Yasemin Celik Altunoglu

A Multidirectional Perspective for Novel Functional Products: In vitro Pharmacological Activities and In silico Studies on Ononis natrix subsp. hispanica

The genus Ononis has important value as traditional drugs and foods. In the present work, we aimed to assess the chemical profiles and biological effects of Ononis natrix subsp. hispanica extracts (ethyl acetate, methanol, and water). For chemical profile, total and individual phenolic components were detected. For biological effects, antioxidant (DPPH, ABTS, CUPRAC, FRAP, phosphomolybdenum, and metal chelating assays), enzyme inhibitory (against cholinesterase, tyrosinase, α-amylase and α-glucosidase), antimicrobial, DNA protection and cytotoxic abilities were tested. The predominant phenolics were apigenin, luteolin, and quercetin in the tested extracts. Generally, the ethyl acetate and methanol extracts were noted as the most active in the antioxidant and enzyme inhibitory assays. Water extract with different concentrations indicated high level of DNA protection activity. Methanol and ethyl acetate extracts showed antibacterial effect against to Staphylococcus aureus and Staphylococcus epidermidis strains. The cytotoxic effects of O. natrix subsp. hispanica extracts on the survival of HeLa and PC3 cells were determined by MTT cell viability assay. Water and methanol extracts caused initiation of apoptosis for PC3 cell line. Furthermore, molecular docking was performed to better understand interactions between dominant phenolic compounds and selected enzymes. Our results clearly indicate that O. natrix subsp. hispanica could be considered a potential candidate for designing novel pharmaceuticals, cosmeceuticals and nutraceuticals.

Novel in vitro and in silico insights of the multi-biological activities and chemical composition of Bidens tripartita L.

Bidens tripartita L. is a traditional phyto-remedy used in several countries, yet there is still a paucity of data on its biological potential. We aimed to provide new insights on the pharmacological potential of extracts prepared from B. tripartita via highlighting its antioxidant, key enzymes inhibitory potency, and DNA protecting effects. Phytochemical profile was established using High-Performance Liquid Chromatography with Diode-Array Detection (HPLC-DAD) and bioactive compound(s) docked against target enzymes using in silico methods. Cytotoxicity against three cancer cell lines was assessed using the methylthiazolyldiphenyl-tetrazolium bromide (MTT) cell viability test. The main compounds were luteolin-7-glucoside (cynaroside), chlorogenic acid, and epicatechin in the extracts. The methanol extract exhibited the highest radical scavenging activity. Ethyl acetate extract showed strongest α-amylase inhibitory activity, while the best α-glucosidase inhibitory effect recorded for the methanol extract. Molecular docking showed that cynaroside strongly interact to α-glucosidase cavity by establishing six hydrogen bonds. B. tripartita extracts were found to protect supercoiled form of pUC19 plasmid (>70%) and also showed anti-proliferative properties. Results amassed in the present study add on to a growing body of literature on the multi-pharmacological potency of B. tripartita which can be applied to bio-products development geared towards management of common diseases.

Immune responses to methanolic extract of black cumin (Nigella sativa) in rainbow trout (Oncorhynchus mykiss)

Abstract The immune stimulating effects of the methanolic extract of black cumin (Nigella sativa) in rainbow trout (Oncorhynchus mykiss) was evaluated. Variable concentrations of black cumin methanolic extract [0 (Control), 0.1 and 0.5 g kg−1 of feed] were individually added to the basal diet and rainbow trout was fed for 30 days to assess the innate immune responses and growth performance. Feed conversion ratio significantly decreased in the group fed with 0.5 g kg−1 black cumin extract. Respiratory burst activity was observed to be the highest in the 0.5 g kg−1 black cumin extract fed group. Lysozyme and myeloperoxidase activities were significantly increased in fish of experimental groups compared to control (P < 0.05). TGF‐&bgr; gene expression increased in black cumin 0.5 g kg−1 treated group. IL‐1&bgr; and TGF‐&bgr; gene expressions decreased in black cumin 0.1 g kg−1 administered group. Expression of IL‐12 gene diminished in both the experimental groups. There was no significant difference in survival rates between black cumin extract treated fish groups and control (P > 0.05) after challenged with Aeromonas hydrophila. The results indicate that the methanolic extract of black cumin is a stimulator of some innate humoral immune responses, but it is ineffective for cytokine‐related gene trancriptions in rainbow trout. HighlightsWe tested the effects of black cumin methanolic extracts in Oncorhyncus mykiss.Growth and survival were not affected in black cumin extract treated fish groups.Cytokine gene expressions were not elevated by black cumin extract treatment.Phagocytic activity was not increased in treated fish.Methanolic extract of black cumin is ineffective to rainbow trout immune system.

Identification, molecular characterization and expression analysis of RPL24 genes in three Cucurbitaceae family members: cucumber, melon and watermelon

Ribosomal proteins are crucial for the proper growth and development of any organism, including plants. The ribosomal protein L24 (RPL24) is found in the large subunit of the ribosome and is responsible for the stabilization of the peptidyl transferase activity. Although RPL24 gene has been individually identified in different organisms, little is known about the genome-wide survey and expression patterns of R24 genes in Cucurbitaceae family members. We identified seven Cucurbitaceae RPL24 genes from cucumber, melon and watermelon. They were phylogenetically clustered into seven major groups. Gene structure and motif composition are relatively conserved in each group. Three-dimensional homology modelling of RPL24 proteins was performed with higher confidence level. CmRPL24-01 was isolated from melon and characterized at a molecular level. The regulation of ribosomal proteins in melon under drought stress conditions was also studied. The expression of CmRPL24-01 gene increased in melon leaf tissue at 3 h upon polyethylene glycol treatment and showed a gradual induction after 12 h. Our study provided a very useful reference for identification and functional analysis of RPL24 protein members in different plants. In addition, this research indicated a potential usage of ribosomal proteins in response to drought stress.

Analysis of DNA protection, interaction and antimicrobial activity of isatin derivatives

Isatin, thiosemicarbazone and their derivatives have been widely used in biological applications such as antimicrobial, antiviral and anticancer therapies. Herein, eight isatin and thiosemicarbazone derivative compounds were re-synthesized and evaluated for DNA binding analysis including DNA protection studies using plasmid DNA (pUC19) and DNA interaction experiments using calf thymus DNA (CT-DNA). All compounds were also utilized in vitro assay to assess the antimicrobial activity of compounds against different pathogenic bacterial strains. All isatin and thiosemicarbazone derivative compounds exhibited DNA protection activity which ranged from 23.5 to 59.5%. Among them, I3-(N-2-MP)-TSC had the greatest DNA protective activity. For DNA binding analysis, all compounds had the same constant concentration (40 μM), which interacts with CT-DNA. It was also observed that DNA interactions gave a high intrinsic binding constant (Kb = 1.72 × 104 M-1-9.73 × 105 M-1). Besides, several derivatives of isatin thiosemicarbazone exhibited significant and selective antibacterial activity with low concentration. These compounds primarily affected Gram-positive bacteria, but were not effective against P. vulgaris and E. coli. The Gram-positive methicillin-resistant S. aureus ATCC 43300 (MRSA) was the most influenced strain by these compounds. It was found that methyphenyl group at isatin was essential for its antibacterial activity for MRSA.

Investigations into the therapeutic potential of Asphodeline liburnica roots: In vitro and in silico biochemical and toxicological perspectives

This study aims to establish the biological and chemical profile of Asphodeline liburnica (Scop.) Rchb. root. The antioxidant, antimicrobial, enzyme inhibitory, DNA protection, apoptotic DNA ladder fragmentation analysis, and anti-proliferative of A. liburnica were established using standard assays. In silico study was also performed to understand interactions between quantified anthraquinones and key enzymes of clinical relevance. Total phenolic and flavonoid contents were found to be 9.67 mgGAE/g and 1.48 mgRE/g extract, respectively. Chrysophanol was detected as a major anthraquinone. The extract exhibited radical scavenging ability against DPPH and ABTS with values of 13.23 and 66.99 mgTE/g extract, respectively. Good inhibitory activity against tyrosinase was recorded. In silico experiments showed that the anthraquinones were able to establish coordinative bonds with the copper atoms present in the enzymatic cavity of tyrosinase. MTT cell viability test on MDA-MB-231 cells showed that at 0.1 and 1 μg of extracts induced anti-proliferative effect. Apoptotic DNA fragmentation analysis indicated nuclear condensation resulting in DNA fragmentation, which exhibited apoptotic cell death in the presence of A. liburnica. This study has provided insights on the potential usage of A. liburnica which could open new avenues for research and stimulate future interest for the development of safe novel biopharmaceuticals.

Pharmacological and polyphenolic profiles of Phyllanthus phillyreifolius var. commersonii Müll. Arg: An unexplored endemic species from Mauritius

Phyllanthus phillyreifolius var. commersonii Müll. Arg is an endemic plant of Mauritius. To date, no study has been performed concerning its polyphenolic profile and pharmacological properties. In this study, a decoction (water), ethyl acetate and methanol extracts of the aerial parts of P. phillyreifolius, obtained from different extraction procedures (maceration and Soxhlet), were studied for antibacterial, antioxidant, anticancer, and enzyme inhibitory properties along with their polyphenolic profile. The ethyl acetate macerated extract showed high antibacterial activity against B. cereus (MIC = 0.293 mg/mL) and E. coli (MIC = 0.417 mg/mL) while S. epidermidis was most susceptible to the ethyl acetate-Soxhlet extract (MIC = 0.521 mg/mL). The methanol-Soxhlet extract displayed the most potent cupric and ferric reducing power, and metal chelating effect, while the macerated methanolic extract was the most effective DPPH and ABTS scavenger, and BChE inhibitor. Only the ethyl acetate-Soxhlet extract exhibited α-glucosidase inhibition. All extracts exhibited a strong anti-tyrosinase activity, which was further investigated by molecular docking and molecular dynamic. After 48 h exposure to the extracts for HeLa cell lines, the ethyl acetate-Soxhlet extract showed the highest inhibition (IC50 = 533.1 μg/mL) while the decoction extract was more cytotoxic to MDA-MB-231 cells (IC50 = 337.4 μg/mL). Treatment of cancer cell lines with all P. phillyreifolius extracts resulted in a time-dependent reduction of cell viability for HeLa and dose-and time-dependent reduction for MDA-MB-231. Gene expression ratio of Bcl-2 to Bax was higher for all Soxhlet-extracts. Total phenolics (TPC) and flavonoids (TFC) content were highest in the decoction and methanol-Soxhlet extract, respectively (122.43 mg GAE/g extract and 31.28 mg RE/g extract, respectively). The extracts were abundant in ellagitannins, although phenolic acids and flavonoids were also detected. Granatin B was detected for the first time in Phyllanthus species. Overall, the aerial parts of P. phillyreifolius exemplify a potent reservoir of bioactive phytochemicals for therapeutic applications.

Comparative identification and evolutionary relationship of fatty acid desaturase (FAD) genes in some oil crops: the sunflower model for evaluation of gene expression pattern under drought stress

ABSTRACT Sunflower (Helianthus annuus L.) and some oil crops have considerable economic value, making them important for commercial use. The fatty acid content of their seeds is crucial for proper nutrition. Additionally, the desaturation reaction via fatty acid desaturases (FADs) leads to formation of double bonds in fatty acids and is an essential step in fatty acid biosynthesis. This study aimed to identify and compare some fatty acid desaturase gene members in sunflower and other oil crops. Totally, 33 FAD genes from sunflower, 19 from sesame, 20 from canola, 25 from cacao and 46 from flax were analysed. The FAD members had roles in oxidation–reduction processes, fatty acid biosynthetic processes or lipid metabolic processes. Comparative phylogenetic and motif analysis revealed a conserved structure among FAD members belonging to various oil crops. The average Ka/Ks rates calculated on the basis of orthologous gene analysis were 0.04, 0.05 and 0.05 between sunflower and, respectively, canola, cacao and sesame. The most recent divergence time between sunflower and cacao was estimated at an average of 100 million years ago (MYA). This was followed by flax, sesame and canola with an average of 101 MYA, 114 MYA and 149 MYA, respectively. Alpha-helices were dominant in the predicted 3D structures of FAD proteins. The FAB2 expression levels from a drought tolerant sunflower variety were not affected by drought. The characterisation of desaturase family members in economically valuable oil crops could be useful for functional cloning studies to enhance the unsaturated fatty acid contents of the plants.