Targeting Proteolysis with Cyanogenic Glycoside Amygdalin Induces Apoptosis in Breast Cancer Cells.

BACKGROUND: Breast cancer is the most diagnosed cancer among women, and its incidence and mortality are rapidly growing worldwide. In this regard, plant-derived natural compounds have been shown to be effective as chemotherapeutic and preventative agents. Apricot kernels are a rich source of nutrients including proteins, lipids, fibers, and phenolic compounds and contain the aromatic cyanogenic glycoside amygdalin that has been shown to exert a cytotoxic effect on cancer cells by affecting the cell cycle, inducing apoptosis, and regulating the immune function. METHODS: Here, we describe a previously unexplored proapoptotic mechanism of action of amygdalin in breast cancer (MCF7) cells that involves the modulation of intracellular proteolysis. For comparative purposes, the same investigations were also conducted upon cell treatment with two apricot kernel aqueous extracts from Prunus armeniaca L. RESULTS: We observed that both the 20S and 26S proteasome activities were downregulated in the MCF7 cells upon 24 h treatments. Simultaneously, the autophagy cascade resulted in being impaired due to cathepsin B and L inhibition that also contributed to a reduction in cancer cell migration. The inhibition of these proteolytic systems finally promoted the activation of apoptotic events in the MCF7 cells. CONCLUSION: Collectively, our data unveil a novel mechanism of the anticancer activity of amygdalin, prompting further investigations for potential application in cancer preventative strategies.

In situ genotoxicity assessment in freshwater zooplankton and sediments from different dams, ponds, and temporary rivers in Tunisia.

Tunisia water resources are limited. The country currently has 29 large dams, more than 1000 hill lakes, and 220 small dams which are essential for economic and social development given their contribution to irrigation, drinking water consumption, flooding protection, production of electrical energy, groundwater recharge, and industrial uses. Given the scarcity of these resources, it is crucial to be able to ensure the quality of freshwater environments, particularly those intended for human consumption. In this study, we meant to assess the health status of various freshwater ecosystems in different regions of Tunisia (north and center west) in order to detect genotoxic components in sediments and their potential effect on zooplankton (cladocerans). Sediment and cladoceran species were collected from dams, ponds, and temporary rivers in Tunisia. For each collection site, micronucleus (MN) assay was performed, in triplicates, using a pool of ten specimens of the same cladoceran species. MN occurrence in cladocerans varied from one site to another and MN frequencies varied between 0.67 and 22‰, suggesting the presence of genotoxic substances in certain sites. Sediment genotoxicity and mutagenicity were assessed using the SOS Chromotest and the Ames test. Sediment results showed that genotoxicity varies from one site to another displaying a quantitative and a qualitative variation of pollutant among the sites. These results suggest an urgent need for continuous monitoring of freshwater environments in Tunisia, particularly those intended for drinking water.

Effect of heated naringenin on immunomodulatory properties and cellular antioxidant activity.

Naringenin is one of the most popular flavonoids derived from citrus. It has been reported to be an effective anti-inflammatory compound. Citrus fruit may be used raw, cooked, stewed, or boiled. The present study was conducted to investigate the effect of thermal processes on naringenin in its immunomodulatory and cellular antioxidant activities. The effects of flavonoids on B and T cell proliferation were assessed on splenocytes stimulated or not with mitogens. However, their effects on cytotoxic T lymphocyte (CTL) and natural killer (NK) activities were assessed in splenocytes co-incubated with target cells. The amount of nitric oxide production and the lysosomal enzyme activity were evaluated in vitro on mouse peritoneal macrophages. Cellular antioxidant activity in splenocytes and macrophages was determined by measuring the fluorescence of the dichlorofluorescin (DCF). Our findings revealed that naringenin induces B cell proliferation and enhances NK activity. The highest concentration of native naringenin exhibits a significant proliferation of T cells, induces CTL activity, and inhibits cellular oxidation in macrophages. Conversely, it was observed that when heat-processed, naringenin improves the cellular antioxidant activity in splenocytes, increases the cytotoxic activity of NK cells, and suppresses the cytotoxicity of T cells. However, heat treatment maintains the anti-inflammatory potency of naringenin.

Evaluation of antioxidant and antigenotoxic activity of two flavonoids from Rhamnus alaternus L. (Rhamnaceae): kaempferol 3-O-ß-isorhamninoside and rhamnocitrin 3-O-ß-isorhamninoside.

The antioxidant activity of kaempferol 3-O-ß-isorhamninoside (K3O-ir) and rhamnocitrin 3-O-ß-isorhamninoside (R3O-ir), isolated from the leaves of Rhamnus alaternus L., was determined by the ability of each compound to inhibit NBT photoreduction and to scavenge the free radical ABTS(+)(.). Genotoxic and antigenotoxic activities were assessed using the SOS chromotest. At a concentration of 150 µg/assay the two compounds showed the most potent inhibitory activity against superoxide anion by respectively 80.4% and 85.6%. K3O-ir was a very potent radical scavenger with an IC(50) value of 18.75 µg/ml. Moreover, these two compounds exhibit an inhibitory activity against genotoxicity induced by nitrofurantoine and aflatoxine B1 using the SOS chromotest bacterial assay system in the presence of Escherichia coli PQ37 strain. In this study, we have also evaluated correlation between antigenotoxic and antioxidant effects of K3O-ir and R3O-ir. The highest correlation was showed with R3O-ir (r=0.999).

Antimutagenic and antioxidant potentials of Teucrium ramosissimum essential oil.

The mutagenic and antimutagenic effects of the essential oil extracted from the aerial parts of Teucrium ramosissimum were evaluated by the bacterial reverse mutation assay in Salmonella typhimurium TA98, TA100, and TA1535, with and without exogenous metabolic activation (S9 fraction). The T. ramosissimum essential oil showed no mutagenic effect. In contrast, our results established that it possessed antimutagenic effects against sodium azide (SA), aflatoxin B1 (AFB1), benzo[a]pyrene (B[a]P), and 4-nitro-o-phenylenediamine (NOPD). The antioxidant capacity of the tested essential oil was evaluated using enzymatic, i.e., the xanthine/xanthine oxidase (X/XOD) assay, and nonenzymatic systems, i.e., the nitro-blue tetrazolium (NBT)/riboflavin and the DPPH assays. A moderate free radical-scavenging activity was observed towards DPPH(.) and O2(.-). In contrast, T. ramosissimum essential oil showed no effect for all the tested concentrations in the X/XOD assay.

Phlomis mauritanica extracts reduce the xanthine oxidase activity, scavenge the superoxide anions, and inhibit the aflatoxin B1-, sodium azide-, and 4-nitrophenyldiamine-induced mutagenicity in bacteria.

Four extracts were prepared from the leaves of Phlomis mauritanica: lyophilized infusion, total oligomer flavonoids, methanol, and ethyl acetate extracts. The antimutagenic properties of these extracts were investigated by assessing the inhibition of the mutagenic effects of direct-acting mutagens such as sodium azide and 4-nitrophenylenediamine and indirect-acting mutagens like aflatoxin B1 (AFB1) using the Ames assay. The four extracts prepared from P. mauritanica strongly inhibit the mutagenicity induced by AFB1 in both Salmonella typhimurium TA 100 and TA 98 assay systems. Lyophilized infusion and methanol extracts at the dose of 250 microg per plate reduced AFB1 mutagenicity by 93% and 91%, respectively, in S. typhymurium strain TA 100. We examined also the antioxidant effect of these extracts by the enzymatic xanthine/xanthine oxidase assay. Result indicated that total oligomer flavonoids and ethyl acetate and methanol extracts were potent inhibitors of xanthine oxidase activity. In contrast, lyophilized infusion, total oligomer flavonoids, and methanol extracts exhibited a high degree of superoxide anion scavenging. Our findings emphasize the potential of P. mauritanica extracts to prevent mutations and oxidant effects. Furthermore, the results presented here could be an additional argument to support the use of this species as a medicinal and dietary plant.

Study of genotoxic, antigenotoxic and antioxidant activities of the digallic acid isolated from Pistacia lentiscus fruits.

The digallic acid obtained from the fruit Pistacia lentiscus exhibits an inhibitory activity against nitrofurantoine and B[a]P induced genotoxicity when tested by the SOS chromotest bacterial assay system in the presence of Escherichia coli PQ37 strain. The antioxidant activity of the tested compound was determined by its ability to scavenge the free radical ABTS(+), to inhibit the xanthine oxidase, involved in the generation of free radicals, and to inhibit the lipid peroxidation induced by H(2)O(2) in the K562 cell line. Our results revealed that digallic acid shows an important free radical scavenging activity towards the ABTS(+) radical (99%) and protection against lipid peroxidation (68%).

Leaf extracts from Moricandia arvensis promote antiproliferation of human cancer cells, induce apoptosis, and enhance antioxidant activity.

The in vitro antiproliferative, apoptotic, and antioxidant activities from leaf extracts of Moricandia arvensis, which are used in traditional cooking and medicines, were investigated. The MTT assay revealed that only TOF (total oligomer flavonoids), ethyl acetate (EA), chloroform (Chl), and petroleum ether (PE) extracts inhibited the proliferation of K562 cells. Apoptosis plays a very important role in the treatment of cancer by promoting the apoptosis of cancer cells and limiting the concurrent death of normal cells. Thus, the possible effects of M. arvensis extracts on the induction of apoptosis in human leukemic cells (K562 cells) were investigated. The electrophoretic analysis of DNA fragmentation confirms that TOF, Chl, PE, and EA extracts provoke DNA fragmentation. Using the lipid peroxidation inhibitory assay, the antioxidant capacity of M. arvensis extracts was evaluated by the ability of each extract to inhibit malondialdehyde formation. It was revealed that EA and TOF extracts are the most active in scavenging the hydroxyl radicals.

A comparative evaluation of mutagenic, antimutagenic, radical scavenging and antibacterial activities of essential oils of Pituranthos chloranthus (Coss. et Dur.).

The Salmonella typhimurium/microsome assay is a widely used bacterial genotoxicity assay to test potential carcinogens. The aim of this work was to evaluate the mutagenic and antimutagenic activities with and without the addition of an extrinsic metabolic activation system of essential oils obtained from an aerial part of Pituranthos chloranthus harvested from different stations in Tunisia. The oils showed no mutagenicity when tested with S. typhimurium strains TA98, TA100, and TA1535. On the other hand, we showed that these essential oils reduced significantly Benzo [a] pyrene (B[a] P) and sodium-azide-induced mutagenicity. The scavenging capacity of these essential oils was also estimated by evaluating the inhibition of DPPH radical. Essential oils harvested at Medenine and Gabes in November were more effective in scavenging activity. The essential oils were tested for their antimicrobial properties against five different bacteria, and were found to be weakly active, with MIC and MBC values in the range 0.6-4 and 2.2-5 mg/mL, respectively.

Phytochemistry and biological activities of Phlomis species.

The genus Phlomis L. belongs to the Lamiaceae family and encompasses 100 species native to Turkey, North Africa, Europe and Asia. It is a popular herbal tea enjoyed for its taste and aroma. Phlomis species are used to treat various conditions such as diabetes, gastric ulcer, hemorrhoids, inflammation, and wounds. This review aims to summarize recent research on the phytochemistry and pharmacological properties of the genus Phlomis, with particular emphasis on its ethnobotanical uses. The essential oil of Phomis is composed of four chemotypes dominated by monoterpenes (alpha-pinene, limonene and linalool), sesquiterpenes (germacrene D and beta-caryophyllene), aliphalic compounds (9,12,15-octadecatrienoic acid methyl ester), fatty acids (hexadecanoic acid) and other components (trans-phytol, 9,12,15-octadecatrien-1-ol). Flavonoids, iridoids and phenylethyl alcohol constitute the main compounds isolated from Phlomis extracts. The pharmacological activities of some Phlomis species have been investigated. They are described according to antidiabetic, antinociceptive, antiulcerogenic, protection of the vascular system, anti-inflammatory, antiallergic, anticancer, antimicrobial and antioxidant properties.

Phytochemical, antibacterial, antiproliferative, and antioxidant potentials and DNA damage-protecting activity of Acacia salicina extracts.

The extract enriched in total oligomer flavonoids (TOF), and the aqueous, methanol, and ethyl acetate extracts of Acacia salicina were investigated for their polyphenolic compound content, antioxidative activity in the nitro blue tetrazolium/riboflavin assay system, antibacterial activity against Gram-positive and Gram-negative bacterial reference strains, antigenotoxic activity tested with the Ames assay, and cytotoxic activity against the K562 human chronic myelogenous leukemia cell line and L1210 leukemia murine cells. TOF extract was effective at inhibiting nitro blue tetrazolium reduction by superoxide radical in a nonenzymatic O(2)(*-)-generating system. Significant activity against bacterial reference strains Staphylococcus aureus, Enterococcus faecalis, Salmonella enteritidis, and Salmonella typhimurium was shown with all the tested extracts. These extracts significantly decreased the genotoxicity induced by sodium azide and 4-nitro-o-phenylenediamine. A pronounced cytotoxic effect on both leukemia cell lines was detected in TOF, methanolic and ethyl acetate extracts. The antioxidant, antimicrobial, antigenotoxic, and cytotoxic activities exhibited by A. salicina depended on the chemical composition of the tested extracts.

Cell protection induced by Acacia salicina extracts: inhibition of genotoxic damage and determination of its antioxidant capacity.

Antioxidant activity of Acacia salicina extracts was determined by the ability of each extract to inhibit lipid peroxidation, to protect against DNA strand scission induced by hydroxyl radicals, and to scavenge the free radical, 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS(*+)). The IC(50) values of the inhibitory activity toward lipid peroxidation of total oligomer flavonoids (TOF), methanol, ethyl acetate, and aqueous extracts were respectively 28, 52, 472, and 480 microg/mL. All extracts have the ability to scavenge the ABTS(*+) radical by a hydrogen-donating mechanism and to protect pKS plasmid DNA against hydroxyl radicals- induced DNA damage. An assay for the ability of A. salicina extracts to prevent mutations induced by various mutagens in Salmonella typhimurium TA102 and TA104 cells was conducted. TOF, methanol, ethyl acetate, and aqueous extracts from leaf parts of A. salicina showed no mutagenicity either with or without the metabolic enzymes preparation (S9). Protection against methylmethanesulfonate-induced mutagenicity was observed for TOF, methanol, and ethyl acetate extracts. Likewise, all extracts exhibited a high inhibition level of the Ames response induced by the indirect mutagen, 2-aminoanthracene. The antigenotoxic activity could be ascribed, at least in part, to their antioxidant properties, but we cannot exclude additionally mechanisms. Thus, A. salicina may serve as an ideal candidate for a cost- effective, readily exploitable natural phytochemical compound.

Investigation of biological activity of polar extracts isolated from Phlomis crinita Cav ssp. mauritanica Munby.

The lyophilized infusion, the methanol, the ethyl acetate, and the total oligomer flavonoid (TOF)-enriched extracts prepared from the dried leaves of Phlomis crinita Cav. ssp. mauritanica Munby were investigated for the contents of flavonoids, tannins, coumarines and steroids. Antibacterial activity was investigated toward five bacterial strains. An inhibitory effect was observed against Staphyllococcus aureus and Enterococcus feacalis, and the minimal inhibitory concentrations ranged from 2.5 to 5 mg/mL of extract. The tested extracts exhibit an important free radical scavenging activity toward the 1,1-diphenyl 2-picrylhydrazyl (DPPH) free radical; with IC(50) values of 30.5, 6, 32, and 31.5 microg/mL, respectively, in the presence of lyophilized infusion, the TOF, the methanol, and the ethyl acetate extracts. Genotoxic and antigenotoxic properties of the different extracts were studied by using the SOS chromotest with Escherichia coli PQ37. The lyophilized infusion and TOF extracts obtained from P. crinita ssp. mauritanica showed no genotoxicity, whereas methanol and ethyl acetate extracts are considered as marginally genotoxic. On the other hand, we showed that each extract inhibited the mutagenicity induced by aflatoxin B1 (AFB1) (10 microg/assay) and nifuroxazide (NF) (10 microg/assay). The ethyl acetate extract showed the strongest level of protection toward the genotoxicity induced by both directly and indirectly genotoxic NF and AFB1. These tests proved that the lyophilized infusion possesses an antiradical activity likewise, it showed no genotoxic effect; that is why we choose this extract to assess its antiulcerogenic activity by using an ethanol-induced ulcerogenesis model in the rat. This test demonstrates that 300 mg/kg of a P. crinita ssp. mauritanica lyophilized infusion was more effective than the reference compound, cimetidine.

Moricandia arvensis extracts protect against DNA damage, mutagenesis in bacteria system and scavenge the superoxide anion.

The mutagenic potential of total aqueous, total oligomers flavonoids (TOF), ethyl acetate (EA), chloroform (Chl), petroleum ether (PE) and methanol (MeOH) extracts from aerial parts of Moricandia arvensis was assessed using Ames Salmonella tester strains TA100 and TA1535 with and without metabolic activation (S9), and using plasmid pBluescript DNA assay. None of the different extracts produced a mutagenic effect, except aqueous extract when incubated with Salmonella typhimurium TA100 after metabolic activation. Likewise, the antimutagenicity of the same extracts was tested using the "Ames test". Our results showed that M. arvensis extracts possess antimutagenic effects against sodium azide (SA) in the two tested Salmonella assay systems, except metabolized aqueous and PE extracts when tested with S. typhimurium TA100 assay system. Different extracts were also found to be effective in protecting plasmid DNA against the strand breakage induced by hydroxyl radicals, except PE and aqueous extracts. Antioxidant capacity of the tested extracts was evaluated using the enzymatic (xanthine/xanthine oxidase assay) (X/XOD) and the non enzymatic (NBT/Riboflavine assay) systems. TOF extract was the more effective one in inhibiting both xanthine oxidase activity and NBT reduction.

Acacia salicina extracts protect against DNA damage and mutagenesis in bacteria and human lymphoblast cell K562 cultures.

Three extracts were prepared from the leaves of Acacia salicina: aqueous, methanol, and ethyl acetate extracts. The antigenotoxic properties of these extracts were investigated by assessing the inhibition of mutagenicity of the indirect-acting mutagen benzo[a]pyrene using the Ames assay and the genotoxicity of the direct-acting mutagen, hydrogen peroxide, using the "Comet assay." Aqueous, methanol, and ethyl acetate extracts at doses of 500, 50, and 500 microg per plate reduced benzo[a]pyrene mutagenicity by 95%, 82%, and 40%, respectively, in Salmonella typhimurium TA98 strain and by 91%, 66% and 63%, respectively, at the same doses with a TA97 assay system. Human lymphoblast cells K562 were pretreated with 50% inhibition concentration of each extracts and then treated by H(2)O(2), for the Comet assay. The Comet assay results showed that ethyl acetate and methanol extracts decreased the DNA damage caused by H(2)O(2) by, respectively, 34.8% and 31.3%. We envisaged also the study of the antioxidant effect of these extracts by the enzymatic xanthine/xanthine oxidase assay. Results indicated that methanol and ethyl acetate extracts were potent inhibitors of xanthine oxidase and superoxide anion scavengers. We conclude that these integrated approaches to antigenotoxicity and antioxidant assessment may be useful to help compare the beneficial effects associated with using A salicina as medicinal and dietary plant.

In vitro evaluation of antibacterial, antioxidant, cytotoxic and apoptotic activities of the tubers infusion and extracts of Cyperus rotundus.

The in vitro antibacterial, antioxidant, cytotoxic and apoptotic activities from tubers extracts of Cyperus rotundus (Cyperaceae) were investigated. Antibacterial activity of different extracts was evaluated against five bacterial reference strains. A marked inhibitory effect was observed against Salmonella enteritidis, Staphylococcus aureus and Enterococcus faecalis with total oligomers flavonoids (TOFs) and ethyl acetate extracts. In addition to their antibacterial activity, the same extracts showed a significant ability to inhibit nitroblue tetrazolium reduction by the superoxide radical in a non-enzymatic superoxide generating system. Apoptosis, a highly organized physiological mechanism to eliminate injured or abnormal cells, is also implicated in multistage carcinogenesis. It was observed that TOF and ethyl acetate extracts suppressed growth and proliferation of L1210 cells derived from murine lymphoblastic leukaemia. Morphological features of treated cells and characteristic DNA fragmentation revealed that the cytotoxicity was due to induction of apoptosis. This study confirms that TOF and ethyl acetate extracts of C.rotundus possess antibacterial and antioxidant properties and provoke DNA fragmentation, a sign of induction of apoptosis. These results were correlated with chemical composition of the tested extracts.

Antimutagenic, antigenotoxic and antioxidant activities of Acacia salicina extracts (ASE) and modulation of cell gene expression by H2O2 and ASE treatment.

The total oligomers flavonoids (TOF), chloroform, petroleum ether and aqueous extracts from Acacia salicina, were investigated for the antioxidative, cytotoxic, antimutagenic and antigenotoxic activities. The viability of K562 cells were affected by all extracts after 48 h exposure. Our results showed that A. salicina extracts have antigenotoxic and/or antimutagenic activities. TOF and chloroform extracts exhibit antioxidant properties, expressed by the capacity of these extracts to inhibit xanthine oxidase activity. To further explore the mechanism of action of A. salicina extracts, we characterized expression profiles of genes involved in antioxidant protection and DNA repair in the human lymphoblastic cell line K562 exposed to H2O2. Transcription of several genes related to the thioredoxin antioxidant system and to the DNA base-excision repair pathway was up-regulated after incubation with chloroform, TOF and petroleum ether extracts. Moreover genes involved in the nucleotide-excision repair pathway and genes coding for catalase and Mn-superoxide-dismutase, two important antioxidant enzymes, were induced after incubation with the chloroform extract. Taken together, these observations provide evidence that the chloroform and TOF extracts of A. salicina leaves contain bioactive compounds that are able to protect cells against the consequences of an oxidative stress.

Antioxidant activity and inhibition of aflatoxin B1-, nifuroxazide-, and sodium azide-induced mutagenicity by extracts from Rhamnus alaternus L.

The effect of extracts obtained from Rhamnus alaternus L. leaves on genotoxicity and SOS response induced by aflatoxin B(1) (10 microg/assay) as well as nifuroxazide (20 microg/assay) was investigated in a bacterial assay system, i.e., the SOS chromotest with Escherichia coli PQ37. The evaluation of the mutagenic and antimutagenic actions of the same extracts against the sodium azide (1.5 microg/plate)-induced mutagenicity was assayed using the Salmonella typhimurium assay system. The R. alaternus tested extracts exhibited no genotoxicity either with or without the external S9 activation mixture. However, all the extracts, particularly aqueous extract (A) and its chloroformic fraction (A(2)) significantly decreased the genotoxicity induced by aflatoxin B(1) and nifuroxazide. Moreover, the different extracts showed no mutagenicity when tested with Salmonella typhimurium strains TA1535 and TA1538 either with or without the S9 mix. Aqueous extract as well as its A(2) fraction exhibited the highest level of protection towards the direct mutagen, sodium azide-induced response in TA1535 strain with mutagenicity inhibition percentages of 83.6% and 91.4%, respectively, at a dose of 250 microg/plate. The results obtained by the Ames test assay confirm those of SOS chromotest. These same active extracts exhibited high xanthine oxidase (XOD) inhibiting with respective IC(50) values of 208 and 137 microg/ml, and superoxide anion-scavenging effects (IC(50) values of 132 and 117 microg/ml) when tested in the XOD enzymatic assay system. Our findings emphasize the potential of R. alaternus to prevent mutations and also its antioxidant effect.

Comparative study of Cyperus rotundus essential oil by a modified GC/MS analysis method. Evaluation of its antioxidant, cytotoxic, and apoptotic effects.

Gas chromatography coupled with mass spectrometry (GC/MS), using both electron impact (EI) and chemical ionization (CI) detection modes on apolar and polar stationary phases, led to the determination of the volatile composition of the essential oil obtained from tubers of Cyperus rotundus (Cyperaceae). In this study, more than 33 compounds were identified and then compared with the results obtained in our previous work. Cyperene, alpha-cyperone, isolongifolen-5-one, rotundene, and cyperorotundene were the principal compounds comprising 62% of the oil. An in vitro cytotoxicity assay with MTT indicated that this oil was very effective against L1210 leukaemia cells line. This result correlates with significantly increased apoptotic DNA fragmentation. The oxidative effects of the essential oil were evaluated using the 1,1-diphenyl-2-picrylhydrazyl (DPPH), xanthine/xanthine oxidase assays, and the scavenging of superoxide radical assay generated by photo-reduction of riboflavin. The antimutagenic activity of essential oil has been examined by following the inhibition of H(2)O(2) UV photolysis which induced strand-break formation in pBS plasmid DNA scission assay. Based on all these results, it is concluded that C. rotundus essential-oil composition established by GC/MS analysis, in EI- and CI-MS modes, presents a variety of a chemical composition we were not able to detect with only GC/MS analysis in our previous work. This essential oil exhibited antioxidant, cytotoxic, and apoptotic properties.

Mutagenic, antimutagenic, cytotoxic, and apoptotic activities of extracts from Pituranthos tortuosus.

Mutagenic and antimutagenic activities against direct acting mutagens, nifuroxazide (NF) and sodium azide (SA), and indirect acting mutagen aflatoxin B1 (AFB1) of extracts prepared from aerial parts of Pituranthos tortuosus were investigated in bacterial assay systems (i.e., the Ames test with Salmonella typhimurium TA100, TA98, TA1538, TA1535, and the SOS chromotest with Escherichia coli PQ 37). It was found that all extracts obtained from P. tortuosus decreased the mutagenicity induced by AFB1 (10 microg/assay), SA (1.5 microg/assay), and NF (20 microg/assay). Ethyl acetate, acetone, methanol, and total oligomer flavenoid extracts exhibited the highest inhibition level of mutagenicity induced by the indirect mutagen AFB1. In addition, antiproliferative and apoptotic properties of these extracts have also been reported using two leukemia cell lines, L1210 and K562. The results revealed that all extracts showed a significant cytotoxic effect on these cell lines, and the effect was greater in the presence of human K562 chronic myelogenous leukemia cells, whereas they do not induce apoptosis.