Abstract. Avemar, a derivative of fermented wheat germ extract, is a non-toxic and natural compound that is used as a dietary supplement by cancer patients undergoing chemotherapy and radiotherapy. Avemar has numerous biological activities, and several recent studies have reported that it may also have metastatic and anti-angiogenic effects. In the present study, the mechanism of the anti-angiogenic effect of Avemar on human cancer cells was investigated. The human cell lines NCI-N87 (gastric tubular adenocarcinoma), PC3 (prostate carcinoma), HeLa (endocervical adenocarcinoma) and A549 (lung adenocarcinoma) were treated with various doses (400, 800, 1,600 and 3,200 µg/ml) of Avemar, and the changes in mRNA and protein levels of two important markers of angiogenesis, vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (Cox-2), were assessed by reverse transcription-quantitative polymerase chain reaction and ELISA. VEGF and Cox-2 protein and mRNA levels were significantly lower in Avemar-treated cells than in untreated cells. The data suggest that Avemar may exert an anti-angiogenic effect on cancer cells. Thus, it is suggested to medical doctors as a potential agent for the anti-angiogenic treatment of cancer. IntroductionAngiogenesis, the physiological formation of new blood vessels from pre-existing ones (1), serves a central role in human physiology during fetal development, wound healing, tissue repair following surgery or trauma, menstruation, cancer, and various ischemic and inflammatory diseases (2). However, unregulated angiogenesis may result in angiogenic diseases, including diabetic retinopathy, rheumatoid arthritis, inflammatory diseases, or tumor growth and metastasis (3,4). As cancer growth is associated with angiogenesis, the inhibition of angiogenesis is a promising therapeutic strategy in cancer treatment. Furthermore, understanding the mechanisms of angiogenesis inhibition well enough to manipulate it may lead to numerous therapeutic possibilities.Avemar (fermented wheat germ extract) is produced by the industrial fermentation of wheat germ. Avemar is a completely natural and non-toxic compound that is used clinically as a dietary supplement for cancer patients undergoing chemotherapy and radiotherapy (5-9). It is known to have certain biological effects due its major components, 2-methoxy-benzoquinone and 2,6-dimethoxy-benzoquinone. Additionally, Avemar has been demonstrated to be associated with anaerobic glycolysis, the pentose cycle and ribonucleotide reductase enzymes; to exert significant anti-proliferative effects in a broad spectrum of tumor cell lines; and to possess the ability to kill tumor cells by inducing apoptosis through the caspase-poly ADP-ribose polymerase pathway (5,10). Furthermore, Avemar was reported to be an effective adjuvant agent in cancer treatment for several types of cancer. such as breast, colon, lung and prostate cancer (11). However, the mechanism of the anti-angiogenic effect of Avemar is unclear. Numerous studies have investigated cytotoxic effects o...

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Background:Fabaceae family members are known to possess preventive and therapeutic potentials against various types of cancers.Objective:The aim of this study was to investigate the cytotoxic and apoptotic effects of hydroalcoholic extracts from the aerial parts and roots of an endemic Ebenus species; Ebenus boissieri Barbey in human lung cancer cell line.Materials and Methods:After treatment with hydroalcoholic extracts cytotoxic activities of both extracts were measured by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide assay, whereas caspase-3 activity, tumor necrosis factor-a lpha (TNF-α) and interferon gamma (IFN-γ) releasewere measured by enzyme linked immunosorbent assay.Results:According to in vitro assay results, the increase in all caspases activity suggested that extracts induce cells to undergo apoptosis. Especially, induction in caspase-3 activity was the most remarkable result of this study. Both aerial part and root extracts induced apoptosis by increasing caspase-3 activity, TNF-α and IFN-γ release. When compared to their relative controls, the concentrations of both TNF-α and IFN-γ in extract-treated groups were significantly and dose dependently exalted.Conclusion:Taken together, our results indicate that the hydroalcoholic extracts of E. boissieri can be considered as a source of new anti-apoptotic and therefore anti-carcinogenic agent.

The family Potamogetonaceae has been taxonomically re-evaluated in Turkey, employing traditional morphological as well as molecular approaches (rbcL, ITS). Our fieldwork, herbarium studies and molecular analyses proved the existence of 22 taxa, of which 18 belong to Potamogeton (including 4 interspecific hybrids), 3 to Stuckenia and 1 to monotypic Groenlandia. Morphological re-descriptions of the recognized taxa were prepared, and the information concerning their distribution in Turkey was refined, based on plant material from extensive fieldwork and on specimens stored in herbaria that were previously not examined. Additionally, new identification keys to genera and species, and distribution maps of the species were prepared. Phylogenetic relationships and intraspecific variations were assessed by including samples from other regions. The status of the Zannichelliaceae was investigated using nuclear and chloroplast DNA markers. Our results from ITS sequence divergence corroborate the separation of the two families noted by some authors, and are in accordance with the substantial morphological differences between them. Relatively large genetic distance and non-monophyly indicate that two genotypes of P. gramineus constitute cryptic species, for which the Turkish localities expand upon the previously known distribution areas. We identified P. schweinfurthii as a new species for Turkey and report P. ×angustifolius for the first time for this country. Weak morphological differentiation and high sequence similarity did not permit reliable differentiation between the closely related Stuckenia amblyphylla and S. filiformis.

Background:Ebenus boissieri Barbey is an Antalya, Turkey-endemic plant belonging to Fabaceae family. The aerial parts and the roots of E. boissieri Barbey were used in this study.Objective:In the present study, we have examined the apoptotic effects of hydroalcoholic extracts of E. boissieri Barbey in human cervical cancer cell line HeLa.Materials and Methods:To determine the cytotoxic effect, cells were treated with various concentrations of extracts for 24, 48, and 72 h incubation periods. Cytotoxic effects were examined by Cell Titer 96 aqueous nonradioactive cell proliferation assay and the results were corrected by live/dead viability/cytotoxicity assay and trypan blue exclusion assay. Apoptotic effects were studied with multicaspase kit. Tumor necrosis factor-alpha (TNF-α) and interferon gamma (IFN-γ) release were also measured by enzyme-linked immunosorbent assay.Results:According to the results, E. boissieri Barbey extract caused significant increase in caspase levels. Thus, we suggest that the extract induces cells to undergo apoptosis. Especially, there was a sharp induction in caspase-3 activity. Levels of both TNF-α and IFN-γ in extract-treated groups were significantly and dose dependently exalted as compared to their relative controls.Conclusion:The effects of the extract on caspase-3, TNF-α, and IFN-γ levels mediate the plausible mechanism of apoptosis induction in HeLa. To the best of our knowledge, this is the first report indicating any pharmacological properties of E. Boissieri on HeLa cells.SUMMARY HeLa cell viability was reduced in dose-dependent manner for 72 h with an IC50 of approximate 28.03 μg/mL for aerial and 41.02 μg/mL for rootHeLa cells, exposure to the aerial extract led to 1.9, 3.8, 1.2, 2.4, and 3.45 fold induction of all caspases activities (-2, -3, -6, -8, and -9, respectively)Both 30 μg/mL of aerial and 45 μg/mL of root extracts allowed the production of anticancer cytokines (TNFalpha; IFNgamma) in HeLa cell culture supernatants. Abbreviations used: Tumor necrosis factor-alpha (TNF-α); Interferon gamma (IFN-γ); 3-(4, 5 dimethylthiazol-2-yl)-5-(3- carboxymethoxy-phenyl)-2-(4-sulfonyl)-2H-tetrazolium (MTS); Phosphate-Buffered Saline (PBS); Fetal Bovine Serum (FBS); para-Nitroanilin pNA; Enzyme-Linked ImmunoSorbent Assay (ELISA); Sodium Dodesyl sulphate –Polyacrilamide gel electrophoresis (SDS-PAGE); Tris-Buffered Saline (TBS); Hydocloric acid (HCl); Standart Error of Mean (SEM); National Cancer Institute (NCI); half maximal inhibitory concentration (IC50)