Euphorbiaceae Plants Research Articles

EFFECTS OF TOPICAL TREATMENT WITH EUPHORBIA TIRUCALLI LATEX ON THE SURVIVAL AND INTESTINAL ADHESIONS IN RATS WITH EXPERIMENTAL PERITONITIS.

Background: The use of plants of the family Euphorbiaceae, particularly Euphorbia tirucalli (avelós) has been popularly widespread for treating a variety of diseases of infectious, tumoral, and inflammatory. Aim: To demonstrated antimicrobial and immunomodulatory effects of these extracts, evaluating the effect of a topical treatment with an aqueous solution of avelós latex on the survival and on intestinal adhesions in rats with experimental peritonitis. Methods: Peritonitis was induced in 24 Wistar rats, that were randomized into four groups of six as follows: (1) Control group (n=6), no treatment; (2) Antibiotic group (n=6), treatment with a single intramuscular dose of antibiotic Unasyn; (3) Saline group (n=6), the abdominal cavity was washed with 0.9% saline; and (4) E.tirucalli group (n=6), the abdominal cavity was washed with E. tirucalli at a concentration of 12 mg/ml. The animals that died were necropsied, and the time of death was recorded. The survivors were killed on postoperative day 11, and necropsy was subsequently performed for evaluation of the intestinal adhesions. Results: Significant differences were observed in the control and antibiotic groups (p<0.01) with respect to the survival hours when compared with the saline and E. tirucalli groups. There was no significant difference (p>0.05) in the survival of animals in the saline andE. tirucalli groups; however, one animal died in the saline group. Necropsy of the animals in the saline and E. tirucalligroups showed strong adhesions resistant to manipulation, between the intestinal loops and abdominal wall. The remaining groups did not show any adhesions. Conclusions: Topical treatment with E. tirucalli latex stimulated an increased formation of intestinal adhesions and prevented the death of all animals with peritonitis.

Analysis of ingenol and its conjugates in some species of the Euphorbia genus by ultra-high performance liquid chromatography-tandem mass spectrometry using isotope dilution method

Ingenol is a chemically and biologically important plant-derived diterpenoid compound, first isolated in 1968 from the seed oil of a leafy shrub Croton tiglium belonging to the Euphorbiaceae plant family. Ingenol is the key precursor for the synthesis of ingenol mebutate (angelate), used to treat actinic keratosis. Although, its total synthesis was published lately, isolation from natural raw material is still used for its industrial scale production. Therefore, a rich source of ingenol seems to be of great economic importance. We have developed an ultra-high performance liquid chromatography-tandem mass spectrometry method for screening of ingenol in 38 species of the Euphorbia genus. The highest ingenol concentration (547 mg.kg-1 of dry weight) was found in the lower leafless stems of E. myrsinites. Moreover, three ingenol conjugates (two known: 3-O-deca(2'Z,4'E)-dienoyl ingenol, 20-O-acetyl-3-O-deca(2'Z, 4'E)-dienoyl ingenol and one new: 3-O-octa (2'Z,4'E)-dienoyl ingenol) were successfully identified in E. myrsinites by means of accurate mass determination.

Lysozyme- and chitinase activity in latex bearing plants of genus Euphorbia – A contribution to plant defense mechanism

Occurrence of latices in plants is widespread, there are 40 families of plants characterized to establish lactiferous structures. Latices exhibit a constitutive part of plant defense due to the stickiness. The appearance of proteins incorporated in latices is well characterized, and hydrolytic active proteins are considerable. A lot of plants constitute so-called pathogenesis-related (PR) proteins, to overcome stressful conditions. In our investigation we are focused on latex bearing plants of Euphorbiaceae Juss., and investigated the appearance of chitinase- and lysozyme activity in particular. The present outcomes represent a comprehensive study, relating to the occurrence of lysozyme and chitinase activity of genus Euphorbia at the first time. 110 different species of genus Euphorbia L. were tested, and the appearance of chitinase and lysozyme were determined in different quantities. The appearance itself, and the physicochemical properties of latices indicate an efficient interaction for plant defense against pathogen attack.

Development of Microsatellite Markers in Tung Tree (Vernicia fordii) Using Cassava Genomic Sequences

Tung tree, Vernicia fordii, is a native oil-bearing woody plant of China. Oil extracted from the fruit is an important industrial and biodiesel feedstock. Microsatellite (simple-sequence repeat (SSR)) markers are effective in germplasm evaluation and marker-assisted breeding of plants. The objective of this study was to develop SSR markers in tung tree using cassava genomic sequences and test the effectiveness of the interspecific transferable SSR markers. According to the principle of comparative genomics, SSR markers were designed based on cassava genomic sequences and tested in tung tree and other Euphorbiaceae plant species. Among 255 cassava SSR markers, 104 (41 %) could effectively amplify DNA of tung tree, and a large portion of these transferable SSR markers could work well in other Euphorbiaceae species, Vernicia montana (98 %), castor bean (92 %), and Jatropha curcas (88 %). Genotyping a set of 16 tung tree accessions using the 104 transferable SSRs, a total of 437 alleles were obtained, 162 of which were polymorphic, and the Shannon information index varied from 0.000 to 0.560 with a mean of 0.139. A phylogenetic tree unambiguously distinguished the 32 accessions of the abovementioned five Euphorbiaceae species. Using cassava genomics resources is an effective approach to develop SSR markers for tung tree when its genome sequence is not available. The developed SSR markers should facilitate germplasm evaluation and other molecular-level investigations in tung tree.

Methanol extracts of Euphorbia cooperi inhibit the production of inflammatory mediators by inhibiting the activation of c-Jun N-terminal kinase and p38 in murine macrophages

Numerous Euphorbiaceae plants have been used for the treatment of diseases, including liver diseases, asthma and rheumatism. The present study evaluated the effect of methanol extracts from Euphorbia cooperi (MEC), a member of the Euphorbiaceae plant family, on the production of inflammatory cytokines interleukin (IL)‑6 and tumor necrosis factor (TNF)‑α, nitric oxide (NO) as well as the activation of mitogen‑activated protein kinase and nuclear factor (NF)‑κB signaling. Non‑cytotoxic concentrations of MEC significantly reduced the production of NO and IL‑6, but not TNF‑α, in lipopolysaccharide (LPS)‑stimulated RAW 264.7 macrophages. The decreased production of NO by MEC was due to alleviated expression of inducible NO synthase. Reporter assays with cells treated with MEC demonstrated reduced activator protein‑1 (AP-1) activity, while NF‑κB activity was not reduced. Furthermore, the phosphorylation levels of c‑Jun N‑terminal kinase (JNK) and p38 were suppressed by MEC while phosphorylation levels of inhibitor of κB were not reduced by MEC, suggesting that MEC‑mediated inactivation of JNK and p38 is the underlying regulatory mechanism for inflammatory mediators in LPS‑stimulated RAW 264.7 macrophages.

Proteinase activity in latex of three plants of the family Euphorbiaceae

In the family of Euphorbiaceae,the genera Euphorbia and Sapium are known to contain essentially latex-bearing species. In the present study, the latex of Euphorbia selloi(Klotzsch &amp; Garcke) Boiss., Euphorbia papillosa A.St.-Hil., and Sapium glandulosum (L.) Morong, plants native from Brazil, were examined concerning proteolytic activity. All studied species have proteins with significant proteolytic activity and E. papillosa has the greatest specific activity. Aiming to verify the type of protease present, an assay with different inhibitors was performed. In the three tested plants, the proteolytic activity was significantly inhibited by a serine protease inhibitor 4-(2-aminoethyl)-benzenesulfonyl fluoride hydrochloride (AEBSF). Using techniques of electrophoresis with polyacrylamide gels (SDS-PAGE), the subunits of proteins were separated according to their molecular masses, and the protein activity was visually detected by zymography.

Microbubbles Enhanced Synthetic Phorbol Ester Degradation by Ozonolysis

A phorbol-12-myristate-13-acetate (TPA) is a synthetic analogue of phorbol ester (PE), a natural toxic compound of Euphorbiaceae plant. The oil extracted from plants of this family is useful source for primarily biofuel. However this oil might also be used as a foodstuff due to its significant nutrition content. The limitations for utilizing the oil as a foodstuff are mainly due to a toxicity of PE. Currently, a majority of PE detoxification processes are expensive as include multi steps alcohol extraction sequence. Ozone is considered as a strong oxidative agent. It reacts with PE by attacking the carbon-carbon double bond of PE. This modification of PE molecular structure yields a non toxic ester with high lipid content. This report presents data on development of simple and cheap PE detoxification process with water application as a buffer and ozone as reactive component. The core of this new technique is an application for a new microscale plasma unit to ozone production and the technology permits ozone injection to the water-TPA mixture in form of microbubbles. The efficacy of a heterogeneous process depends on the diffusion coefficient which can be controlled by contact time and interfacial area. The low velocity of rising microbubbles and high surface to volume ratio allow efficient mass transfer to be achieved during the process. Direct injection of ozone is the most efficient way to process with such highly reactive and short lived chemical. Data on the plasma unit behavior are presented and the influence of gas oscillation technology on the microbubble production mechanism has been discussed. Data on overall process efficacy for TPA degradation is shown.

Evaluation of hemostatic activity of latex from three Euphorbiaceae species

Ethnopharmacological relevanceLatices from several plant species of Euphorbiaceae family have been traditionally applied over fresh cuts to stop bleeding and subsequently applied over wounds to enhance healing process. The latex arrested bleeding from fresh wounds by reducing bleeding and whole blood coagulation time which are important indices of hemostatic activity. It has been accepted that hemostatic activity is due to the proteolytic fraction of plant latices. Thus, the present study aimed to assess the clot inducing properties of three Euphorbiaceae plants viz., Euphorbia nivulia Buch.-Ham., Pedilanthus tithymaloides (L.) Poit and Synadenium grantii Hook F. Materials and methodsIn the present study, various proteolytic activities namely protease, gelatinase, milk clotting and whole blood clotting assay of the enzyme fraction of latices of Euphorbia nivulia, Pedilanthus tithymaloides and Synadenium grantii have been investigated. The inhibition profile of protease specific inhibitors was assessed. Also, the effects of protein fractions were studied using bleeding/clotting time test of fresh experimentally-induced wounds in mice. ResultsEuphorbia nivulia latex protease has noticeable blood clotting activity followed by Pedilanthus tithymaloides and Synadenium grantii. Stem latex protease of Pedilanthus tithymaloides exhibits superior procoagulant activity in different mammal's blood samples viz., Capra hircus, Bubalus bubalis, Ovibos moschatus and Bos indicus. Blood sample of ox was the most sensitive to latex protease than other mammal's blood. Concomitantly, the plant latex protease could significantly reduce whole blood clotting time of human and mice blood samples. ConclusionThe protease fraction of latices of Euphorbia nivulia, Pedilanthus tithymaloides and Synadenium grantii possesses phytoconstituents capable of arresting wound bleeding, and accelerating whole blood coagulation process. It suggests good potentiality for use of latex proteases in wound management. Also, the finding of this study showed that the protease enzyme of Pedilanthus tithymaloides has the most potent hemostatic agent.

Both plants Sebastiania chamaelea from Niger and Chrozophora senegalensis from Senegal used in African traditional medicine in malaria treatment share a same active principle

Ethnopharmacological relevanceBased on ethnobotanical data obtained from Nigerien and Senegalese traditional healers, two Euphorbiaceae plants, Sebastiania chamaelea and Chrozophora senegalensis, traditionally used to treat malaria, were selected for further investigations. Materials and methodsPlant extracts were prepared with different solvents and tested both in vitro on several strains of Plasmodium falciparum, and in vivo to evaluate their antiplasmodial properties and isolate their active principles. ResultsWith IC50 values around 6.5µg/ml and no significant cytotoxicity (>50µg/ml), the whole plant aqueous extract from S. chamaelea showed the best in vitro results. In vitro potentiation assays showed strong synergistic activity of S. chamaelea extract with the antiplasmodial drug chloroquine on the chloroquine-resistant P. falciparum strain W2-Indochina. In other respects, the aqueous crude extract of C. senegalensis leaves showed the most significant antiplasmodial activity in vitro (IC50 values less than 2µg/ml). We also demonstrated the prophylactic activity of C. senegalensis in vivo in a murine malaria model. Bioassay-guided fractionation of aqueous extracts of these plants enabled the isolation and identification of ellagic acid (EA, 1) as the main compound responsible for their antiplasmodial activity. Together with EA, other derivatives belonging to different chemical groups were isolated but showed moderate antimalarial activity: gallic acid (2), brevifolin carboxylic acid (3), protocatechuic acid (4), corillagin (5), rutin (6) and 3,4,8,9,10-pentahydroxy-dibenzo(b,d)pyran-6-one (7). The structures were determined by the usual spectroscopic methods and by comparison with published data. Furthermore, we report here the quantification of compound 1 (EA) by RP-HPLC in the dried extracts of these plants, reported for the first time in both these species, and possessing the highest in vitro antiplasmodial activity with IC50 values from 180 to 330nm. ConclusionsThese in vitro and in vivo results support the traditional use in Africa of crude extracts of both S. chamaelea and C. senegalensis as an antimalarial treatment and prove the significant antiplasmodial property of EA.

English

The antibacterial activity of the methanolic and n-hexane extracts of Fagonia cretica L., Peganum harmala L., Tribulus terrestris L., Chrozophora tinctoria (L.) Raf and Ricinus communis L., were investigated against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoni and Salmonella typhi. Phytochemical screening of the methanolic extract of F. cretica, P. harmala and C. tinctoria revealed the presence of saponins, phlobatannins, anthraquinones and alkaloids. Tannins and flavonoids were found in C. tinctoria and R. communis. Glycosides were found in F. cretica and P. harmala, while terpenoids and phenolics occurred in P. harmala. Group tests revealed that the methanolic and n-hexane extract of F. cretica, P. harmala, T. terrestris, C. tinctoria andR. communis had OH, carboxylic and phenolic groups. The methanolic and n-hexane extracts of F. cretica, P. harmala, T. terrestris, C. tinctoria and R. communis showed the absence of aldehyde and ketone group. The ability of the methanolic and n-hexane extracts of F. cretica, P. harmala, T. terrestris, C. tinctoria and R. communis to inhibit the growth of different bacteria has shown their broad spectrum antibacterial potential, which may be used in the management of microbial infections. Key words: Zygophyllaceae, Euphorbiaceae, antibacterial activity, extract, phytochemical screening.

Biosynthesis of Silver Nanoparticles Using Latex from Few Euphorbian Plants and Their Antimicrobial Potential

The synthesis of well-dispersed and ultrafine metal nanoparticles has great interest due to their distinctive physicochemical properties and biomedical applications. This study is the first report of one-step solvent-free synthesis of AgNPs using Euphorbiaceae plant latex. Among evaluated eight latex-producing plants, four (Jatropha curcas, Jatropha gossypifolia, Pedilanthus tithymaloides, and Euphorbia milii) showed high potential to produce physicochemically distinct, small-sized and bactericidal AgNPs. Phytochemical screening showed presence of rich amount of biochemicals in these plants. J. gossypifolia showed uniformly dispersed comparatively small-sized AgNPs. Dose-dependent growth inhibition of bacterial pathogens Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus epidermis, and Micrococcus luteus was observed for J. gossypifolia latex-synthesized AgNPs with minimum inhibitory concentration values 30, 40, 70, 60, and 60 ppm, respectively, after 24 h. Possible mode of action of AgNPs against pathogens was confirmed by analyzing enzymes and cell leakage.

Bioactive components in the fruits of Ziziphus jujuba Mill. against the inflammatory irritant action of Euphorbia plants

Chinese jujube (also known as Chinese date) is the fruit of Ziziphus jujuba Mill. (Rhamnaceae). As a famous folk medicine, it is used as antidote in traditional Chinese formula, Shi Zao Decoction, to relieve the drastic inflammatory irritant nature of Euphorbia species. The irritant activities may cause serious adverse effects in clinical practices. This study aimed to investigate the active components of Z. jujuba through the inhibitory effects on the inflammatory cells activated by Euphorbia kansui and prostratin, a phorbol ester isolated from Euphorbia fischeriana. Peritoneal macrophage of rat and splenic lymphocyte (splenocyte) of mouse were selected to evaluate these actions in vitro. Nitric oxide (NO) release of macrophage and the proliferation of splenocyte were examined through Griess method and MTT assay. TNF-α, as an important pro-inflammatory cytokines, was detected with enzyme-linked immunosorbent assay (ELISA) method. Six fractions extracted from Z. jujuba were evaluated and fraction F (triterpene acids fraction) was demonstrated to be the most active part, and then, 21 compounds isolated from Z. jujuba were tested at the concentrations range from 1μg/ml to 100μg/ml. The results show that 7 compounds of them are likely to be active compounds concerning to their pronounced inhibitory action on the activated inflammatory cells. These effects might be helpful to attenuate the irritant action of Euphorbiaceae plants and protect the gastrointestinal tissue from potent inflammatory injury, which should be beneficial to some diseases, like inflammatory bowel disease.

First Report of Powdery Mildew on Euphorbia pekinensis Caused by Podosphaera euphorbiae-helioscopiae in China.

Euphorbia pekinensis Rupr, the Peking spurge, is a perennial grass that is planted throughout northeast China. Its roots can be used as a Chinese medicine for diuretic or purgative purposes. Some diterpenoids isolated from Euphorbia spp. show strong antitumor, antivirus, skin-irritating, and tumor-promoting activities. Some species contain chemicals that can be used as pesticides. In September and October 2009, spurge plants were seriously infected by powdery mildew fungi in Changchun Agri-Expon, Jilin Province, China. White mildew colonies appeared in early autumn and initial colonies coalesced to cover the entire upper sides of leaves, stems, and petals 3 to 4 weeks after the disease was noticed. Until late October, chasmothecia were observed on leaves and stems and the infected areas also became dark brown. Mycelium was amphigenous, in patches, white, and persistent. Conidia with distinct fibrosin bodies were ellipsoid-cylindrical, 21.6 to 33.8 × 12 to 15.6 μm, and produced in chains. Germ tubes arose near a pole of the conidia and terminated in simple, unlobed apices. Conidiophores were 59.8 to 139.9 μm; foot cells were straight to slightly flexuous, followed by one to three cells, and 24 to 48 × 8.4 to 10.8 μm. Hyphal appressoria were papillate. Chasmothecia were 70 to 95 μm in diameter, averaging 79.4 μm, and were immersed in mycelia. Mycelioid appendages (5 to 12) occurred in the lower part of the chasmothecia and were flexuous, curved, thick walled, and strongly undulate. Appendages were long (349-) 490 to 1,000 (-1,300) μm, averaging 759.4 μm. They were often densely crowded around the chasmothecia. Asci were single, sessile or short stalked, 60 to 80 × 45 to 70 μm, and eight spored. Ascospores were ellipsoid-ovoid and 16.8 to 28.8 × 10.8 to 18 μm. The fungus was identified as Podosphaera euphorbiae-helioscopiae on the basis of morphological characteristics of the anamorphic and teleomorphic stages. An herbarium specimen was deposited in the Herbarium of Mycology of Jilin Agriculture University (CJLCC100). Genomic DNA was extracted from the chasmothecia, and the internal transcribed spacer region (ITS) and the D1/D2 domains of the 28S rDNA were amplified and sequenced (GenBank Nos. JF795490 and JF795491). The ITS sequence has 98% identity with those of Podosphaera lini, P. macularis, and P. spiraeae, and the D1-D2 sequence of 28S rDNA has 99% identity with those of P. negeri and P. spiraeae, which indicated that this fungus belongs to the genus Podosphaera, even though there is no direct sequence evidence that it is P. euphorbiae-helioscopiae. P. euphorbiae-helioscopiae has only been previously reported in Japan (2), occurring on Euphorbia helioscopia. This species is unique because of the long and curved, thick-walled appendages, and there is no comparable species (1). P. euphorbiae-hirtae and P. euphorbiae are reported to infect Euphorbiaceae plants. Both species differ from P. euphorbiae-helioscopiae in their conspicuously shorter appendages (1,3). To our knowledge, this is the first reported occurrence of powdery mildew on E. pekinensis, and P. euphorbiae-helioscopiae is new to China. This is also the first description of the anamorph of the fungus since there is no record in the Erysiphales Database.

Cross-genera transferability of (simple sequence repeat) SSR markers among cassava (Manihot esculenta Crantz), rubber tree (Hevea brasiliensis Muell. Arg.) and physic nut (Jatropha curcas L.)

Cross-genera transferability of simple sequence repeat (SSR) markers among three economically important plants of family Euphorbiaceae has been proposed. A set of SSR loci generated from cassava (199), rubber tree (49) and physic nut (42) were used to determine transferability with five accessions each of cassava, rubber tree and physic nut. The results revealed that cross-genera transferability among these species was observed. Of the 290 markers, 144 could amplify DNA of at least one nondonor species and 34 markers could amplify DNA of all tested species. A total of 57, 120 and 59 alleles were detected in cassava, rubber tree and physic nut, respectively, by transferable markers. The highest transferability (59.18%) was observed from cassava to rubber tree, followed by from rubber tree to cassava. Low transfer rates were found between cassava and physic nut, and between rubber tree and physic nut. These identified transferable markers for cassava, rubber tree and physic nut (37, 61 and 46, respectively) will be useful for comparative mapping and genomic studies. In addition, this finding is an important initial knowledge on cross-genera transferability of SSR markers in these three commercial species. Key words: Microsatellites, transferability, Euphorbiaceae, cassava, rubber tree, physic nut.

Toxicological assessment of Ricinus communis Linn root extracts

Ricinus communis Linn (Euphorbiaceae) plant parts are claimed to be used as carminative, asthma, bronchitis, leprosy, anti-inflammatory, cathartic, and aphrodisiac. The toxicological study was carried out in the root part of the plant. The collected root was extracted with methanol and water. The extracts were vacuum-dried to yield the respective aqueous (AE) and methanol (ME) extracts. Toxicological assessment sought to determine the safety of Ricinus communis root extracts. The extracts were evaluated in the acute toxicity study (OECD-423 guidelines) and 90 days repeated dose toxicological assessment in Wistar albino rats. The acute oral toxicity of the aqueous (AE) and methanol (ME) extracts did not produce any toxic symptoms or mortality at the dose level of 2000 mg/kg in rats. In the 90 days (sub-chronic toxicity) repeated dose toxicity study the extracts (AE and ME) were administered 1000 mg/kg daily through oral route. The sub-chronic toxicity study demonstrated no significant changes in body weight, food, and water intake. Hematology parameters RBC, WBC, DLC, Hb, blood clotting time, and the biochemical parameters glucose, blood urea nitrogen, creatinine, total cholesterol, total protein, total bilirubin AST, ALT, and ALP were estimated. Histopathology observation of the major vital organs (liver, kidney, heart, spleen, lungs, ovary, testis, and brain) were tested. The hematology, biochemical and histopathology evaluations did not show any adverse effects in any of the organs tested. These results demonstrate the non-toxic nature of the root extracts AE and ME can be used for long-term usage in clinical practice.

New tigliane diterpenes from Euphorbia grandicornis

Phorbol esters are specific acylated polyhydroxy Δ1,6-tigliane diterpenes, which occur in plants of Euphorbiaceae and Thymelaeaceae family. Phorbol derivatives are promising candidates of drug development for HIV therapy, because they reactivate HIV-1 latency by protein kinase C dependent NF-κB activation, and avoid the new infection of CD4+ cells by down-regulating the expression of the HIV 1 receptors [1,2]. Recently we reported the isolation and structure determination of three phorbol analogues from Euphorbia grandicornis Goebel, a succulent cactiform South African plant whose phytochemical investigation has not been reported previously [3]. In continuation of our investigations we report now on the isolation and structural characterization of further six diterpene polyesters. The chloroform-soluble phase of a methanol extract of the fresh aerial parts of E. grandicornis was fractionated by column chromatography on polyamide, then by vacuum liquid chromatography on silica gel. Selected fractions from these separations were further purified by preparative TLC and HPLC to yield six pure compounds, including three new natural products. The structure elucidation was performed by means of HRESIMS and advanced two-dimensional NMR methods, including 1H NMR, JMOD, 1H-1H COSY, NOESY, HSQC, and HMBC experiments. The three known compounds were identified as 12 deoxyphorbol mono- and diesters, acylated with acetic, isobutyric and 2-methylbutyric acids. All of the three new compounds are 12-deoxy-16-hydroxyphorbols; one of them was found to have an unusual parent alcohol with 5-ene-7-ol function.

New promising Euphorbiaceae extracts with activity in human lymphocytes from primary cell cultures

Context: Euphorbiaceae plants exhibit anti-inflammatory and immunomodulatory properties.Methods: We evaluated the activity of 14 extracts from seven Euphorbiaceae plants on primary immune cell cultures from healthy individuals. Peripheral blood mononuclear cells (PBMC) were exposed to the extracts w/o phytohaemagglutinin A or cycloheximide as agents that induce proliferation or apoptosis in PBMC, respectively.Results: We found that five up to 14 Euphorbiaceae’s extracts had the ability to modulate at least one of the immune parameters evaluated in this study. However, only the latex extracts of Euphorbia cotinifolia and Euphorbia tirucalli strongly induced both proliferation and apoptosis in PBMC. These extracts were further subfractioned by silica gel column chromatography. Two subfractions with enhanced activity in comparison to the crude extracts were obtained. Although these subfractions induced proliferation on both CD3+ and CD3− cells, the most prominent effects were observed in the former subpopulation. Interestingly, the subfraction from E. tirucalli induced lymphocyte proliferation without the need of accessory cells; this ability was not inhibited by the carbohydrates d-galactose and α-Methyl-d-Mannopyranoside.Conclusions: Altogether, these results reveal the presence of novel candidates within the Euphorbia plants to induce proliferation and apoptosis in human lymphocytes, mainly in CD3+ T cells.

Cloning of casbene and neocembrene synthases from Euphorbiaceae plants and expression in Saccharomycescerevisiae

A large number of diterpenes have been isolated from Euphorbiaceae plants, many of which are of interest due to toxicity or potential therapeutic activity. Specific Euphorbiaceae diterpenes of medical interest include the latent HIV-1 activator prostratin (and related 12-deoxyphorbol esters), the analgesic resiniferatoxin, and the anticancer drug candidate ingenol 3-angelate. In spite of the large number of diterpenes isolated from these plants and the similarity of their core structures, there is little known about their biosynthetic pathways. Other than the enzymes involved in gibberellin biosynthesis, the only diterpene synthase isolated to date from the Euphorbiaceae has been casbene synthase, responsible for biosynthesis of a macrocyclic diterpene in the castor bean ( Ricinus communis). Here, we have selected five Euphorbiaceae species in which to investigate terpene biosynthesis and report on the distribution of diterpene synthases within this family. We have discovered genes encoding putative casbene synthases in all of our selected Euphorbiaceae species and have demonstrated high-level casbene production through expression of four of these genes in a metabolically engineered strain of Saccharomyces cerevisiae. The only other diterpene synthase found among the five plants was a neocembrene synthase from R. communis (this being the first report of a neocembrene synthase gene). Based on the prevalence of casbene synthases, the lack of other candidates, and the structure of the casbene skeleton, we consider it likely that casbene is the precursor to a large number of Euphorbiaceae diterpenes. Casbene production levels of 31 mg/L were achieved in S. cerevisiae and we discuss strategies to further increase production by maximizing flux through the mevalonate pathway.

Larvicidal activity of some Euphorbiaceae plant extracts against Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae)

Larvicidal activity of ethyl acetate, butanol, and petroleum ether extracts of five species of Euphorbiaceae plants, Jatropha curcas, Pedilanthus tithymaloides, Phyllanthus amarus, Euphorbia hirta, and Euphorbia tirucalli, were tested against the early fourth instar larvae of Aedes aegypti L. and Culex quinquefasciatus (Say). The larval mortality was observed after 24 h of exposure. All extracts showed low larvicidal effects; however, the highest larval mortality was found in petroleum ether extract. The LC50 value of petroleum ether extracts of J. curcas, P. tithymaloides, P. amarus, E. hirta, and E. tirucalli were 8.79, 55.26, 90.92, 272.36, and 4.25 ppm, respectively, against A. aegypti and 11.34, 76.61, 113.40, 424.94, and 5.52 ppm, respectively, against C quinquefasciatus. Of the various ratios tested, the petroleum ether extracts of J. curcas and E. tirucalli were observed to be more efficient than the other plant extracts. It is, therefore, suggested that E. tirucalli can be applied as an ideal potential larvicide against A. aegypti and C. quinquefasciatus. This is an ideal ecofriendly approach for the control of the dengue vector, A. aegypti, and the lymphatic filariasis vector, C. quinquefasciatus.

Toxische Phytodermatitis nach Kontakt mit Euphorbia helioscopia L. (Sonnenwend-Wolfsmilch)

A 6-year-old patient presented with linear erythema and bullae on the face. The lesions developed after playing with plants the day before. The plant was identified as Euphorbia helioscopia L. (sun spurge). The sun spurge belongs to the Euphorbiaceae plant family. These plants produce a typical milky juice that causes toxic reactions following contact with skin and mucous membranes. In the literature several cases of toxic dermatitis caused by plants of the Euphorbiaeae family have been described. The most important differential diagnosis of these skin lesions is the bullous phototoxic dermatitis caused by psoralens. Plant-induced toxic dermatitis is of increasing importance in dermatology. The exact determination of the causative plants is a prerequisite for the diagnosis of phytodermatitis.