Classification of buckwheat honey produced in Kazakhstan according to their biochemical ingredients and bioactivities by chemometric approach.

Herein, nineteen buckwheat honey samples collected from 19 stations of different ecological zones of Kazakhstan were analysed for their pollen density, physicochemical properties, chemical composition, antioxidant, anticholinesterase, tyrosinase inhibitory, and urease inhibitory activities with chemometric approaches. Twelve phenolic compounds and fumaric acid were identified using HPLC-DAD, and mainly fumaric, p-hydroxybenzoic, p-coumaric, trans-2-hydroxy cinnamic acids, and chrysin were detected in all samples. The honey samples collected from the Northern zone exhibited best antioxidant activity in lipid peroxidation inhibitory (IC50:8.65 ± 0.50 mg/mL), DPPH• (IC50:17.07 ± 1.49 mg/mL), ABTS•+ (IC50:8.90 ± 0.65 mg/mL), CUPRAC (A0.50:7.51 ± 0.30 mg/mL) and metal chelating assay (IC50:10.39 ± 0.71 mg/mL). In contrast, South-eastern zone samples indicated better acetylcholinesterase (55.57 ± 0.83%), butyrylcholinesterase (49.59 ± 1.09%), tyrosinase (44.40 ± 1.21%), and moderate urease (24.57 ± 0.33%) inhibitory activities at 20 mg/mL. The chemometric classification of nineteen buckwheat honey was performed using PCA and HCA techniques. Both were supported by correlation analysis. Thirteen compounds contributed significantly to the clustering of buckwheat honey based on geographical origin.

Anatoluin A and B isolated from medicinal Tricholoma anatolicum are new cytotoxic ergostanoids against the most common cancers.

Tricholoma anatolicum is an edible mushroom from the matsutake group growing under Cedar trees. Bioactivity-guided fractionation of Tricholoma anatolicum afforded two new (1 and 2), three known ergosterols (3-6), and four known (6-9) compounds. Structures were identified as anatoluin A (1), anatoluin B (2), 5α,6α-epoxy-ergosta-7,22-dien,3ß-ol (3), ergosterol-endoperoxide (4), ergosterol,3ß-ol (5), 3,5-dihydroxyfuran-2(5H)-one (6), mannitol (7), turanose (8), fumaric acid (9) using spectroscopic techniques. The cytotoxic activity of extract and isolated compounds was performed using MTT assay against MCF7, HT29, H1299, and HeLa cancerous cell lines while toxicity against PDF and L929 fibroblast healthy cell lines. The lipid peroxidation inhibitory and ABTS•+ scavenging activities were used to determine antioxidant activity. The polar extracts exhibited significant cytotoxic activity. The more perfect is that the extracts and isolated compounds (1-5) were inactive against PDF and L929 healthy cell lines. Compounds 1-3 and 4 exhibited noticeable cytotoxic activity, while 1-5 moderately inhibited lipid peroxidation.

Preclinical study of the medicinal plants for the treatment of malignant melanoma.

Melanoma is the most aggressive type of skin cancer and originates from pigment-containing cells called melanocytes. The incidence of melanoma has been increasing worldwide. In the current study, the cytotoxic and photo-cytotoxic activities of different medicinal plants from Lamiaceae (Salvia cedronella, Salvia chionantha, and Salvia adenophylla), Asteraceae (Klasea kurdica, Klasea bornmuelleri, and Achillea millefolium), Apiaceae (Cuminum cyminum, and Anethum graveolens), and Polygonaceae (Rumex crispus) families were studied against HT 144 (Human malignant melanoma) cancer cell lines. The activities were performed by employing the MTT assay. Moreover, the apoptotic effects of the plant extracts were investigated by flow cytometry with annexin V/PI dual staining technique. The production of intracellular ROS by DCFH-DA technique and the effects of TNF-α secretion on apoptosis were also investigated. All plant extracts exhibited cytotoxic, and photo-cytotoxic effects against HT 144 cancer cells. Salvia species and Klasea species induced apoptosis via intracellular ROS generation secreted by TNF-α. On the other hand, A. millefolium, C. cyminum, A. graveolens, and R. crispus extracts induced apoptosis due to the intracellular generation of ROS, but, via the different pathway. In conclusion, this study indicates that the tested medicinal plant extracts have the potential in the treatment of melanoma.

The anticancer activity of visnagin, isolated from Ammi visnaga L., against the human malignant melanoma cell lines, HT 144.

Melanoma is a cancer of melanocyte cells and has the highest global incidence. There is a need to develop new drugs for the treatment of this deadly cancer, which is resistant to currently used treatment modalities. We investigated the anticancer activity of visnagin, a natural furanochromone derivative, isolated from Ammi visnaga L., against malignant melanoma (HT 144) cell lines. The singlet oxygen production capacity of visnagin was determined by the RNO bleaching method while cytotoxic activity by the MTT assay. Further, HT 144 cells treated with visnagin were also exposed to visible light (λ ≥ 400 nm) for 25 min to examine the illumination cytotoxic activity. The apoptosis was measured by flow cytometry with annexin V/PI dual staining technique. The effect of TNF-α secretion on apoptosis was also investigated. In standard MTT assay, visnagin (100 µg/mL) exhibited 80.93% inhibitory activity against HT 144 cancer cell lines, while in illuminated MTT assay at same concentration it showed lesser inhibitory activity (63.19%). Visnagin was induced apoptosis due to the intracellular generation of reactive oxygen species (ROS) and showed an apoptotic effect against HT 144 cell lines by 25.88%. However, it has no effect on TNF-α secretion. Our study indicates that visnagin can inhibit the proliferation of malignant melanoma, apparently by inducing the intracellular oxidative stress.

Effect of Sideritis leptoclada against HT-144 human malignant melanoma.

Sideritis leptoclada O. Schwarz et P.H. Davis extracts were evaluated for its singlet oxygen production capacity using spectrophotometric method. The extracts producing singlet oxygen were then evaluated for cytotoxicity against malignant melanoma cancer (HT-144) and fibroblast (3T3) cell lines using the 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay. The photocytotoxicity against the HT-144 human melanoma cell line in the presence of illumination (∼≥400 nm) was also evaluated. In the standard MTT assay, the ethanol extract of S. leptoclada (100 µg/ml) showed 83.49±3.33% inhibition of HT-144 cancer cells, whereas in the illuminated MTT assay, it showed 77.46±1.97% inhibition of HT-144 cancer cells. The effects of ethanol extract on reactive oxygen species production, apoptosis, and tumor necrosis factor-α secretion were also evaluated on HT-144 cell lines. The extract triggered an increase in intracellular reactive oxygen species production and tumor necrosis factor-α secretion compared with the respective controls. Thus, the ethanol extract may cause apoptosis. The LC-MS/MS analyses of S. leptoclada ethanolic extract showed that it has quinic acid (137213±11.25 µg/g extract), malic acid (1468±0.16 µg/g extract), chlorogenic acid (881.7±0.06 µg/g extract), and apigetrin (223.2±0.13 µg/g extract) as major constituents. The ethanolic extract of S. leptoclada should be further investigated as a potential treatment for malignant melanoma cancer.

Evaluation of fruit extracts of six Turkish Juniperus species for their antioxidant, anticholinesterase and antimicrobial activities.

BACKGROUND: Juniperus L. (Cupressaceae) species are mostly spread out in the Northern Hemisphere of the world, and some of them are used as folkloric medicines. The fruits of some species are eaten. Since oxidative stress is one of the reasons for neurodegeneration and is associated with the Alzheimer's disease (AD), the extracts prepared from the fruits of six Juniperus species were screened for their antioxidant activity. Therefore, the extracts were also evaluated against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), which are chief enzymes in the pathogenesis of AD. In addition, antimicrobial activity was also evaluated. RESULTS: In the ß-carotene-linoleic acid assay, acetone extracts of J. oxycedrus subsp. oxycedrus, J. sabina and J. excelsa, and methanol extracts of J. phoenicea and J. sabina, effectively inhibited oxidation of linoleic acid. The hexane extracts of J. oxycedrus subsp. oxycedrus, J. foetidissima and J. phoenicea showed remarkable inhibitory effect against AChE and BChE. CONCLUSION: Because of their high antioxidant activity, J. excelsa, J. oxycedrus subsp. oxycedrus, J. sabina and J. phoenicia might be used in the food industry as preservative agents or extension of the shelf-life of raw and processed foods. Since the hexane extracts of J. oxycedrus subsp. oxycedrus and J. foetidissima demonstrated significant anticholinesterase activity they should be considered as a potential source for anticholinesterase agents.

GC-MS analysis and antimicrobial activity of essential oil of Stachys cretica subsp. smyrnaea.

The essential oil from the aerial parts of Stachys cretica L. subsp. smyrnaea Rech. fil. (Lamiaceae), endemic to Turkey, was investigated by using GC and GC-MS. Thirty-four of 37 components, represented 99.7% of the total oil, were identified. The major components of the essential oil were trans-beta-caryophyllene (51.0%), germacrene-D (32.8%), a-humulene (3.1%), delta-cadinene (2.1%) and delta-elemene (2.1%). The antimicrobial activity of the essential oil, trans-beta-caryophyllene and five different extracts of the aerial parts of S. cretica L. subsp. smyrnaea were investigated by the standard disc diffusion method. The essential oil and trans-beta-caryophyllene exhibited antibacterial and antifungal activities. The activity increased with increasing concentrations of the essential oil and the extracts. The essential oil showed antimicrobial activity, particularly against Pseudomonas aeruginosa and Bacillus subtilis. The extracts exhibited either moderate or no activity.