Total Content of Saponins, Phenols and Flavonoids and Antioxidant and Antimicrobial Activity of In Vitro Culture of Allochrusa gypsophiloides (Regel) Schischk Compared to Wild Plants.

Allochrusa gypsophiloides is a rare Central Asian species, a super-producer of triterpene saponins with pharmacological and technical value. In this work, a comparative evaluation of the in vitro culture of adventitious roots (ARs), in vitro adventitious microshoots (ASs), natural roots and aboveground parts of wild plants from Kazakhstan to define the total saponin (TS), phenol (TP) and flavonoid (TF) content, as well as antioxidant (AOA) and antimicrobial activity, is presented for the first time. In the AR culture, growth index (GI), TS, TP and TF were evaluated on days 25, 45 and 60 of cultivation on ½ MS medium without (control) and with auxin application. It was found out that TS and TF were higher in the in vitro AR culture. The amount of TP and TF are higher in the aerial part of vegetative plants with maximum AOA. The concentration of the extract required to inhibit 50% of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical formation (ICO50) in extracts from natural material negatively correlated with TS, TP, TF and in the in vitro AR culture with TF. Control extracts from the in vitro AR culture with high TS levels showed growth-inhibitory activity against S. thermophillus, S. cerevisiae and C. albicans. The influence shares of medium composition factor, cultivation duration factor and their interaction with GI, TS, TP and TF were determined. The in vitro AR culture is promising for obtaining triterpene saponins TSR with high antibacterial and antifungal activity, and the in vitro ASs culture-for shoot multiplication with antioxidant properties.

Antioxidant, Cytotoxicity, Apoptotic Properties of Extracts of Andricus sternlichti galls and Their Phenolic Characterisation by HPLC.

Quercus L. galls have been used in Western and Eastern cultures for various diseases in traditional medicine. Galls are also used in the East for many purposes, including consumption as food, commercial inks, leather tanning. In the current study, Andricus sternlichti Bellido, Pujade-Villar & Melika, 2003 galls were extracted in different solvents. The possible antioxidant effects of gall extracts were determined using 7 different methods (ß-carotene-linoleic acid assay, Phosphomolybdenum assay, DPPH and ABTS radical scavenging activity, CUPRAC and FRAP assay, Metal Chelating activity) to support each other. Total phenolic, flavonoid and tannin amounts of extracts are calculated by using standard curves. In addition, HPLC method used to characterize the phenolic component with 15 different standards. The MIA PaCa-2 cell lines was preferred to identify possible cytotoxic activities of galls. Expression of some genes (Bax, Bcl-2, FAS, BID, caspase-3, caspase-8, caspase-9, caspase-10, FADD, TRADD) role in the apoptosis was determined to investigate apoptotic effects of extracts. According the results, the gall extracts of A. sternlichti may be considered as a potential source of biological agents for their antioxidant capacity and rich bioactive compounds. The gall extracts exhibit antiproliferative activity via regulating expressions of apoptotic genes.

Assessment of Antioxidant and Cytotoxic Activities and Identification of Phenolic Compounds of Centaurea solstitialis and Urospermum picroides from Turkey

HIGHLIGHTS The phenolic composition, antioxidant activity and cytotoxic potential of the extracts of C. solstitialis and U. picroides were investigated. Caffeic acid was found as the most abundant phenolic compound in the extracts. Both species showed promising antioxidant activity towards different assays. The highest cytotoxic potential was observed in the extract of C. solstitialis.

Abstract It is known that some genera of the Asteraceae family are commonly used in Turkish folk medicine. Several studies have investigated the biological effects of different extracts of Centaurea and Urospermum species, but studies involving the phenolic composition of C. solstitialis and U. picroides extracts are very limited. This study aimed to investigate the phenolic composition and antioxidant activity of C. solstitialis and U. picroides and evaluate their possible cytotoxic effect. RP-HPLC analysis was used to elucidate the phenolic profiles of the ethanolic extracts of flowering parts of C. solstitialis and U. picroides.The both ethanolic extracts were assessed for their antioxidant properties using DPPH, FRAP, phosphomolybdenum and metal chelating assays. Furthermore, the effect of the extracts on cell viability was evaluated against MCF-7 and PC-3 cancer cells and HEK293 cell line using the MTT assay. The most abundant phenolic compound in both extracts was determined to be caffeic acid, and the amount of this compound was 24078.03 and 14329.59 µg g-1 in the extracts of C. solstitialis and U. picroides, respectively. The antioxidant activity of the extracts was found similar. Compared with U. picroides extract, C. solstitialis extract had higher potential on the inhibition of cell viability. The IC50 value of C. solstitialis on MCF cells was found as 58.53 µg mL-1. These data suggest that the extracts of C. solstitialis and U. picroides may be considered as novel and alternative natural antioxidant and anticancer sources.

The Effects of Injected Geophyte Plant (Sternbergia candida) Extract on the Hemato-Immunological Parameters in Gilthead Seabream, Sparus aurata (L. 1758)

Abstract This study investigated effects of a geophyte plant extract (Sternbergia candida) on some hemato-immunological parameters in gilthead seabream, Sparus aurata (L. 1758). To serve this purpose, the plant was collected and extracted in ethanol solvent. A total 300 fish were used in experiment. Three groups were designed including control group, 0.5 mg/fish and 2 mg/fish and plant extract was given by intraperitoneal injection. Following injections on the 1st, 7th, 14th, 21st, 28th and 35th days, blood samples were collected from each group. Some of the hemato-immunological parameters in serum and blood samples such as, phagocytic activity (NBT (+) cells), total leukocyte level, lysozyme activity, total protein, hematocrit level and the differential leucocyte were examined. Present results show that intraperitoneal injection of S. candida plant extract increases some non-specific immune parameters. In the determination of the non-specific immune responses of the gilthead seabream, blood samples collected from 2mg/fish group indicated that maximum levels of pre-determined parameters were reached on days 7, 14, and 21 following the injection. According to these finding, administration of geophyte plant is suggested in seabream to improve non-specific immune response and health status.

Antioxidant activity, phytochemical composition of Andricus tomentosus and its antiproliferative effect on Mia-Paca2 cell line.

Plant derived products are widely used in cancer treatment. Gall species has been preferred for treatment various diseases in folk medicine, but there are few studies on the anticancer effects of gall species. The present study reports the antioxidant activity and total secondary metabolites of extracts of A. tomentosus galls. The antioxidant potency of galls was carried out using different in-vitro model systems. Their cytotoxic efficacy on Mia-Paca cell line was also explored. Gall extract was found to contain a large amount of phenolic acids. The extract potently scavenged free radicals including DPPH (IC50 9.56 ± 1.08 µg/mL), ABTS (IC50 18.51 ± 0.25 µg/mL). It can be seen as a potential source of antioxidants according to ß-carotene/linoleic acid method (%92.58 ± 0.92) and Phosphomolybdenum assays (104.36 ± 4.95 mgAE/g). Gall extract also posses ability of metal chelating (%40.07 ± 2.30) and Fe3+ (184.01 ± 2.83 mgTE/g) and Cu2+ (89.81 ± 0.96 mgTE/g) reducing activity. According to total secondary metabolites tests, gall extract showed high total phenolic, total flavonoid and total tannin amount. HPLC analysis of the extract suggested it to contain caffeic acid (424.068.479 µg/g), ellagic acid (187.696.132 µg/g). XTT assay revealed gall extract to enhance percent survival of Mia-Paca2 cell line exposed A. tomentosus extracts. The best cytotoxic effect was determined in acetone extracts (IC50: 124.7 µM). Expression of some genes (Bax, Bcl-2, FAS, BID, caspase-3, caspase-8, caspase-9, caspase-10, FADD, TRADD) in the apoptosis pathway was determined to invastigate apoptosis inducing activity. These results indicate that A. tomentosus galls possess potent antioxidant activity, when tested both in chemical as well as biological models.

Chemical composition and bioactivity of essential oils and Ethanolic extracts of Ocimum basilicum L. and Thymus algeriensis Boiss. & Reut. from the Algerian Saharan Atlas.

BACKGROUND: There is increasing interest in the pharmaceutical and food industries to substitute synthetic chemicals with naturally occurring compounds possessing bioactive properties. Plants are valuable sources of bioactive compounds. The present study investigates the chemical composition and antioxidant, antimicrobial, and anticancer activities of ethanolic extracts (EEs) and essential oils (EOs) from two species in the Lamiaceae family, Ocimum basilicum L. and Thymus algeriensis Boiss. & Reut., cultivated in the Algerian Saharan Atlas. METHODS: The total flavonoid contents of the plants' ethanolic extracts were determined by the aluminium chloride method, while the total phenols were determined using the Folin-Ciocalteu method. Essential oils were obtained by hydrodistillation of the aerial parts of the plants and were analysed by GC-MS. The free radical-scavenging ability and antioxidant potential of the plants' EEs and EOs were probed using the 2, 2-diphenyl-picrylhydrazyl radical-scavenging, ABTS radical-scavenging, ferric-reducing power and phosphomolybdenum assays. The antimicrobial activities were evaluated against several pathogens characteristic of gram-negative bacteria (three species), gram-positive bacteria (three species) and fungi (two species). The microdilution method was used to estimate the minimum inhibitory concentrations (MICs). The oils' anticancer potential against several cancer types was also studied using the MTT assay and reported as the toxic doses that resulted in a 50% reduction in cancer cell growth (LD50). RESULTS: Phenolic compounds in the EEs from both plants were analysed by HPLC and demonstrated a rich flavonoid content. Chemical analysis of the essential oil from Ocimum basilicum revealed 26 unique compounds, with linalool (52.1%) and linalyl acetate (19.1%) as the major compounds. A total of 29 compounds were identified in the essential oil from Thymus algeriensis, with α-terpinyl acetate (47.4%), neryl acetate (9.6%), and α-pinene (6.8%) as the major compounds. The ethanolic extracts and essential oils from both plants exhibited moderate antioxidant activities and moderate to weak antimicrobial activities. Furthermore, anticancer activities against the examined human cancer cell lines were associated with only the EOs from both plants, with LD50 values ranging between 300 and 1000 µg/mL. CONCLUSION: The results suggest that the bioactive compounds found in the ethanolic extracts and essential oils from Ocimum basilicum and Thymus algeriensis, with diverse antioxidant, antimicrobial and anticancer activities, may have beneficial applications in nutraceutical and pharmaceutical technologies.

The determination of the potential anticancer effects of Coriandrum sativum in PC-3 and LNCaP prostate cancer cell lines.

Coriander (Coriandrum sativum L.) is such an herb from the Apiaceae family, used both for its medicinal and nutritional properties for many centuries. In this study, the effects of C. sativum extract on gene expression, viability, colony formation, migration, and invasion of PC-3 and LNCaP prostate cancer cell lines have been investigated. The half maximal inhibitory concentration (IC50 ) dose in PC-3 and LNCaP cells was detected to be 2 and 5 mg/mL at the 24th hour, respectively. C. sativum extracts have been observed to cause a significant decrease in the expression of Akt and Bcl-2 in the PC-3 cells and just Akt in LNCaP cells while increasing in the expression of p53, caspase-9, caspase-10, PTEN, DR5, TRADD, PUMA, and NOXA. DR4 expression was increased in LNCaP cell line but not PC-3, and APAF and BID had increased expression in PC-3 but not the LNCaP cells. Our observations have shown that C. sativum extract decreased colony formation while inhibiting cell invasion and migration. Cell migration was hindered in PC-3 but not the LNCaP cells. In conclusion, this data present a valuable addition to the very limited data available out there on the potential use of C. sativum in prostate cancer treatment.

Evaluation of Antioxidant Properties, Phenolic Compounds, Anthelmintic, and Cytotoxic Activities of Various Extracts Isolated from Nepeta cadmea: An Endemic Plant for Turkey.

Nepeta cadmea Boiss. is a species endemic to Turkey that belongs to the Nepeta genus. Several species of this genus are used in folk medicine. This study was designed to investigate the phenolic compounds, antioxidant, anthelmintic, and cytotoxic activities of various extracts (ethanol, methanol, acetone, and water) of N. cadmea. The antioxidant activities of these extracts were analyzed using scavenging methods (DPPH, ABTS, and H2 O2 scavenging activity), the ß-carotene/linoleic acid test system, the phosphomolybdenum method, and metal chelating activity. Among the 4 different extracts of N. cadmea that were evaluated, the water extract showed the highest amount of radical scavenging (DPPH, 25.54 µg/mL and ABTS, 14.51 µg/mL) and antioxidant activities (ß-carotene, 86.91%). In the metal chelating and H2 O2 scavenging activities, the acetone extract was statistically different from the other extracts. For the phosphomolybdenum method, the antioxidant capacity of the extracts was in the range of 8.15 to 80.40 µg/mg. The phenolic content of the ethanol extract was examined using HPLC and determined some phenolics: epicatechin, chlorogenic, and caffeic acids. With regard to the anthelmintic properties, dose-dependent activity was observed in each of the extracts of N. cadmea. All the extracts exhibited high cytotoxic activities. The results will provide additional information for further studies on the biological activities of N. cadmea, while also helping us to understand the importance of this species. Furthermore, based on the results obtained, N. cadmea may be considered as a potentially useful supplement for the human diet, as well as a natural antioxidant for medicinal applications. PRACTICAL APPLICATION: The plants of the Nepeta genus have been extensively used as traditional herbal medicines. Nepeta cadmea Boiss., one of the species belonging to the Nepeta genus, is a species endemic to Turkey. In our study, we demonstrated the antioxidant capacities, total phenolic, flavonoid, tannin content, anthelmintic, and cytotoxic activities of various extracts of Nepeta cadmea. The present study could well supply valuable data for future investigations and further information on the potential use of this endemic plant for humans, in both dietary and pharmacological applications.

Anti-invasive effect of Cyclamen pseudibericum extract on A549 non-small cell lung carcinoma cells via inhibition of ZEB1 mediated by miR-200c.

Scientists are increasingly focusing attention on natural products of plant origin for use as agents in cancer protection and treatment. Cyclamen L. tuber extracts contain saponin glycosides that have been shown to have anti-cancer and other biological activities. The epithelial-to-mesenchymal transition (EMT) is thought to enhance malignant tumour progress. The transcriptional repressor zinc-finger E-box binding homeobox 1 (ZEB1) is an important inducer of EMT in different human tumours and has recently been shown to boost invasion by tumour cells. In this study, we investigated the effects of endemic Cyclamen pseudibericum (CP) saponin-rich tuber extract on the capacity of non-small cell lung cancer line A549 cells to proliferate, invade and migrate and also examined the expression levels of several invasion-migration-related microRNAs (miRNAs) to identify those which directly targeted ZEB1. The cytotoxicity effect of the CP extract on the A549 cancer cells was determined by the luminometric method. The half-minimal (50%) inhibitory concentration dose in the A549 cells was determined to be 41.64 ± 2.35 µg/mL. Using the Matrigel invasion chamber system and the wound healing assay we observed that the CP extract suppressed the invasion and migration capacity of A549 cells, respectively. The expression of miRNAs in A549 cells was evaluated by real-time PCR. Our data showed that overexpression of miRNA miR-200c hindered the EMT by increasing the expression of E-cadherin and decreasing the expression of both N-cadherin and vimentin through the direct targeting of ZEB1. These findings suggest that the saponin-rich tuber extract of CP may have considerable anti-cancer properties in lung cancer. Further studies are required to examine in detail the molecular-based mechanism involved in the EMT process of the extract along with isolation and identification of active saponin components.

Screening of some biological activities of Alyssum fulvescens var. fulvescens known as ege madwort.

In this research, the phenolic composition, antioxidant, antibacterial and cytotoxic activities of the methanolic extracts obtained from Alyssum fulvescens var. fulvescens aerial parts known as Ege kuduzotu in western Turkey, were firstly investigated. The antioxidant activity of the extract was determined by DPPH, metal chelating, phosphomolybdenum, ß-carotene/linoleic acid and ferric reducing power assays. Moreover, total phenolic and flavonoid contents in the extract were investigated. The brine shrimp (Artemia salina L.) lethality test was used to investigate for the possible cytotoxic activity of the extract. Microdilution broth method was used to study antibacterial potency of extract against Gram-positive and Gram-negative bacteria. The extract exhibited good biological activities. Total phenolic and flavonoid contents in the extract were significantly correlated with antioxidant potentials. HPLC analysis showed that chlorogenic acid was the major phenolic in extract tested. The results indicated that the extract of A. fulvescens var. fulvescens may be considered as a potential source of biological agents and in vivo investigations are needed to test the biological effects of A. fulvescens var. fulvescens.

Cytotoxic Activities of Certain Medicinal Plants on Different Cancer Cell Lines.

OBJECTIVES: In recent years, the use of plants for the prevention and treatment of cancer is gaining more attention due to their diverse range of phytochemical constituents and fewer adverse effects. In this study, four medicinal plant species from the Kars province of Turkey were investigated for their cytotoxic potential against six different cancer cell lines and one normal cell line. MATERIALS AND METHODS: MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-dipenyltetrazolium bromide] assay was performed to assess cytotoxic activity and apoptotic effect was determined using flow cytometry and caspase-3 analyses. RESULTS: Significant cytotoxicity (≥70%) was observed with the leaf extract of Artemisia absinthium on A-549, CCC-221, K-562, MCF-7, PC-3 cells, whereas seed extracts caused significant cytotoxicity (≥70%) on CCC-221, K-562, MCF-7, PC-3 cells. Selective cytotoxicity was obtained with leaf extract on A-549 and K-562 cells; and with seed extract on K-562, MCF-7 and PC-3 cells compared with normal Beas-2B cells. The levels of cytotoxicity for both extracts were time- and dose-dependent at lower concentrations. Moreover, selective cytotoxicity (78%) was detected on A-549 cells with the seed extract of Plantago major. Cytotoxicity of extracts from Hyoscyamus niger and Amaranthus retrosa ranged between 10% and 30%. CONCLUSION: A. absinthium extracts and P. major seed extract have potential for development as therapeutic agents for cytotoxicity on certain cancer cells following further investigation.

ASSESSMENT OF SOME BIOLOGICAL ACTIVITIES OF ALYSSUM L. KNOWN AS MADWORT.

In this research the phenolic composition, and antioxidant, antibacterial and cytotoxic activities of the methanolic extracts obtained from three Alyssum L. taxa (A.foliosum var. megalocaipum, A. simplex and A. stHgosum subsp. stfigosum) were investigated for the first time. The antioxidant activity of the extracts was evaluated by DPPH, metal chelating, phosphomolybdenum, P-carotene/linoleic acid and ferric reducing power assays. In addition, total phenolic and flavonoid contents in the extracts were determined. The brine shrimp lethality test was used to screen for possible cytotoxic activity of the extracts. Also, the extracts were evaluat- ed for cytotoxicity against three human cancer cell lines: MCF-7, H1299 and HeLa. HPLC analysis showed that cinnamic acid, chlorogenic acid and rutin were the major phenolics in extracts tested. Among the three Alyssum extracts evaluated, the highest biological activities were obtained from A. simplex compared to other two Alyssum L. taxa. Based upon its name, future studies will be targeted at investigating which components can be caused neurorehabilitation in Alyssum.

Theoretical calculations of a compound formed by Fe(+3) and tris(catechol).

Phenolic compounds generally have special smell, easily soluble in water, organic solvents (alcohols, esters, chloroform, ethyl acetate), in aqueous solutions of bases, colorless or colorful, crystalline and amorphous materials. Phenols form colorful complexes when they form compounds with heavy metals. In this study, the structural properties of a compound formed by catechol and Fe(+3) are investigated theoretically. The electronic and thermodynamic properties of the complex were also investigated in gas phase and organic solvents at B3LYP/6-31+G(d,p) and B3LYP/6-311++G(d,p) basis set. The formation of Fe-tris(catechol) complex compound is exothermic, and it is difficult to obtain the complex as the temperature increases. The observed and calculated FT-IR and geometric parameters spectra are in good agreement with empirical.

Larvicidal activity of Cyclamen (Myrsinaceae) extracts against the larvae of West Nile virus vector Culex pipiens L. (Diptera: Culicidae).

OBJECTIVE: To evaluate the larvicidal activity of the tuber extracts isolated from two species of Cyclamen Cyclamen mirabile Hildebr. and Cyclamen alpinum Dammann ex. Sprenger) against Culex pipiens L. (Diptera: Culicidae). METHODS: Twenty young (first-second) or older (third-fourth) instars of the Culex pipiens were exposed to various concentrations (100 to 1000 ppm) of the extracts. Mortality was recorded after 12-, 24-, 48-, 72- and 96-h of exposure. RESULTS: We determined that the young larval stages were more susceptible to the extracts in comparison with the older larval stages. A comparison of LC 50 and LC 90 values has shown that Cyclamen mirabile was more active than Cyclamen alpinum. CONCLUSIONS: This study is the first to report on the larvicidal activity of the extracts of Cyclamen species against Culex pipiens. More studies are needed to isolate and identify the active components involved, their mode of action, and effects on other mosquito and pest species.

The effects of Crataegus aronia var. dentata Browicz extract on biochemical indices and apoptosis in partially hepatectomized liver in rats.

Crataegus species have been widely used in herbal medicine, especially for the hearth diseases. In the present study, the effect of Crataegus aronia var. dentata Browicz extract on partially hepatectomized rats was investigated with biochemical and TUNEL apoptosis assays. The extracts of the plant at the concentrations of 0.5 and 1 ml/100 g body weight/day were administered orally to the two experimental groups including partially hepatectomized rats for 42 days. At the end of the experimental period, animals were sacrificed, blood was collected for the assessment of serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and gamma-glutamyltransferase (GGT), and the liver tissue was used for TUNEL assay. In biochemical assay, it was found a significant decrease in the levels of serum ALT and AST in the experimental groups. On the other hand, the plant extract did not cause any significant changes in the level of GGT in these groups. In apoptosis assay, TUNEL positive hepatocytes could not be detected in both experimental groups. The present findings can suggest that Crataegus aronia var. dentata Browicz extract can decrease the levels of serum ALT and AST and play a role in apoptosis of hepatocytes in the liver of partially hepatectomized rats. However, further studies are required to confirm the effects of the plant extract on hepatoprotection and apoptosis in the regenerating liver after partial hepatectomy in animal models.

Convolvulus galaticus, Crocus antalyensis, and Lilium candidum extracts show their antitumor activity through induction of p53-mediated apoptosis on human breast cancer cell line MCF-7 cells.

Conventional and newly emerging treatment procedures such as chemotherapy, catalytic therapy, photodynamic therapy, and radiotherapy have not succeeded in reversing the outcome of cancer diseases to any drastic extent, which has led researchers to investigate alternative treatment options. The extensive repertoire of traditional medicinal knowledge systems from various parts of the world are being re-investigated for their healing properties. It has been reported that several members of the Convolvulaceae, Iridaceae, and Liliaceae families have antitumor activity against some tumor cell lines. Here we first report that Convolvulus galaticus, Crocus antalyensis, and Lilium candidum species have cytotoxic activity on human breast cancer cell line MCF-7 cells. Plant samples were collected and identified, and their cytotoxic effects on the MCF-7 cell line were examined at different concentrations of methanol extracts. We found that all three plants have cytotoxic effects on MCF-7 cells but that C. galaticus has the strongest cytotoxic effect even in the lowest extract concentration tested (0.32 µg/mL). Our results indicate that these plant extracts have cytotoxic effects on human breast carcinoma cell line MCF-7 cells and that this cytotoxic effect comes from p53-mediated stimulation of apoptosis.