The effect of oral administration of Allium sativum extracts on lead nitrate induced toxicity in male mice

Lead is a common environmental occupational toxic metal, known to have indirect oxidative effects. Considering the antioxidant properties of garlic, this study was undertaken to evaluate the therapeutic efficacy of garlic extracts in terms of normalization of altered hematological, biochemical and immunological parameters, and depletion of inorganic lead burden in blood, kidney and brain tissues. Chronic lead nitrate ingestion showed a significant decline in total erythrocyte count, total leukocyte count, hemoglobin concentration, lymphocyte and monocyte content, while neutrophil content increased in lead nitrate treated group. Pb(NO₃)₂ exposure elicited a significant escalation in thiobarbituric acid reactive substances level and depletion in reduced glutathione content and antioxidant enzymes namely, superoxide dismutase and catalase in kidney and brain. Activities of aspartate transaminase, alanine transaminase, acid phosphatase and alkaline phosphatase augmented significantly in kidney and brain of lead exposed mice. Lead nitrate treatment decreased protein content while cholesterol and lead burden increased significantly. A decrease in viability of macrophage, phagocytic index, immunoglobulin level and plaque count were the salient features observed in lead exposed animals. However, oral administration of garlic extracts to Pb(NO₃)₂ treated groups attenuated the deranged parameters to some extent. This indicates that garlic can be a protective regimen for lead toxicity.     

Aquilegia vulgaris L. extract counteracts oxidative stress and cytotoxicity of fumonisin in rats

Exposure to fumonisins (FB) is known to have toxic and carcinogenic effects in different animal species, and to express toxicity in cells via the induction of oxidative stress. The aim of the current study was to evaluate the protective effects of the ethanol extract of Aquilegia vulgaris L. against the oxidative stress and the genotoxicity using micronucleus assay and random amplified polymorphism DNA (RAPD-PCR) in FB-treated rats. Sixty mature female Sprague-Dawley were divided into six treatment groups and treated for 4 weeks as follow: the control group, the group fed FB-contaminated diet (200 mg/kg diet), the groups treated orally with the extract (5 and 10 mg/kg bw) and the groups fed FB-contaminated diet and treated with the extract at the two doses. The results showed that treatment with FB alone disturbed lipid profile in serum, increases Sa/So ratio, induces micronucleated polychromatic erythrocytes (Mn-PCEs) in bone marrow, increases DNA and RNA in liver accompanied with significant changes in histological picture The extract alone at the two tested doses did not induce any significant changes in the biochemical or histological picture. The combined treatment showed significant improvements in all biochemical, cytogenetic parameters tested and histological pictures in the liver tissues. Moreover, this improvement was more pronounced in the group received the high dose of the extract. It could be concluded that the ethanol extract of A. vulgaris induced its protective effect via the increase in the antioxidant capacity, inhibition of lipid peroxidation and scavenging of free radicals.     

Mechanisms of thiosulfinates from Allium tuberosum L.-induced apoptosis in HT-29 human colon cancer cells

This study was performed to elucidate the apoptotic pathways by thiosulfinates, major biologically active components of Allium tuberosum L., in HT-29 human colon cancer cells. Thiosulfinates significantly induced cell death in dose- and time-dependent manners in HT-29 cells, which is associated with apoptosis. Thiosulfinates activated the initiator caspase-8, and -9, and the effector caspase-3. In the present study, thiosulfinates were found to stimulate Bid cleavage, indicating that the apoptotic action of caspase-8-mediated Bid cleavage leads to the activation of caspase-9. Thiosulfinates down-regulated the expression of the anti-apoptotic protein Bcl-2, and up-regulated the expression of the pro-apoptotic protein Bax. We also found that thiosulfinates increased the expression of AIF, a caspase-independent mitochondrial apoptosis factor, and induced DNA fragmentation and chromatin condensation in HT-29 cells. These results indicate that thiosulfinates from A. tuberosum L. inhibited cell proliferation and activated both the caspase-dependent and caspase-independent apoptotic pathways in HT-29 cells.     

Composition,antimicrobial activity and in vitro cytotoxicity of essential oil from Cinnamomum zeylanicum Blume (Lauraceae)

The essential oil from the bark of Cinnamomum zeylanicum Blume was analyzed by GC–MS and bioassays were carried out. Nine constituents representing 99.24% of the oil were identified by GC–MS. The major compounds in the oil were (E)-cinnamaldehyde (68.95%), benzaldehyde (9.94%) and (E)-cinnamyl acetate (7.44%). The antimicrobial activity of the oil was investigated in order to evaluate its efficacy against 21 bacteria and 4 Candida species, using disc diffusion and minimum inhibitory concentration methods. The essential oil showed strong antimicrobial activity against all microorganisms tested. The cytotoxic and apoptotic effects of the essential oil on ras active (5RP7) and normal (F2408) fibroblasts were examined by MTT assay and acridine orange/ethidium bromide staining, respectively. The cytotoxicity of the oil was quite strong with IC₅₀ values less than 20 μg/mL for both cell lines. 5RP7 cells were affected stronger than normal cells. Morphological observation of apoptotic cells indicated the induction of apoptosis at the high level of the oil, especially in 5RP7 cells. The present study showed the potential antimicrobial and anticarcinogenic properties of the essential oil of cinnamon bark, indicating the possibilities of its potential use in the formula of natural remedies for the topical treatment of infections and neoplasms.     

Subchronic toxicity of Citrus aurantium L. (Rutaceae) extract and p-synephrine in mice

Extracts of Citrus aurantium L. (Rutaceae) unripe fruits have gained popularity for the treatment of obesity. Due to the wide use of C. aurantium/p-synephrine-containing products, this research was undertaken to evaluate its subchronic toxicity in mice and their actions in oxidative stress biomarkers. Groups of 9–10 mice received for 28 consecutive days a commercial C. aurantium dried extract (containing 7.5% p-synephrine) 400, 2000 or 4000 mg/kg and p-synephrine 30 or 300 mg/kg by oral gavage. There was a reduction in body weight gain of animals treated with both doses of p-synephrine. Organs relative weight, biochemical and hematological parameters were not altered in all treated mice. There was an increase in reduced glutathione (GSH) concentration in groups treated with C. aurantium 4000 mg/kg and p-synephrine 30 and 300 mg/kg. In glutathione peroxidase (GPx), there were an inhibition of the activity in C. aurantium 400 and 2000 mg/kg and p-synephrine 30 and 300 mg/kg treated animals, respectively, and was no alteration in malondialdehyde (MDA) levels. Thus, the results indicate a low subchronic toxicity of the tested materials in mice and a possible alteration in the oxidative metabolism. However, further tests are required to better elucidate the effects of these compounds in the antioxidant system.     

Cardio protective effect of Coriandrum sativum L. on isoproterenol induced myocardial necrosis in rats

The preventive effect of Coriandrum sativum L. (CS) on cardiac damage was evaluated by Isoproterenol (IP) induced cardiotoxicity model in male Wistar rats. Rats were pretreated with methanolic extract of CS seeds at a dose of 100, 200 or 300 mg/kg orally for 30 days and they were subsequently administered (s.c.) with IP (85 mg/kg body weight) for the last two days. IP treated rats showed increased LPO, decreased levels of endogenous antioxidants and ATPases in the cardiac tissue together with increased plasma lipids and markers of cardiac damage. TTC staining showed increased infarct areas while HXE staining showed myofibrillar hypertrophy and disruption. CS (200 and 300 mg/kg body weight) pretreatment significantly prevented or resisted all these changes. Our results show that methanolic extract of CS is able to prevent myocardial infarction by inhibiting myofibrillar damage. It is also concluded that, the rich polyphenolic content of CS extract is responsible for preventing oxidative damage by effectively scavenging the IP generated ROS.     

Protective effects of Coriandrum sativum extracts on carbon tetrachloride-induced hepatotoxicity in rats

Oxidative damage is implicated in the pathogenesis of various liver injuries. The study was aimed to investigate the antioxidant activity of Coriandrum sativum on CCl₄ treated oxidative stress in Wistar albino rats. CCl₄ injection induced oxidative stress by a significant rise in serum marker enzymes and thiobarbituric acid reactive substances (TBARS) along with the reduction of antioxidant enzymes. In serum, the activities of enzymes like ALP, ACP and protein and bilirubin were evaluated. Pretreatment of rats with different doses of plant extract (100 and 200 mg/kg) significantly lowered SGOT, SGPT and TBARS levels against CCl₄ treated rats. Hepatic enzymes like SOD, CAT, GPx were significantly increased by treatment with plant extract, against CCl₄ treated rats. Histopathological examinations showed extensive liver injuries, characterized by extensive hepatocellular degeneration/necrosis, inflammatory cell infiltration, congestion, and sinusoidal dilatation. Oral administration of the leaf extract at a dose of 200 mg/kg body weight significantly reduced the toxic effects of CCl₄. The activity of leaf extract at the dose of 200 mg/kg was comparable to the standard drug, silymarin. Based on these results, it was observed that C. sativum extract protects liver from oxidative stress induced by CCl₄ and thus helps in evaluation of traditional claim on this plant.     

Polyphenolics from various extracts/fractions of red onion (Allium cepa) peel with potent antioxidant and antimutagenic activities

In order to determine antioxidant activity, the five extracts/fractions of red onion peel were studied for their total content of phenolics (TPC), flavonoids (TFC), antioxidant activity (AOA), free radical scavenging activity (FRSA), assayed by DPPH radical in the terms of anti-radical power (ARP) and reducing power (RP), expressed as ascorbic acid equivalents (ASE)/ml. High TPC (384.7 ± 5.0 mg GAE/g), TFC (165.2 ± 3.2 mg QE/g), AOA (97.4 ± 7.6%), ARP (75.3 ± 4.5) and RP (1.6 ± 0.3 ASE/ml) were found for the ethyl acetate (EA) fraction. EA fraction had markedly higher antioxidant capacity than butylated hydroxytoluene (BHT) in preventive or scavenging capacities against FeCl₃-induced lipid peroxidation, protein fragmentation, hydroxyl (site-specific and non-site-specific), superoxide anion and nitric oxide radicals. EA fraction also showed dose dependent antimutagenic activity by following the inhibition of tobacco-induced mutagenicity in Salmonella typhimurium strains (TA102) and hydroxyl radical-induced nicking in plasmid pUC18 DNA. HPLC and MS/MS analysis showed the presence of ferulic, gallic, protocatechuic acids, quercetin and kaempferol. The large amount of polyphenols contained in EA fraction may cause its strong antioxidant and antimutagenic properties. This information shows that EA fraction of red onion peel can be used as natural antioxidant in nutraceutical preparations.     

Nutritional and nutraceutical potential of rape (Brassica napus L. var. napus) and “tronchuda” cabbage (Brassica oleraceae L. var. costata) inflorescences

Two traditional cultivated vegetables highly consumed among Northern Portuguese regions were tested for their chemical composition, nutritional profile and in vitro antioxidant properties using four assays: 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals scavenging activity, reducing power, inhibition of β-carotene bleaching and inhibition of lipid peroxidation by thiobarbituric acid reactive substances (TBARS) assay. The studied varieties of two Brassica species, locally known as “grelos” (rape) and “espigos” (“tronchuda” cabbage) are nutritionally well-balanced vegetables; particularly “tronchuda” cabbage revealed the highest levels of moisture, proteins, fat, energy, β-carotene and vitamin C; rape gave the highest contents of ash, carbohydrates, sugars (including fructose, glucose, sucrose and raffinose), essential n-3 fatty acid α-linolenic acid, and the best ratios of PUFA/SFA and n-6/n-3 fatty acids, tocopherols, lycopene, chlorophylls, phenolics, flavonoids, and also the highest antioxidant properties. The health benefits associated to the antioxidant properties reinforce their contribution to a healthy and balanced diet, highlight the interest of their consumption, validate the empirical use and add new values to traditional/regional products which have been used for a long time.     

New antibacterial and cytotoxic activities of falcarindiol isolated in Crithmum maritimum L. leaf extract

Bioassay-guided fractionation of a chloroformic extract obtained from Crithmum maritimum leaves led to the chemical isolation of falcarindiol, a polyacetylene widely distributed within the Apiaceae family. Structure of this compound was confirmed by NMR and activity was tracked using a screening microplate bioassay. Falcarindiol strongly inhibited the growth of Micrococcus luteus and Bacillus cereus, with a minimum inhibitory concentration (MIC) value of 50 μg ml⁻¹. Moreover, this compound showed cytotoxicity against IEC-6 cells with an IC₅₀ value of 20 μM after 48 h of exposition. These results suggest that Crithmum maritimum could be potentially used in food manufactures and cosmetology as preservative agents and biopesticides, or in medicine as new antibiotics, confirming the interest in studying halophytic species as sources of bioactive compounds.     

Hematological cell findings in bone marrow and peripheral blood of rabbits after experimental acute exposure to Loxosceles intermedia (brown spider) venom.

The purpose of this work was to find out the cellular changes occurring in bone marrow and peripheral blood after acute exposure to the venom of Loxosceles intermedia. Doses of 40 microg of venom were injected intradermally into five rabbits, and five rabbits receiving only phosphate-buffered saline (PBS) were used as controls. Bone marrow and peripheral blood samples were obtained before the envenomation and 4, 8, 12, 24 and 48 h, and 5, 10, 15, 20 and 30 days after envenomation. In bone marrow samples we assessed cellularity, nucleated red cells, megakaryocytes and neutrophils, and in peripheral blood we assessed red cells (red cell concentration, hemoglobin and hematocrit), leukocytes, neutrophils and platelets. Our objective was to find out if the venom has a direct effect on bone marrow and peripheral blood or if changes in both of them are secondary to the needs of tissues, and if there is a good correlation between histopathological and hematological findings. We found that the red cell parameters were not affected by the venom, except for nucleated red cells which decreased after venom exposure. The depression of megakaryocyte numbers and thrombocytopenia showed a strong correlation with the histopathologic changes observed in skin biopsies obtained from the rabbits. The changes in cellularity and neutrophils of bone marrow were strongly correlated with those in peripheral blood and skin. The thrombocytopenia and neutropenia in peripheral blood are due to marrow depression, which may be a consequence of an extensive migration of platelets and neutrophils to the necrotic lesion or the marrow depression may be a transitory effect of evenoming by L. intermedia.     

Fermentation enhances the in vitro antioxidative effect of onion (Allium cepa) via an increase in quercetin content

Yellow onion (Allium cepa) extract showed enhanced antioxidative effects in 2,2-diphenyl-1-picrylhydrazyl (DPPH), Trolox equivalent antioxidant capacity (TEAC) and 5-(and-6)-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate and acetyl ester (CM-H₂DCFDA) assay after being treated with a crude enzyme extract from soybean paste fungi, Aspergillus kawachii. HPLC analysis showed two increased and two decreased peaks after enzyme treatment. The decreased peaks were identified as quercetin-3,4′-di-O-β-d-glucoside (1) and quercetin-4′-O-β-d-glucoside (2), and peaks that increased were quercetin-3-O-β-d-glucoside (3) and quercetin (4), respectively. It was expected that 3 and 4 were originated from the glucosidic cleavage of their glucosides, 1 and 2. Among the increased compounds, only quercetin (4) showed strong antioxidative activity in the DPPH assay. In addition, the protective effect against glutamate-induced neurotoxicity in HT22 cells was increased when treated with 25 μg/ml of fermented onion. The enhanced neuroprotective effect was also originated from the increased quercetin content. As a consequence, fermentation raised the quercetin content in onion, and subsequently increased the antioxidative and neuroprotective activities.     

In vitro cytokine release from human peripheral blood mononuclear cells in the assessment of the immunotoxic potential of chemicals

Alternative methods to the use of animals in testing of chemicals are needed. We investigated if the immunotoxic potential of 12 dietary toxicants could be predicted from effects on cytokine release from human peripheral blood mononuclear cells (PBMC) after in vitro exposure. Nine cytokines were selected to reflect different types of immune responses. The toxicants were classified as immunotoxic or non-immunotoxic substances according to the published in vivo data.Isolated human PBMC were exposed for 20 h to three concentrations of each of the 12 substances in the presence of human liver S9 fraction. After further incubation of PBMC in fresh medium containing the mitogen phytohemagglutinin (PHA, 10 μg/ml) for 48 h, release of the nine selected cytokines into the supernatant as well as cell proliferation were measured by Luminex technology™ and the BrdU incorporation assay, respectively.All 12 substances investigated affected the release of one or more cytokines, and each of the substances showed different cytokine release patterns. Within the limitations of the study design, the present study suggests that the effect of the substances on mitogen-induced cytokine release from PBMC cannot predict their immunotoxic potential, but may be useful in mechanistic studies.     

Biological potentials and cytotoxicity of various extracts from endemic Origanum minutiflorum O. Schwarz & P.H. Davis

The aim of the present study was to investigate antioxidative, antimicrobial and cytotoxic effects of endemic Origanum minutiflorum (O. Schwarz & P.H. Davis). Antioxidant activities of the extracts from O. minutiflorum were evaluated by using DPPH radical scavenging, β-carotene bleaching and metal chelating activity assays. In addition, the amounts of total phenol components in the plant extracts were determined. In the β-carotene bleaching test, the extracts exhibited in the range of 58.1 ± 0.2% – 98.2 ± 0.3% inhibition against linoleic acid oxidation. The antimicrobial efficiency of the plant was evaluated according to agar well diffusion and microdilution broth methods. The n-hexane extract of O. minutiflorum having an inhibition zone of 20.2 ± 0.2 mm, had the maximum antibacterial efficiency against Shigella sonnei RSKK 878. Cytotoxic effects of the extracts were determined by MTT assay. O. minutiflorum extracts (at concentration of 10–100 μg/ml) did not show any cytotoxic effect on baby hamster kidney fibroblast cell line. The results showed that O. minutiflorum could be used as a natural source in food industry.     

Cytotoxicity,cell cycle arrest,and apoptosis in breast cancer cell lines exposed to an extract of the seed kernel of Mangifera pajang (bambangan)

An extract of Mangifera pajang kernel has been previously found to contain a high content of antioxidant phytochemicals. The present research was conducted to investigate the anticancer potential of this kernel extract. The results showed that the kernel crude extract induced cytotoxicity in MCF-7 (hormone-dependent breast cancer) cells and MDA-MB-231 (non-hormone dependent breast cancer) cells with IC50 values of 23 and 30.5 μg/ml, respectively. The kernel extract induced cell cycle arrest in MCF-7 cells at the sub-G1 (apoptosis) phase of the cell cycle in a time-dependent manner. For MDA-MB-231 cells, the kernel extract induced strong G2-M arrest in cell cycle progression at 24 h, resulting in substantial sub-G1 (apoptosis) arrest after 48 and 72 h of incubation. Staining with Annexin V-FITC and propidium iodide revealed that this apoptosis occurred early in both cell types, 36 h for MCF-7 cells and 24 h for MDA-MB-231cells, with 14.0% and 16.5% of the cells respectively undergoing apoptosis at these times. This apoptosis appeared to be dependent on caspase-2 and -3 in MCF-7 cells, and on caspase-2, -3 and -9 in MDA-MB-231 cells. These findings suggest that M. pajang kernel extract has potential as a potent cytotoxic agent against breast cancer cell lines.     

Metabolic profiling and biological capacity of Pieris brassicae fed with kale (Brassica oleracea L. var. acephala)

Phenolic and organic acid profiles of aqueous extracts from Pieris brassicae material and the host kale (Brassica oleracea L. var. acephala) leaves were determined by HPLC/UV–DAD/MSⁿ-ESI and HPLC–UV, respectively. The identified phenolics included acylated and nonacylated flavonoid glycosides, hydroxycinnamic acyl gentiobiosides, and sulphate phenolics. Kale exhibited the highest content (11 g/kg lyophilized extract), while no phenolics were identified in the butterflies or exuviae. Nine different organic acids were characterized in the materials, with kale showing the highest amount (112 g/kg lyophilized extract). With the exception of the exuviae extract, the rest were screened for bioactivity. Using spectrophotometric microassays, all exhibited antiradical capacity against DPPH and NO in a concentration-dependent way, whereas only kale and excrement extracts were active against superoxide. All displayed activity on intestinal smooth muscle, albeit with distinct relaxation–contraction profiles. Larvae and butterfly extracts were more efficacious for intestinal relaxation than was kale extract, whereas excrement extract evoked only contractions, thus evidencing their different compositions. Collectively, these results show that P. brassicae sequesters and metabolizes kale’s phenolic compounds. Moreover, the extract’s bioactivities suggest that they may constitute an interesting source of bioactive compounds whose complex chemical structures preclude either synthesis or isolation.     

Toxigenicity of enniatins from Western Australian Fusarium species to brine shrimp (Artemia franciscana)

The high prevalence (14 of 24 isolates) of enniatin-producing isolates from Western Australian Fusarium species isolated from pasture legumes associated with sheep feed refusal and rat deaths, and the high toxicity of their crude extracts to brine shrimp (Artemia franciscana) from a previous study warranted further investigation of this class of mycotoxin. Crude extracts from Fusarium acuminatum, Fusarium avenaceum, Fusarium tricinctum and Fusarium sambucinum, along with enniatins A, A1, B and B1 purified from a Western Australian strain of F. acuminatum using semi-preparative HPLC, were bioassayed using brine shrimp. All Fusarium isolates produced both enniatins B and B1, except for F. tricinctum WAC 8019, and 11 of the 17 isolates produced enniatin A1. Overall, all of the F. avenaceum isolates produced high amounts of enniatins, in particular enniatin B. One isolate of F. acuminatum (WAC 5715) and of F. tricinctum (WAC 11486) also produced high amounts of both enniatins B and B1. Only F. acuminatum WAC 5715 produced enniatin A among the tested isolates. All four purified enniatins A, A1, B, B1, individually and in combination, caused brine shrimp toxicity after 6 h of exposure, implicating that this emerging class of mycotoxin as a cause of the acute toxicity to brine shrimp observed. The mixture of all four enniatins was the most toxic to brine shrimp compared to purified individual enniatins, where the relative toxicity order was B > B1 > A1 > A. Enniatin B was the individual most toxic enniatin with some bioactivity at 5 μg/mL and almost 100% brine shrimp death at 50 μg/mL after 24 h of exposure. This study is the first report to confirm the acute toxicity of enniatins A, A1, B and B1 to brine shrimp, and also highlights the need for further investigation of the potential toxicity of these cyclic hexadepsipeptides to animals and humans.     

Coronatin-1 isolated from entomopathogenic fungus Conidiobolus coronatus kills Galleria mellonella hemocytes in vitro and forms potassium channels in planar lipid membrane

Entomopathogenic fungi are important natural regulatory factors of insect populations and have potential as biological control agents of insect pests. The cosmopolitan soil fungus Conidiobolus coronatus (Entomopthorales) easily attacks Galleria mellonella (Lepidoptera) larvae. Prompt death of invaded insects is attributed to the action of toxic metabolites released by the invader. Effect of fungal metabolites on hemocytes, insect blood cells involved in innate defense response, remains underexplored to date.C. coronatus isolate 3491 inducing 100% mortality of G. mellonella last instar larvae exposed to sporulating colonies, was cultivated at 20 °C in minimal medium. Post-incubation filtrates were used as a source of fungal metabolites. A two-step HPLC (1 step: Shodex KW-803 column eluted with 50 mM KH₂PO₄ supplemented with 0.1 M KCl, pH 6.5; 2 step: ProteinPak™ CM 8HR column equilibrated with 5 mM KH₂PO₄, pH 6.5, proteins eluted with a linear gradient of 0.5 M KCl) allowed the isolation of coronatin-1, an insecticidal 36 kDa protein showing both elastolytic and chitinolytic activities.Addition of coronatin-1 into primary in vitro cultures of G. mellonella hemocytes resulted in rapid disintegration of spherulocytes freely floating in culture medium and shrinkage of plasmatocytes adhering to the bottom of culture well. Coronatin-1 stimulated pseudopodia atrophy and, in consequence, disintegration of nets formed by cultured hemocytes.After incorporation of coronatin-1 into planar lipid membrane (PLM) ion channels selective for K⁺ ions in 50/450 mM KCl solutions were observed. Potassium current flows were recorded in nearly 70% of experiments with conductance from 300 pS up to 1 nS. All observed channels were active at both positive and negative membrane potentials. Under experimental conditions incorporated coronatin-1 exhibited a zero current potential (E_rev) of 47.7 mV, which indicates K⁺-selectivity of this protein.The success of the purification of coronatin-1 will allow further characterization of the mode of action of this molecule, including ability of coronatin-1 to form potassium channels in immunocompetent hemocytes.     

Cytotoxic and antioxidative activities of Plantago lagopus L. and characterization of its bioactive compounds

Bioactivity guided isolation and characterization of phytoconstituents from the aerial parts of Plantago lagopus L. were performed to give a new insight into the usage of Plantago species in traditional medicine. The extract showed strong radical scavenging effects against DPPH, nitric oxide (NO) and superoxide (SO) radicals comparable to that of known antioxidants 3-t-butyl-4-hydroxyanisole, ascorbic acid (vitamin C), and quercetin in addition to its cytotoxic activities against HEP-2 (human larynx epidermoid carcinoma) and RD (human rhabdomyosarcoma) cell lines based on MTT assay for growth inhibition. The gallic acid equivalent total phenolic content of the plant was found to be 79.94 mg/g dry extract. Phenylethanoid glycosides, verbascoside and calceorioside A were isolated from the most active fraction and both compounds showed strong radical scavenging activity against tested radicals and cytotoxicity against HEP-2, RD and MCF-7 (human breast adenocarcinoma) cell lines. In addition apoptotic cell death was observed in histological analysis. Taken together, these findings suggest that verbascoside and calceorioside A may be used in cancer prevention.     

Potential analgesic,anti-inflammatory and antioxidant activities of hydroalcoholic extract of Areca catechu L. nut

The hydroalcoholic extract of Areca catechu L. (ANE) nut was screened for its analgesic, anti-inflammatory and in vitro antioxidant potential. Three doses of ANE (250, 500 and 1000 mg/kg orally) were tested for analgesic and anti-inflammatory activities. Evaluation of analgesic activity of ANE was performed using hot plate and formalin test in mice. ANE showed maximum increase in hot plate reaction time (56.27%, p < 0.01), while reduced the duration of licking/biting behaviors in first (39.45%, p < 0.05) and second (92.71%, p < 0.01) phases of the formalin test indicating significant analgesic activity. ANE reduced the paw edema considerably (86.79% inhibition after 24 h, p < 0.01) in dose-dependent manner compared to carrageenan-induced rat. In addition, in vitro antioxidant activity of ANE was investigated by total phenolic content (TPC) and hydrogen peroxide assay. The IC₅₀ observed in hydrogen peroxide assay was 83.14 μg/ml and TPC 120.56 ± 21.09 mg QE/g. Altogether, these results suggest that the hydroalcoholic extract of Areca catechu could be considered as a potential analgesic, anti-inflammatory and antioxidant agent.