[Medicinal and poisonous plants. VIII. Bryonia alba and Bryonia dioica].
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Bryonia↗
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Evaluation of anti-inflammatory activity of chloroform extract of Bryonia laciniosa in experimental animal models.
The anti-inflammatory effect of the leaves of Bryonia laciniosa was evaluated using carrageenan, dextran, histamine, serotonin induced rat paw oedema and cotton pellet induced granuloma (chronic) models in rats. In mice, carrageenan peritonitis test was performed for the extract by oral administration. The chloroform extract of Bryonia laciniosa (CEBL) exhibited significant anti-inflammatory effect at the dose 50, 100 and 200 mg/kg. Maximum inhibition (52.4%) was noted at the dose of 200 mg/kg after 3 h of drug treatment in carrageenan induced paw oedema, whereas the indomethacin (standard drug) produced 62.1% of inhibition. The extract exhibited significant anti-inflammatory activity in dextran induced paw oedema in a dose dependent manner. The extract also exhibited significant inhibition on the hind paw oedema in rats caused by histamine and serotonin respectively. In the chronic model (cotton pellet induced granuloma) the CEBL (200 mg/kg) and standard drug showed decreased formation of granuloma tissue by 50.1 and 57.3% (p<0.001) respectively. The extract also inhibited peritoneal leukocyte migration in mice. Thus, the present study revealed that the chloroform extract of Bryonia laciniosa exhibited significant anti-inflammatory activity in the tested models.
Animals↗
Isolation, sequencing and analysis of the expression of Bryonia calmodulin after mechanical perturbation.
A cDNA clone (Bc329) encoding calmodulin was isolated from a Bryonia cDNA library by screening with cloned Arabidopsis calmodulin cDNA. The cDNA Bc329 was 899 bp full-length clone. The predicted amino acid sequence consists of 149 residues and reveals a high homology with other known plant calmodulins (91 to 99% identity). Genomic southern blot suggests that Bryonia calmodulin is encoded by a single-copy gene. The Bc329 clone was used as a probe to study the expression of calmodulin mRNA after a mechanical stimulus applied on young Bryonia internodes. The steady-state of calmodulin mRNA reached a maximum 30 min after the treatment before it progressively decreased. The role of calcium and calmodulin as second messengers is discussed with regard to environmental changes.
Amino Acid Sequence↗
AFLP analysis of a worldwide collection of Didymella bryoniae.
Didymella bryoniae (anamorph Phoma cucurbitacearum) is an ascomycete that causes gummy stem blight, a foliar disease that occurs on cucurbits in greenhouses and fields throughout the world. In a previous study using RAPD analysis, little genetic diversity was found among isolates of D. bryoniae from New York and South Carolina, USA. Here we report the use of amplified fragment length polymorphism (AFLP) analysis to assess the genetic variation within a worldwide collection of D. bryoniae, 102 field and greenhouse isolates from ten states in the USA (California, Delaware, Florida, Georgia, Indiana, Maryland, Michigan, Oklahoma, South Carolina, and Texas) and seven other countries (Australia, Canada, China, Greece, Israel, Sweden, and The Netherlands) were examined. Seven different AFLP primer-pair combinations generated 450 bands, of which 134 were polymorphic (30%). Using cluster analysis, two groups and a total of seven subgroups were delineated. Representative isolates varied in their virulence on muskmelon and watermelon seedlings, but the degree of virulence was not strongly associated with AFLP groupings. However, isolates from the northern USA grouped separately from isolates originating from the southern USA.
Ascomycota↗
Evaluation of analgesic, antipyretic activity and toxicity study of Bryonia laciniosa in mice and rats.
Analgesic, antipyretic activity and toxicity study of the leaves of Bryonia laciniosa Linn. (Family: Cucurbitaceae) was evaluated in the standard animal models. The methanol extract of Bryonia laciniosa (MEBL) was evaluated by hot plate and acetic acid-induced writhing methods to assess analgesic activity. The antipyretic activity of the extract was also evaluated by normal body temperature and yeast-induced hyperpyrexia. The extract showed significant analgesic and antipyretic activity. The MEBL was further evaluated for toxicity at the doses of 125 and 250 mg/kg administered orally for 14 days in rats. At the end of experiments, the blood, liver function and kidney metabolism were observed. The hematological profile and different biochemical parameters such as SGOT, SGPT and ALP were estimated. The present study revealed that MEBL exhibited significant analgesic and antipyretic activity in the tested experimental animal models. The toxicity study indicates that the extract is not toxic at the tested doses.
Alanine Transaminase↗
Cytological indication of the involvement of calcium and calcium-related proteins in the early responses of Bryonia dioica to mechanical stimulus.
The distribution of membrane-bound calcium, activated calmodulin, and callose synthesis was visualized in Bryonia dioica internodes before and after mechanical stimulus, using fluorescent probes, respectively, chlorotetracycline, fluphenazine, and aniline blue. Bright chlorotetracycline fluorescence remains localized in the plasma membrane of control cells, 30 s after stimulation calcium left the plasmalemma. A delocalization of activated calmodulin was observed after wounding and deposition of callose, which could not be detected before, appeared in the same times in most cells. The callose formation and the decrease in membrane-associated calcium suggest a rapid influx of calcium in the cytosol and an intervention of this ion in the cascade of the early events underlying Bryonia dioica thigmomorphogenesis.
Calcium↗
Restoration of the disordered glucose-fatty acid cycle in alloxan-diabetic rats by trihydroxyoctadecadienoic acids from Bryonia alba, a native Armenian medicinal plant.
We studied how administration of trihydroxyoctadecadienoic acids obtained from the roots of the native Armenian plant Bryonia alba L. (0.05 mg/kg/day for 15 days. i.m.) restores the disordered lipid metabolism of alloxan-diabetic rats. Diabetes was accompanied by an increase in total non-esterified fatty acid content of blood together with decreases in muscle and adipose tissue of total non-esterified fatty acids and triglycerides, together with marked alterations of phospholipid fatty acid distribution within the membranes from muscle, including increased short chain fatty acid and decreased arachidonate content. All these metabolic changes induced in diabetes were significantly restored by trihydroxyoctadecadienoic acid treatment towards their normal values (P < 0.005) with the exception of diminished triglyceride content of muscle which was not restored. In experiments on rat neutrophil leukocytes in vitro, it was found that the standardised preparation of Bryonia C-18 fatty acids (a mixture of four diastereoisomeric forms of the positional isomers I 12,13,16-trihydroxy-9Z,14E-octadecadienoic acid, II 12,15,16-trihydroxy-9Z,13E-octadecadienoic acid, III 9,10, 13-trihydroxy-11E,15Z-octadecadienoic acid and IV 9,12,13-trihydroxy-10E,14Z-octadecadienoic acid) had no effect on granular enzyme secretion or 5-lipoxygenase activity at 5-50 micrograms/ml, but dose-dependently reduced thromboxane B2 generation, with a corresponding increase in prostaglandin E2 release. We conclude that trihydroxyoctadecadienoic acids from B. alba can correct major metabolic abnormalities typical of severe diabetes mellitus, and that they can influence the profile of the formation of stable prostaglandins by actions downstream of prostaglandin endoperoxides.
Alloxan↗
Specific Inhibition of Lignification in Bryonia dioica: Effects on Thigmomorphogenesis.
Rubbing-induced lignification in Bryonia dioica internodes is significantly impaired by N(o-hydroxyphenyl) and N(o-aminophenyl) sulfinamoyl tertiobutyl acetate, specific inhibitors of cinnamyl alcohol dehydrogenase, an enzyme which is strictly associated with lignin monomer synthesis. Along with the reduction of lignification, these inhibitors counteract the inhibition of elongation due to rubbing. These results indicate that lignification participates in the thigmomorphogenetic growth response of Bryonia dioica internodes. In a general way, the data point to the causal role of lignification in the limitation of plant growth.
Journal Article↗
[Effect of Bryonia cucurbitacins on the biosynthesis of eicosanoids in human leukocytes].
2 beta,25-di (beta-D-glucopyranosyl)-16 alpha,20-dihydroxy-3,11,22- trioxocucurbit-5-en and 2 beta-(beta-D-glucopyranosyl)-16 alpha,20,25-trihydroxy-3,11,22-trioxocucurbit-5-en isolated from bryonia (Bryonia alba L.) roots have been demonstrated to inhibit in vitro the [1-14C]arachidonic acid release from neutrophils. Aglicon 2 beta,16 alpha,20,25-tetrahydroxy-3,11,22-trioxocucurbit-5-en is much less active. When the cells are stimulated by calcium ionophore A23187, the aglycon potentiates the release of arachidonic acid. In these conditions the glucosides show little activity. Both the glucosides and their aglycon suppress the biosynthesis of 5S,12R-dihydroxy-6,8,10,14(Z, E, E, Z)-eicosatetraenoic acid (LTB4) and 5S,12S-dihydroxy-6,8,10,14(E, Z, E, Z)-eicosatetraenoic acid (5S,12S-DHETE). Inhibition of the biosynthesis of these compounds by 2 beta,16 alpha,20,25-tetrahydroxy-3,11,22-trioxocucurbit-5-en also takes place on incubation of human neutrophils with exogenous arachidonic acid. The formation of other products of cycloxygenase and lipoxygenase oxidation pathways remains practically unchanged.
Arachidonic Acids↗
Changes in plasma membrane fluidity of Bryonia dioica internodes during thigmomorphogenesis.
Fluidity changes in plasma membrane (PM) lipid extracts or native membranes isolated from Bryonia dioica internodes after a mechanical stimulation were monitored by steady-state fluorescence polarization with 1,6-diphenyl-1,3,5-hexatriene as a probe. The signal was shown to rapidly induce an increase in the bulk lipid fluidity. This event was closely related to a relative enrichment in some phospholipid species (PC, PG and PS) as well as a significant increase in the unsaturation index of total fatty acyl chains. Free sterols and protein content did not appear to be involved into this process. After 48 h, lipids from rubbed internodes became less fluid than PM lipids from control internodes.
Cell Membrane↗
Structural revision of bryonoside and structure elucidation of minor saponins from Bryonia dioica.
The structure of bryonoside, which has previously been isolated from the roots of Bryonia dioica, has been revised. In addition, three new cucurbitacin saponins, named brydioside A, B and C have also been obtained. Their structures were established on the basis of spectral and chemical evidence.
Carbohydrate Sequence↗
A toxic protein from Bryonia dioica Jacq. fruits: the brydiofin.
A toxic protein isolated from Bryonia dioica Jacq. fruits is described. The fruit extract, obtained in phosphate buffer saline, in mild conditions, showed a strongly toxic activity after intraperitoneal injection of 0.4 mg per Balb/c mouse that was killed in eighteen minutes. The extract, filtered through a Bio-Gel P-60 column, presented a main peak that showed similar toxicity. When the material from this peak was analyzed in polyacrylamide slab gels, a single band corresponding to a molecular weight of about 66,000 was observed. The lethal effect of B. dioica fruit could be attributed to a protein that we called Brydiofin. The extracts from roots and leaves that did not present this toxicity showed no band in this molecular weight region.
Animals↗
Modulation of the ER Ca2+ channel BCC1 from tendrils of Bryonia dioica by divalent cations, protons and H2O2.
Electrical properties of the ER Ca2+ channel BCC1 from tendrils of Bryonia dioica were analyzed after incorporation of BCC1 into black lipid bilayers. Single channel current fluctuations were modulated by divalent cations, protons and H2O2. Whereas the channel is permeable for Ca2+, Ba2+ and Sr2+, its conductance is strongly reduced in solutions containing MgCl2. Cu2+ and Zn2+ are potent inhibitors of BCC1 in micromolar concentrations. The open channel conductance of BCC1 increases with acidification of the electrolyte solution. H2O2 shows strong inhibitory effects on BCC1. The channel is almost completely closed at submillimolar concentrations of H2O2. The effects of pH and H2O2 on channel properties are directional and affect BCC1 at the Ca exit side, but not on the entry site. Thus, cytosolic pH and H2O2 levels may play an important role in the modulation of the cytoplasmic free calcium concentration through BCC1.
Calcium↗
Cycloartane triterpenoids from the aerial part of Bryonia dioica.
A novel triterpenoid isolated from the non-saponifiable lipid of the aerial part extract of white bryony (Bryonia dioica) was established to be 3beta-hydroxy-27-norcycloartan-24-one based on spectroscopic methods. Isolation and identification of other known cycloartane triterpenoids and sterols also are described.
Journal Article↗
Characterization of ribosome-inactivating proteins isolated from Bryonia dioica and their utility as carcinoma-reactive immunoconjugates.
Two ribosome-inactivating proteins (RIPs) were isolated and characterized from the roots of Bryonia dioica. One of these was a novel 27-kDa protein termed bryodin 2 (BD2), while the second was a previously reported RIP, referred to here as bryodin 1 (BD1). The amino-terminal sequence obtained for BD2 was similar, but distinct from BD1, ricin A chain, trichosanthin, and momorcharin. BD2-specific monoclonal antibodies were generated and found not to react with BD1 or ricin A chain. Purified BD1 and BD2 RIP inhibited protein synthesis in a cell-free in vitro translation assay at EC50 values of 7 and 9 pM, respectively. Intravenous administration of BD1 was less toxic to mice than BD2, with LD50 values of > 40 for BD1 and 10-12 mg/kg for BD2. Primary human endothelial cells were 5-8-fold less sensitive to BD1 and BD2 than compared to ricin A chain. BD1 and BD2 were constructed as immunoconjugates with the chimeric form of BR96 (chiBR96), a carcinoma-reactive, internalizing antibody. ChiBR96-BD1 and chiBR96-BD2 were found to bind to and kill BR96 antigen-positive carcinoma cells while not killing antigen-negative carcinoma cells. Bryodins represent RIPs that may be useful in constructing immunotoxin conjugates with reduced toxicity and vascular sensitivity, as compared to ricin A chain immunotoxins.
Amino Acid Sequence↗
Molecular, biological, and preliminary structural analysis of recombinant bryodin 1, a ribosome-inactivating protein from the plant Bryonia dioica.
Bryonia dioica (Cucurbitaceae family) produces at least two type I ribosome-inactivating proteins, bryodin 1 (BD1) and bryodin 2 (BD2). A cDNA sequence encoding BD1 was isolated from B. dioica leaf mRNA using degenerative oligonucleotides and codes for a 22 amino acid signal peptide followed by a protein of 267 residues. Expression of two recombinant BD1 (rBD1) forms in Escherichia coli yielded proteins of 267 (to the natural stop codon) and 247 amino acids (to the putative cleavage site yielding the mature protein) that had identical protein synthesis inhibition activity as compared to native BD1. The substitution of Lys for Glu at position 189 near the active site reduced the ability of rBD1 to inhibit protein synthesis by 10-fold. Toxicologic analysis showed that rBD1 was well tolerated in rodents with LD50 values of 40 mg/kg in mice and >25 mg/kg in rats. A crystal of mature rBD1 protein was used to collect X-ray diffraction data to 2.1 A resolution. The rBD1 crystal structure was solved and showed extensive homology with other type I RIPs and A chains of type II RIPs. The studies described here demonstrate that rBD1 retains full biologic activity and serve as a guide for using this potent, yet nontoxic, RIP in the construction of single-chain immunotoxin fusion proteins.
Amino Acid Sequence↗
Bryodin, a ribosome-inactivating protein from the roots of Bryonia dioica L. (white bryony).
Bryodin is a strongly basic (pI greater than or equal to 9.5) glycoprotein (neutral sugar content 6.3%) with Mr 30,000, purified from the roots of Bryonia dioica (white bryony). This protein inhibits protein synthesis by a rabbit reticulocyte lysate with and ID50 (concentration causing 50% inhibition) of 0.12 nM (3.6 ng/ml) and has much less effect on protein synthesis by whole cells, with ID50 values ranging from 46 nM to 2.27 microM (1.4-67 micrograms/ml). Bryodin acts by inactivating ribosomes, with a less-than-equimolar ratio, which suggests a catalytic action. Bryodin decreases the number of local lesions induced by tobacco mosaic virus in the leaves of Nicotiana glutinosa. From all its properties, bryodin can be considered to be a ribosome-inactivating protein, similar to those already known [reviews: Barbieri & Stirpe (1982) Cancer Surveys 1, 489-520; Stirpe & Barbieri (1986) FEBS Lett. 195, 1-8].
DNA↗
Effects of a mechanical stimulation of localization of annexin-like proteins in Bryonia dioica internodes.
Mechanical stimulation exerted by rubbing a young internode of Bryonia dioica plants inhibits its growth. Previous cellular and biochemical studies showed that this growth inhibition is associated with Ca(2+) redistribution and profound modifications of plasma membrane characteristics. We extracted and purified Ca(2+)-dependent phospholipid-binding proteins from B. dioica internodes. Two main proteins, p33 and p35, and other minor bands were isolated and identified as annexin-like proteins because of their biochemical properties and their cross-reactions with antibodies against maize (Zea mays L.) annexins. Rabbit antiserum was obtained by injection of B. dioica p35. This antiserum was used for the immunocytolocalization of annexin-like proteins in internode parenchyma cells. It appeared that the distribution of annexin-like proteins was different before and 30 min after the mechanical stimulation. Western analysis of proteins in membrane fractions after separation by free-flow electrophoresis showed that p35 was present in most fractions, whereas p33 appeared mainly in plasmalemma-enriched fractions after the mechanical stimulation. It is hypothesized that a subcellular redistribution of these proteins might be involved in growth inhibition by mechanical stress.
Animals↗