxygen levels and gradients were accurately predicted by mathematical simulation. It is demonstrated that Dex-HI hydrogels provide prolonged hypoxic conditions up to 12 h. The Dex-HI hydrogel offers an innovative approach to delineate not only the mechanism by which hypoxia regulates cellular responses, but may facilitate the discovery of new pathways involved in the generation of hypoxic and oxygen gradient environments.

Plant regeneration and biochemical accumulation of hydroxybenzoic and hydroxycinnamic acid derivatives in Hypoxis hemerocallidea organ and callus cultures.

Micropropagation of Hypoxis hemerocallidea Fisch. and C.A. Mey was used as a model system to study the influence of cytokinins (CKs) on plant regeneration and biochemical accumulation of hydroxybenzoic and hydroxycinnamic acid derivatives in organ and callus cultures and their antioxidant activity. Fourteen free phenolic acids were detected using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) while antioxidant activity was evaluated using oxygen radical absorbance capacity (ORAC) and 2,2-diphenyl-1-picryl hydrazyl (DPPH) radical scavenging activity. Cytokinins had a significant effect on the biochemical accumulation of hydroxybenzoic and hydroxycinnamic acid derivatives in H. hemerocallidea organ cultures. In particular, meta-topolin-treated organ cultures produced high concentrations of gallic, protocatechuic, gentisic, p-hydroxybenzoic, m-hydroxybenzoic, salicylic, chlorogenic and trans-cinnamic acids. The isoprenoid CK, N(6)-(2-isopentenyl)-adenine significantly increased the accumulation of hydroxycinnamic acid derivatives, namely, caffeic, p-coumaric, sinapic and ferulic acids. Cytokinin-treated organ cultures exhibited a significant increase in antioxidant activity, particularly in the ORAC model. In callus cultures, CKs decreased the concentrations of hydroxycinnamic acid derivatives and antioxidant activity when compared to the control. Overall, both CK type and concentration had a significant effect on plant regeneration, callus proliferation, biochemical accumulation of free phenolic acids and antioxidant activity of the resultant extracts.

Macro, minor and toxic elemental uptake and distribution in Hypoxis hemerocallidea, "the African Potato"--an edible medicinal plant.

The elemental uptake and distribution, in various parts of the admired herbal plant, Hypoxis hemerocallidea, the 'African potato' and its ability to accumulate elements in response to the growth soil quality are investigated. The total and exchangeable concentrations of twelve elements in the growth soils and their distribution in the roots, potato bulb and leaves of the plants grown under four different settings were compared. The typical concentrations of the twelve selected elements, in the bulb and leaves of the plant grown in a nursery pot (site 2) were (in microg g(-1)dry weight) Ca (8430 and 27075), Mg (2113 and 1566), Fe (66 and 150), Al (10 and 368), Zn (105 and 6.1), Mn (42 and 51), Cu (7.2 and 20.8), Ba (0.23 and 4.44), Co (0.20 and 0.42), As (2.05 and 24.56), Hg (0.92 and 1.82) and Cr (0.13 and 0.33). Except for Ca, Mg, Zn and Mn, the exchangeable cation concentrations in all the growth soils were low. Ca, Mg, Mn, Zn and As had bioaccumulation factors >1. Fe, Al and Co concentrations were high in the roots with little in the rest of the plant. High concentration of arsenic (approximately 13 microg g(-1) dry weight) with bioaccumulation factors of 7 and 20 were observed in the roots and leaves of the plant respectively (site 2), but the concentration of mercury in bulb was very low (0.92 microg g(-1) dry weight).

Role of membranes on the antibacterial and anti-inflammatory activities of the bioactive compounds from Hypoxis rooperi corm extract.

Hypoxis rooperi corm extract ('African potato') is known for its traditional and ethnomedical uses in the treatment of a large variety of diseases. Its main bioactive compound hypoxoside (HYP) and its aglycone derivative rooperol (RO) were isolated and the interaction of these compounds with several types of model membranes was studied in order to contribute to the understanding of their molecular mechanism. The results show that RO abolishes the main transition phase and perturb the van der Waals interactions between phospholipid acyl chains in a stronger way than HYP in dimiristoylphosphatidylcholine (DMPC), dielaidoylphosphatidylethanolamine (DEPE) and dimiristoylphosphatidylglycerol membranes (DMPG), probably indicating that this molecule inserts into the bilayer. This effect decreases as the acyl chain length of the phospholipid increases. RO also promoted the formation of hexagonal H(II) phases at lower temperatures compared to pure DEPE. On the contrary, HYP showed a shallow interaction with phospholipids. This compound promoted the formation of gel-fluid like intermediate structures with isotropic motion in phosphatidylglycerol membranes at physiological pH, and affected the phospholipid/water interface probably through the variation of the surface charge of the phospholipid phosphate groups. Moreover, RO inhibited Staphylococcus aureus in a stronger manner than Escherichia coli and promoted a higher leakage level in E. coli, PG and PE-containing synthetic membranes. Furthermore, RO showed a significant degree of inhibition of cyclooxygenase-2 (COX-2) and cyclooxygenase-1 (COX-1) evidencing an approximate COX-2/COX-1 IC50 ratio of 1.9, therefore this compound may be responsible for the anti-inflammatory activity of H. rooperi corm extract. These results may contribute to understand the molecular mechanism of the antibacterial and/or anti-inflammatory properties of the bioactive compounds deriving from the African potato corm extract.

In vitro evaluation of human cytochrome P450 and P-glycoprotein-mediated metabolism of some phytochemicals in extracts and formulations of African potato.

African potato (Hypoxis hemerocallidea, AP) is a traditional herbal medicine widely used as an immune booster and also for the treatment of various ailments such as urinary diseases, prostrate hypertrophy and cancer. Amongst the chemical components contained in AP, the norlignan glycoside, hypoxoside (HYP) is purported to be the most important phytochemical in terms of AP's medicinal value. Additional constituents in AP include the sterols, beta-sitosterol (BSS), stigmasterol (STG), and the stanol, stigmastanol (STN). The potential of extracts of AP, AP formulations as well as HYP, its aglycone rooperol (ROP) and the sterols to inhibit in vitro metabolism of drug marker substrates by human cytochrome P450 (CYP) enzymes such as CYP3A4, 3A5 and CYP19 were investigated. Samples were also assessed for their effect on drug transport proteins such as P-glycoprotein (P-gp). The effects on CYP-mediated metabolism were studied by fluorometric microtitre plate assay. The potential interaction with P-gp was investigated by measuring the efflux of the fluorescent dye rhodamine 123 (Rh 123) in the CaCo-2 (colon carcinoma) cell line. Various extracts of AP, AP formulations, only STG and the norlignans, in particular the aglycone ROP, exhibited inhibitory effects on CYP3A4-, 3A5- and 19-mediated metabolism. The extracts and the formulations that contained a significant amount of HYP showed high induction of P-gp compared to the positive control, ritonavir. Whilst extrapolation of the current in vitro findings to clinical effects may well be considered speculative, these in vitro data should be heeded as a signal of possible in vivo interactions. Appropriate measures are therefore necessary to explore the possibility of such in vitro-in vivo correlations.