Effect of consumption of organically and conventionally produced apples on antioxidant activity and DNA damage in humans

The present study was performed to compare the effects on antioxidant activity and on DNA damage of organic and conventionally produced apples grown under controlled conditions in human peripheral blood lymphocytes. Six healthy volunteers consumed either organically or conventionally grown apples (Golden Delicious, 1000 g) from two neighboring commercial farms in a double-blinded, randomized, cross-over study. The average content of total identified and quantified polyphenols in the organically and conventionally produced apples was 308 and 321 microg/g fresh weight, respectively. No statistically significant differences in the sum of phenolic compounds or in either of the polyphenol classes were found between the agricultural methods. Consumption of neither organically nor conventionally grown apples caused any changes in antioxidant capacity of low-density lipoproteins (lag time test), endogenous DNA strand breaks, Fpg protein-sensitive sites, or capacity to protect DNA against damage caused by hydrogen peroxide. However, a statistically significant decrease in the levels of endonuclease III sensitive sites and an increased capacity to protect DNA against damage induced by iron chloride were determined 24 h after consumption in both groups of either organic or conventionally grown apples, indicating the similar antigenotoxic potential of both organically and conventionally grown apples.     

A vestige of Earth's oldest ophiolite

A sheeted-dike complex within the approximately 3.8-billion-year-old Isua supracrustal belt (ISB) in southwest Greenland provides the oldest evidence of oceanic crustal accretion by spreading. The geochemistry of the dikes and associated pillow lavas demonstrates an intraoceanic island arc and mid-ocean ridge-like setting, and their oxygen isotopes suggest a hydrothermal ocean-floor-type metamorphism. The pillows and dikes are associated with gabbroic and ultramafic rocks that together make up an ophiolitic association: the Paleoarchean Isua ophiolite complex. These sheeted dikes offer evidence for remnants of oceanic crust formed by sea-floor spreading of the earliest intact rocks on Earth.     

Crypthecodinium cohnii Growth and Omega Fatty Acid Production in Mediums Supplemented with Extract from Recycled Biomass

Crypthecodinium cohnii is a marine heterotrophic dinoflagellate that can accumulate high amounts of omega-3 polyunsaturated fatty acids (PUFAs), and thus has the potential to replace conventional PUFAs production with eco-friendlier technology. So far, C. cohnii cultivation has been mainly carried out with the use of yeast extract (YE) as a nitrogen source. In the present study, alternative carbon and nitrogen sources were studied: the extraction ethanol (EE), remaining after lipid extraction, as a carbon source, and dinoflagellate extract (DE) from recycled algae biomass C. cohnii as a source of carbon, nitrogen, and vitamins. In mediums with glucose and DE, the highest specific biomass growth rate reached a maximum of 1.012 h⁻¹, while the biomass yield from substrate reached 0.601 g·g⁻¹. EE as the carbon source, in comparison to pure ethanol, showed good results in terms of stimulating the biomass growth rate (an 18.5% increase in specific biomass growth rate was observed). DE supplement to the EE-based mediums promoted both the biomass growth (the specific growth rate reached 0.701 h⁻¹) and yield from the substrate (0.234 g·g⁻¹). The FTIR spectroscopy data showed that mediums supplemented with EE or DE promoted the accumulation of PUFAs/docosahexaenoic acid (DHA), when compared to mediums containing glucose and commercial YE.     

Kinetic and Stoichiometric Modeling-Based Analysis of Docosahexaenoic Acid (DHA) Production Potential by Crypthecodinium cohnii from Glycerol,Glucose and Ethanol

Docosahexaenoic acid (DHA) is one of the most important long-chain polyunsaturated fatty acids (LC-PUFAs), with numerous health benefits. Crypthecodinium cohnii, a marine heterotrophic dinoflagellate, is successfully used for the industrial production of DHA because it can accumulate DHA at high concentrations within the cells. Glycerol is an interesting renewable substrate for DHA production since it is a by-product of biodiesel production and other industries, and is globally generated in large quantities. The DHA production potential from glycerol, ethanol and glucose is compared by combining fermentation experiments with the pathway-scale kinetic modeling and constraint-based stoichiometric modeling of C. cohnii metabolism. Glycerol has the slowest biomass growth rate among the tested substrates. This is partially compensated by the highest PUFAs fraction, where DHA is dominant. Mathematical modeling reveals that glycerol has the best experimentally observed carbon transformation rate into biomass, reaching the closest values to the theoretical upper limit. In addition to our observations, the published experimental evidence indicates that crude glycerol is readily consumed by C. cohnii, making glycerol an attractive substrate for DHA production.     

Cloudy apple juice is more effective than apple polyphenols and an apple juice derived cloud fraction in a rat model of colon carcinogenesis

As recently shown, a cloudy apple juice (CloA) was effective to modulate colon cancer associated parameters in rats treated with 1,2-dimethylhydrazine (DMH). To identify the bioactive substance classes in CloA, we fractionated CloA to yield a total polyphenol (PF) and a cloud (CF) fraction consisting of proteins, fatty acids, polyphenols, and cell wall polysaccharides. Rats received water (control (Cont)) or CloA, PF, and CF separate or combined (PF-CF) ad libitum for 7 weeks starting one week before the first DMH-injection. As determined by comet assay, the DMH-induced genotoxicity in colonocytes of controls (Cont/DMH: 7.7 +/- 0.5%) was significantly reduced by CloA (3.3 +/- 0.3%) but not by any of the fractions. The crypt cell proliferation induced by DMH (Cont/NaCl: 7.5 +/- 0.6%; Cont/DMH: 14.9 +/- 0.8%) was significantly decreased by CloA (9.4 +/- 0.4%), PF (12.4 +/- 0.7%), CF (11.6 +/- 0.4%), and PF-CF (12.4 +/- 0.6%). Although not statistically significant, CloA tended to reduce the number of large aberrant crypt foci (ACF) (Cont/DMH: 19.0 +/- 3.7; CloA/DMH: 12.3 +/- 1.9), while none of the fractions affected ACFs. Neither CloA nor the fractions changed mRNAs of colonic cyclooxygenases (COX-1, COX-2), glutathione-associated enzymes (GST-M2, gamma-GCS, GST-P), the splenocyte CD4/CD8 ratio, natural killer cell activity, and plasma antioxidant status. These results demonstrate that CloA had a higher cancer-preventive potential than the fractions and further, besides PF, identified CF as an additional bioactive fraction of CloA.     

Phytoestrogens modulate binding response of estrogen receptors alpha and beta to the estrogen response element

Binding of estrogen receptor (ER) to estrogen response element (ERE) induces gene activation and is an important step in estrogen-induced biological effects. Here, we investigated the effects of some dietary phytoestrogens such as the isoflavones genistein and daidzein, its metabolite equol, and the coumestane coumestrol on the binding rate of ERalpha and ERbeta to ERE by a nonradioactive real-time method, the Biacore Technology. ERalpha and ERbeta were able to bind to ERE immobilized on the surface of a sensor chip even in the absence of estrogens. 17beta-Estradiol and phytoestrogens induced an increase in ER binding to ERE in a concentration-dependent manner. 17beta-Estradiol was a more potent activator of binding than the phytoestrogens studied. The concentrations of 17beta-estradiol inducing an increase in the binding response of ERalpha and ERbeta to ERE by 50% (EC(50)) as compared to unliganded ER were 0.03 and 0.01 microM, respectively. Regarding the efficacy of activation of ERalpha, from the most to the least effective compound, the sequence and the EC(50) were as follows: 17beta-estradiol (0.03 microM) > coumestrol (0.2 microM) > equol (3.5 microM) > genistein (15 microM) > daidzein (>300 microM) and for ERbeta 17beta-estradiol (0.01 microM) > coumestrol (0.025 microM) > genistein (0.03 microM) > daidzein (0.35 microM) > equol (0.4 microM). The ratios EC(50)alpha/EC(50)beta were calculated to be for 17beta-estradiol, 3; coumestrol, 8; equol, 8.8; genistein, 500; daidzein > 850. These ratios indicate that genistein and daidzein preferentially activate the binding of ERbeta to ERE. The endogenous hormone 17beta-estradiol as well as coumestrol and daidzein metabolite equol activate the binding of ERbeta to ERE only slightly more effectively than the binding of ERalpha to ERE. Thus, the effect of daidzein can be changed from a specific activator of ERbeta to an activator of both ER isotypes alpha and beta in humans who are able to convert daidzein to equol. While the results of the measurements with ERalpha were in line with the binding affinities of compounds tested for ER, there was a distinct difference between our results and the binding affinities of phytoestrogens for the ERbeta. This leads to the conclusion that phytoestrogens differ not only in their binding affinities for the ER, but also in their potential to increase the rate of receptor binding to the ERE.     

Piceid (resveratrol glucoside) synthesis in stilbene synthase transgenic apple fruit

A stilbene synthase gene along with the selectable marker gene bar for herbicide resistance was transferred via Agrobacterium tumefaciens mediated transformation into apple (Malus domesticaBorkh.) cvs. 'Elstar' and 'Holsteiner Cox'. The stilbene synthase catalyzes the conversion of 1 molecule of p-coumaroyl-CoA and 3 molecules of malonyl-CoA into 3,4',5-trihydroxystilbene, commonly known as resveratrol. This phytoalexin has implications in both phytopathology and human health. Greenhouse-grown transgenic and nontransformed control plants were grafted onto dwarfing rootstock M27. Flowering and fruiting occurred within the following years, offering the opportunity to analyze transgenic apple fruit and fertility of transgenic plants as well as inheritance of the transgenes into the seedling progeny. Molecular analysis revealed that the stilbene synthase is expressed in transgenic plants and in the skin and flesh of transgenic apple fruit. After formation, resveratrol is modified by the addition of a hexose sugar. The resulting component was characterized as piceid. With the aim of characterizing the influence of the novel biosynthetic pathway on the accumulation of other phenolic compounds naturally present in apple fruit, the amounts of flavanols, flavonols, phloretin derivatives and hydroxycinnamic acids in wild type and transgenic fruit were determined by HPLC. In all investigated transformed lines that accumulated piceid, no negative correlation between levels of piceid and the above-mentioned compounds was observed, except for the flavonol contents, which slightly decreased. Inheritance of the transgenes was confirmed in the seedling progeny, which were obtained after pollination of transgenic plants with nontransgenic pollen and vice versa after pollination of nontransgenic plants with pollen obtained from transgenic plants. The fertility of stilbene synthase transgenic plants was demonstrated. To the authors' knowledge this is the first time that data are available on piceid synthesis in transgenic apple fruit and the effects of its accumulation on levels of other phenolic compounds present in the fruit.     

Early life recorded in archean pillow lavas

Pillow lava rims from the Mesoarchean Barberton Greenstone Belt in South Africa contain micrometer-scale mineralized tubes that provide evidence of submarine microbial activity during the early history of Earth. The tubes formed during microbial etching of glass along fractures, as seen in pillow lavas from recent oceanic crust. The margins of the tubes contain organic carbon, and many of the pillow rims exhibit isotopically light bulk-rock carbonate delta13C values, supporting their biogenic origin. Overlapping metamorphic and magmatic dates from the pillow lavas suggest that microbial life colonized these subaqueous volcanic rocks soon after their eruption almost 3.5 billion years ago.     

Cellular uptake of carotenoid-loaded oil-in-water emulsions in colon carcinoma cells in vitro

Oil-in-water emulsions allow the preparation of lipophilic compounds such as carotenoids in the liquid form. Here, the effect of a combination of some emulsifiers, such as two whey protein isolates (BiPro and BioZate), sucrose laurate (L-1695), and polyoxyethylene-20-sorbitan-monolaurate (Tween 20), on the stability of lycopene and astaxanthin in emulsions, droplet size, and cellular uptake of these carotenoids has been investigated. The degradation of lycopene was slightly more pronounced than that of astaxanthin in all emulsions. The concentration of lycopene and astaxanthin decreased by about 30% and 20%, respectively, in all emulsions after 3 weeks of storage in the dark at 4 degrees C. The kind of emulsifiers or their combinations have played an important role in the cellular uptake by the colon carcinoma cells line HT-29 and Caco-2.