DNA Protection Activity Research Articles

Determination of antioxidant, DNA protection, enzyme inhibition potential and molecular docking studies of a biomarker ursolic acid in Nepeta species

Ursolic acid (UA), which has many biological properties such as anti-cancer, anti-inflammatory and antioxidant, and regulates some pharmacological processes, has been isolated from the flowers, leaves, berries and fruits of many plant species. In this work, UA was purified from the methanol-chloroform crude extract of Nepeta species (N. aristata, N. baytopii, N. italica, N. trachonitica, N. stenantha) using a silica gel column with chloroform or ethyl acetate solvents via bioactivity-guided isolation. The most active sub-fractions were determined under bioactivities using antioxidant and DNA protection activities and enzyme inhibitions. UA was purified from these fractions and its structure was elucidated by NMR spectroscopy techniques. The highest amount of UA was found in N. stenantha (8.53 mg UA/g), while the lowest amount of UA was found in N. trachonitica (1.92 mg UA/g). The bioactivities of UA were evaluated with antioxidant and DNA protection activities, enzyme inhibitions, kinetics and interactions. The inhibition values (IC50) of α-amylase, α-glucosidase, urease, CA, tyrosinase, lipase, AChE, and BChE were determined between 5.08 and 181.96 µM. In contrast, Ki values of enzyme inhibition kinetics were observed between 0.04 and 0.20 mM. In addition, Ki values of these enzymes for enzyme-UA interactions were calculated as 0.38, 0.86, 0.45, 1.01, 0.23, 0.41, 0.01 and 2.24 µM, respectively. It is supported that UA can be widely used as a good antioxidant against oxidative damage, an effective DNA protector against genetic diseases, and a suitable inhibitor for metabolizing enzymes. Communicated by Ramaswamy H. Sarma

Phytochemical analysis, DNA barcoding and DNA protective activity of Ferulago cassia Boiss.

Phytochemical analysis, DNA barcoding and DNA protective activity of Ferulago cassia Boiss.

Koelreuteria paniculata Seed Oil-A Rich Natural Source of Unsaturated Fatty Acids and Phytocompounds with DNA Protective Potential.

The present work is focused on the physicochemical characteristics, chemical composition, and some biological activities of Koelreuteria paniculata seed oil. The glyceride oil, obtained with a Soxhlet apparatus by extraction with hexane, was characterized by a relatively high oil content (over 20%), and it is defined as a non-drying oil (iodine value-44 gI2/100 g) with good oxidative stability (over 50 h). There were identified 11 fatty acids, 6 sterols, 3 tocopherols, and 6 phospholipids, as the last group was reported for the first time. The major components among them were-monounsaturated eicosenoic and oleic acids, β-sitosterol, β-tocopherol, and phosphatidylcholine. The in vitro tests demonstrated DNA protective activity and a lack of cytotoxicity of the oil, data that has been reported for the first time. The in vitro MTT test of the oil on HT-29 and PC3 cell lines did not indicate antitumor activity. The seed oil studied contains valuable bio-components, which have proven benefits for human health, and that is why it could be used in food, cosmetic, and pharmaceutical products.

Antimicrobial, Antioxidant, Cytotoxic, DNA Protective Activities, and Molecular Docking Studies of the Methanolic Extract of Salvia siirtica Kahraman, Celep & Dogan sp. Nov. (Lamiaceae)

Background: Medicinal plants have long been utilized in traditional medicine in Turkey and worldwide. The flora of Turkey is so rich it needs to be discovered, documented, and explored. This plant species is the origin of many valuable medicinal chemicals. Salvia species was used in folk medicine for many years in Turkey. One of the new Salvia species is Salvia siirtica. Methods: The antimicrobial, cytotoxic, antioxidant, DNA restriction inhibition, and DNA protective activities of methanolic extract of S. siirtica were studied. Results: The extract has better activity than the standard antibiotics against P. aeruginosa, meanwhile, showed high DPPH scavenging activity with an IC50 value of 28.72 ± 8.02 µg/ml. The extract exhibited good cytotoxic potential with IC50 value 0.31 ± 0.07 mg/ml towards H1299 cell line. The restriction enzyme inhibition in presence of plant extract proves that extract can interact with restriction endonuclease enzymes. Apigenin and Luteolin 7- glycoside are two high components of S. siirtica were docked against potential intracellular targets to evaluate their inhibition capability. Luteolin its derivative luteolin-7-glycoside showed the highest potential in molecular docking studies. Conclusions: Determining the bioactivity of these components by isolating them from the plant may pave the way for their use as supportive agents in the fight against many diseases.

Synthesis and anti-Alzheimer potential of novel α-amino phosphonate derivatives and probing their molecular interaction mechanism with acetylcholinesterase

Pleiotropic interference may be a prerequisite for the efficient limitation of the progression of multi-factorial diseases such as Alzheimer's disease (AD). Concept of designing the single chemical entity acting on two or more targets of interest has potential advantage in AD therapy. In line with this, rational design and synthesis of frame work of hybrids bearing 2,3-disubstituted quinazolinone, vanillin and α-amino phosphonate scaffolds (5a─v) were carried out. A congeneric set of twenty-two synthetic derivatives (5a─v) were evaluated for their cholinesterase inhibitory, antioxidant, DNA nicking, DNA protection, neuroprotective and Aβ aggregation modulatory activities. Amongst tested activities, the most significant and worth mentioning is that the analogues 5m, 5p and 5u were found to be the most potent, selective, and mixed type inhibitors of EeAChE with IC50 values of 0.296 ± 0.030, 0.289 ± 0.027, and 0.306 ± 0.028 μM, respectively. Further, the biophysical approaches indicated that the compounds 5m, 5p, and 5u have a strong binding affinity towards AChE. Kinetic and Molecular docking studies have revealed that the most active congeners were well oriented in the AChE active site by interacting with both catalytic active site (CAS) and peripheral anionic site (PAS). A few parameters derived from molecular dynamics (MD) simulation trajectories emphasized the stability of AChE-5p and 5m complexes throughout the 100 ns simulations, and the local conformational changes of the residues of AChE validate the stability of AChE-5p and 5m complexes. Further, these derivatives significantly impacted ABTS radical scavenging capacities and maximal DNA protection activity. Importantly, Thioflavin T (ThT) assay and FE-SEM study demonstrated compounds 5m, 5p and 5u as effective Aβ1−42 fibril modulators at molecular level by the formation of micro size co-assembled mature structures, thus efficiently abolishing the cytotoxicity of Aβ1–42. Finally, these active compounds are determined to be non-toxic and highly neuroprotective against H2O2-induced cell death in SK-N-SH cell lines. Furthermore, in silico ADMET prediction studies have revealed that the targeted analogues satisfied most of the characteristics of CNS acting drugs. These multi-functional efficacies indicated worthiness of these α-amino phosphonate derivatives being chosen for further pharmacokinetics, toxicity, and behavioral research to test their potential for AD treatment.

DNA protection, molecular docking, antioxidant, antibacterial, enzyme inhibition, and enzyme kinetic studies for parietin, isolated from Xanthoria parietina (L.) Th. Fr.

Parietin was isolated from Xanthoria parietina (L.) Th. Fr.’ (methanol:chloroform) extract, using a silica column. 13 C NMR and 1H NMR were used to confirm the structure of the isolated parietin. For the first time, parietin was investigated for its antioxidant, antibacterial and DNA protective activities. Molecular docking was carried out to determine the binding affinity and interactions between the enzymes and our molecule. Inhibition and kinetic mechanism studies for the action of the enzymes were performed too. Parietin exhibited high metal chelating activity. The MIC values of parietin were sufficient to inhibit different bacterial strains; E. coli, P. aeruginosa, K. pneumoniae and S. aureus. Molecular docking applications exhibited that acetylcholinesterase (AChE), butyrylcholinesterase (BChE), lipase, and tyrosinase have high potential for binding with the parietin. Especially, the parietin’s highest binding affinity was recorded with AChE and tyrosinase. These results were confirmed by the inhibition and kinetics results, where, parietin observed a potent inhibition with an IC50 values between 0.013-0.003 µM. Moreover, parietin acts’ as a non-competitive inhibitor against AChE, BChE, and lipase, and as a competitive inhibitor against tyrosinase with a high rate of inhibition stability. The promising biological properties of parietin revealed its effectiveness in terms of suitability in the food and pharmaceutical industries. Communicated by Ramaswamy H. Sarma

Evaluation of antioxidant potential, DNA damage and hepatoprotective properties of Lagenaria siceraria plant against acetaminophen induced hepatotoxicity

Introduction: Lagenaria siceraria has various pharmacological effects like antioxidant, anti-hyperglycemic, anti-ulcer, anti-cancer, hepatoprotective, antimicrobial, anti-hyperlipidemic, anti-inflammatory and cardioprotective activities and is used to treat various diseases like diabetes, jaundice, piles, ulcers, hypertension, colitis, insanity, and skin diseases.Context and purpose of the study: The aim of the present study was to evaluate the phytochemical content, antioxidant activity, DNA damage and hepatoprotective activities of Lagenaria siceraria (LS)plant’s parts to explore their nutraceutical value.Methods: Phytochemical content was measured by the estimation of total phenolics, carotenoids, ascorbic acid, protein and carbohydrates while antioxidant activities were investigated by assaying the parameters of free radical scavenging activity, lipid peroxidation, superoxide anion radical scavenging activity, reducing power, and DNA damage protection assay. Hepatoprotective activity of LS leaf was measured by assaying cell viability, superoxide dismutase activity, malondialdehyde and nitric oxide formation.Results: Among the tested plant samples, LS leaves showed high contents of total phenolics, carotenoids, ascorbic acid, and protein contents whereas LS fruits have high carbohydrate contents. Evaluation of antioxidant activities in different parts of the LS plant showed that the LS leaf has high antioxidant properties as compared to other plant parts. The L. siceraria leaf also showed DNA protection activities against Fenton’s reagent as well as UV induced hydroxyl radical’s damage. To test the hepatoprotective activity of LS leaf against acetaminophen, induced hepatocellular toxicity and leaf extract concentrations of 2.5, 5.0 and 7.5 μg/104 cells were administered to cultured hepatocytes before acetaminophen exposure. The experimental results showed that LS leaf extract restores superoxide dismutase activity and reduces malondialdehyde and nitric oxide formation as compared to acetaminophen alone treated cultured cells.Conclusion: Our findings suggest that leaves of L. siceraria plants have high antioxidant potential and DNA damage-protective and hepatoprotective activities. Hence, the leaf part of this plant may be used in different food applications to provide nutritional and health benefits, as well as in various liver care herbal formulations.Keywords: Antioxidant, Hepatoprotective activity, Hepatocytes, Lipid peroxidation, Total phenolics

Antioxidant and DNA-Protective Activity of an Extract Originated from Kalamon Olives Debittering.

Table olives are a major component of the Mediterranean diet and are associated with many beneficial biological activities, which are mainly related to their phenolic compounds. Olive fruit debittering process defines the quantitative and qualitative composition of table olives in biophenols. The aim of the present study was to evaluate the in vitro antioxidant capacity and DNA-protective activity of an extract originated from brine samples, according to the Greek style debbitering process of Kalamon olive fruits. The main phenolic components determined in the brine extract were hydroxytyrosol (HT), verbascoside (VERB) and tyrosol (T). The in vitro cell-free assays showed strong radical scavenging capacity from the extract, therefore antioxidant potential. At cellular level, human endothelial cells (EA.hy296) and murine myoblasts (C2C12) were treated with non-cytotoxic concentrations of the brine extract and the redox status was assessed by measuring glutathione (GSH), reactive oxygen species (ROS) and lipid peroxidation levels (TBARS). Our results show cell type specific response, exerting a hormetic reflection at endothelial cells. Finally, in both cell lines, pre-treatment with brine extract protected from H2O2-induced DNA damage. In conclusion, this is the first holistic approach highlighted table olive wastewaters from Kalamon- Greek style debittering process, as valuable source of bioactive compounds, which could have interesting implications for the development of new products in food or other industries.

Ethyl Acetate Extract of Colletotrichum gloeosporioides Promotes Cytotoxicity and Apoptosis in Human Breast Cancer Cells.

Fungal endophytes are known to be a paragon for producing bioactive compounds with a variety of pharmacological importance. The current study aims to elucidate the molecular alterations induced by the bioactive compounds produced by the fungal endophyte Colletotrichum gloeosporioides in the tumor microenvironment of human breast cancer cells. GC/MS analysis of the ethyl acetate (EA) extract of C. gloeosporioides revealed the presence of bioactive compounds with anticancer activity. The EA extract of C. gloeosporioides exerted potential plasmid DNA protective activity against hydroxyl radicals of Fenton's reagent. The cytotoxic activity further revealed that MDA-MB-231 cells exhibit more sensitivity toward the EA extract of C. gloeosporioides as compared to MCF-7 cells, whereas non-toxic to non-cancerous HEK293T cells. Furthermore, the anticancer activity demonstrated by the EA extract of C. gloeosporioides was studied by assessing nuclear morphometric analysis and induction of apoptosis in MDA-MB-231 and MCF-7 cells. The EA extract of C. gloeosporioides causes the alteration in cellular and nuclear morphologies, chromatin condensation, long-term colony inhibition, and inhibition of cell migration and proliferation ability of MDA-MB-231 and MCF-7 cells. The study also revealed that the EA extract of C. gloeosporioides treated cells undergoes apoptosis by increased production of reactive oxygen species and significant deficit in mitochondrial membrane potential. Our study also showed that the EA extract of C. gloeosporioides causes upregulation of pro-apoptotic (BAX, PARP, CASPASE-8, and FADD), cell cycle arrest (P21), and tumor suppressor (P53) related genes. Additionally, the downregulation of antiapoptotic genes (BCL-2 and SURVIVIN) and increased Caspase-3 activity suggest the induction of apoptosis in the EA extract of C. gloeosporioides treated MDA-MB-231 and MCF-7 cells. Overall, our findings suggest that the bioactive compounds present in the EA extract of C. gloeosporioides promotes apoptosis by altering the genes related to the extrinsic as well as the intrinsic pathway. Further in vivo study in breast cancer models is required to validate the in vitro observations.

Yerli ve ithal Hypericum perforatum sulu ekstraktlarının kimyasal içeriği ve biyoaktif kapasitesinin karşılaştırılması

Hypericum perforatum, known as St. John's Wort and most used ethnaformacolgically, belongs to the Hypericaceae family. In this work, we aimed to evaluate the antioxidant, antimicrobial, DNA protective activities, and enzyme inhibitor properties of domestic and import H. perforatum aqueous extracts. The total phenolic and flavonoid contents were also determined for both samples. Results exhibited a high phenolic content for Domestic H. perforatum aqueous extract (DHPE) and Import H. perforatum aqueous extract (IHPE) samples. Meanwhile, inhibition activity levels for α-glucosidase were remarkable, the IC50 value of DHPE was 16.35±0.07 µg/mL, and the IC50 value of IHPE was 15.05±1.36 µg/mL, both samples demonstrated almost twice of the standard inhibitor effect (IC50: 30.62±2.07 µg/mL). Moreover, the inhibition activity of both samples against α-amylase was more effective than acarbose. The antibacterial results were as follows; the highest MIC value detected by IHPE was 64 µg/mL against B. cereus. In addition, IHPE was shown to be more effective than the domestic sample against all microorganisms. Furthermore, DHPE exhibited good protective activity from oxidative H2O2-induced DNA damage. Both tested samples had an abundance of phenolic content and were high in inhibitory activity against diabetic enzymes. Generally, bioactivity tests’ results appeared quite effective for both samples compared to the standards.

The Investigation of the Effect of the Salicylic Acid on the Antioxidant Potential, Vitamin C Content and DNA Protection Activity of Wheatgrass (Triticum aestivum L.)

Wheatgrass is considered as a superfood because of its high antioxidant potential and beneficial ingredients. Especially in recent years, wheatgrass juice and its powder have been tested in vivo animal models and clinical studies against some diseases. Salicylic acid (SA) one of the important phytohormones controlling plant growth is used as an exogenous elicitor to increase plant bioactive compounds. The aim of this study was to investigate antioxidant potential, vitamin C content and DNA protection of wheatgrass grown from SA pre-treated seeds. For this purpose, total antioxidant statue, total oxidant statue, oxidative stress index and vitamin C level were determined. Additionally, pUC19 plasmid was incubated with Fenton’s solution to determine DNA protection activity of lyophilized wheatgrass extract. Bread wheat caryopses were imbibed in different concentrations of SA for 2 hours. Wheatgrass grown from seeds pre-treated with 10-8 M SA had significantly higher total antioxidant statue, vitamin C and soluble protein content than control. It was observed that wheatgrass extracts had a DNA protective role against hydroxyl radicals. It was concluded that SA pretreatment of seeds could be a good approach to increase their antioxidant potential, soluble protein content and vitamin C level of plants used as antioxidant sources by people.

Evaluation of DNA Protective and Antimicrobial Properties of some Cladonia Species

The present study evaluated the DNA protective properties and antimicrobial activities of the methanol extracts of nine Cladonia species, namely C. pocillum, C. subulata, C. pyxidata, C. coniocraea, C. foliacea, C. firma, C. furcata, C. fimbriata and C. rangiformis collected in Turkey. DNA protection properties efficiency of Cladonia extracts was evaluated using pBR322 plasmid DNA. In vitro antimicrobial activities of methanol extracts against two Gram-negative bacteria (Escherichia coli and Proteus mirabilis), three Gram-positive bacteria (Staphylococcus aureus, Micrococcus luteus and Bacillus subtilis) and two fungal strains (Candida glabrata and Candida albicans) were examined using the disc diffusion method and through the determination of minimal inhibitory concentrations (MIC). DNA protective studies, all Cladonia extracts protected pBR322 plasmid DNA against damage caused by the hydrogen peroxide (H2O2) with ultraviolet (UV). The results demonstrated that the inhibition zones in the disc diffusion method ranged from 6.5 to 19.0 mm. MIC results were ranged from 3.12 to 6.25 mg/mL. Cladonia extracts show a better antimicrobial effect against bacterial strains than fungal strains. The highest antimicrobial effect among lichen species was demonstrated by Cladonia pocillum. Our results demonstrated that tested Cladonia extracts had strong antibacterial and DNA protective effects. This is the first comprehensive study to evaluate the DNA protective properties activity of Cladonia extracts.

Thioredoxin domain-containing protein 12 (TXNDC12) in red spotted grouper (Epinephelus akaara): Molecular characteristics, disulfide reductase activities, and immune responses

Thioredoxins are small ubiquitous redox proteins that are involved in many biological processes. Proteins with thiol-disulfide bonds are essential regulators of cellular redox homeostasis and diagnostic markers for redox-dependent diseases. Here, we identified and characterized the thioredoxin domain-containing protein 12 (EaTXNDC12) gene in red spotted grouper (Epinephelus akaara), evaluated transcriptional responses, and investigated the activity of the recombinant protein using functional assays. EaTXNDC12 is a 19.22-kDa endoplasmic reticulum (ER)-resident protein with a 522-bp open reading frame and 173 amino acids, including a signal peptide. We identified a conserved active motif (66WCGAC70) and ER retention motif (170GDEL173) in the EaTXNDC12 amino acid sequence. Relative EaTXNDC12 mRNA expression was analyzed using 12 different tissues, with the highest expression seen in brain tissue, while skin tissue showed the lowest expression level. Furthermore, mRNA expression in response to immune challenges was analyzed in the head kidney, blood, and gill tissues. EaTXNDC12 was significantly modulated in response to bacterial endotoxin lipopolysaccharide (LPS), nervous necrosis virus (NNV), and polyinosinic:polycytidylic acid (poly(I:C)) challenges in all of the tested tissues. Recombinant EaTXNDC12 (rEaTXNDC12) displayed antioxidant ability in an insulin reductase assay, and a capacity for free radical inhibition in a 2,2-diphenyl-1-picryl-hydrazyl-hydrate assay. In addition, a DNA nicking assay revealed that purified rEaTXNDC12 exhibited concentration-dependent DNA protection activity, while results from 2-hydroxyethyl disulfide and L-dehydroascorbic assays indicated that rEaTXNDC12a possesses reducing ability. Furthermore, fathead minnow (FHM) cells transfected with EaTXNDC12-pcDNA demonstrated significantly upregulated cell survival against H2O2-induced apoptosis. Collectively, the results of this study strengthen our knowledge of EaTXNDC12 with respect to cellular redox hemostasis and immune regulation in Epinephelus akaara.

An exopolysaccharide from Leuconostoc mesenteroides showing interesting bioactivities versus foodborne microbial targets

An exopolysaccharide (EPS_B3) produced by a Leuconostoc mesenteroides strain isolated from a semi-hard Italian cheese was chemically and biologically characterized. HPLC-SEC, NMR, FT-IR and monosaccharide composition experiments were performed. Antimicrobial, antibiofilm, bifidogenic, antioxidant, and DNA-protective activity of EPS_B3 were also studied. Results revealed that EPS_B3 was a mixture of two high-molecular-weight dextran with low branching degree. Moreover, EPS_B3 displayed significant antibacterial activity against eight foodborne pathogens and inhibited biofilm formation by Listeria monocytogenes. EPS_B3 also evidenced bifidogenic activity, stimulating the growth of three probiotic bifidobacteria, and improving the tolerance of Bifidobacterium animalis subsp. lactis to oxygen stress. It also protected plasmid DNA from hydrogen peroxide damage. Only limited antioxidant capacity was observed.In conclusion, data suggest that EPS_B3 might be exploited in the context of functional foods especially for its marked antimicrobial activity as well as for the ability to improve the viability of bifidobacteria in probiotic foods. However, further studies should be carried out to assess the ability of EPS_B3 to reach intact the target site (i.e., gastrointestinal tract) to consider the possibility of use it as a new functional ingredient in foods.

Bioprospecting roots, stem and leaves extracts of Berberis baluchistanica Ahrendt. (Berberidaceae) as a natural source of biopharmaceuticals

ABSTRACT The present study was designed to explore the biological potential of aerial parts of Berberis baluchistanica. HPLC analysis revealed the highest berberine content in leaves (19453.15 ppm) followed by roots (3225.12ppm) and stem (29.11ppm). Phenolics, flavonoids, tannins, DPPH inhibition activity and total antioxidant capacity are reported in root, leaf and stem extracts. Antihaemolytic assay displayed the highest (97%) inhibition potential of extracts against H2O2-induced haemolysis than heat-induced haemolysis (75%). The highest DNA protection activity was observed in root and leaves extracts while the antimicrobial potential was recorded highest in the leaves extract against Staphylococcus aureus (23.00 ± 1.0 mm) and Klebsiella pneumonia (19.67 ± 0.2 mm). Similarly, brine shrimp cytotoxicity potential was observed highest in root extract (LC50 – 10.04 ppm) and the allelopathic potential was confirmed by radish seed phytotoxicity assay. Overall, results suggest a remarkable nutritional and biological potential B. baluchistanica, especially areal parts can help to preserve the endemic plants by discouraging the use of roots.

Phenolic profile, antioxidant, DNA protection, acetylcholinesterase, butyrylcholinesterase and urease inhibition activities of Coriandrum sativum L. leaf, seed and flower extracts

In this study, chemical content, antioxidant activities, enzyme inhibition activities, and DNA protection activities of extracts obtained from different solvents of the Coriandrum sativum leaf, flower and seed were determined. Total phenol and flavonoid contents of C. sativum leaf, seed and flower hexane extracts were higher than water, methanol, ethanol and ethyl acetate extracts. The highest anthocyanin content was found in the flower part of C. sativum. From the antioxidant activity tests, total antioxidant activity of flower aqueous extract, reducing power of seed aqueous extract, H2O2 scavenging activity of leaf ethyl acetate extract, OH˙ radical scavenging activity of leaf hexane extract, free radical scavenging activity of flower methanol extract, metal chelate activity of leaf ethyl acetate extract, superoxide anion scavenging activity of leaf aqueous extract and lipid peroxidation inhibition activity of the leaf ethyl acetate extract had the highest. It was found that the urease inhibition activity of the seed methanol extract and the acetylcholinesterase and butyrylcholinesterase inhibition activities of the seed ethanol extract presented effective inhibition activity as 80.30±0.20%, 112.83±10.75 µg/mL, and 334.28±23.09 µg/mL, respectively. The leaf hexane, flower ethyl acetate, and leaf methanol extracts showed the highest DNA protection activities with values of 71.86%, 70.89%, and 69.38%, respectively. According to the phytochemical content and biochemical activity results, this study is a valuable report proving that the C. sativum plant is a natural effective product.

Investigation of the effects of different extraction methods on some biological activities of Dactylorhiza romana subsp. georgica (Klinge) Soó ex Renz & Taubenheim

Dactylorhiza romana subsp. georgica is found in Anatolia and is traditionally used by local people both as food and to alleviate various ailments. Although various biological activities of the plant have been partially investigated, these properties have not been comprehensively revealed in the light of scientific data. In this study, it was aimed to elucidate the antioxidant, antimicrobial, enzyme inhibition (alpha amylase, alpha glucosidase, and tyrosinase) and DNA protective activity and phenolic content of the extracts obtained from orchid tubers using different extraction methods. For this, the fresh and dry tubers of the plant were extracted with ethanol at different times in an ultrasonic water bath, shaker and their combinations with each other. ABTS, DPPH, and FRAP methods were used to determine the antioxidant properties of plant tubers. In addition, the phosphomolybdate method was used to determine the total antioxidant capacity. The spectrophotometric method was preferred to reveal the enzyme inhibition activity. The antimicrobial activity of the extracts was investigated by microdilution method. In addition, the phenolic content of the plant was analysed by HPLC. The DNA protective activity of the extracts was determined by using the pUC 19 plasmid. According to the results obtained, it was revealed that each extract exhibited various biological activities. According to the HPLC results, the highest phenolic component in the extracts was determined as p-hydroxybenzoic acid. As a result, it was observed that the biological activities of orchid tubers changed according to different extraction methods. Accordingly, due to the high biological activities of orchid tubers, they have a high potential for use in the food and pharmaceutical industries.

Characterization of peroxiredoxin from Neocaridina denticulata sinensis and its antioxidant and DNA protection activity analysis

Peroxiredoxin (Prx) is an antioxidant protein that widely exists in various organisms. To further investigate the role of Prx in the antioxidant and immune responses of Neocaridina denticulata sinensis, the full-length cDNA sequence of a Prx gene (Nd-Prx) from N. denticulata sinensis was obtained. The open reading frame (ORF) of Nd-Prx is 597 bp and encodes 198 amino acids. Amino acid similarity alignment showed that Nd-Prx contained a conserved sequence region “FYPLDFTFVCPTEI”. qRT-PCR assay showed that Nd-Prx was expressed in all tested tissues and its expression was highest in the ovary. Nd-Prx was most highly expressed at 36 h after copper stimulation. Nd-Prx expression levels in hepatopancreas were significantly upregulated after Vibrio parahaemolyticus challenge (P < 0.05). In addition, the recombinant Nd-Prx was prepared and its enzyme activity was most stable at 70 °C with pH of 6.0. The antioxidant activity and DNA protection of recombinant Nd-Prx was also demonstrated. In summary, this study investigated the role of Prx in antioxidant and immune responses of N. denticulata sinensis, which might provide a foundation for further exploring Prx in immune system of crustaceans and for the application in disease control.

Development and biological evaluation of protective effect of kidney targeted N-acetylated chitosan nanoparticles containing thymoquinone for the treatment of DNA damage in cyclophosphamide-induced haemorrhagic cystitis

Thymoquinone (TQ), the most prominent constituent of Nigella sativa seeds, essential oil, is reported to possess an organ protective effect via Nrf2 expression and activation of Phase-II antioxidant enzymes. Haemorrhagic cystitis is the sudden onset of haematuria combined with bladder pain and irritable bladder symptoms are the known toxic effects of cyclophosphamide (CYP) chemotherapy. The objective of the present study was to investigate and compare the protective effect of thymoquinone (TQ) and thymoquinone nanoparticles (TQ-NP) in the kidney against CYP-induced haemorrhagic cystitis. Primarily, TQ-NP was fabricated by synthesis of N-acetylated chitosan and nanoparticle preparation by the ionic gelation technique. They were characterized by particle size, polydispersive index (PDI), zeta potential, entrapment efficiency (EE), SEM, and dynamic scattering calorimetry (DSC). Moreover, fluorescein isothiocyanate (FITC) labeled NPs were prepared for biodistribution studies. The protective mechanisms of TQ-NP included its anti-inflammatory activity, inhibitory effects on cytokine levels, and protection against the DNA damage in the bladder epithelium. The cystitis was induced in rats by orally administering 200 mg/kg of CYP. The dose-dependent protective effect of the TQ-NP was determined by intravenously administering 1, 2, and 5 mg/kg of the TQ-NP to CYP-treated rats. The present study revealed that the TQ-NP prepared by ionic gelation method provides kidney targeted delivery of TQ as compared to TQ solution. The mean particle size, PDI, and %EE of TQ-NP were 272.6 nm, 0.216, 70.81 ± 0.12% respectively. The zeta potential of thymoquinone-loaded nanoparticles was found to be −20.7 mV and − 22.6 mV respectively before and after lyophilization. SEM study also confirmed the small size and spherical shape. Pharmacokinetic studies revealed the improvement in half-life and prolonged action of the TQ-NP as compared to the TQ solution. Also, TQ-NP administration showed more protection against the characteristic histological alterations in the bladder in comparison to TQ solution. The present study indicates that TQ-NP exerts potent anti-oxidant, DNA protective and cytokine inhibitory activity at considerably lower concentrations as compared to plain TQ solution. The nano formulation of TQ using N-acetylated chitosan provides effective kidney targeted delivery of TQ, which in turn improves its retention and protective efficacy against CYP-induced haemorrhagic cystitis.

Antioxidant and DNA-Protective Potentials, Main Phenolic Compounds, and Microscopic Features of Koelreuteria paniculata Aerial Parts.

Interest in plant extracts as a natural source of antioxidants has grown significantly in recent years. The tree species Koelreuteria paniculata deserves attention due to its wide distribution, good adaptability, and growth to the degree of invasiveness in a number of European countries. The purpose of the present study was to analyze flavonoids and phenolic acids of the ethanol extracts from aerial parts of K. paniculata and to screen their antioxidant and DNA-protective activity. HPLC profiling revealed the presence of five flavonoids, with rutin (4.23 mg/g DW), hesperidin (2.97 mg/g DW), and quercetin (2.66 mg/g DW) as the major ones in the leaves, and (−)-epicatechin (2.69 mg/g DW) in the flower buds. Among the nine phenolic acids identified, rosmarinic, p-coumaric, salicylic, vanillic, and gallic acids were the best represented. All the extracts tested showed in vitro antioxidant activity that was determined by DPPH, ABTS, FRAP, and CUPRAC assays. The highest activity was recorded in the flower parts (in the range from 1133 to 4308 mmol TE/g DW). The DNA-protective capacity of the flower and stem bark extracts from the in vitro nicking assay performed, as well as the main diagnostic microscopic features of the plant substances, are given for the first time. According to the results obtained, the aerial parts of K. paniculata could be valuable sources of natural antioxidants.