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Chemosystematic investigations of irregular diterpenes in Anisotome and related New Zealand Apiaceae.

A chemosystematic HPLC-UV and HPLC-MS investigation of New Zealand members of the Apiaceae was performed. Diterpenes were identified and quantified in methanolic extracts from subaerial parts of 28 taxa and 54 samples of Aciphylla, Anisotome, Apium, Gingidia, Lignocarpa, Oreomyrrhis, and Scandia. Six diterpenes (1-2, 4-7) and four polyacetylenes (8-11) were identified. The known compounds were the diterpenes anisotomenoic acid 1, anisotomene-1-ol 2, 16-acetoxyanisotomenoic acid 4 and anisotomene-1,12-diol 5; and the polyacetylenes falcarinol 8, falcarindiol 9, (+)-9(Z),17-octadecadiene-12,14-diyne-1,11,16-triol 10, and (+)-9(Z),17-octadecadiene-12,14-diyne-1,11,16-triol 1-acetate 11. New irregular diterpenes 13,14-dihydroanisotom-12E-ene-1,14-diol 6 and 14-methoxy-13,14-dihydroanisotom-12E-ene-1-ol 7 were isolated from A. haastii. Isomers of the new semi-synthetic diterpene 16-hydroxyanisotomenoic acid 3 were detected in extracts of Anisitone flexuosa. Structure elucidation was performed by HR mass spectrometry and 1D and 2D NMR spectroscopy. In crude extracts, compounds were identified by their HPLC retention times and their on-line HPLC-UV and MS spectra. Anisotomene diterpenes occurred in eight out of 16 species of the genus Anisotome, but were not detected in any of the other genera. In contrast, polyacetylenes were present in all the genera investigated.

Apiaceae↗

Modulation of phosphatidylcholine biosynthesis in celery by exogenous fatty acids.

The effects of C16 and C18 fatty acids on the synthesis of phosphatidylcholine were studied in Apium graveolens cell suspension cultures and postmitochondrial supernatants. When cells were exposed to exogenous oleic acid, the rate of phosphatidylcholine biosynthesis increased 1.4-fold within 5 min of the addition of the fatty acid to the culture medium. The sensitivity of microsomal CTP:cholinephosphate cytidylyltransferase (EC 2.7.7.15) to saturated and unsaturated fatty acids was monitored through the addition of unesterified fatty acids to postmitochondrial supernatants. The saturated fatty acids, palmitic and stearic, appeared to have little effect on CTP:cholinephosphate cytidylyltransferase activity, whereas exposure to oleic, linoleic and cis-vaccenic acids resulted in significant increases in enzyme activity. Optimal microsomal CTP:cholinephosphate cytidylyltransferase activities were achieved by the incubation of postmitochondrial supernatants with 500 microM oleate. The exogenous fatty acids were found to be incorporated into microsomal membranes in their unesterified form. Removal of unesterified fatty acids by incubation of microsomal membranes with defatted bovine serum albumin resulted in the reduction of microsomal CTP:cholinephosphate cytidylyltransferase activity; demonstrating that the enzyme requires unesterified unsaturated fatty acids.

Animals↗

Anti-nociceptive and anti-inflammatory effects of some Jordanian medicinal plant extracts.

The anti-nociceptive effect of ethanolic extract of 11 traditionally used Jordanian plants was studied by using the acetic acid-induced writhing and hot-plate test in mice. The anti-inflammatory effect of these plants was determined by xylene-induced ear oedema in mice and cotton pellet granuloma test in rats. Mentha piperita, Cinnamomum zeylanicum, Apium graveolens, Eucalyptus camaldulentis, and Ruta graveolens possess an anti-nociceptive effect against both acetic acid-induced writhing and hot plate-induced thermal stimulation. M. piperita, Jasminum officinale, Commiphora molmol, and Beta vulgaris possess an anti-inflammatory effect against acute (xylene-induced ear oedema) and chronic (cotton-pellet granuloma) inflammation. The anti-nociceptive and anti-inflammatory effects were dose dependent. These data affirm the traditional use of some of these plants for painful and inflammatory conditions.

Analgesics↗

Assessment of the transfer of 137Cs in three types of vegetables consumed in Hong Kong.

A dynamic food chain model has been built for the modeling of the transfer of 137Cs in three types of vegetables consumed in Hong Kong, namely, white flowering cabbage (Brassica chinensis), head lettuce (Lactuca sativa) and celery (Apium graveolens). Some parameters have been estimated from the experimental data obtained in this work. The experimental data include the transfer factors of 137Cs from soil to the different vegetable species which are determined through high resolution gamma spectrometry, maximum crop biomasses for the vegetable species, the dry-to-fresh ratios for the vegetable species, the bulk density of soil layers and the average concentration of 137Cs in air. The derived parameters include the deposition rate and the root uptake rate, information for tillage, the logistic growth model and radionuclide concentrations in vegetables. The dynamic food chain model is solved by the Birchall-James algorithm to give the 137Cs concentration in subsurface soil, from the 0.1-25 cm soil layer, and the 137Cs concentration in harvested and unwashed vegetables. As validation of the model and parameters, the concentrations obtained experimentally and from the model are compared and are found to be in good agreement.

Apiaceae↗

Alpha-tocopherol content in 62 edible tropical plants.

Vitamin E was determined by the high-performance liquid chromatography (HPLC) method. All the plants tested showed differences in their alpha-tocopherol content and the differences were significant (p < 0.05). The highest alpha-tocopherol content was in Sauropus androgynus leaves (426.8 mg/kg edible portion), followed by Citrus hystrix leaves (398.3 mg/kg), Calamus scipronum (193.8 mg/kg), starfruit leaves Averrhoa belimbi (168.3 mg/kg), red pepper Capsicum annum (155.4 mg/kg), local celery Apium graveolens (136.4 mg/kg), sweet potato shoots Ipomoea batatas (130.1 mg/kg), Pandanus odorus (131.5 mg/kg), Oenanthe javanica (146.8 mg/kg), black tea Camelia chinensis (183.3 mg/kg),papaya Carica papaya shoots (111.3 mg/kg), wolfberry leaves Lycium chinense (94.4 mg/kg), bird chili Capsicum frutescens leaves (95.4 mg/kg), drumstick Moringa oleifera leaves (90.0 mg/kg), green chili Capsicum annum (87 mg/kg), Allium fistulosum leaves (74.6 mg/kg), and bell pepper Capsicum annum (71.0 mg/kg). alpha-Tocopherol was not detected in Brassica oleracea, Phaeomeria speciosa, Pachyrrhizus speciosa, Pleurotus sajor-caju, and Solanum melongena.

Chromatography, High Pressure Liquid↗

Phosphatidylinositol phosphodiesterase in higher plants.

1. The lower regions of the stem of celery (Apium graveolens L.) contain a soluble enzyme that hydrolyses phosphatidylinositol. 2. The lipoidal product of hydrolysis is diacylglycerol, and the water-soluble products are 1:2-cyclic phosphoinositol and phosphoinositol in the approximate proportions of 60% and 40% respectively: this indicates that a phosphodiesterase (phospholipase C-like) activity is cleaving the phosphatidylinositol. 3. The enzyme requires a bivalent cation, Ca2+ being the most effective activator. 4. The enzyme has a pH optimum, depending on conditions of assay, of pH 5.9-6.6 and in this pH range shows no detectable activity against phosphatidylcholine or phosphatidylethanolamine. 5. The activity is stimulated by phosphatidic acid and slightly inhibited (30% at concentrations equimolar with phosphatidylinositol) by phosphatidylcholine. 6. The phosphodiesterase was also detected (but not quantified) in the tips of the flowers in cauliflowers, in outer leaves of onion and in the elongating stem of daffodils. 7. The enzyme's properties are compared with equivalent mammalian enzymes, and its possible role in the catabolism of phosphatidylinositol in higher plants is discussed.

Cations↗

Correlating IgE reactivity with three-dimensional structure.

This Commentary discusses the work of Neudecker et al. in this issue of the Biochemical Journal in which site-directed mutagenesis and NMR spectroscopy have been used to analyse in detail the IgE-binding capacity of two cross-reactive allergens: Apg1.0101 from celery ( Apium graveolens ) and Pru av 1 from cherry ( Prunus avium ), which are both members of the pathogenesis-related allergen family. The study, showing that the IgE-binding epitopes are highly patient specific, will have a profound impact on our understanding of conformational IgE-binding epitopes, raising serious questions about the therapeutic usefulness of conventional site-directed-mutagenic approaches for the production of hypo-allergenic protein variants.

Allergens↗

NBPA: a cerebral ischaemic protective agent.

1. NBPA is a derivative of 3-n-butylpathalide isolated from Apium granolens Linn. 2. At concentrations ranging from 6 x 10(-6) to 10(-6) mol/L, NBPA inhibited the L-type calcium current in guinea-pig myocardial cells and cultured human neuroblastoma cells. 3. At 10(-6) mol/L, NBPA markedly inhibited calcium-dependent and -independent release of glutamate from synaptosomes. 4. The [31P] nuclear magnetic resonance spectrum has shown that pretreatment with NBPA at 15 mg/kg, i.p., improved energy metabolism. 5. In situ hybridization has shown that 10 and 20 mg/kg, i.p., NBPA prior to cerebral artery occlusion can accelerate the expression of heat shock protein 70 mRNA and inhibit c-fos mRNA expression. 6. It has been shown that NBPA decreases the nitric oxide content and bc nitric oxide synthase (NOS) activity in the global cerebral ischaemia-reperfusion model in rats. In addition, it has been shown that NBPA significantly inhibits the expression of inducible NOS protein.

Animals↗

Sequence analysis of a mannitol dehydrogenase cDNA from plants reveals a function for the pathogenesis-related protein ELI3.

Mannitol is the most abundant sugar alcohol in nature, occurring in bacteria, fungi, lichens, and many species of vascular plants. Celery (Apium graveolens L.), a plant that forms mannitol photosynthetically, has high photosynthetic rates thought to results from intrinsic differences in the biosynthesis of hexitols vs. sugars. Celery also exhibits high salt tolerance due to the function of mannitol as an osmoprotectant. A mannitol catabolic enzyme that oxidizes mannitol to mannose (mannitol dehydrogenase, MTD) has been identified. In celery plants, MTD activity and tissue mannitol concentration are inversely related. MTD provides the initial step by which translocated mannitol is committed to central metabolism and, by regulating mannitol pool size, is important in regulating salt tolerance at the cellular level. We have now isolated, sequenced, and characterized a Mtd cDNA from celery. Analyses showed that Mtd RNA was more abundant in cells grown on mannitol and less abundant in salt-stressed cells. A protein database search revealed that the previously described ELI3 pathogenesis-related proteins from parsley and Arabidopsis are MTDs. Treatment of celery cells with salicylic acid resulted in increased MTD activity and RNA. Increased MTD activity results in an increased ability to utilize mannitol. Among other effects, this may provide an additional source of carbon and energy for response to pathogen attack. These responses of the primary enzyme controlling mannitol pool size reflect the importance of mannitol metabolism in plant responses to divergent types of environmental stress.

Amino Acid Sequence↗

A vesiculo-bullous disease in pigs resembling foot and mouth disease. I. Field cases.

Eleven incidents of bullae and vesicles on the snouts and less frequently the feet of white-skinned pigs on seven farms are described. Bullous and vesicular lesions up to 5 cm in diameter and containing clotted gelatinous fluid were located on the dorsal aspect of the snout, behind the flange. Lesions ruptured, became ulcerated, developed scabs and healed within three weeks. There was no transmission to other pigs or ruminants. The condition was associated with contact with green vegetable material containing parsnips (Pastinaca sativa) or celery (Apium graveolens), followed by exposure to periods of extended sunshine. Parsnips and celery are known to contain furocoumarins, potent phototoxic compounds. It is suggested that absorption of furocoumarins on the skin of the snout and feet after contact with parsnips and celery and exposure to ultraviolet light caused the lesions in the cases reported. It is postulated that a similar condition may have been responsible for national foot and mouth disease scares in pigs at Warkworth and Temuka in New Zealand and at Legana in Tasmania.

Journal Article↗

A vesiculo-bullous disease in pigs resembling foot and mouth disease. II. Experimental reproduction of the lesion.

Vesiculo-bullous dermatitis of pigs characterised by presence of vesicles and bullae on the snout and feet of white skinned pigs was reproduced experimentally. Leaves of parsnips (Pastinaca sativa), or celery (Apium graveolens) infected with the fungus Sclerotinia sclerotiorum were fed or rubbed on the snouts and feet of white skinned pigs. Pigs were then exposed to sunlight or to UV light of intensity approximately 212 m W/M2 at a wavelength 340-360 nm for eight hours per day until vesicles developed. All treated pigs developed lesions on the snouts, and less frequently on the feet. Lesions were characterised by the appearance of erythema at 24 hours after treatment. Vesicles developed at 48 hours and became maximal by 72 hours. Pigs treated with plant material without exposure to UV light or exposed to UV light without contact with plant material did not develop lesions. The experimental lesions closely resemble those observed in several field cases in 1984 and 1985 in New Zealand and to lesions present in three well publicized foot and mouth disease scares at Warkworth, and Temuka in New Zealand and Legana in Tasmania.

Journal Article↗

Chemoprevention of benzo[a]pyrene-induced forestomach cancer in mice by natural phthalides from celery seed oil.

Bioassay-directed fractionation of celery seed oil from the plant Apium graveolens (Umbelliferae) led to the isolation of five natural products, including d-limonene, p-mentha-2,8-dien-1-ol, p-mentha-8(9)-en-1,2-diol, 3-n-butyl phthalide, and sedanolide. Of these compounds p-mentha-2,8-dien-1-ol,3-n-butyl phthalide, and sedanolide exhibited high activities to induce the detoxifying enzyme glutathione S-transferase (GST) in the target tissues of female A/J mice. 3-n-Butyl phthalide and sedanolide (20 mg/dose every two days for a total of 3 doses) increased GST activity 4.5-5.9 and 3.2-5.2 times over the controls in the mouse liver and small intestinal mucosa, respectively. At the same dose, p-mentha-2,8-dien-1-ol induced GST activity about 3.7-fold above that of the controls. Thus, these compounds were further tested for their ability to inhibit benzo[a]pyrene- (BP) induced tumorigenesis in mice. After treatment with 3-n-butyl phthalide and sedanolide, the tumor incidence was reduced from 68% to 30% and 11%, respectively. About 67% and 83% reduction in tumor multiplicity was also observed with 3-n-butyl phthalide and sedanolide. p-Mentha-2,8-dien-1-ol produced only a small or no significant reduction of forestomach tumor formation. The data indicating that 3-n-butyl phthalide and sedanolide were both active in tumor inhibition and GST assays suggested a correlation between the inhibitory activity and the GST-inducing ability. The phthalides are known to determine the characteristic odor of celery. The results suggest that phthalides, as a class of bioactive natural products occurring in edible umbelliferous plants, may be effective chemopreventive agents.

Animals↗

Study of stereoselective synthesis of (+/-)-neocnidilide.

Neocnidilide 1, isolated from Apium graveleues L (Umbelliferae), has shown activity to inhibit the growth of mycotoxin-producing fungi. An efficient method for the synthesis of the racemic neocnidilide by the stereoselective reaction of hemiacetal 6 with n-BuMgBr has been developed.

Benzofurans↗

Tissue Printing as a Tool for Observing Immunological and Protein Profiles in Young and Mature Celery Petioles.

Tissue printing onto membranes such as nitrocellulose is a technique employed to study the localization of proteins, nucleic acids, and soluble metabolites from freshly cut tissue slices. We probed tissue prints of young and mature celery (Apium graveolens) petioles with antibodies raised against two proteins, spinach ribulose-1,5-bisphosphate carboxylase and tomato fruit catalase. The purposes of this study were to determine if these proteins are developmentally regulated and to determine if the patterns and intensities of cross-reactivity of antibodies on tissue blots corresponded only to the presence of specific epitopes or was related to the amount of protein present in any given area on the tissue prints. Different and distinct cross-reactivity patterns were observed with each of the two antibodies used. Tissue prints from young and mature tissues also showed differences in antibody cross-reactivity. Comparison of Coomassie blue staining patterns with antibody reactivity patterns showed that there is little relationship between tissue protein concentration and antibody reactivity.

Journal Article↗

Effect of Different Carbon Sources on Relative Growth Rate, Internal Carbohydrates, and Mannitol 1-Oxidoreductase Activity in Celery Suspension Cultures.

Little information exists concerning the biochemical route of mannitol catabolism in higher plant cells. In this study, the role of a recently discovered mannitol 1-oxidoreductase (MDH) in mannitol catabolism was investigated. Suspension cultures of celery (Apium graveolens L. var dulce [Mill.] Pers.) were successfully grown on nutrient media with either mannitol, mannose, or sucrose as the sole carbon source. Cell cultures grown on any of the three carbon sources did not differ in relative growth rate, as measured by packed cell volume, but differed drastically in internal carbohydrate concentration. Mannitol-grown cells contained high concentrations of mannitol and extremely low concentrations of sucrose, fructose, glucose, and mannose. Sucrose-grown cells had high concentrations of sucrose early in the growth cycle and contained a substantial hexose pool. Mannose-grown cells had a high mannose concentration early in the cycle, which decreased during the growth cycle, whereas their internal sucrose concentrations remained relatively constant during the entire growth cycle. Celery suspension cultures on all three carbon substrates contained an NAD-dependent MDH. Throughout the growth cycle, MDH activity was 2- to 4-fold higher in mannitol-grown cells compared with sucrose- or mannose-grown cells, which did not contain detectable levels of mannitol, indicating that MDH functions pre-dominantly in an oxidative capacity in situ. The MDH activity observed in celery cells was 3-fold higher than the minimum amount required to account for the observed rate of mannitol utilization from the media. Cultures transferred from mannitol to mannose underwent a decrease in MDH activity over a period of days, and transfer from mannose to mannitol resulted in an increase in MDH activity. These data provide strong evidence that MDH plays an important role in mannitol utilization in celery suspension cultures.

Journal Article↗

Mannitol Metabolism in Celery Stressed by Excess Macronutrients.

The effect of excess macronutrients in the root environment on mannitol and sucrose metabolism was investigated in celery (Apium graveolens L. var dulce [Mill.] Pers.). Plant growth was inhibited progressively as macronutrient concentration in the media, as measured by electrical conductivity (E.C.), increased from 1.0 to 11.9 decisiemens m-1. Plants grown for 35 d at higher E.C. had a lower water content but similar dry weight in their roots, leaves, and petioles compared to plants grown at lower E.C. Macronutrient concentrations of leaves, roots, and petioles were not affected by the imposed stress, indicating that the macronutrient stress resulted in a water-deficit stress response rather than a salt-specific response. Mannitol accumulated in sink tissues and was accompanied by a drastic decrease in activity of mannitol-1-oxidoreductase. Sucrose concentration and activities of sucrose-metabolizing enzymes in sink tissues were not affected by the macronutrient stress. Mature leaves exhibited increased concentrations of both mannitol and sucrose, together with increased activity of mannose-6-phosphate reductase and sucrose phosphate synthase, in response to macronutrient stress. Thus, mannitol accumulation in osmotically stressed celery is regulated by diminished catabolism in sink tissues and increased capacity for mannitol biosynthesis in source leaves.

Journal Article↗

Purification of NAD-dependent mannitol dehydrogenase from celery suspension cultures.

Mannitol dehydrogenase, a mannitol:mannose 1-oxidoreductase, constitutes the first enzymatic step in the catabolism of mannitol in nonphotosynthetic tissues of celery (Apium graveolens L.). Endogenous regulation on the enzyme activity in response to environmental cues is critical in modulating tissue concentration of mannitol, which, importantly, contribute to stress tolerance of celery. The enzyme was purified to homogeneity from celery suspension cultures grown on D-mannitol as the carbon source. Mannitol dehydrogenase was purified 589-fold to a specific activity of 365 mumol h-1 mg-1 protein with a 37% yield of enzyme activity present in the crude extract. A highly efficient and simple purification protocol was developed involving polyethylene glycol fractionation, diethylaminoethyl-anion-exchange chromatography, and NAD-agarose affinity chromatography using NAD gradient elution. Sodium dodecylsulfate gel electrophoresis of the final preparation revealed a single 40-kD protein. The molecular mass of the native protein was determined to be approximately 43 kD, indicating that the enzyme is a monomer. Polyclonal antibodies raised against the enzyme inhibited enzymatic activity of purified mannitol dehydrogenase. Immunoblots of crude protein extracts from mannitol-grown celery cells and sink tissues of celery, celeriac, and parsley subjected to sodium dodecyl sulfate gel electrophoresis showed a single major immuno-reactive 40-kD protein.

Amino Acid Sequence↗

Immunolocalization of mannitol dehydrogenase in celery plants and cells.

Immunolocalization of mannitol dehydrogenase (MTD) in celery (Apium graveolens L.) suspension cells and plants showed that MTD is a cytoplasmic enzyme. MTD was found in the meristems of celery root apices, in young expanding leaves, in the vascular cambium, and in the phloem, including sieve-element/companion cell complexes, parenchyma, and in the exuding phloem sap of cut petioles. Suspension cells that were grown in medium with mannitol as the sole carbon source showed a high anti-MTD cross-reaction in the cytoplasm, whereas cells that were grown in sucrose-containing medium showed little or no cross-reaction. Gel-blot analysis of proteins from vascular and nonvascular tissues of mature celery petioles showed a strong anti-MTD sera cross-reactive band, corresponding to the 40-kD molecular mass of MTD in vascular extracts, but no cross-reactive bands in nonvascular extracts. The distribution pattern of MTD within celery plants and in cell cultures that were grown on different carbon sources is consistent with the hypothesis that the Mtd gene may be regulated by sugar repression. Additionally, a developmental component may regulate the distribution of MTD within celery plants.

Animals↗