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Acute and chronic antiinflammatory effects of Hedera colchica in rats.

Hedera helix and Hedera colchica are the members of Araliaceae family. In the present study, we tested the possible antiinflammatory effects of crude (CSE) and purified (SPE) extracts of Hedera colchica in carrageenan- and cotton pellet-induced acute and chronic inflammation models in rats. Both CSE and SPE of Hedera colchica were found to have antiinflammatory effects. The most potent drug was found as indomethacin (90%) in the 4 h measurements, while SPE (83%) and CSE (55%) of Hedera colchica in 100 mg/kg doses were found to have quite potent effects in acute phase of inflammation, in respect to control values. For testing chronic antiinflammatory (antiproliferative) effects, cotton pellet granuloma test was conducted. SPE was found as the most potent drug in chronic phase of inflammation with an effect of 64.32%. Indomethacin was found more potent than the CSE of Hedera colchica, and potency was found as 61.2 and 58%, respectively.

Acute Disease↗

Chloroplast and nuclear evidence for multiple origins of polyploids and diploids of Hedera (Araliaceae) in the Mediterranean basin.

Chloroplast (trnT-L) and nuclear rDNA (ITS) sequence analyses of the Araliaceae provide strong molecular evidence for the monophyly of the genus Hedera. Phylogenetic reconstructions suggest multiple origins and an active polyploidization process not only in the formation of tetraploids (2n = 96), hexaploids (2n = 144), and octoploids (2n = 192), but also of diploids (2n = 48). A high basic chromosome number of x = 24, extensive polyphyly in widespread diploids, and terminal placement of Hedera in phylogenies of the Araliaceae reveal that extant diploid taxa may be, in fact, assemblages of ancestral polyploids from plants of n = 12. Four major lineages containing four types of chloroplast (chlorotypes I, II, III, and IV), which are defined by different trnT-L nucleotide substitutions and two large insertions (50- and 30-bp), provide evidence for evolutionary processes and historical biogeography in Hedera. We propose a scenario where an initial colonization in the Mediterranean basin by Asian ancestors (carrying the ancestral Araliaceae chlorotype I) is followed by differentiation into the four chlorotypes of the Mediterranean region, and then recolonization of Asia and northern Europe only by chlorotype III. The Macaronesian taxa (Hedera azorica, Hedera maderensis ssp. maderensis, and Hedera canariensis) appear to have originated from a single-colonization event to each archipelago with no further contact either with continental or insular species.

Base Sequence↗

Acute and chronic antiinflammatory profile of the ivy plant, Hedera helix, in rats.

Hedera helix is a plant well-known as ivy or English ivy, and a member of the Araliaceae family. In the present study, we tested the possible antiinflammatory effects of a crude saponin extract (CSE) and a saponin's purified extracts (SPE) of Hedera helix in carrageenan- and cotton-pellet-induced acute and chronic inflammation models in rats. Both the CSE and SPE of Hedera helix were found to have antiinflammatory effects. The most potent drug screened was indomethacin (89.2% acute antiinflammatory effect), while the most potent extract screened was the CSE of Hedera helix at 100 and 200 mg/kg body wt. doses with 77% acute antiinflammatory effects. For testing chronic antiinflammatory (antiproliferative) effects, the cotton-pellet-granuloma test was conducted. Indomethacin was found to be the most potent drug in the chronic phase of inflammation, with 66% effect. The SPE of Hedera helix was more potent than the CSE in its chronic antiinflammatory effect (60% and 49%, respectively).

Animals↗

Phylogeography of the common ivy (Hedera sp.) in Europe: genetic differentiation through space and time.

We studied the phylogeography of ivy (Hedera sp.), a liana widespread in Europe, throughout its natural range. The populations sampled belong to four closely related species differing by ploidy levels and morphological characters. Chloroplast (cp) markers were used and 13 haplotypes were detected, usually shared across species, contrary to ribosomal internal transcribed spacer (ITS) variants. We demonstrated the existence of a strong overall cpDNA phylogeographical structure. Several methods of data analysis were conducted to describe how this structure and the genetic diversity change through space and time. Southern populations, especially those from Spain, are the most divergent. Pairwise estimates of differentiation point to isolation by distance, and the existence of a latitudinal gradient of divergence was demonstrated using a regression procedure. Similarly, latitudinal differences in haplotype richness and diversity exist, as shown by population permutations ('differentiation through space'). Finally, we measured differentiation by taking into account successive levels of divergence between haplotypes ('differentiation through time'). Genetic differentiation turns out to be much greater when differences between closely related haplotypes are not considered. Further, these results indicate that the phylogeographical structure is essentially due to the relative distribution of the most similar haplotypes. Diversity decreases from south to north, whereas haplotype frequencies change longitudinally. It appears that Hedera survived in Spanish and Balkan refugia during the last ice age. A third refugium must have been present in the Alps or in Italy. During the northward expansion, the decrease in overall diversity was attenuated by some mixing of lineages at intermediate latitudes, resulting in comparatively higher levels of differentiation in the south.

Biological Evolution↗

An unusual case of death: suffocation caused by leaves of common ivy (Hedera helix). Detection of hederacoside C, alpha-hederin, and hederagenin by LC-EI/MS-MS.

We report one fatal case of asphyxia caused by leaves of common ivy. Macroscopic examination of the corpse during the autopsy disclosed an incredible quantity of leaves of Hedera helix in the mouth and throat of the decedent. In order to rule out the possibility of poisoning by the toxic saponins contained in the plant, we have developed an efficient LC-EI/MS-MS assay of hederacoside C, alppha-hederin, and hederagenin in biological fluids and plant material. Sample cleanup involved solid-phase extraction of the toxins on C18 cartridges followed by LC analysis under reversed-phase conditions in the gradient elution mode. Solute identification was performed using full scan MS-MS spectrum of the analytes. Oleandrine was used as internal standard. Under these conditions, saponins in powdered dried leaves of Hedera helix were measured at a concentration of 21.83 mg/g for hederacoside C, 0.41 mg/g for alpha-hederin and 0.02 mg/g for hederagenin. No toxin was detected in cardiac blood, femoral blood, or urine of the deceased, but hederacoside C was quantitated at 857 ng/mL in the gastric juice. These findings led us to conclude that the man committed suicide and that the death was caused by suffocation by leaves of common ivy.

Adult↗

Triterpenoid saponins from leaves of Hedera pastuchowii.

Five new triterpenoid saponins, pastuchoside A (1), B (3), C (5), D (7) and E (9), were isolated from the leaves of Hedera pastuchowii. They have oleanolic acid or hederagenin as aglycone. The structures were established by NMR spectroscopy including gs (gradient selected)-COSY, gs-HSQC, gs-HSQC-TOCSY and gs-HMBC experiments, and mass spectrometry (ESI-HR-MS). Heptaoside saponins, compounds 1 and 3, are described for the first time in the genus Hedera.

Hedera↗

Anti-elastase and anti-hyaluronidase activities of saponins and sapogenins from Hedera helix, Aesculus hippocastanum, and Ruscus aculeatus: factors contributing to their efficacy in the treatment of venous insufficiency.

Triterpene and steroid saponins and sapogenins of medicinal plants (Aesculus hippocastanum L., Hedera helix L., Ruscus aculeatus L.) are claimed to be effective for the treatment/prevention of venous insufficiency. In this work we evaluated the inhibitory effects of these plant constituents on the activity of elastase and hyaluronidase, the enzyme systems involved in the turnover of the main components of the perivascular amorphous substance. The results evidence that for Hedera helix L., the sapogenins only non-competitively inhibit hyaluronidase activity in a dose-dependent fashion, showing comparable IC50 values (hederagenin IC50 = 280.4 microM; oleanolic acid IC50 = 300.2 microM); both the saponins hederacoside C and alpha-hederin are very weak inhibitors. The same behaviour is observed for serine protease porcine pancreatic elastase: the glycosides are devoid of inhibitory action, while genins are potent competitive inhibitors (oleanolic acid IC50 = 5.1 microM; hederagenin IC50 = 40.6 microM). Constituents from Aesculus hippocastanum L. show inhibitory effects only on hyaluronidase, and this activity is mainly linked to the saponin escin (IC50 = 149.9 microM), less to its genin escinol (IC50 = 1.65 mM). By contrast, ruscogenins from Ruscus aculeatus L., ineffective on hyaluronidase activity, exhibit remarkable anti-elastase activity (IC50 = 119.9 microM; competitive inhibition). The mechanism of elastase inhibition by triterpene and steroid aglycones, with a nitroanilide derivative as substrate, is discussed.

Animals↗

Ultrastructure of minor-vein phloem and assimilate export in summer and winter leaves of the symplasmically loading evergreens Ajuga reptans L., Aucuba japonica Thunb., and Hedera helix L.

Minor-vein ultrastructure and sugar export were studied in mature summer and winter leaves of the three broadleaf-evergreen species Ajuga reptans var. artropurpurescens L., Aucuba japonica Thunb. and Hedera helix L. to assess temperature effects on phloem loading. Leaves of the perennial herb Ajuga exported substantial amounts of assimilates in form of raffinose-family oligosaccharides (RFOs). Its minor-vein companion cells represent typical intermediary cells (ICs), with numerous small vacuoles and abundant plasmodesmal connectivity to the bundle sheath. The woody plants Hedera and Aucuba translocated sucrose as the dominant sugar species, and only traces of RFOs. Their minor-vein phloem possessed a layer of highly vacuolated cells (VCs) intervening between mesophyll and sieve elements. Depending on their location and ontogeny, VCs were classified either as companion or parenchyma cells. Both cell types showed symplasmic continuity to the adjacent mesophyll tissue although at a lower plasmodesmal frequency compared to the Ajuga ICs. p-Chloromercuribenzenesulfonic acid did not reduce leaf sugar export in any of the plants, indicating a symplasmic mode of phloem loading. Winter leaves did not show symptoms of frost injury, and the vacuolar pattern in ICs and VCs was equally prominent in both seasons. Starch accumulation as a result of reduced phloem loading was not observed to be triggered by low temperature. In contrast, high amounts of starch were found in mesophyll and bundle-sheath cells of summer leaves. Physiological data on season-dependent leaf exudation showed the maintenance of sugar export in cold-acclimated winter leaves.

Biological Transport↗

Allergic and irritant contact dermatitis from falcarinol and didehydrofalcarinol in common ivy (Hedera helix L.).

Experimental and chemical investigations revealed that common ivy (Hedera helix susp. helix) contains 3 compounds which are powerful irritants and moderate sensitizers. Only 2 of these constituents, falcarinol and didehydrofalcarinol, are present in the plant during the whole year. Besides Panax ginseng and Schefflera arboricola, this is the third species of the Araliaceae in which these polyacetylenic sensitizers have been found. Falcarinol and didehydrofalcarinol also occur in Hedera helix subsp. canariensis. 4 patients have been patch tested. Even in low concentrations (0.03%), the main allergen falcarinol elicited strong reactions in all of them. One of the authors became sensitized during the investigations.

Adult↗

Triterpenoid saponins from berries of Hedera colchica.

Fifteen triterpenoid saponins were isolated from the berries of Hedera colchica and their structures established on the basis of chemical and spectroscopic evidence. Among them, two are new compounds: coichiside A (3) and colchiside B (15) and four are described for the first time in the berries of Hedera colchica (compounds 1, 8, 9 and 11).

Magnetic Resonance Spectroscopy↗

[Contact dermatitis allergy to common ivy (Hedera helix L.)].

A case of allergic contact dermatitis to common ivy (Hedera helix) is presented. The patient is a 16 year old female gardner, who developed severe blistering dermatitis of the hands, forearms and face after frequent contact with Hedera helix. We wish to draw attention to common ivy as a sensitizer and to demonstrate the severity of the reaction in a sensitized person.

Adolescent↗

Mechanisms of cuticular uptake of xenobiotics into living plants: 1. Influence of xenobiotic dose on the uptake of three model compounds applied in the absence and presence of surfactants into Chenopodium album, Hedera helix and Stephanotis floribunda leaves.

This study determined the uptake of three model compounds, applied in the presence and absence of surfactants, into the leaves of three plant species (Chenopodium album L, Hedera helix L and Stephanotis floribunda Brongn). The results with 2-deoxy-D-glucose, 2,4-dichlorophenoxyacetic acid and epoxiconazole in the presence ofsurfactants (the polyethylene glycol monododecyl ethers C12EO3, C12EO6, C12EO10 and a trisiloxane ethoxylate with mean EO of 7.5, all used at one equimolar concentration and therefore different percentage concentrations) illustrate that the initial dose (nmol mm(-2)) of xenobiotic applied to plant foliage is a strong positive determinant of uptake. This held true for all the xenobiotics studied over a wide concentration range in the presence of these surfactants. Uptake on a unit area basis (nmol mm(-2)) could be related to the initial dose of xenobiotic applied per unit area (ID) by an equation of the form: Uptake = a [ID]b at time t = 24h. ID is given by the mass of xenobiotic applied, M divided by the droplet spread area, A. Total mass uptake is then calculated from an equation of the form: Total Uptake = a [ID]b x A.

2,4-Dichlorophenoxyacetic Acid↗

The antioxidant activity of a triterpenoid glycoside isolated from the berries of Hedera colchica: 3-O-(beta-D-glucopyranosyl)-hederagenin.

The antioxidant activity of a triterpenoid glycoside [3-O-(beta-D-glucopyranosyl)-hederagenin; OGH] isolated from the berries of Hedera colchica, an ivy species endemic in Georgia, was investigated. The antioxidant properties of OGH were evaluated using different antioxidant assays: 1,1-diphenyl-2-picryl-hydrazyl free radical (DPPH.) scavenging, total antioxidant activity, reducing power, superoxide anion radical (O2*-)) scavenging, hydrogen peroxide (H2O2) scavenging and metal chelating activities. The total antioxidant activity was measured according to the ferric thiocyanate method. alpha-Tocopherol and trolox, a water-soluble analogue of tocopherol, were used as reference antioxidant compounds. At a 30 microg/mL concentration, the inhibitory effects of OGH on the peroxidation of linoleic acid emulsion was found to be 95.3%, whereas alpha-tocopherol and trolox exhibited 88.8% and 86.2% inhibition of peroxidation in the system, respectively. In addition, OGH had effective DPPH. scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, total reducing power and metal chelation of ferrous ions activities. These various antioxidant activities were compared with alpha-tocopherol and trolox.

Antioxidants↗

[Allergic contact dermatitis to common ivy (Hedera helix L.)].

Common ivy (Hedera helix L.) is a ubiquitous plant in Europe whose major allergen falcarinol has moderate allergic potential. It is not related to poison ivy (Toxicodendron spp.). There are no cross reactions between the allergens of common ivy (falcarinol) and poison ivy (urushiol). Contact with common ivy or falcarinol may lead to sensitization and then a delayed hypersensitivity reaction. There are only few cases described in the literature. We report on a male hobby gardener with appropriate clinical history and positive patch test. The pathogenic mechanism is a type IV reaction following a sensitization exposure. Gardeners and landscape architects with frequent exposure to common ivy and thus a high risk of sensitization should wear appropriate protective clothing.

Adult↗

Host-related variability in arbuscular mycorrhizal fungal structures in roots of Hedera rhombea, Rubus parvifolius, and Rosa multiflora under controlled conditions.

The arbuscular mycorrhizal (AM) morphology of three host plant species inoculated with single and mixed fungal culture and the distribution of AM fungal species in roots of the hosts treated with a mixed culture of AM fungi were determined. The aim was to investigate the effect of host plants and AM fungi on AM morphology of coexisting plant species. Noncolonized rooted cuttings of Hedera rhombea (Miq) Bean, Rubus parvifolius L., and Rosa multiflora Thunb. were inoculated with five fungal species as single and mixed culture inocula. The fungal species used were Gigaspora rosea and Scutellospora erythropa, previously isolated from H. rhombea; Acaulospora longula and Glomus etunicatum from R. parvifolius; and Glomus claroideum from both plant species. A few hyphal and arbusculate coils were seen in the mixed culture-inoculated roots of R. parvifolius; all fungal treatments produced this Paris-type AM in H. rhombea and Arum-type AM in R. parvifolius, and R. multiflora indicates that AM morphology is strongly controlled by the identity of the host plants used in this study. AM fungal rDNA was extracted separately from roots of each replicate plant species inoculated with the mixed fungal culture, amplified, cloned, sequenced, and analyzed to determine the AM fungal species and their respective proportions in roots of each plant species. Glomus etunicatum and G. claroideum of the family Glomaceae generally occurred more frequently in R. parvifolius and R. multiflora, which form Arum-types, whereas S. erythropa, of the family Gigasporaceae, was the most frequently detected species in H. rhombea, which produced Paris-type AM. Although the genotype of the plant species used appears to determine the AM morphologies formed, there was preferential association between the hosts and AM fungal inoculants.

Hedera↗

In vitro activity of hederacolchisid A1 compared with other saponins from Hedera colchica against proliferation of human carcinoma and melanoma cells.

Hederacolchisid A1, a new oleanolic acid monodesmoside, isolated from Hedera colchica K. Koch, an ivy species endemic in Georgia, was evaluated in vitro for antiproliferative activity on cancer cells versus normal cells in comparison to cisplatin. Investigations were made on six human cell lines (colon adenocarcinoma DLD-1, ovarian teratocarcinoma PA 1, lung carcinoma A 549, breast adenocarcinoma MCF7, prostatic adenocarcinoma PC 3 and malignant melanoma M4 Beu) versus normal human fibroblasts, by assaying both cellular metabolic activity (RTT test) and DNA content in living cells (test with Hoechst 33,342) after 48 h continuous contact. Results demonstrated the strong cytotoxicity of hederacolchisid A 1 on all cancer cells (IC50 from 4.5 to 12 microM). The antiproliferative effects on malignant melanoma M4 Beu (IC50 ca 4.5 microM) versus normal cells (IC50 ca 7.5 microM) suggests that, despite a lack of specificity for cancer cells, hederacolchisid A1 has potential anti-tumor applications. Comparison of the cytotoxicity of hederacolchisid A 1 with that of five other saponins from H. colchica, offers some new information about structure-activity relationships. It was observed that i) for a same sugar sequence, monodesmosides with oleanolic acid as aglycone exhibit higher cytotoxicity than those containing hederagenin, ii) the sugar sequence O-alpha-L-rhamnopyranosyl (1 --> 2)-alpha-L-arabinopyranoside at C3 induces strong cytotoxicity and might be identified as a basic sequence for anti-tumor activity of oleanolic acid monodesmosides. iii) a complementary glucopyranosyl moiety branched at C1 of arabinose increases the cytotoxicity against malignant melanoma M4 Beu, prostatic adenocarcinoma PC 3 and normal fibroblasts in a different manner for each type of monodesmoside. A slight increase whose amplitude was quite similar on cancers and normal cells, was observed with oleanolic acid monodesmoside. This increase was much higher with hederagenin monodesmoside and markedly elevated in normal cells than in cancer cells.

Antineoplastic Agents↗

A multifunctional acyl-acyl carrier protein desaturase from Hedera helix L. (English ivy) can synthesize 16- and 18-carbon monoene and diene products.

A desaturase with 83% sequence identity to the coriander delta(4)-16:0-ACP desaturase was isolated from developing seeds of Hedera helix (English ivy). Expression of the ivy desaturase in Arabidopsis resulted in the accumulation of 16:1delta(4) and its expected elongation product 18:1delta(6) (petroselinic acid). Expression in Escherichia coli resulted in the accumulation of soluble, active protein that was purified to apparent homogeneity. In vitro assays confirmed delta(4) desaturation with 16:0-ACP; however, with 18:0-acyl acyl carrier protein (ACP) desaturation occurred at the delta(9) position. The ivy desaturase also converted 16:1delta(9)-ACP and 18:1delta(9)-ACP to the corresponding delta(4,9) dienes. These data suggest at least two distinct substrate binding modes, one placing C4 at the diiron active site and the other placing C9 at the active site. In the latter case, 18:0 would likely bind in an extended conformation as described for the castor desaturase with 9-carbons accommodated in the cavity beyond the dirron site. However, delta(4) desaturation would require the accommodation of 12 carbons for C16 substrates or 14 carbons for C18 substrates. The amino acids lining the substrate binding cavity of ivy and castor desaturases are conserved except for T117R and P179I (castor/ivy). Paradoxically, both substitutions, when introduced into the castor desaturase, favored the binding of shorter acyl chains. Thus, it seems likely that delta(4) desaturation would require a non-extended, perhaps U-shaped, substrate conformation. A cis double bond may facilitate the initiation of such a non-extended conformation in the monounsaturated substrates. The multifunctional properties of the ivy desaturase make it well suited for further dissection of the determinants of regiospecificity.

Amino Acid Sequence↗

A new technique for measurement of water permeability of stomatous cuticular membranes isolated from Hedera helix leaves.

Transpiration of cuticular membranes isolated from the lower stomatous surface of Hedera helix (ivy) leaves was measured using a novel approach which allowed a distinction to be made between gas phase diffusion (through stomatal pores) and solid phase diffusion (transport through the polymer matrix membrane and cuticular waxes) of water molecules. This approach is based on the principle that the diffusivity of water vapour in the gas phase can be manipulated by using different gases (helium, nitrogen, or carbon dioxide) while diffusivity of water in the solid phase is not affected. This approach allowed the flow of water across stomatal pores ('stomatal transpiration') to be calculated separately from the flow across the cuticle (cuticular transpiration) on the stomatous leaf surface. As expected, water flux across the cuticle isolated from the astomatous leaf surface was not affected by the gas composition since there are no gas-filled pores. Resistance to flux of water through the solid cuticle on the stomatous leaf surface was about 11 times lower than cuticular resistance on the astomatous leaf surface, indicating pronounced differences in barrier properties between cuticles isolated from both leaf surfaces. In order to check whether this difference in resistance was due to different barrier properties of cuticular waxes on both leaf sides, mobility of 14C-labelled 2,4-dichlorophenoxy-butyric acid 14C-2,4-DB) in reconstituted cuticular wax isolated from both leaf surfaces was measured separately. However, mobility of 14C-2,4-DB in reconstituted wax isolated from the lower leaf surface was 2.6 times lower compared with the upper leaf side. The significantly higher permeability of the ivy cuticle on the lower stomatous leaf surface compared with the astomatous surface might result from lateral heterogeneity in permeability of the cuticle covering normal epidermal cells compared with the cuticle covering the stomatal cell surface.

Algorithms↗