Characterization of nutrients uptake and enzymes activity in Khatouni melon (Cucumis melo var. inodorus) seedlings under different concentrations of nitrogen,potassium and phosphorus of nutrient solution

A nutrient solution experiment was done to evaluate effects of different concentrations of nitrogen (N), phosphorus (P) and potassium (K) on leaf mineral concentrations and some enzymes activity of melon seedlings (Cucumismelo var. inodorus subvar. Khatouni). Different levels of these nutrients including 0, 53, 105, 158 and 210?mg L^?1 N; 0, 8, 16, 23 and 31?mg L^?1 P; 0, 59, 118, 176 and 235?mg L^?1 K, all corresponding to 0, 25, 50, 75 and 100% of their concentrations in Hoagland nutrient solution, were applied to plants. The results showed that the highest leaf nitrate reductase (NR) activity was observed at highest N and P levels, whereas the three highest K levels showed the highest NR activity. The highest leaf peroxidase activity was observed at 8?mg L^?1 P, 59?mg L^?1 K and 158?mg L^?1 N. The leaf catalase activity was highest at zero concentration of P, 158?mg L^?1 N and 176?mg L^?1 K; however, catalase activity was decreased by increasing P levels. Leaf protein content showed an increasing trend with increasing N, P and K levels of nutrient solution, while there was no significant difference between 158 and 210?mg L^?1 N. The highest leaf concentrations of N, P, K and Mg were observed at highest nitrogen, potassium and phosphorus levels of nutrient solution, whereas the highest leaf concentration of Ca were obtained at 53 or 105?mg L^?1 N, 176?mg L^?1 K and 23–31?mg L^?1 P. The highest iron concentration of leaves was obtained from 23 to 31?mg L^?1 P, 176?mg L^?1 K and 210?mg L^?1 N.     

On the occurrence, distribution, taxonomy and genepool relationship of Cucumis callosus (Rottler) Cogn., the wild progenitor of Cucumis melo L. from India

Recent research point to the Indian wild taxa of Cucumis callosus (Rottler) Cogn. as the wild progenitor of melon (C. melo L.). Overlapping distribution with cultivated and weedy and feral forms of melo and normal fertility of F₁ and BC₁ generations of its cross with cultivated melon indicate its progenitor status. A perusal of herbarium data indicate its natural distribution pattern in the region comprising Vindhya Hills and Aravalli mountain ranges extending northwards to Indo-Gangetic plains and southwards to the Deccan plateau touching rain shadow areas of Western Ghats. Characterised by drought tolerance and field resistance to a host of pests and diseases, it is conspicuously absent in the high rainfall areas of Western Ghats and upper Himalayan region. Based on its morphological distinction and F₁ and BC₁ fertility with C. melo, a subspecific rank within C. melo is postulated. First-hand information on its occurrence, distribution and crossability relationship with other Indian taxa of Cucumis are given.     

Effect of caffeic acid phenethyl ester, an antioxidant from propolis, on inducing apoptosis in human leukemic HL-60 cells.

Caffeic acid phenethyl ester (CAPE) is an active component isolated from propolis. The aim of this study was to investigate the mechanism of CAPE-induced apoptosis in human leukemic HL-60 cells. It was found that CAPE entered HL-60 cells very quickly and then inhibited their survival in a concentration- and time-dependent manner. CAPE induced characteristic DNA fragmentation and morphological changes typical of apoptosis in these cells. Estimation of the apoptotic percentage showed a time-dependent increase after CAPE (6 microg/mL) treatment (up to 66.7 +/- 2.0% at 72 h). Treatment with CAPE caused rapid activation of caspase-3 after 4 h, down-regulation of Bcl-2 expression after 6 h, and up-regulation of Bax expression after 16 h. These results suggest that CAPE is a potent apoptosis-inducing agent; its action is accompanied by activation of caspase-3, down-regulation of Bcl-2, and up-regulation of Bax in human leukemic HL-60 cells.     

热水结合果蜡处理抑制振动引起哈密瓜衰老的机理

远距离运销是哈密瓜流通过程中的重要过程,其中运输过程中的振动是影响哈密瓜衰老的重要逆境。为了减少运输振动导致的哈密瓜的成熟衰老。该文以哈密瓜为原料,利用模拟振动台对哈密瓜进行振动处理,并利用热水结合果蜡对哈密瓜进行保鲜。随后在模拟销售条件的室温条件下（23℃）对哈密瓜的细胞膜完整性及相关的指标进行了检验,同时还利用透射电镜对新鲜和贮藏结束后哈密瓜组织细胞微观结构变化进行了检测。结果表明,运输振动胁迫加速了哈密瓜细胞的微观变化及细胞膜完整性丧失,在经过28 d的模拟销售条件的室温条件下,对照组（未经处理）的相对电导率分别是振动处理组的95%和热水果蜡处理组的1.15倍,且三者间区别显著（P<0.05）。这主要是由于运输振动增加了哈密瓜果实的呼吸作用,伴随较高的丙二醛产生,较低的超氧化物歧化酶,过氧化氢酶和过氧化物酶等抗氧化酶活性。而热水结合果蜡处理抑制了运输振动引起的哈密瓜细胞膜脂质的氧化及与相关的变化,延缓了哈密瓜的衰老,保持了贮藏过程中哈密瓜的品质。研究结果可以为提高采后哈密瓜的运销品质和保鲜效果提供一定的参考。     

Polyphenols from evening primrose ( Oenothera paradoxa ) defatted seeds induce apoptosis in human colon cancer Caco-2 cells

Polyphenols extracted from evening primrose seeds (industrial waste product) were studied as apoptosis inducers in human colorectal adenocarcinoma Caco-2 and HT-29 cell lines and in rat normal intestinal IEC-6 cells. The extract dose-dependently inhibited the growth of Caco-2, HT-29, and IEC-6 cells. However, nuclear DNA fragmentation characteristic of apoptosis was observed only in Caco-2. After 72 h of incubation with the extract at 150 μM gallic acid equivalents (44.1 μg extract/mL), Caco-2 cell numbers decreased to 19% of control and 48.8% of the cells were identified by flow cytometry as apoptotic. Under the same conditions only 8% of HT-29 cells and 12.6% of IEC-6 cells exhibited hypodiploid DNA content. The effects of the extract and its fractions on phosphatidylserine exposure and cell membrane integrity were assessed by high content screening image cytometry. The fractions strongly and dose-dependently reduced Caco-2 cell numbers, whereas HT-29 and IEC-6 cells were affected to lesser extents.     

Structural determination of hypnosin, a spore germination inhibitor of phytopathogenic Streptomyces sp. causing root tumor in melon (Cucumis sp.)

The structure of a germination inhibitor, hypnosin, isolated from phytopathogenic Streptomyces sp. causing root tumor of melon was determined to be 3-acetylaminopyrazine-2-carboxylic acid (1) by mass spectrometry, computational chemical prediction of UV spectrum, and synthesis of candidates. The structure-activity relationship of hypnosin and anthranilic acid was examined, and it was concluded that pyrazinecarboxylic acid or pyridine-2-carboxylic acid was the fundamental structure with activity, that methylation of the carboxyl group or decarboxylation destroyed activity, and that the presence of an amino group was inhibitory to the activity, whereas acetylation or deletion of an amino group enhanced activity. Hypnosin inhibited spore germination of some Streptomyces spp. in addition to the species with which it was isolated.     

Induction of cell death in Caco-2 human colon carcinoma cells by ellagic acid rich fractions from muscadine grapes (Vitis rotundifolia)

Possible anticancer mechanisms exerted by polyphenolic compounds contained in fruits and vegetables include antioxidant activity, the inhibition of proliferation, and the induction of apoptosis in cancer cells. This study examined the effects of four isolated polyphenolic extracts from red muscadine grapes (Vitis rotundifolia) on vital cell parameters and the induction of apoptosis in Caco-2 colon carcinoma cells. The magnitude of effects in cell culture was then correlated to polyphenolic composition and antioxidant capacity. Whereas anticancer effects of individual polyphenolic compounds have been demonstrated multiple times, information relating to anticancer effects of polyphenolic extracts is not available in abundance. All four extracts induced apoptosis, decreased cell number, and caused alterations in cell cycle kinetics in a concentration-dependent manner. The efficacy of the polyphenolics on vital cell parameters correlated well to the presence of ellagic acid glycosides and flavonoids and also to the antioxidant capacity. This study demonstrated the anticancer properties of ellagic acid rich extracts from red muscadine juice.     

Polyozellin isolated from Polyozellus multiplex induces phase 2 enzymes in mouse hepatoma cells and differentiation in human myeloid leukaemic cell lines

Induction of cellular phase 2 detoxifying enzymes is associated with cancer preventive potential. Quinone reductase (QR) has been used as a prototype for anticarcinogenic phase 2 enzymes because of its widespread distribution in mammalian systems, large amplitude of inducer response, and ease of measurement in murine hepatoma cells. Methanol extract of Polyozellus multiplex, which shows a strong QR induction potential, was subjected to bioassay-guided fractionation, and polyozellin (PM1) appeared to be a major active component. In in vitro cultured cells (hepa1c1c7 and BPRc1 cells), polyozellin enhanced QR, glutathione S-transferase (GST) activities, and glutathione (GSH) content in a dose-dependent manner. The compound also significantly promoted differentiation of HL-60 human promyelocytic emia cells. In conclusion, polyozellin deserves further in vivo study to evaluate its potential as a cancer preventive agent.     

Propolin G, a prenylflavanone, isolated from Taiwanese propolis, induces caspase-dependent apoptosis in brain cancer cells

We have previously shown that six propolins, A-F, could be isolated from Taiwanese propolis (TP) and that they exerted a broad spectrum of biological activities. Recently, we isolated a seventh compound, propolin G. Its chemical structure has been identified by NMR and fast atom bombardment-mass spectrometry spectra and was found to be identical to a known compound, nymphaeol C. We used high-performance liquid chromatography to determine the relative contents of propolins C, D, F, and G in TP collected in various seasons and regions and found them to be relatively higher in TPs collected from May to July than from September to October. In our present study, we were interested in the various biological activities of TP extract as well as in propolin G as a pure compound. We found that propolin G could efficiently induce apoptosis in brain cancer cell lines (glioma and glioblastoma). The apoptosis might have been through a mitochondrial- and caspase-dependent pathway. This result demonstrated that the TP collection season was more an important factor than the geographical region. Propolis has been suggested to possess a potent antioxidant activity. We further evaluated the antioxidant property of propolin G using DPPH (1,2-diphenyl-2-picryhydrazyl). Our results indicate that propolin G does possess free radical scavenging activity. We also evaluated the neuroprotective action of propolin G, TP, and BP (Brazilian propolis) extracts against oxidative stress in rat primary cortical neurons. Our data demonstrate that propolin G and TP extracts have a marked neuroprotective effect that is greater than BP extract. In conclusion, the isolation and characterization of propolin G from TP have demonstrated for the first time that this compound is a potent inducer of apoptosis in brain cancer cells and that this compound and TP extract exhibit a protective effect against oxidative stress in rat cortical neurons.     

Carryover of perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) from soil to plant and distribution to the different plant compartments studied in cultures of carrots (Daucus carota ssp. Sativus), potatoes (Solanum tuberosum), and cucumbers (Cucumis Sativus)

A vegetation study was carried out to investigate the carryover of Perfluorooctanoic Acid (PFOA) and Perfluorooctane Sulfonate (PFOS) from soil mixed with contaminated sewage sludge to potato, carrot, and cucumber plants. Analysis was done by liquid-extraction using acetonitrile with dispersive SPE cleanup and subsequent HPLC-MS/MS. In order to assess the transfer potential from soil, transfer factors (TF) were calculated for the different plant compartments: TF = [PFC](plant (wet substance))/[PFC](soil (dry weight)). The highest TF were found for the vegetative plant compartments with average values for PFOS below those for PFOA: cucumber, 0.17 (PFOS), 0.88 (PFOA); potato, 0.36 (PFOS), 0.40 (PFOA); carrot, 0.38 (PFOS), 0.53 (PFOA). Transfer of PFOA and PFOS into potato peelings (average values of TF: PFOA 0.03, PFOS 0.04) exceeded the carryover to the peeled tubers (PFOA 0.01, PFOS < 0.01). In carrots, this difference did not occur (average values of TF: PFOA 0.04, PFOS 0.04). Transfer of PFOS into the unpeeled cucumbers was low and comparable to that of peeled potatoes (TF < 0.01). For PFOA, it was higher (TF: 0.03).     

Interaction among free-living N-fixing bacteria isolated from Drosera villosa var. villosa and AM fungi (Glomus clarum) in rice (Oryza sativa)

Rice is usually grown in N-deficient soils, demanding that the element be supplied to the field by commercially available N fertilizers. Unfortunately, a substantial amount of the urea-N or NO₃-N applied as fertilizers is lost through different mechanisms, causing environmental pollution problems. Utilization of biological N₂ fixation (BNF) technology can decrease the application of N fertilizers, reducing environmental risks. This study evaluated the effects of four free-living N-fixing bacterial species, isolated from oligotrophic soil conditions, as single inoculants or combined with arbuscular mycorrhizal fungi (Glomus clarum), on the development of rice plants grown as flooded or upland rice, in the greenhouse. Upland rice roots were inoculated with Methylobacterium sp., Burkholderia sp. and Sphingomonas sp., whereas the species Burkholderia sp., Pseudomonas sp. and Sphingomonas sp., were inoculated on flooded rice. Inoculants consisted of individual bacterial species or their mixtures, with or without G. clarum. Controls included non-bacteria/non-AM fungi, and AM fungi alone. Experiments were carried out in five replicates. The presence of G. clarum decreased or did not significantly affect plant growth under the different culture conditions. The presence of AM fungi stimulated the N-fixing bacterial population of upland rice. Bacterial species had different effects, under both culture conditions, and some genera of N-fixing bacteria increased root and shoot growth at different plant growth stages. The level of mycorrhiza colonization had no influence on plant growth     

Mumefural, citric acid derivative improving blood fluidity from fruit-juice concentrate of Japanese apricot (Prunus mume Sieb. et Zucc).

The effects of food components on blood fluidity were studied by in vitro assay using a dedicated microchannel instrument for model capillaries. We found that the fruit-juice concentrate of the Japanese apricot (Prunus mume Sieb. et Zucc), a traditional Japanese food, markedly improved the fluidity of human blood. Using HPLC, we isolated the active compounds and characterized them using UV, MS, IR, and NMR. They included a novel compound, 1-[5-(2-formylfuryl)methyl] dihydrogen 2-hydroxypropane-1,2, 3-tricarboxylate (mumefural), and a related compound, 5-hydroxymethyl-2-furfural (HMF). Mumefural markedly improved blood fluidity in all subjects, while HMF worked differently in different individuals. The flow rate of blood spiked with mumefural or HMF was compared to that of the two predominant organic acids in the fruit. Citric acid, malic acid, and furfuryl alcohol also improved fluidity in all subjects. The activity of P. mume is derived from not only artifacts produced during thermal processing, such as mumefural, but also from endogenous organic acids.     

Antiallergic tea catechin, (-)-epigallocatechin-3-O-(3-O-methyl)-gallate,suppresses FcepsilonRI expression in human basophilic KU812 cells

We previously found that the O-methylated derivative of (-)-epigallocatechin-3-O-gallate (EGCg), (-)-epigallocatechin-3-O-(3-O-methyl)-gallate (EGCG' '3Me), has potent antiallergic activity. The high-affinity IgE receptor, FcepsilonRI, is found at high levels on basophils and mast cells and plays a key role in a series of acute and chronic human allergic reactions. To understand the mechanism of action for the antiallergic EGCG' '3Me, the effect of EGCG' '3Me on the cell surface expression of FcepsilonRI in human basophilic KU812 cells was examined. Flow cytometric analysis showed that EGCG' '3Me was able to decrease the cell surface expression of FcepsilonRI. Moreover, immunoblot analysis revealed that total cellular expression of the FcepsilonRI alpha chain decreased upon treatment with EGCG' '3Me. FcepsilonRI is a tetrameric structure comprising one alpha chain, one beta chain, and two gamma chains. The level of mRNA production of each subunit in KU812 cells was investigated. EGCG' '3Me reduced FcepsilonRI alpha and gamma mRNA levels. The cross-linkage of FcepsilonRI causes the activation of basophils, which leads to the secretion of inflammatory mediators including histamine. EGCG' '3Me treatment inhibited the FcepsilonRI cross-linking-induced histamine release. These results suggested that EGCG' '3Me can negatively regulate basophil activation through the suppression of FcepsilonRI expression.     

Tea polyphenol (-)-epigallocatechin 3-gallate suppresses heregulin-beta1-induced fatty acid synthase expression in human breast cancer cells by inhibiting phosphatidylinositol 3-kinase/Akt and mitogen-activated protein kinase cascade signaling

Tumor-associated fatty acid synthase (FAS) is implicated in tumorigenesis and connected to HER2 (human epidermal growth factor receptor 2) by systemic analyses. Suppression of FAS in cancer cells may lead to growth inhibition and cell apoptosis. Our previous study demonstrated that (-)-epigallocatechin 3-gallate (EGCG), the green tea catechin, could down-regulate FAS expression by suppressing EGFR (epidermal growth factor receptor) signaling and downstream phosphatidylinositol 3-kinase (PI3K)/Akt activation in the MCF-7 breast cancer cell line. Herein, we examined the effects of EGCG on FAS expression modulated by another member of the erbB family, that is, HER2 or HER3. We identified that heregulin-beta1 (HRG-beta1), a HER3 ligand, stimulated dose-dependent FAS expression in breast cancer cell lines MCF-7 and AU565, but not MDA-MB-453. The time-dependent increase in FAS expression after HRG-beta1 stimulation was also observed in MCF-7 cells, and this up-regulation was de novo RNA synthesis dependent. Treatment of MCF-7 cells with EGCG markedly inhibited HRG-beta1-dependent induction of mRNA and protein of FAS. EGCG also decreased the phosphorylation of Akt and extracellular signal-regulated kinase 1/2 that were demonstrated as selected downstream HRG-beta1-responsive kinases required for FAS expression using dominant-negative Akt, PI3K inhibitors (LY294002 and wortmannin), or MEK inhibitor (PD98059). FAS induction by HRG-beta1 was also blocked by AG825, a selective HER2 inhibitor, and by genistein, a selective tyrosine kinase inhibitor, indicating the formation of a heterodimer between HER2 and HER3, and their tyrosine kinase activities are essential for HRG-beta1-mediated elevation of FAS. Additionally, growth inhibition of HRG-beta1-treated cells was parallel to suppression of FAS by EGCG. Taken together, these findings extend our previous study to indicate that EGCG may be useful in the chemoprevention of breast carcinoma in which FAS overexpression results from HER2 or/and HER3 signaling.     

Quantitation of silibinin, a putative cancer chemopreventive agent derived from milk thistle (Silybum marianum), in human plasma by high-performance liquid chromatography and identification of possible metabolites

Silibinin has recently received attention as a potential cancer chemopreventive agent because of its antiproliferative and anticarcinogenic effects. A simple and specific reversed-phase high-performance liquid chromatography method was developed and validated for the quantitation of silibinin in human plasma. Sample preparation involved simple protein precipitation, and separation was achieved on a Waters Atlantis C18 column with flow rate of 1.0 mL/min at 40 degrees C and UV detection at 290 nm. Silibinin was detected as two peaks corresponding to trans-diastereoisomers. The peak area was linear over the investigated concentration range (0-5000 ng/mL). The limits of detection were 2 and 1 ng/mL for the two diastereoisomers (d1 and d2), with a recovery of 53-58%. This method was utilized to detect silibinin in plasma of colorectal patients after 7 days of treatment with silipide (silibinin formulated with phosphatidyl choline).     

Estrogenic effects of extracts from cabbage, fermented cabbage, and acidified brussels sprouts on growth and gene expression of estrogen-dependent human breast cancer (MCF-7) cells

Cruciferous vegetable extracts from freeze-dried cabbage (FDC), freeze-dried fermented cabbage (FDS), and acidified Brussels sprouts (ABS) were prepared by exhaustive extraction with ethyl acetate. Estrogenic and antiestrogenic effects of these extracts were analyzed. To identify whether the extracts are potential estrogen receptor (ER) ligands that can act as agonists or antagonists, the binding affinity of extracts for the ER was measured using a competitive radiometric binding assay. The extracts bound with low affinity to the ER, and the relative binding affinity is estradiol > FDS > FDC > ABS. These extracts were evaluated for their estrogenic and antiestrogenic activities in estrogen-dependent human breast cancer (MCF-7) cells using as endpoints proliferation and induction of estrogen-responsive pS2 gene expression, which was analyzed using Northern blot assay. At low concentrations (5-25 ng/mL) all of the extracts reduced 1 nM estradiol-induced MCF-7 cell proliferation. Extracts at 25 ng/mL also inhibited estradiol-induced pS2 mRNA expression. At higher extract concentrations (50 ng/mL-25 microg/mL), however, increased proliferation in MCF-7 cells was observed. Similarly, expression of the pS2 gene was induced by higher extract concentrations (0.25-25 microg/mL). The pure estrogen antagonist, ICI 182,780, suppressed the cell proliferation induced by the extracts as well as by estradiol and also the induction of pS2 expression by the extracts. The ER subtype-selective activities of FDC and FDS were analyzed using a transfection assay in human endometrial adenocarcinoma (HEC-1) cells. FDS acted as an ERalpha-selective agonist while FDC fully activated both ER-alpha and ER-beta. Growth of the ER-negative MDA-231 cells was not affected by the extracts or by estradiol. This study demonstrates that cruciferous vegetable extracts act bifunctionally, like an antiestrogen at low concentrations and an estrogen agonist at high concentrations.     

A mixture of trans, trans conjugated linoleic acid induces apoptosis in MCF-7 human breast cancer cells with reciprocal expression of Bax and Bcl-2

The growth inhibitory effect of a mixture of trans, trans conjugated linoleic acid isomers (t, t CLA) was investigated in a human breast cancer cell line, MCF-7, with references to c9, t11 CLA, t10, c12 CLA, and linoleic acid. The t, t CLA treatment effectively induced a cytotoxic effect in a time-dependent (0-6 days) and concentration-dependent (0-40 microM) manner, as compared to the reference and control treatments. The apoptotic parameters were measured on cells treated with 40 microM t, t CLA for 4 days. The occurrence of the characteristic morphological changes and DNA fragmentation confirmed apoptosis. The t, t CLA treatment led to an increase in the level of p53 tumor suppressor protein and Bax protein, but suppressed the expression of Bcl-2 protein. In addition, cytochrome c was released from the mitochondria into the cytosol, and the activation of caspase-3 led to the cleavage of poly(ADP-ribose) polymerase (PARP). Moreover, the composition of the linoleic and arachidonic acids was decreased in cellular membranes. These findings suggest that incorporation of t, t CLA in the membrane induces a mitochondria-mediated apoptosis that can enhance the antiproliferative effect of t, t CLA in MCF-7 cells.