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BACKGROUND: Guaiacol peroxidases (GP) are haem-containing enzymes participating in many physiological processes in plants. The expression pattern of these enzymes is organ-specific and developmentally regulated. METHODS: The presence of GP activity in extract samples, prepared from Crocus sativus L. corms that were either dormant or rooting for 3, 6 and 10 days, was investigated. RESULTS: Kinetic studies revealed a significant similarity among GP activities detectable in the corm at different stages of development: in all extract samples, the activity was maximal at pH 7.5 and after preincubation at 30-40 degrees C. When guaiacol was used as the varying substrate, Michaelis-Menten kinetics behavior was observed in all extract samples and resulted in similar KM values; catalytic efficiencies were also very similar. The corm GP activity was inhibited by cyanide, azide and ascorbate. The GP activities from different extract samples had the same sensitivities to azide, cyanide and ascorbate and the type of inhibition by azide and cyanide was competitive and uncompetitive, respectively, while ascorbate inhibited the GP activity non-competitively. Corm extract samples from different stages of rooting similarly responded to temperature treatment and a biphasic Arrhenius plot resulted for each extract sample studied. When dormant, 3-, 6- and 10-days-rooting corm extracts were submitted to non-denaturing polyacrylamide gel electrophoresis, the GP-specific activity staining revealed one band on the gel, with the same migrating distances. CONCLUSION: This finding in combination with kinetic studies demonstrated that at least one form of GP, with an apparent molecular weight of 68 kDa, was expressed during development of Crocus sativus L. corm. 相似文献
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Aparicio-Fernández X García-Gasca T Yousef GG Lila MA González de Mejia E Loarca-Pina G 《Journal of agricultural and food chemistry》2006,54(6):2116-2122
The antiproliferative effects of 100% methanol crude extract and of Toyopearl and silica gel fractions from the seed coats of black Jamapa beans (Phaseolus vulgaris L.) were evaluated using HeLa, human adenocarcinoma cells, and HaCaT, human premalignant keratinocytes. The 100% methanol crude extract [172.2 microM equiv of (+)-catechin] increased adhesion of HeLa cells; however, 3- and 5-fold higher concentrations decreased the number of cells attached as a function of the treatment time. The highest concentration tested diminished the cell adhesion until 40% (after 24 h) to almost 80% (after 72 h). The IC50 values showed that the 100% methanol crude extract was the most effective inhibitor of HeLa cell proliferation, even when it was dissolved in dimethylsulfoxide (DMSO) [34.5 microM equiv of (+)-catechin] or in medium [97.7 microM equiv of (+)-catechin]. The Toyopearl 5 (TP5) fraction and silica gel 2 (SG2) fraction inhibited 60% of the HeLa cell proliferation. The IC50 was 154 microM equiv of (+)-catechin of the 100% methanol crude extract on HaCaT cells. Toyopearl fractions TP4 and TP6 significantly inhibited HaCaT cell proliferation, but the silica gel fractions did not have a significant effect. The 100% methanol crude extract (35 microg of dry material/mL) decreased the number of HeLa cells in the G0/G1 phase from 68.9% (for control cells) to 51.4% (for treated cells) and increased apoptosis (2.9 and 21.2% for control and treated cells, respectively). The results indicated that black Jamapa beans could be a source of polyphenolic compounds, which have an inhibitory effect toward HeLa cancer cells but are less aggressive on HaCaT premalignant cells. 相似文献
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Phytochemical composition and metabolic performance-enhancing activity of dietary berries traditionally used by Native North Americans 总被引：1，自引：0，他引：1
Burns Kraft TF Dey M Rogers RB Ribnicky DM Gipp DM Cefalu WT Raskin I Lila MA 《Journal of agricultural and food chemistry》2008,56(3):654-660
Four wild berry species, Amelanchier alnifolia, Viburnum trilobum, Prunus virginiana, and Shepherdia argentea, all integral to the traditional subsistence diet of Native American tribal communities, were evaluated to elucidate phytochemical composition and bioactive properties related to performance and human health. Biological activity was screened using a range of bioassays that assessed the potential for these little-known dietary berries to affect diabetic microvascular complications, hyperglycemia, pro-inflammatory gene expression, and metabolic syndrome symptoms. Nonpolar constituents from berries, including carotenoids, were potent inhibitors of aldose reductase (an enzyme involved in the etiology of diabetic microvascular complications), whereas the polar constituents, mainly phenolic acids, anthocyanins, and proanthocyanidins, were hypoglycemic agents and strong inhibitors of IL-1beta and COX-2 gene expression. Berry samples also showed the ability to modulate lipid metabolism and energy expenditure in a manner consistent with improving metabolic syndrome. The results demonstrate that these berries traditionally consumed by tribal cultures contain a rich array of phytochemicals that have the capacity to promote health and protect against chronic diseases, such as diabetes. 相似文献
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Lu CH Engelmann NJ Lila MA Erdman JW 《Journal of agricultural and food chemistry》2008,56(17):7710-7714
Radioisotope-labeled lycopene is an important tool for biomedical research but currently is not commercially available. A tomato cell suspension culture system for the production of radioisotope-labeled lycopene was previously developed in our laboratory. In the current study, the goal was to optimize the lycopene extraction efficiency from tomato cell cultures for preparatory high-performance liquid chromatography (HPLC) separation. We employed response surface methodology (RSM), which combines fractional factorial design and a second-degree polynomial model. Tomato cells were homogenized with ethanol, saponified by KOH, and extracted with hexane, and the lycopene content was analyzed by HPLC-PDA. We varied five factors at five levels: ethanol volume (1.33-4 mL/g); homogenization period (0-40 s/g); saturated KOH solution volume (0-0.67 mL/g); hexane volume (1.67-3 mL/g); and vortex period (5-25 s/g). Ridge analysis by SAS suggested that the optimal extraction procedure to extract 1 g of tomato cells was at 1.56 mL of ethanol, 28 s homogenization, 0.29 mL of KOH, 2.49 mL of hexane, and 17.5 s vortex. These optimal conditions predicted by RSM were confirmed to enhance lycopene yield from standardized tomato cell cultures by more than 3-fold. 相似文献
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Campbell JK Rogers RB Lila MA Erdman JW 《Journal of agricultural and food chemistry》2006,54(3):747-755
This work describes the development and utilization of a plant cell culture production approach to biosynthesize and radiolabel phytoene and phytofluene for prostate cancer cell culture studies. The herbicide norflurazon was added to established cell suspension cultures of tomato (Lycopersicon esculentum cv. VFNT cherry), to induce the biosynthesis and accumulation of the lycopene precursors, phytoene and phytofluene, in their natural isomeric forms (15-cis-phytoene and two cis-phytofluene isomers). Norflurazon concentrations, solvent carrier type and concentration, and duration of culture exposure to norflurazon were screened to optimize phytoene and phytofluene synthesis. Maximum yields of both phytoene and phytofluene were achieved after 7 days of treatment with 0.03 mg norflurazon/40 mL fresh medium, provided in 0.07% solvent carrier. Introduction of 14C-sucrose to the tomato cell culture medium enabled the production of 14C-labeled phytoene for subsequent prostate tumor cell uptake studies. In DU 145 prostate tumor cells, it was determined that 15-cis-phytoene and an oxidized product of phytoene were taken up and partially metabolized by the cells. The ability to biosynthesize, radiolabel, and isolate these carotenoids from tomato cell cultures is a novel, valuable methodology for further in vitro and in vivo investigations into the roles of phytoene and phytofluene in cancer chemoprevention. 相似文献
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Bibalani GH Bazhrang Z Mohsenifar H Shibaei N Joodi L 《Pakistan journal of biological sciences: PJBS》2008,11(8):1126-1131
A pulling effect by side roots is one way in which roots help to side in-plane strong of a little depth soil mass. In contrast to the effect of vertically-enlarge roots, whereby soil is strengthened by an increase in its shear strength, the pulling effect strengthens the soil by increasing the tensile strength of the rooted soil zone. To verify whether or not a pulling effect exists in the root system of Prunus avium in the Roudsar, North Iran and to study the importance and size of this effect, a direct in situ test was led at a site in the Chaboksar Forests. The results from the site showed that, in the surface soil (0-30 cm), Side roots can provide a pull force of up to 490-712 N (Newtons) over a vertical cross-section area of 20-50 cm2, or an enhance in the pulling stability of the rooted soil by about 48.1%. The test results suggest that, together with the Prunus avium vertical roots, which keep the little depth rooted soil zone to the deep and more stable soil mass, the side roots of the Prunus avium, with their pulling effect, are able to make less against little depth instability in the forest slopes, such as little depth slide, to a certain degree. 相似文献
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Aparicio-Fernández X Reynoso-Camacho R Castaño-Tostado E García-Gasca T González de Mejía E Guzmán-Maldonado SH Elizondo G Yousef GG Lila MA Loarca-Pina G 《Plant foods for human nutrition (Dordrecht, Netherlands)》2008,63(1):35-40
Jamapa bean is a black Phaseolus vulgaris variety rich in condensed tannins, anthocyanins and flavonols with interesting biological activities. The objective of this
work was to evaluate the antiradical capacity (ARC) of a Jamapa bean methanolic extract (BME) and some of the proanthocyanidin-rich
fractions derived from it, using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. The effect of the BME on some proteins involved
in apoptosis on HeLa cells was also evaluated. A strong correlation between proanthocyanidin concentration in BME and antiradical
capacity was found, suggesting that these compounds contribute significantly to antiradical activity. BME was a better radical
scavenger than butylated hydroxytoluene (45.6 and 33.9% ARC at 400 μM, respectively). Two proanthocyanidin-rich fractions
obtained after a preliminary separation of the BME using Toyopearl (TP4 and TP6) exhibited a higher antiradical activity than
the parent extract. The treatment of HeLa cells with 35 μg BME/ml/24 h increased the expression of Bax and Caspase-3, pro-apoptotic
proteins (6.13 and 1.2 times for Caspase-3 and Bax, respectively). The mechanism of action of some proteins involved in apoptosis
was also evaluated, and the results suggest that black Jamapa bean could be an important source of polyphenolic compounds
with potential biological use as antioxidant and anticancer agents. 相似文献
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Boulekbache-Makhlouf L Meudec E Chibane M Mazauric JP Slimani S Henry M Cheynier V Madani K 《Journal of agricultural and food chemistry》2010,58(24):12615-12624
A method based on high-performance liquid chromatography coupled with diode array detection and electrospray ionization mass spectrometry (HPLC-DAD-ESI-MS) following fractionation by chromatography on a Sephadex LH-20 column has been developed to determine the phenolic composition of fruit of Eucalyptus globulus growing in Algeria. The presence of 18 gallotannins, 26 ellagitannins, and 2 flavonols was established. Tentative identification is provided for these compounds on the basis of UV-visible spectra and mass spectrometry data. Most compounds described in this study have not previously detected in fruit of E. globulus. Moreover, this is the first report of methyl digalloyl diglucose, 3,3'-O-dimethylellagic acid 4-O-β-glucopyranoside, ellagic acid hexose, methyl ellagic acid pentose, methyltetragalloylglucose, and valoneic acid isomers (sanguisorbic, flavogallic acid dilactone) in the genus Eucalyptus. Quantitatively, ellagic acid and its derivatives, including ellagitannins, are largely predominant. 相似文献
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Effects of a twin strain of saccharomyces cerevisiae live cells on mixed ruminal microorganism fermentation in vitro 总被引：5，自引：0，他引：5
Lila ZA Mohammed N Yasui T Kurokawa Y Kanda S Itabashi H 《Journal of animal science》2004,82(6):1847-1854
This experiment was designed to investigate the effects of different concentrations (0, 0.33, 0.66, 0.99, and 1.32 g/L) of a twin-strain of Saccharomyces cerevisiae live cells on in vitro mixed ruminal microorganism fermentation of corn starch, soluble potato starch, and sudangrass hay (60.5%, DM basis) plus concentrate mixture (39.5%, DM basis). Ruminal fluid was collected from two dairy cows, mixed with phosphate buffer (1:2), and incubated (30 mL) anaerobically at 38 degrees C for 6 and 24 h with or without yeast supplement, using 200 mg (DM basis) of each substrate. Medium pH, ammonia-N, and numbers of protozoa were unaffected (P = 0.38) by yeast cells in all substrates. Molar proportion of acetate was unchanged (P = 0.56) with cornstarch and soluble potato starch, but increased quadratically (P = 0.02) with hay plus concentrate by treatment. Addition of yeast cells caused a linear increase of total VFA (P = 0.008) in all substrates. Excluding the soluble potato starch, supplementation of S. cerevisiae resulted in a quadratic increase of propionate (P = 0.01), with a quadratic decrease (P = 0.04) of acetate:propionate. When soluble potato starch was used as a substrate, a linear increase (P = 0.006) of the molar proportion of propionate and a quadratic decrease (P = 0.007) in acetate:propionate was observed by treatment. Molar proportion of butyrate was unchanged (P = 0.35) with cornstarch and soluble potato starch, whereas it decreased linearly (P = 0.007) with hay plus concentrate by yeast cell supplementation. When cornstarch and soluble potato starch were used as a substrate, minor VFA were decreased (P = 0.05) by treatment. Accumulation of lactate was linearly decreased by treatment (P = 0.007) in all substrates. During incubation with hay plus concentrate, IVDMD was linearly increased (P = 0.006), whereas production of methane (linear; P = 0.02) and accumulation of hydrogen was decreased (quadratic; P = 0.005) by treatment after 24 h. These results showed that a twin strain of S. cerevisiae live cells stimulated in vitro mixed ruminal microorganism fermentation with decreased lactate, and a small decrease of methane and hydrogen with hay plus concentrate. 相似文献