Genotoxic and cytotoxic properties of two medical plants (Teucrium arduini L.and Teucrium flavum L.) in relation to their polyphenolic contents.

A large number of species belonging to the genus Teucrium are used in pharmacy and traditional medicine for the treatment of different diseases. This study aimed to evaluate the polyphenolic composition as well as genotoxic and cytotoxic effects of methanolic extracts from T. arduini and T. flavum, two native species found in Montenegro. We determined the total phenolic and flavonoid contents of these plants using spectrophotometric methods; the qualitative content of polyphenolic compounds was investigated by high-performance liquid chromatography (HPLC). Genotoxicity in cultured human lymphocytes was measured in the cytokinesis-block micronucleus assay (CBMN) and comet assay in the range between 125 and 1000 µg/mL. Cytotoxicity was assessed by the MTT viability assay in normal human MRC-5 fibroblasts and MDA-MB-231 breast carcinoma cells. The content of total phenolics and flavonoids in T. arduini extract was higher than in T. flavum (200.35 mg GA/g vs. 171.08 mg GA/g; 96.32 mg RU/g vs. 78.14 mg RU/g). The polyphenolic composition of both extracts was qualitatively similar and eight phenol compounds were identified. The most commonly present phenol was caffeic acid and among four flavonoids, the most common was quercetin. Both plant extracts were genotoxic in both the CBMN and comet assays at concentrations of 250, 500 and 1000 µg/mL. After 72 h of exposure, the extracts of T. arduini and T. flavum were found to induce cytotoxicity in MRC-5 fibroblasts but not in MDA-MB-231 breast cancer cells. The results suggest that the constituents of both plant species are genotoxic and cytotoxic, therefore these extracts warrant additional evaluation to be safely applied in humans.

Evaluation of the cytotoxicity, oral toxicity, genotoxicity, and mutagenicity of the latex extracted from Himatanthus drasticus (Mart.) Plumel (Apocynaceae).

ETHNOPHARMACOLOGICAL RELEVANCE: Himatanthus drasticus is a tree popularly known as janaguba. Endemic to Brazil, it is found in the Cerrado and Caatinga biomes, rock fields, and rainforests. Janaguba latex has been used in folk medicine for its antineoplastic, anti-inflammatory, analgesic, and antiallergic activities. However, studies investigating the safety of its use for medicinal purposes are limited. AIM OF THE STUDY: This study aimed to evaluate the toxicity of the latex extracted from H. drasticus. MATERIALS AND METHODS: The latex was extracted from H. drasticus specimens by removing a small area of bark (5 × 30 cm) and then dissolving the exudate in water and lyophilizing it. Phytochemical screening was performed by TLC and GC-MS, protein, and carbohydrate levels. Cell viability was performed by the MTT method. Acute oral toxicity, genotoxicity, and mutagenicity assays were performed in mice. RESULTS: TLC showed the presence of saponins and reducing sugars, as well as steroids and terpenes. The GC-MS analysis of the nonpolar fraction identified lupeol acetate, betulin, and α/ß-amyrin derivatives as the major compounds. The latex was toxic to S-180 cells at 50 and 100 µg/mL. No signals of toxicity or mutagenicity was found in mice treated with 2000 mg/kg of the latex, but genotoxicity was observed in the Comet assay. CONCLUSIONS: H. drasticus latex showed toxicity signals at high doses (2000 mg/kg). Although the latex was not mutagenic to mice, it was genotoxic in the Comet assay in our experimental conditions. Even testing a limit dose of 2000 mg/kg, which is between 10 to 35-fold the amount used in folk medicine, caution must be taken since there is no safe level for genotoxic compounds exposure. Further studies on the toxicological aspects of H. drasticus latex are necessary to elucidate its possible mechanisms of genotoxicity.

Preclinical safety evaluation of the ethanolic extract from Campomanesia pubescens (Mart. ex DC.) O.BERG (guavira) fruits: analysis of genotoxicity and clastogenic effects.

Genotoxicity studies of plants with medicinal and nutritional properties are recommended by international regulatory agencies as part of the risk assessment. Due to their consumption as food, nutraceutical use and ethnopharmacological relevance, Campomanesia pubescens represents one of these plants to be studied. The aim of the present study was to evaluate the genotoxic, cytotoxic potential and clathogenic effects of the ethanolic extract obtained from the pulp of C. pubescens (EEFCP) fruits on rats submitted to experimental genotoxicity models and through the SMART test performed in Drosophila melanogaster. The comet assay and the micronucleus test were performed on peripheral and bone marrow blood, respectively, of Wistar rats orally treated with EEFCP at doses of 125, 250, 500 and 1000 mg per kg per bw for 28 days. In the SMART test, the standard cross between three mutant D. melanogaster strains was used. Larvae were treated with EEFCP at different concentrations and the wings of adult flies were evaluated for the presence/frequency of mutant spots and compared to the negative control group. Phytochemical analysis of EEFCP indicated high levels of flavonoids. The tests performed in rats showed that EEFCP did not present significant genotoxic or clastogenic effects. The biotransformation metabolites of EEFCP did not present genotoxic activity, as demonstrated by the SMART test. Together, all results indicate that, under the experimental conditions used, EEFCP did not reveal any preclinical genetic toxicity. Therefore, the safe consumption can be fomented increasing, consequently, the economic liquidity in the industrial market from the fruits of guavira.

The mutagenic and antimutagenic activity of Sutherlandia frutescens extracts and marker compounds.

BACKGROUND: Sutherlandia frutescens (L.) R. Br is endemic to Southern Africa where it has been traditionally used for cancer and diabetes. In recent times it has been marketed for its reputed (but not proven) anticancer, antidiabetic and anti-HIV properties. Little is known about the mutagenic and antimutagenic potential of extracts and common marker compounds of Sutherlandia frutescens. Therefore this study aimed to investigate the putative efficacy and possible long-term adverse effects of using this herb. METHODS: Ethylacetate (EA) and 50% Methanol (MeOH) extracts were screened for mutagenic and antimutagenic activity using the Ames assay utilising TA97a, TA98, TA100 and TA102 in the presence and absence of metabolic activation. Four compounds, L-arginine, L-canavanine, GABA and D-pinitol known to occur in sutherlandia were also included. The total polyphenolic content of the both extracts was determined using the Folin-Ciocalteau method and FRAP and ABTS were used to determine the anti-oxidant potential of the extracts. RESULTS: The extracts and the standards did not show any cytotoxicity except in TA97a. The EA extract exhibited antimutagenicity against all the bacterial strains at all concentrations tested. The MeOH extract showed both pro-mutagenic and antimutagenic activities with 2-acetamidofluorene and aflatoxin B1 in the presence of metabolic activation of TA98 and TA100, respectively. All compounds, except L-canavanine exhibited antimutagenic activity against all strains. L-canavanine, on the other hand showed co-mutagenicity with 9-aminoacridine on TA97a, at all test concentrations. The extracts and pure compounds exhibited their antimutagenic activity in a dose response manner. L-arginine and GABA showed an some antimutagenic response. EA extract had three times the total phenolic content (12.56 µg GE / mg) observed in the MeOH extract. There was correlation between total phenolic content, antioxidant potential and antimutagenicity. CONCLUSION: Both extracts exhibited a protective effect, with the EA extract exhibiting greater potency. L-canavanine acted as a co-mutagen in a dose response manner without metabolic activation. It is suggested that the EA extract be priotized for future development work as it showed a better risk profile and activity.

Mutagenicity evaluation of Anastatica hierochuntica L. aqueous extract in vitro and in vivo.

Anastatica hierochuntica L. ( A. hierochuntica), a folk medicinal plant, was evaluated for mutagenic potential via in vitro and in vivo assays. The in vitro assay was conducted according to modified Ames test, while the in vivo study was performed according to Organisation for Economic Co-operation and Development guideline for mammalian erythrocyte micronucleus assay. Four groups ( n= 5 males and 5 females per group) Sprague Dawley rats were randomly chosen as the negative control, positive control (received a single intramuscular injection of cyclophosphamide 50 mg/kg), 1000 and, 2000 mg/kg A. hierochuntica aqueous extracts. All groups except the positive control were treated orally for three days. Findings of the in vitro assay showed mutagenic potential of AHAE at 0.04 and 0.2 mg/ml. However, no mutagenic effect was demonstrated in the in vivo study up to 2000 mg/kg. No significant reduction in the polychromatic and normochromatic erythrocytes ratio was noted in any of the groups. Meanwhile, high micronucleated polychromatic erythrocytes frequency was seen in cyclophosphamide-treated group only. These findings could perhaps be due to insufficient dosage of A. hierochuntica aqueous extracts to cause genetic damage on the bone marrow target cells. Further acute and chronic in vivo toxicity studies may be required to draw pertinent conclusion on the safety aspect of A. hierochuntica aqueous extracts consumption. Impact statement In this paper, we report on the mutagenicity evaluation of Anastatica hierochuntica aqueous extract. This is a significant research in view of the popularity of this herb consumption by the people across the globe despite of limited scientific evidence on its toxicity potential. This study is intended to encourage more extensive related research in order to provide sufficient evidence and guidance for determining its safe dosage.

Genotoxic and cytotoxic potential of Alternanthera Bettzickiana, an important ethno-medicinal plant.

The present study was carried out to investigate the mutagenic and cytotoxic potential of n-hexane and aqueous-methanolic whole plant extracts of Alternanthera bettzickiana. Aqueous-methanolic and n-hexane extracts of Alternanthera bettzickiana extracts were assessed for the mutagenic potential with Salmonella tester strains TA-100 and TA-102 in the presence and absence of the rodent enzyme activation system and cytotoxic potential was assessed by MTT assay. Aqueous-methanolic extract showed the presence of saponins, tannins, terpenoids, flavonoids and glycosides. However n-hexane extract revealed the presence of tannins and terpenoids only. It was found that a concentration as low as 15mg/mL of both extracts was more mutagenic to the TA 102 tester strain than TA-100. Hexane whole plant extract of Altenanthera bettzickiana was more mutagenic than aqueous-methanolic extract considering revertant colonies of TA 100 strain. Aqueous-methanolic and n-hexane whole plant extracts of Altenanthera bettzickiana showed higher mutagenic potential in the presence of the enzyme activation system. Mutagenicity of aqueous-methanolic extract increased with an enzyme activation system in case of TA 100 whereas mutagenicity of n-hexane extract decreased in the presence of the enzyme activation system with TA 100 and TA 102 strains. Aqueous-methanolic and n-Hexane whole plant extracts of Alternanthera bettzickiana showed an IC-50 of 493 and 456 µg/mL in BHK-21 cells respectively. It can be concluded that Altenanthera bettzickiana exhibited mutagenic activity in a bacterial reverse mutation assay with and without enzyme activation systems. However, it showed limited cytotoxicity to BHK-21 cells.

Mutagenicity and oxidative damage induced by an organic extract of the particulate emissions from a simulation of the deepwater horizon surface oil burns.

Emissions from oil fires associated with the "Deepwater Horizon" explosion and oil discharge that began on April 20, 2010 in the Gulf of Mexico were analyzed chemically to only a limited extent at the time but were shown to induce oxidative damage in vitro and in mice. To extend this work, we burned oil floating on sea water and performed extensive chemical analyses of the emissions (Gullett et al., Marine Pollut Bull, in press, ). Here, we examine the ability of a dichloromethane extract of the particulate material with an aerodynamic size ≤ 2.5 µm (PM2.5 ) from those emissions to induce oxidative damage in human lung cells in vitro and mutagenicity in 6 strains of Salmonella. The extract had a percentage of extractable organic material (EOM) of 7.0% and increased expression of the heme oxygenase (HMOX1) gene in BEAS-2B cells after exposure for 4 hr at 20 µg of EOM/ml. However, the extract did not alter mitochondrial respiration rate as measured by extracellular flux analysis. The extract was most mutagenic in TA100 +S9, indicative of a role for polycyclic aromatic hydrocarbons (PAHs), reflective of the high concentrations of PAHs in the emissions (1 g/kg of oil consumed). The extract had a mutagenicity emission factor of 1.8 ± 0.1 × 105 revertants/megajoulethermal in TA98 +S9, which was greater than that of diesel exhaust and within an order of magnitude of open burning of wood and plastic. Thus, organics from PM2.5 of burning oil can induce oxidative responses in human airway epithelial cells and are highly mutagenic. Environ. Mol. Mutagen. 58:162-171, 2017. © 2017 Wiley Periodicals, Inc.

Antioxidant and Genotoxic Properties of Hispidin Isolated from the Velvet-Top Mushroom, Phaeolus schweinitzii (Agaricomycetes).

Antioxidant and genotoxic properties of hispidin isolated from the Phaeolus schweinitzii mushroom were evaluated with various assays. Hispidin demonstrated strong free radical scavenging, oxygen radical absorbance capacity, and ferric-reducing antioxidant power; in all applied assays, hispidin exhibited antioxidant capacity similar to or higher than that of the reference antioxidant Trolox. Genotoxic activity of hispidin was assessed using different end points: chromosome aberrations, micronuclei, sister chromatid exchanges, and primary DNA damage (detected by the comet assay) in human lymphocytes in vitro, and gene mutations in the Salmonella/microsome test. Hispidin did not increase the frequency of chromosome aberrations, micronuclei, or primary DNA damage in human lymphocytes in vitro and did not produce reverse mutation in bacterial cells. However, we identified in human lymphocytes a statistically significant dose-dependent increase in sister chromatid exchange frequency and a decrease in replication index and nuclear division index values.

The mutagenic, antimutagenic and antioxidant properties of Hypericum lydium.

CONTEXT: There is a growing market demand for Hypericum sp., a pharmacologically active plant that has been traditionally used to treat various ailments. However, there have been limited studies on the extract or essential oil of Hypericum lydium Boiss (Hypericaceae). OBJECTIVE: This study investigates for the first time the antioxidant, mutagenic and antimutagenic activity of an ethanol extract of H. lydium. MATERIAL AND METHODS: Ethanol extract from aerial parts of H. lydium harvested from Turkey were tested for this mutagenic and antimutagenic activities (2.0-0.002 mg/plate) using Ames Salmonella/microsome test system. 4-Nitro-o-phenylenediamine (4-NPD) (3 µg/plate) for the Salmonella typhimurium TA98 and sodium azide (NaN3) (8 µg/plate) for the S. typhimurium TA100 were used as positive controls. The antioxidant activity, total antioxidant activity and phenolic constituent of the extract (2.0-0.002 mg/mL) was determined by the inhibition of 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), ß-carotene-linoleic acid model and by means of Folin-Ciocalteu reagent, respectively. RESULTS: The extract showed no sign of mutagenicity at the tested concentrations (0.002-2.0 mg/mL), and showed concentration-dependent antimutagenic activity against NaN3 and 4-NPD ranging from 26.8 to 81.5%. The extract was found to be an efficient scavenger of DPPH (IC50 0.165 ± 0.23 mg/mL) and to inhibit ß-carotene-linoleic acid bleaching (IC50 0.39 ± 0.11 mg/mL). DISCUSSION AND CONCLUSION: These findings indicate ethanol extract of H. lydium to be a safe and effective agent that may be incorporated into new strategies for the prevention of cancer and mutagenesis.

Assessment in vitro of the genotoxicity, antigenotoxicity and antioxidant of Ceratonia siliqua L. extracts in murine leukaemia cells L1210 by comet assay.

Genotoxicity of Ceratonia siliqua extracts, was investigated by assessing their capacity to induce nucleus DNA degradation of murine leukaemia cells L1210, using the "Comet assay". The ability of total oligomer flavonoids (TOF) and aqueous extracts to protect cell DNA against oxidative stress induced by H2O2, was performed by pre- co or post-treatment of cells with the before mentioned extracts for different periods preceding exposure to H2O2 stress. No significant genotoxic effect was detected at different exposure times, except at the lowest concentration of TOF extract (16.25 µg/ml). It appears that extracts decreased DNA damage, induced by H2O2. Both of TOF and aqueous extracts exhibited cellular antioxidant capacity, with EC50 values of respectively <16.25 and < 35 µg/ml, as well as, a protective capacity against lipidperoxidation inducing using L1210 cells line as a cellular model. MDA inhibition percentages reached 88.43% and 90.52% with respectively 35.5 µg/ml of TOF extract and 70 µg/ml of aqueous extract. Antioxidant properties of carob leaf extracts revealed by our study make a good antioxidant protection and thus a good candidate as food addition component.

Cytotoxic and genotoxic effects of ethanolic extract of Euphorbia hyssopifolia L. on HepG2 cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Euphorbia hyssopifolia L. is a weed with recognized antimicrobial potential employed in Indian, Asian and Latin-American popular medicine. However, little is known with regard to its toxic potential. The present study aimed to investigate the cytotoxic and genotoxic effects of ethanolic extract of E. hyssopifolia in HepG2 cell culture. MATERIALS AND METHODS: Phytochemical screening of ethanolic extract was carried out to determine the presence of active secondary plant metabolites. Six concentrations (0.00001, 0.0001, 0.001, 0.01, 0.1 and 1.0mg/mL) of ethanolic extract were tested by the MTT assay to verify cytotoxicity. Then, genotoxic evaluations (alkaline comet assay and cytokinesis-block micronucleus assay - CBMN) were carried out in HepG2 cells with extract concentrations of 0.01, 0.1 and 1.0mg/mL. RESULTS: Mono and sesquiterpenes, triterpenes and steroids, and flavonoids were the main classes found in the phytochemical screening. Extract concentrations used in the MTT assay showed no cytotoxic activity. On the other hand, genotoxic activity was verified at 0.1 and 1.0mg/mL in the alkaline comet assay. Additionally, the 1.0mg/mL concentration induced severe cell damage leading to death in the CBMN assay, indicating a cytotoxic effect for this concentration in the latter method. CONCLUSION: The use of E. hyssopifolia extract for medicinal purposes should be avoided, because concentrations above 0.01mg/mL may pose risk to human health due to cytotoxic and/or genotoxic effects.

Investigations into the genotoxic potential of olive extracts.

The phenolic anti-oxidant 3-hydroxytyrosol (HT) is a major constituent of olives and olive oil. Published data showed it was negative in the Ames test at concentrations up to 5 µL per plate, but did induce chromosomal aberrations in human lymphocytes. HIDROX, an olive extract containing approximately 2.4% HT, was reported as both positive and equivocal in an Ames test in different papers from the same laboratory. Negative results for micronucleus induction in vivo in both an acute study and as part of a 90-day rat toxicity study were also reported for HIDROX. Given the widespread use and consumption of olives, olive oil and olive extracts, it was important to obtain more data. Here we confirm that pure HT, and an olive extract containing 15% HT, both induced micronuclei in cultured cells in vitro, but show that these responses were either due to high levels of cytotoxicity or to reaction of HT with culture medium components to produce hydrogen peroxide. Another extract (H40) containing 40% HT also induced micronuclei in vitro, probably via the same mechanism. However, both extracts were negative in robust Ames tests. The 15% HT formulated extract did not induce micronuclei in rat bone marrow after 4 weeks of dosing up to 561 mg HT/kg/day. H40 produced increased rat bone marrow micronucleus frequencies at 250 and 500 mg HT/kg/day in a 90-day toxicity study, but the results were questionable for various reasons. However, when two different batches of this extract were tested in acute micronucleus studies at doses up to 2000 mg HT/kg, giving plasma exposures that exceeded those in the 90-day study, negative results were obtained. Based on weight of evidence it is concluded that the olive extracts tested are not genotoxic at high doses in vivo, and any genotoxic risks for human consumers are negligible.

Genotoxic and biochemical changes in Baccharis trimera induced by coal contamination.

The processing and combustion of coal in thermal power plants release anthropogenic chemicals into the environment. Baccharis trimera is a common plant used in folk medicine that grows readily in soils degraded by coal mining activities. This shrub bioaccumulates metals released into the environment, and thus its consumption may be harmful to health. The purpose of this study was to investigate the phytochemical profile, antioxidant capacity (DPPH), genotoxic (comet assay) and mutagenic potential (CBMN-cyt) in V79 cells of B. trimera aqueous extracts in the coal-mining region of Candiota (Bt-AEC), and in Bagé, a city that does not experience the effects of exposure to coal (Bt-AEB, a reference site). In the comet assay, only Bt-AEC was genotoxic at the highest doses (0.8mg/mL and 1.6mg/mL), compared to the control. For extracts from both areas, mutagenic effects were observed at higher concentrations compared to the control. The cell damage parameters were significantly high in both extracts; however, more striking values were observed for Bt-AEC, up to the dose of 0.8mg/mL. In chemical analysis, no variation was observed in the contents of flavonoids and phenolic compounds, neither the antioxidant activity, which may suggest that DNA damage observed in V79 cells was induced by the presence of coal contaminants absorbed by the plant.

Investigation on the toxic potential of Tribulus terrestris in vitro.

CONTEXT: Tribulus terrestris L. (Zygophyllaceae) has been commonly used to energize, vitalize, and improve sexual function and physical performance in men. OBJECTIVE: This study investigates the potential cytotoxic and genotoxic, and endocrine disrupting activities of T. terrestris in vitro. MATERIALS AND METHODS: The whole T. terrestris plant was extracted with water, methanol, and chloroform. The genotoxic potential of T. terrestris extracts at 3-2400 µg/mL was assessed by Comet assay in a rat kidney cell line (NRK-52E) and by Ames assay in Salmonella typhimurium TA98 and TA100 strains. Endocrine disrupting effects of the extracts at concentrations of 0.22-25 000 µg/mL were assessed by YES/YAS assay in Saccharomyces cerevisiae. Cytotoxic activity of the extracts was determined by the MTT test in NRK-52E cells. The different exposure times were used for four tests (3-48 h). RESULTS: The methanol extract of T. terrestris IC50 value was 160 µg/mL. The other extracts did not show cytotoxic effects. In the Comet and Ames genotoxicity assays, none of the extracts possessed genotoxic activities at concentrations of 0-2400 µg/mL. Only the water extract of T. terrestris induced frame shift mutations after metabolic activation. The water extract also showed estrogenic activity by YES/YAS assay in S. cerevisiae at concentrations ≥27 µg/mL (≥2.6-fold), while the other T. terrestris extracts had anti-estrogenic properties. CONCLUSION: Tribulus terrestris had estrogenic and genotoxic activities. The study was useful in determining its toxicological effects and the precautions regarding consumption.

Antifungal activity against Cryptococcus neoformans strains and genotoxicity assessment in human leukocyte cells of Euphorbia tirucalli L.

In the last times, focus on plant research has increased all over the world. Euphorbia tirucalli L., a plant known popularly as Aveloz, and originally used in Africa, has been drawing attention for its use in the United States and Latin America, both for use as an ornamental plant and as a medicinal plant. E. tirucalli L. is a member of the family Euphorbiaceae and contains many diterpenoids and triterpenoids, in particular phorbol esters, apparently the main constituent of this plant, which are assumed to be responsible for their activities in vivo and in vitro. The in vitro antifungal activities of Euphorbia tirucalli (L.) against opportunistic yeasts were studied using microbroth dilution assay. The results showed that aqueous extract and latex preparation were effective against ten clinical strains of Cryptococcus neoformans in vitro (Latex and extract MIC range of 3.2 - > 411 µg/mL). Aiming the safe use in humans, the genotoxic effects of E. tirucalli were evaluated in human leukocytes cells. Our data show that both aqueous extract and latex preparation have no genotoxic effect in human leukocytes cells in vitro. Although the results cannot be extrapolated by itself for use in vivo, they suggest a good perspective for a therapeutic application in future. In conclusion, our results show that the aqueous extract and latex preparation from E. tirucalli L. are antifungal agents effectives against several strains of C. neoformans and do not provoke DNA damage in human leukocyte cells, considering the concentrations tested.

Preparation of nano/submicrometer Ganoderma tsugae and its mutagenic potencies and cytotoxicity.

This study explored the feasibility of preparing nano/submicrometer particles from Ganoderma tsugae to enhance the contents of bioactive compounds and to assess its mutagenic potencies and cytotoxicity. Hot-water extract, a common product, was employed as a reference. After 3 h of media milling, almost all of the particles were smaller than 1 µm with a number-mean diameter of 0.11 µm. There were about 62% particles smaller than 0.1 µm in terms of number of particles. Scanning electron microscopy (SEM) confirmed the presence of particles at nano/submicrometer scale. The content of 1→3-ß-D-glucan in nano/submicrometer G. tsugae was 3.5 times of that in hot-water extract. Both nano/submicrometer and hot-water extract G. tsugae exhibited no mutagenic potential to Salmonella Typhimurium tester strains. Cell toxicity test also confirmed the safety of both nano/submicrometer and hot-water extract G. tsugae. The effect of media milling on the structural change of hyphae was also discussed.

Antifungal activity against Cryptococcus neoformans strains and genotoxicity assessment in human leukocyte cells of Euphorbia tirucalli L.

In the last times, focus on plant research has increased all over the world. Euphorbia tirucalli L., a plant known popularly as Aveloz, and originally used in Africa, has been drawing attention for its use in the United States and Latin America, both for use as an ornamental plant and as a medicinal plant. E. tirucalli L. is a member of the family Euphorbiaceae and contains many diterpenoids and triterpenoids, in particular phorbol esters, apparently the main constituent of this plant, which are assumed to be responsible for their activities in vivo and in vitro. The in vitro antifungal activities of Euphorbia tirucalli (L.) against opportunistic yeasts were studied using microbroth dilution assay. The results showed that aqueous extract and latex preparation were effective against ten clinical strains of Cryptococcus neoformans in vitro (Latex and extract MIC range of 3.2 - > 411 µg/mL). Aiming the safe use in humans, the genotoxic effects of E. tirucalli were evaluated in human leukocytes cells. Our data show that both aqueous extract and latex preparation have no genotoxic effect in human leukocytes cells in vitro. Although the results cannot be extrapolated by itself for use in vivo, they suggest a good perspective for a therapeutic application in future. In conclusion, our results show that the aqueous extract and latex preparation from E. tirucalli L. are antifungal agents effectives against several strains of C. neoformans and do not provoke DNA damage in human leukocyte cells, considering the concentrations tested.

Isolation of acetylated bile acids from the sponge Siphonochalina fortis and DNA damage evaluation by the comet assay.

From the organic extracts of the sponge Siphonochalina fortis, collected at Bahía Bustamante, Chubut, Argentina, three major compounds were isolated and identified as deoxycholic acid 3, 12-diacetate (1), cholic acid 3, 7, 12-triacetate (2) and cholic acid, 3, 7, 12-triacetate. (3). This is the first report of acetylated bile acids in sponges and the first isolation of compound 3 as a natural product. The potential induction of DNA lesions by the isolated compounds was investigated using the comet assay in lymphocytes of human peripheral blood as in vitro model. The results showed that the administration of the bile acid derivatives would not induce DNA damages, indicating that acetylated bile acids are nontoxic metabolites at the tested concentrations. Since the free bile acids were not detected, it is unlikely that the acetylated compounds may be part of the sponge cells detoxification mechanisms. These results may suggest a possible role of acetylated bile acids as a chemical defense mechanism, product of a symbiotic relationship with microorganisms, which would explain their seasonal and geographical variation, and their influence on the previously observed genotoxicity of the organic extract of S. fortis.

Insecticidal and genotoxic activity of Psoralea corylifolia Linn. (Fabaceae) against Culex quinquefasciatus Say, 1823.

BACKGROUND: Indiscriminate use of synthetic insecticides to eradicate mosquitoes has caused physiological resistance. Plants provide a reservoir of biochemical compounds; among these compounds some have inhibitory effect on mosquitoes. In the present study the larvicidal, adulticidal and genotoxic activity of essential oil of Psoralea corylifolia Linn. against Culex quinquefasciatus Say was explored. METHODS: Essential oil was isolated from the seeds of P. corylifolia Linn. Larvicidal and adulticidal bioassay of Cx. quinquefasciatus was carried out by WHO method. Genotoxic activity of samples was determined by comet assay. Identification of different compounds was carried out by gas chromatography- mass spectrometry analysis. RESULTS: LC50 and LC90 values of essential oil were 63.38±6.30 and 99.02±16.63 ppm, respectively against Cx. quinquefasciatus larvae. The LD50 and LD90 values were 0.057±0.007 and 0.109±0.014 mg/cm2 respectively against adult Cx. quinquefasciatus,. Genotoxicity of adults was determined at 0.034 and 0.069 mg/cm2. The mean comet tail length was 6.2548±0.754 µm and 8.47±0.931 µm and the respective DNA damage was significant i.e. 6.713% and 8.864% in comparison to controls. GCMS analysis of essential oil revealed 20 compounds. The major eight compounds were caryophyllene oxide (40.79%), phenol,4-(3,7-dimethyl-3-ethenylocta-1,6-dienyl) (20.78%), caryophyllene (17.84%), α-humulene (2.15%), (+)- aromadendrene (1.57%), naphthalene, 1,2,3,4-tetra hydro-1,6-dimethyle-4-(1-methyl)-, (1S-cis) (1.53%), trans- caryophyllene (0.75%), and methyl hexadecanoate (0.67%). CONCLUSION: Essential oil obtained from the seeds of P. corylifolia showed potent toxicity against larvae and adult Cx. quinquefasciatus. The present work revealed that the essential oil of P. corylifolia could be used as environmentally sound larvicidal and adulticidal agent for mosquito control.

Isolation of some luteolin derivatives from Mentha longifolia (L.) Hudson subsp. longifolia and determination of their genotoxic potencies.

This study was designed to evaluate the mutagenic and antimutagenic activities of luteolin derivatives (luteolin 7-O-glucoside, luteolin 7-O-rutinoside and luteolin 7-O-glucuronide) isolated from Mentha longifolia (L.) Huds. subsp. longifolia by using Ames Salmonella test (TA 1535 and TA1537 strains). In the antimutagenicity assays, luteolin 7-O-glucoside, luteolin 7-O-rutinoside and luteolin 7-O-glucuronide showed antimutagenic effects on TA1537 and TA1535 strains. The highest inhibition rates for luteolin 7-O-glucoside, luteolin 7-O-rutinoside and luteolin 7-O-glucuronide on TA1537 strain were 84.03%, 87.63% and 67.77%, respectively. Similarly, in the antimutagenicity assays performed with the TA1535 strain, the inhibition rates for luteolin 7-O-glucoside and luteolin 7-O-rutinoside were 23.86% and 23.76% respectively. Our findings showed that the antimutagenic properties of luteolin derivatives on TA1537 and TA1535 strains have been found to be structure dependent. The clarification of differences in antimutagenic potency of these luteolin derivatives based on their structures has been demonstrated in this study.