Chemical characteristics and biological activity screening of Pistacia lentiscus mastic gum and leaves from Türkiye.

BACKGROUND: Mastic gum is a resin that is produced by Pistacia lentiscus. It has many traditional uses, dating from ancient times, such as the treatment of gastrointestinal disorders and as a food additive. In this study, the leaves and mastic gum of trees of different ages from Karaburun and the Cesme peninsula in Türkiye were examined chemically and biologically. Flavonoids, and phenolic and fatty acid components were evaluated by a liquid chromatography system coupled with high resolution mass spectrometry (LC-HRMS) and gas chromatography with flame ionization detection (GC-FID). Cytotoxicity was screened against several cancer and healthy cell lines using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Inducible nitric oxide synthase (iNOS) inhibition was determined on lipopolysaccharide (LPS)-induced murine macrophage cell line (RAW 264.7) cells. Antiviral activity was measured against avian coronavirus using an in ovo virucidal antiviral activity assay. RESULTS: The main phenolic constituents of the gum were found to be salicylic, rosmarinic, and caffeic acids whereas the most abundant compounds detected were flavonoids in the leaf extracts. The most abundant fatty acids in hexane extracts were palmitic and oleic acids. All gum extracts except 3-year-old gum had significant cytotoxic activity on HeLa (IC50 1.74 ± 0.03-4.76 ± 0.95) and PC-3 (0.64 ± 0.25-6.22 ± 1.40) cells. Moreover, reducing virus activity by fivefold or sixfold logarithmically between the range of 5-10 µg g-1 of 30-year-old gum extracts underscored the biological activity. CONCLUSION: In ovo antiviral activity studies on the P. lentiscus were conducted for the first time. The mastic gum and leaves obtained from P. lentiscus may have strong potential in terms of their chemical content and antiviral and cytotoxic activity. As a consequence of these properties, it is a sustainable, renewable natural resource that can be used as an additive and flavoring in the food and pharmaceutical industries. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Anthemis tricolor Containing Unusual Totarol with Cytotoxic and Acetylcholinesterase-Inhibitory Activity.

Anthemis tricolor is an endemic species of Cyprus, and there is very limited information on its chemistry and pharmacological activities. The study aims to identify the in-vitro cytotoxic and acetylcholinesterase activities of Anthemis tricolor. The compounds responsible for the activities were also identified. Potent extracts of A. tricolor were subjected to preparative isolation and spectral structure determination studies. The chloroform extract contained many components, and due to the small quantity of extract available, enough pure compound(s) cannot be obtained for structure determination studies, though the n-hexane extract afforded two known compounds, totarol (1) and taraxasterol (2). The structures of the compounds (1 and 2) were determined by 1 H and 13 C NMR experiments. The pure compounds were also tested for their acetylcholinesterase inhibitory properties. For compound 1, the IC50 value was found to be 87.88 µg/mL. However, no inhibition was seen for 2. Anthemis tricolor was established to be a valuable source of pharmacologically active compounds and requires further studies.

Assessment of in vitro Cytotoxic, iNOS, Antioxidant and Photodynamic Antimicrobial Activities of Water-soluble Sulfonated Phthalocyanines.

In recent years, much effort has been devoted to the development of effective anticancer agents. In this manner, the utilization of water-soluble sulfonated phthalocyanines is crucial for many cancer cell lines. In this study, phthalonitrile and metallophthalocyanine compounds linked by benzenesulfonic acid groups have been prepared. Antimicrobial behaviors of those compounds were investigated by performing disk diffusion and photodynamic assays on gram-positive and negative bacteria. Indium phthalocyanine (InClPc) (3) showed inhibition activity against B. cereus, B. subtilis and S. aureus with disk diffusion assay. Also, gallium and indium phthalocyanines (2 and 3) exhibited inhibitory activity on both gram-positive and -negative microorganisms after light activation. Increasing the inhibitor concentration and light exposure time increased the inhibition activity for both molecules. GaClPc (2) demonstrated the maximum reducing power capacity among studied compounds, and CoPc (4) showed even better DPPH radical scavenging ability than the standard molecule Trolox at 2000 µg mL-1 concentration. The dose-dependent effect of compounds on cytotoxicity was studied against cancer cells PANC-1, MDA-MB-231, HepG2, A549, HeLa, CaCo-2 and non-tumorigenic cells HEK-293. All compounds showed no significant cytotoxic effect on any cell line up to the highest treated concentration at 50 µg mL-1 . However, all phthalocyanines had significant nitric oxide inhibition activity, and only in copper phthalocyanine (CuPc) (5), the MTT IC50 value was reached on LPS-activated RAW 264.7 macrophage cells. The lowest inducible nitric oxide synthase (iNOS) IC50 values were defined as 6 ± 1 µg mL-1 and 7 ± 0.5 µg mL-1 for CuPc (5) and InClPc (3), respectively.

Investigation of the supportive therapy potential of propolis extract and Lactobacillus acidophilus LA-5 milk combination against breast cancer in mice.

Immunotherapy has been applied in cancer treatments for many years as an alternative treatment method to radiotherapy, chemotherapy. It is well known that immunotherapy could suppress tumor formation by modulating the immune system of the host. The aim of the study is to investigate supportive therapy potential of acidophilus milk (AS) and propolis extract (PE) in the mouse xenograft breast cancer model. For this purpose, firstly cytotoxic effect of PE was determined by MTT assay against 4 T1 mouse breast cancer cells. Apoptotic effect of PE analyzed by flow cytometry. The antibacterial activity of PE was determined by the 96-well microplate broth-dilution method on Lactobacillus acidophilus LA-5. Then, Balb/c mice were injected subcutaneously with 4 T1 cells (2x105 cells/mouse) and also mice were given daily oral gavage with PE (66 mg/kg/day) and/or acidophilus milk (108 CFU/mL/mouse/day) for 14 days. The Balb/c mice were weighed throughout the study, and the tumor sizes were measured by caliper at the 14th day. The proliferation of splenocytes which collected spleen from mice was measured by MTT. CD8 + T cell response was analyzed by flow cytometry and results were evaluated in comparison with control and tumor control groups. The IC50 value for PE on 4 T1 cells was determined as 129.25 ± 1.90 µg/mL. The apoptotic effect of PE at IC50 concentration was determined as 3.3% of cells to late-apoptosis, 4.3% of cells to pro-apoptosis and 2.5% of cells to necrosis. The MIC and MBC values for PE on L. acidophilus LA-5 were 5000 ppm. The treatment of PE, AS and the combination of PE and AS were inhibited the tumor volumes by 59.16%, 28.29% and 63.39%, respectively. Acidophilus milk and PE combination significantly enhanced the ConA-, LPS- and PHA-induced splenocyte proliferation (P < 0.05). The acidophilus milk and PE combination were also found to stimulate IFN- γ production. In conclusion, the best anti-tumor effect was obtained by the combination of acidophilus milk and propolis.

Chemical Characterization and Biological Activity of the Mastic Gum Essential Oils of Pistacia lentiscus Var. Chia from Turkey.

The essential oils (EOs) were isolated by hydrodistillation from wild and cultivated Pistacia lentiscus L. var. chia-mastic gum tree (Anacardiaceae) from two natural habitats, namely from Cesme-Uzunkoy (1) and Mordogan (2), and one cultivated source, Cesme-Germiyan (3), in Izmir, Turkey. This comparative study evaluated the chemical composition and biological activity of mastic gum essential oils (MGEOs). For this purpose, MGEOs 1-3 were analyzed by gas chromatography-flame ionization detection (GC-FID), gas chromatography-mass spectrometry (GC-MS), and chiral GC for α-pinene. Laboratory assays were conducted to assess for potential in vitro cytotoxicity (multiple in vitro cancer cell lines), antimicrobial properties (five bacterial species and yeast), anti-inflammatory activity (inhibition of inducible nitric oxide synthase, iNOS), and the attraction of Ceratitis capitata (Mediterranean fruit fly, medfly), respectively. Chemical analysis indicated that MGEOs 1 and 2 were rich in α-pinene (56.2% and 51.9%), myrcene (20.1% and 18.6%), and ß-pinene (2.7% and 3.1%), respectively; whereas MGEO-3 was characterized by a high level of α-pinene (70.8%), followed by ß-pinene (5.7%) and myrcene (2.5%). Chiral GC analyses showed that concentration ratios between (-)/(+)-α-pinene and (-)-α-pinene/myrcene allowed for differentiation between wild and cultivated MGEO sources. In biological assays, MGEOs 1-3 did not exhibit significant antimicrobial effects against the pathogens evaluated and were not strong attractants of male medflies; however, all three MGEOs displayed a dose-dependent inhibition of iNOS, and MGEOs 1 and 2 exhibited selective in vitro cytotoxicity against human cancer cells. These results suggest that wild-type mastic gum oils from Cesme and Mordogan (MGEOs 1 and 2) are potential sources of beneficial products and warrant further investigation.

Triterpene glycosides from Silene odontopetala.

Silene species are known for their use in traditional medicine in treating several diseases. To the authors' knowledge there is no report on the chemical composition of S. odontopetala. Therefore, the phytochemical investigation of the methanol extract of S. odontopetala was carried out, leading to the isolation of six undescribed oleanane-type glycosides along with the known saponin azukisaponin IV. Their structures were elucidated by the analysis of 1D and 2D-NMR experiments, along with mass spectrometry analysis. The cytotoxic activity of oleanane-type saponins was evaluated against a small panel of cancer cell lines, including PC-3 (prostate carcinoma cells), MCF-7 (breast cancer cells), A549 (alveolar basal carcinoma cells), and HeLa (cervical carcinoma cells). Furthermore, the activity of isolated compounds against a normal cell line HEK-293, used for assessing their cytotoxicity, was evaluated.

Cytotoxic, antimicrobial and nitric oxide inhibitory activities of supercritical carbon dioxide extracted Prunus persica leaves.

Different parts of Prunus persica as fruits, flowers, leaves and kernels have been consumed with dietary and therapeutic purposes traditionally. During fruit production, remarkable amount of leaves which can hold important bioactive groups as phenolics, have been left unutilized. The aim of this study was to investigate cytotoxic, antimicrobial and nitric oxide inhibitory activities of supercritical carbondioxide extracts of Prunus persica leaves. Among studied cell lines, supercritical carbon dioxide extract which was processed at 150 bar, 60 °C, and 6% co-solvent ethanol, exhibited remarkable cytotoxic activity against HeLa, MPanc-96 and MCF-7 cell lines with IC50 values of 12.22 µg/ml, 28.17 µg/ml and 35.51 µg/ml respectively, whereas IC50 value of conventional solvent extract was above 50 µg/ml. Minimum inhibitory concentration values determined for antibacterial and antifungal activities against Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Enterococcus faecium and Candida albicans were found as 62.50 µg/ml. Strong nitric oxide inhibition was achieved with IC50 of 9.30 µg/ml. The promising results revealed that Prunus persica leaves may have remarkable potential as supplement both for drug and food industries. This study is the first report revealing cytotoxic, antimicrobial and nitric oxide inhibitory activity of supercritical carbon dioxide extract of Prunus persica leaves.

Development of adjuvant nanocarrier systems for seasonal influenza A (H3N2) vaccine based on Astragaloside VII and gum tragacanth (APS).

Adjuvants are chemical/biological substances that are used in vaccines to increase the immunogenicity of antigens. A few adjuvants have been developed for use in human vaccines because of their limitations including lack of efficacy, unacceptable local or systemic toxicity, the difficulty of manufacturing, poor stability, and high cost. For that reasons, novel adjuvants/adjuvant systems are under search. Astragaloside VII (AST-VII), isolated from Astragalus trojanus, exhibited significant cellular and humoral immune responses. The polysaccharides (APS) obtained from the roots of Astragalus species have been used in traditional Chinese medicine and possess strong immunomodulatory properties. In the present study, the immunomodulatory effects of a newly developed nanocarrier system (APNS: APS containing carrier) and its AST-VII containing formulation (ANS: AST-VII + APNS), on seasonal influenza A (H3N2) vaccine were investigated. Inactivated H3N2 alone or its combinations with test compounds/formulations were intramuscularly injected into Swiss albino mice. Four weeks after immunization, the immune responses were evaluated in terms of antibody and cytokine responses as well as splenocyte proliferation. APNS demonstrated Th2 mediated response by increasing IgG1 antibody titers, whereas ANS showed response towards Th1/Th2 balance and Th17 by producing of IFN-γ, IL-17A and IgG2a. Based on these results, we propose that APNS and ANS are good candidates to be utilized in seasonal influenza A vaccines as adjuvants/carrier systems.

Anti-inflammatory properties of Centaurea calolepis Boiss. and cnicin against Macrovipera lebetina obtusa (Dwigubsky, 1832) and Montivipera xanthina (Gray, 1849) venoms in rat.

Macrovipera lebetina obtusa (Dwigubsky, 1832) and Montivipera xanthina (Gray, 1849) (Ottoman Viper) are viper snakes from Viperidae family and found in various locations in Anatolia. Both snakes are responsible for major snake bite cases in Turkey Their venoms cause necrosis, hemorrhage, pain and local edema. Centaurea L. (Asteraceae) species draw attention as potential anti-inflammatory sources due to their traditional uses and accomplished studies on this field. C. calolepis Boiss. is an endemic taxon distributed in Aegean and Antalya regions in Turkey. Chloroform extract of C. calolepis and its major compound cnicin, a sesquiterpene lactone, are reported to have strong anti-inflammatory activities in-vitro, by previous studies. In the present study, in-vivo anti-inflammatory activities of C. calolepis chloroform extract and the sesquiterpenoid cnicin against edema induced by Macrovipera lebetina obtusa and Montivipera xanthina venoms were evaluated in the rat model. Protein contents and induction doses of the venoms were determined. Carrageenan and snake venoms were used as inducing agents in paw edema tests. Extract demonstrated strong inhibition on edema at all doses and hours against M. xanthina venom and carrageenan. Inhibition ratio of extract at 25 mg/kg dose (84.13% inhibition) after 0.5 h M. xanthina venom injection was more than indomethacin's value (45.4% inhibition). The extract also showed significant effect also on inflammation caused by M. lebetina obtusa venom at all doses. However, 2.5 mg/kg cnicin was more effective than total extract of C. calolepis against rat paw edema induced by (27.31%) M. lebetina obtusa venom. This is the first study reported therapeutic potential of C. calolepis, an endemic plant of Turkey, in case of snake-bites cause inflammation by venomous species in natural fauna of Anatolia.

Immunomodulatory, hemolytic and cytotoxic activity potentials of triterpenoid saponins from eight Cephalaria species.

BACKGROUND: Saponins isolated from a number of plants possess a broad spectrum of biological and pharmacological activities by using in vitro and in vivo bioassays. The recent investigations and findings in biological activity studies of saponins have mostly focused on immunomodulatory, hemolytic and cytotoxic properties. HYPOTHESIS/PURPOSE: Considering the great potential of saponins as bioactive agents, we investigated the cytotoxic, hemolytic and immunomodulatory activities of nineteen triterpenoid saponins from the aerial parts of eight Cephalaria species from Turkey. STUDY DESIGN: The isolated oleanane type saponins from Cephalaria species were tested for their hemolytic, cytotoxic and immunomodulatory activities through anin vitro cell based assay systems. METHODS: HeLa, A549, and a normal cell line HEK293 were used for testing cytotoxicity using MTT method. Immunomodulatory activity was performed in stimulated whole blood cells by PMA plus ionomycin. Hemolytic activity was assessed on human erythrocytes. RESULTS: Aristatoside C (3) and davisianoside B (16) displayed significant inhibitory effects on cancerous A549 and HeLa cells, and non-cancerous HEK293 cells with IC50 values of 3.52 ± 0.11, 35.69 ± 0.50, 8.96 ± 0.62 µM and 4.08 ± 0.06, 11.74 ± 0.82, 20.43 ± 3.21 µM, respectively. Thirteen saponins out of the nineteen tested increased IL-1ß concentrations considerably, while six compounds changed IL-2 or IFN-γ levels slightly. Almost all of the saponins showed noticeable hemolysis in human erythrocyte cells based on the sugar units. CONCLUSION: In this study, almost all saponins with oleanolic acid as aglycones exhibited significant hemolysis, monodesmosidic saponins with hederagenin as aglycone were the most active compounds against lung cancer cells with greater activity than standard commercial chemotherapy drug doxorubicin and some of the hederagenin type saponins induced remarkable IL-1ß secretion.

Secondary metabolites from Astragalus karjaginii BORISS and the evaluation of their effects on cytokine release and hemolysis.

A new cycloartane sapogenol and a new cycloartane xyloside were isolated from Astragalus karjaginii BORISS along with thirteen known compounds. The structures of the new compounds were established as 3-oxo-6α,16ß,24(S),25-tetrahydroxycycloartane (1) and 6-O-ß-d-xylopyranosyl-3ß,6α,16ß,24(S),25-pentahydroxycycloartane (2) by 1D- and 2D-NMR experiments as well as ESIMS and HRMS analyses. The presence of the keto function at position 3 was reported for the first time for cyclocanthogenol sapogenin of Astragalus genus. In vitro immunomodulatory effects of the new compounds (1 and 2) along with the n-BuOH and MeOH extracts of A. karjaginii at two different doses (3 and 6µg) were tested on human whole blood for in vitro cytokine release (IL-2, IL-17A and IFN-γ) and hemolytic activities. The results confirmed that compound 2, a monodesmosidic saponin, had the strongest effect on the induction of both IL-2 (6µg, 6345.41±0.12pg/mL (×5), P<0.001) and a slight effect upon IL-17A (3µg, 5217.85±0.72pg/mL, P<0.05) cytokines compared to the other test compounds and positive controls (AST VII: Astragaloside VII; and QS-21: Quillaja saponin 21). All tested extracts and molecules also induced release of IFN-γ remarkably ranging between 5031.95±0.05pg/mL, P<0.001 for MeOH extract (6µg) and 5877.08±0.06pg/mL, P<0.001 for compound 1 (6µg) compared to QS-21 (6µg, 5924.87±0.1pg/mL, P<0.001). Administration of AST VII and other test compounds did not cause any hemolytic activity, whereas QS-21 resulted a noteworthy hemolysis.

Chemical constituents and in vitro biological activities of Eremurus spectabilis leaves.

In the present study, phytochemical investigation of the young leaves of Eremurus spectabilis let to the isolation six compounds (1-6). Their structure was determined by spectroscopic analysis and comparison with the literature data. All compounds were isolated for the first time from E. spectabilis. Isolated compounds and prepared extracts did not show cytotoxic activity against HeLa (Human cervix adenocarcinoma), A-549 (Human alveolar adenocarcinoma), MCF-7 (Human breast adenocarcinoma), mPANC96 (Human pancreas adenocarcinoma), U87MG (Glioblastoma), PC3 (Prostate), CaCo-2 (Colon), HEK293 (Human embryonic kidney cells) cell lines. RAW 264.7 (murine macrophage cell lines) was used to determine the inhibition levels of inducible nitric oxide synthase (iNOS).The hexane extract was found to be active in terms of inhibition of iNOS (IC50: 25 µg/ml).

Immunomodulatory, hemolytic properties and cytotoxic activity potent of triterpenoid saponins from Cephalaria balansae.

Phytochemical investigations on n-butanol extract of Cephalaria balansae Raus. (Caprifoliaceae) led to the isolation of four previously undescribed triterpenoid saponins based on hederagenin type aglycone, namely, balansoides A-D, along with ten known compounds. Their structures were proposed based on 1D and 2D NMR spectroscopic data, HRESIMS analysis and chemical evidence as 3-O-α-L-rhamnopyranosyl-(1 â†’ 3)-ß-D-xylopyranosyl-(1 â†’ 3)-α-L-rhamnopyranosyl-(1 â†’ 2)-α-L-arabinopyranosyl hederagenin, 3-O-α-L-rhamnopyranosyl-(1 â†’ 3)-ß-D-xylopyranosyl-(1 â†’ 3)-α-L-rhamnopyranosyl-(1 â†’ 2)-α-L-arabinopyranosyl hederagenin 28-O-ß-D-glucopyranosyl ester, 3-O-α-L-rhamnopyranosyl-(1 â†’ 3)-ß-D-xylopyranosyl-(1 â†’ 3)-α-L-rhamnopyranosyl-(1 â†’ 2)-α-L-arabinopyranosyl hederagenin 28-O-ß-D-galactopyranosyl-(1 â†’ 6)-ß-D-glucopyranosyl ester and 3-O-α-L-rhamnopyranosyl-(1 â†’ 3)-ß-D-xylopyranosyl-(1 â†’ 3)-α-L-rhamnopyranosyl-(1 â†’ 2)-α-L-arabinopyranosyl hederagenin 28-O-ß-D-glucopyranosyl-(1 â†’ 6)-ß-D-glucopyranosyl ester. The four saponins were evaluated for their potential cytotoxic activity against HEK-293, A-549, HeLa cells and for hemolytic properties on human blood cells. Balansoide A displayed significant inhibitory effects on cancerous A-549 and HeLa cells, and non-cancerous HEK-293 cells with IC50 values of 12, 15 and 8 µM, respectively. Balansoides A-D together with n-butanol extract exhibited considerable hemolysis in human erythrocyte cells. Immunomodulatory properties of balansoides A-D were also evaluated in activated whole blood cells by PMA plus ionomycin. While balansoides A-C increased IL-1ß concentration with values of 1004.47, 991.57 and 966.50 pg/ml, only balansoide B augmented a slight IFN-γ secretion with value of 5219.14 pg/ml. None of the compounds changed IL-2 levels significantly.

Cytotoxic and Nitric Oxide Inhibition Activities of Propolis Extract along with Microencapsulation by Complex Coacervation.

In this study, cytotoxicity of ethanol extract of propolis (EEP) originating from Sivas, Turkey was screened against several cancer cell lines, namely PC-3, U87MG, A-549, mPANC96, CaCo-2, MCF-7, HeLa, MDA-MB-231 and a non-tumor cell line HEK293 by MTT assay. The inhibition levels of inducible nitric oxide synthase (iNOS) were also determined by using RAW 264.7 macrophage cells following lipopolysaccharide (LPS) treatment. EEP exhibited significant cytotoxic nitric oxide inhibition activities with an IC50 value of 0.1 ± 0.1 µg/ml indicating a high potential as an anti-inflammatory agent. In spite of these promising results and the fact that propolis is a highly nutritive substance, its low solubility and bitter taste limit the applications as a natural supplement. Encapsulation might serve as a good strategy in order to overcome these problems. Complex coacervation was applied where the main focus was on surfactant type, polymer ratio (alginate:gelatin), stirring rate and concentration of core material. The mean particle size of unloaded microparticles were 22.62 µm obtained with gelatin:alginate ratio of 1:1 at a stirring rate of 1400 rpm with 2 ml of 1 % (w/v) sodium carboxymethyl cellulose (Na-CMC), whereas addition of EEP at a concentration of 100 mg/ml increased the mean particle size to 36.44 µm and yielded an encapsulation efficiency of 98.77 %. The cytotoxicities of EEP loaded microparticles were also assessed both on MCF-7 and MDA-MB-231 where similar results were achieved as free EEP which can enhance the possible use of propolis extract in the industry as a natural supplement.

The Induction of Growth Inhibition and Apoptosis in HeLa and MCF-7 Cells by Teucrium sandrasicum, Having Effective Antioxidant Properties.

The hidromethanolic (Met/W), ethyl acetate (EA(EA/W)), and water (W(EA/W)) extracts from Teucrium sandrasicum leaves (L) and flowers (F) were investigated for antioxidant properties and antiproliferative effects on HeLa, MCF-7, and L929. The highest DPPH scavenging, metal chelating capacities, and total phenolic and flavonoid contents were observed in Met/WL. The highest hydroxyl scavenging and reducing power capacities were found in EA(EA/W)L. Met/WL, EA(EA/W)L and EA(EA/W)F inhibited cancer cell growths, while they did not show significant cytotoxicity on L929. While the reactive oxygen species (ROS) levels were generally close to controls in HeLa, they were induced in MCF-7 with the treatment of Met/WL, EA(EA/W)L, and EA(EA/W)F and acted as antioxidant for L929. The highest apoptosis inductions were observed in Met/WL-treated HeLa and EA(EA/W)L-treated MCF-7, which were supported with the changes in mitochondrial membrane potentials. The highest caspase-9 activities were found in Met/WL-treated HeLa and EA(EA/W)F-treated MCF-7. Caspase-3 activity was only induced in EA(EA/W)F-treated HeLa.

Essential Oil Composition of Pimpinella cypria and its Insecticidal, Cytotoxic, and Antimicrobial Activity.

A water-distilled essential oil (E) from the aerial parts of Pimpinella cypria Boiss. (Apiaceae), an endemic species in northern Cyprus, was analyzed by GC- FID and GC-MS. Forty-five compounds were identified in the oil, which comprised 81.7% of the total composition. The compound classes in the oil were oxygenated sesquiterpenes (33.9%), sesquiterpenes (22.0%), monoterpenes (11.4%), oxygenated monoterpenes (2.6%), and phenylpropanoids (7.5%). The main components of the oil were (Z)-ß-farnesene (6.0%), spathulenol (5.9%), ar-curcumene (4.3%), and 1,5-epoxy-salvial(4)14-ene (3.8%). The P. cypria EO deterred yellow fever mosquitoes (Aedes aegypti) from biting at a concentration of 10 µg/cm2 in in vitro bioassays. The oil was tested for repellency in assays using human volunteers. The oil had a minimum effective dosage (MED) for repellency of 47 ± 41 µg/cm² against Ae. aegypti, which was less efficacious than the positive control NN-diethyl-3-methylbenzamide (DEET). In larval bioassays, P. cypria EO showed an LC50 value of 28.3 ppm against 1st instar Ae. aegypti larvae. P. cypria EO demonstrated dose dependent repellency against nymphs of the lone star tick, Amblyomma americanum. Between 45.0% and 85.0% repellency was observed at concentrations ranging from 26 to 208 µg/cm². However, P. cypria EO was less effective compared with DEET in the tick bioassays. Cytotoxicity assays showed that the P. cypria EO did not exhibit significant effects up to the maximum treatment concentration of 50 µg/mL on HEK293, PC3, U87MG, and MCF cells. P. cypria EO also demonstrated moderate antimicrobial activity against Gram-negative and -positive bacteria with MICs ranging from 15.6 to 62.5 µg/mL, except for Candida albicans, which showed the same MIC value of 7.8 µg/mL as the positive control, flucytosine. This is the first report on the chemical composition of P. cypria EO and its insecticidal, toxicant, cytotoxic, and antimicrobial activity.

In vitro synergistic efficacy of conjugated linoleic acid, oleic acid, safflower oil and taxol cytotoxicity on PC3 cells.

The aim of this study was to determine in vitro synergistic efficacy of conjugated linoleic acid (CLA), oleic acid (OLA), safflower oil and taxol (Tax) cytotoxicity on human prostate cancer (PC3) cell line. To determine synergistic efficacy of oil combinations, PC3 treated with different doses of compounds alone and combined with 10 µg/mL Tax. The MTT results indicated that OLA-Tax combinations exhibited cytotoxicity against PC3 at doses of 30 nM+10 µg-Tax, 15 nM+5 µg-Tax and 7.5 nM+2.5 µg-Tax. The treatment of OLA or Tax did not show significant inhibition on PC3, while OLA-Tax combinations showed effective cytotoxicity at treated doses. CLA-Tax combinations demonstrated the same effect on PC3 as combined form with 45.72% versus the alone form as 74.51% viability. Cytotoxic synergy between Tax, OLA and CLA shows enhanced cytotoxicity on PC3 which might be used in the therapy of prostate cancer.

Bioactivity screening of five Centaurea species and in vivo anti-inflammatory activity of C. athoa.

CONTEXT: Centaurea L. (Asteraceae) species used as herbal remedies in Turkish traditional medicine have shown several biological properties. OBJECTIVE: Extracts obtained from the aerial parts of Centaurea aphrodisea Boiss., Centaurea athoa DC., Centaurea hyalolepis Boiss., Centaurea iberica Trev. and Centaurea polyclada DC. were evaluated for their antioxidant, cytotoxic and anti-inflammatory activities. MATERIALS AND METHODS: Extracts of Centaurea species were tested for their antioxidant activity in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) screening assays and for in vitro anti-inflammatory activity by Nf-κB and iNOS inhibition assays. The extracts were tested for their in vitro cytotoxicities against a panel of human solid tumor cell lines (SK-MEL: malignant melanoma, KB: oral epidermal carcinoma, BT-549: breast ductal carcinoma and SK-OV-3: ovary carcinoma) as well as non-cancerous kidney fibroblast (Vero) and kidney epithelial cells (LLC-PK1) by Neutral Red assay. In vivo anti-inflammatory activity of C. athoa was evaluated by the carrageenan-induced paw edema test in rats. RESULTS: Antioxidant activities were observed for methanol extracts of plants. C. polyclada had the strongest effect on BT-549, KB and SK-OV-3 cell lines (30, 33 and 47 µg/ml, respectively). Nf-κB inhibition of chloroform extract of C. athoa was determined equivalent to positive control parthenolide (IC50: 6 µg/ml). This extract also showed anti-inflammatory activity by the carrageenan-induced paw edema test in rats, in all hours at a dose of 50 mg/kg compared to the control group. DISCUSSION AND CONCLUSION: C. athoa is suggested to be a potential source of lead compounds for inflammatory diseases due to the significant in vitro and in vivo anti-inflammatory results.

Viper venom induced inflammation with Montivipera xanthina (Gray, 1849) and the anti-snake venom activities of Artemisia absinthium L. in rat.

The present study was conducted to explore the characterization of Montivipera xanthina crude venom partially by in vitro and in vivo and the anti-snake venom activities of Artemisia absinthium L. in comparison with carrageenan-induced acute inflammation model in rats. The LD50 value was estimated as 8.78 mg/kg within 24 h by different venom doses administrated intraperitoneally in mice. The IC50 value was 0.43 ± 0.18 µg/ml after 48 h treatment while the calculated value was 0.73 ± 0.10 µg/ml for the culture media totally refreshed after 2 h treatment with venom. Wistar rats were treated intraperitoneally with A. absinthium extract, 30 min before venom or carrageenan was injected subplantarly into the left hind paw. Intraperitoneal administration of 25 and 50 mg/kg extract was inhibited venom induced paw swelling at 0.5, 1, 2 and 3 h (p < 0.05) while 12.5, 25 and 50 mg/kg extract treatment was inhibited carrageenan-induced paw swelling at 2, 3, 4 and 5 h (p < 0.05). In conclusion, the in vivo toxicity and inflammatory actions and in vitro cytotoxic actions of crude M. xanthina venom were performed as a first report and inhibition of venom-induced inflammation by methanolic extract of A. absinthium was described.

Evaluation of the immunomodulatory properties in mice and in vitro anti-inflammatory activity of cycloartane type saponins from Astragalus species.

ETHNOPHARMACOLOGICAL RELEVANCE: Astragalus roots are used to treat leukemia and for their wound healing properties in Southeast Anatolia-Turkey. MATERIALS AND METHODS: In vivo studies to investigate the effects of two Astragalus saponins were carried out on the immune response cytokines by using six to eight weeks old male Swiss albino mice. The production of IL-1ß, TGF-1ß, TNF-α, IL-2, IL-4 and IFN-γ cytokines was determined by ELISA. The spleen and lymph nodes, isolated from the mice subjects, were analyzed to realize induction of the surface antigen productions for IL-2Rα (CD25) and CD69. In addition, their effects on the targets of inflammation such as NF κB, iNOS and NAG-1 were investigated in cell-based assays. RESULTS: The results suggested that AST VII and Mac B had positive effect on Th1 cytokine release (IL-2 and IFN-γ), and suppression on Th2 cytokine production (IL-4). The immunohistochemical results exhibited induction of both IL-Rα (CD25) and CD69 surface receptors justifying the Th1 cytokine release. The compounds did not affect NF-κB or NAG-1 activity but iNOS activity was inhibited by Mac B with an IC(50) of 156 µg/ml. CONCLUSIONS: The results show that Ast VII and Mac B create powerful immunoregulatory effects without the stimulation of inflammatory cytokines in mice, and have no significant effect on the inflammatory cellular targets in vitro.