Oleuropein Has Modulatory Effects on Systemic Lipopolysaccharide-Induced Neuroinflammation in Male Rats.

BACKGROUND: Neuroinflammation induced by systemic inflammation is a risk factor for developing chronic neurologic disorders. Oleuropein (OLE) has antioxidant and anti-inflammatory properties; however, its effect on systemic inflammation-related neuroinflammation is unknown. OBJECTIVES: This study aimed to determine whether OLE protects against systemic lipopolysaccharide (LPS)-induced neuroinflammation in rats. METHODS: Six-wk-old Wistar rats were randomly assigned to 1 of the following 5 groups: 1) control, 2) OLE-only, 3) LPS + vehicle, 4) OLE+LPS (O-LPS), and 5) a single-dose OLE + LPS (SO-LPS group). OLE 200 mg/kg or saline as a vehicle was administered via gavage for 7 d. On the seventh day, 2.5 mg/kg LPS was intraperitoneally administered. The rats were decapitated after 24 h of LPS treatment, and serum collection and tissue dissection were performed. The study assessed astrocyte and microglial activation using glial fibrillary acidic protein (GFAP) and CD11b immunohistochemistry, nod-like receptor protein-3, interleukin (IL)-1ß, IL-17A, and IL-4 concentrations in prefrontal and hippocampal tissues via enzyme-linked immunosorbent assay, and total antioxidant/oxidant status (TAS/TOS) in serum and tissues via spectrophotometry. RESULTS: In both the O-LPS and SO-LPS groups, LPS-related activation of microglia and astrocytes was suppressed in the cortex and hippocampus (P < 0.001), excluding cortical astrocyte activation, which was suppressed only in the SO-LPS group (P < 0.001). Hippocampal GFAP immunoreactivity and IL-17A concentrations in the dentate gyrus were higher in the OLE group than those in the control group, but LPS-related increases in these concentrations were suppressed in the O-LPS group. The O-LPS group had higher cortical TAS and IL-4 concentrations. CONCLUSIONS: OLE suppressed LPS-related astrocyte and microglial activation in the hippocampus and cortex. The OLE-induced increase in cortical IL-4 concentrations indicates the induction of an anti-inflammatory phenotype of microglia. OLE may also modulate astrocyte and IL-17A functions, which could explain its opposing effects on hippocampal GFAP immunoreactivity and IL-17A concentrations when administered with or without LPS.

Synthesis, Antimicrobial Activities, and Molecular Modeling Studies of Agents for the Sortase A Enzyme.

Sortase A (SrtA) is an attractive target for developing new anti-infective drugs that aim to interfere with essential virulence mechanisms, such as adhesion to host cells and biofilm formation. Herein, twenty hydroxy, nitro, bromo, fluoro, and methoxy substituted chalcone compounds were synthesized, antimicrobial activities and molecular modeling strategies against the SrtA enzyme were investigated. The most active compounds were found to be T2, T4, and T19 against Streptococcus mutans (S. mutans) with MIC values of 1.93, 3.8, 3.94 µg/mL, and docking scores of -6.46, -6.63, -6.73 kcal/mol, respectively. Also, these three active compounds showed better activity than the chlorohexidine (CHX) (MIC value: 4.88 µg/mL, docking score: -6.29 kcal/mol) in both in vitro and in silico. Structural stability and binding free energy analysis of S.mutans SrtA with active compounds were measured by molecular dynamic (MD) simulations throughout 100 nanoseconds (ns) time. It was observed that the stability of the critical interactions between these compounds and the target enzyme was preserved. To prove further, in vivo biological evaluation studies could be conducted for the most promising precursor compounds T2, T4, and T19, and it might open new avenues to the discovery of more potent SrtA inhibitors.

What Does a Community Think About Aromatherapy?

The awareness and usage of aromatherapy applications are increasing day by day in society. The current study aims to evaluate the thoughts and attitudes of individuals toward aromatherapy in Türkiye and to determine factors affecting the use of aromatherapy. This community-based descriptive study is conducted throughout Türkiye. Nine hundred eighty-five questionnaires were suitable for data quality. SPSS 23.0 statistical package program was used, and the statistical significance level was accepted as P value of less than .05. Approximately 72.4% of the participants have heard of aromatherapy. The rate of hearing about the concept of aromatherapy was found to be statistically significantly higher among women (P = .000), those residing in a province/district (P = .011), and those working in an income-generating positions (P = .003). It was determined that 58.9% of the participants used any essential oil. The 5 most commonly used essential oils are lavender, tea tree, peppermint, rose, and thyme, respectively. The rate of essential oil use in women is higher than in men (P = .000). The conditions in which essential oils are most frequently used are to provide beauty to the skin (51.7%), skin diseases (51.0%), and respiratory tract diseases (46.0%), respectively. The most common sources of information about the essential oils used by the participants are the Internet/social media (57.6%) and pharmacists (53.4%). The results of this research revealed that the awareness and use of aromatherapy practice in the community are high according to the literature. Considering the high tendency to use these products among the public, physicians, pharmacists, and nurses must provide consultancy services on the use of these products.

Pharmacy Students' Knowledge and Attitudes about Homeopathy: A Descriptive Survey Conducted in Turkey.

BACKGROUND: Homeopathy belongs to "Traditional and Complementary Medicine" (TCM) and is defined in the TCM regulation in Turkey as a holistic practice method that aims to improve health status with personalized medicines. The international and national literature includes a limited number of studies that examine individuals' knowledge, attitudes and behaviors toward homeopathy. Although in Turkey the Regulation on Pharmacists and Pharmacies states that the sale of homeopathic medicines is allowed only in pharmacies, no study was found that evaluated awareness about homeopathy among pharmacy students. OBJECTIVE: The present study aims to assess pharmacy students' knowledge and attitudes toward homeopathy and to identify the factors that influence them. METHODS: This descriptive study was conducted between December 1, 2021 and February 1, 2022 among the students of the Faculty of Pharmacy, Karadeniz Technical University (KTU), through a face-to-face survey. In total, 418 questionnaires suitable for data quality were included in the study. The SPSS 23.0 statistical program was used to analyze the data and the statistical significance level was taken as p < 0.05. RESULTS: It was determined that 73% of the participants had heard about TCM practices and 55% had heard of homeopathy. Students in the fifth grade (p = 0.0001) and those working in an income-generating job (p = 0.026) were found to be those most aware of homeopathy. The students of the Faculty of Pharmacy correctly knew the basic working principles of homeopathy (p = 0.002). The source from which students obtained the most information about homeopathy was undergraduate courses. 80.4% of the participants thought that homeopathy should be applied by pharmacists. 47.0% of the students wanted to learn more about homeopathy or to carry out studies in the field of homeopathy in their careers. CONCLUSION: The results of this research revealed a high awareness amongst KTU pharmacy students about the practice of homeopathy.

A descriptive survey study in endocrinology clinic: evaluation of the use of natural products.

OBJECTIVE: The use of herbal products/dietary supplements (HP/DS) in endocrinal chronic diseases is growing. However, no studies have evaluated their use in patients who present to endocrinology and metabolic diseases clinics. This descriptive study aims to investigate the rate of HP/DS use and the factors affecting this in patients who presented to Karadeniz Technical University (KTU) Farabi Hospital Endocrinology and Metabolic Diseases Clinic, Türkiye between 01.11.2021 and 01.05.2022. METHODS: Five hundred six questionnaires with acceptable data quality were included this investigation. The data were analyzed on SPSS version 23.0 software. The factors with the greatest effect on the use of HP/DS were determined using binary logistic regression analysis. RESULTS: Analysis showed that 49.4% of the participants used HP/DS. The main factors affecting the use of herbal products were age, diagnosis of the disease, and treatment compliance problems. The most frequently used products were lemon, cinnamon, black cumin, ginger, turmeric, and dill. The participants main sources of information about HP/DS were friends/relatives, the internet/social media, and television, respectively. 74.8% of the participants using HP/DS did not inform their physcisian/pharmacist about such use, although 81.8% of these nevertheless wished to receive information from these occupational groups. CONCLUSION: Herbal product monitoring in patients should be performed in collaboration with pharmacists, herbal product use should be investigated, and counseling services should be made available in order to maintain and promote public health.

In vitro tyrosinase inhibitory, DNA interaction studies, and LC-HRMS analysis of Ficus carica leaves.

Turkey is the world's leading producer of figs, a typical Mediterranean fruit. The fig, Ficus carica L. (Moraceae), has been widely cultivated since ancient times due to the nutritional value of its fruits. It was aimed to investigate the phytochemical characterization and biological properties of F. carica leaf extracts in order to determine their potential for use in the treatment of various diseases. F. carica leaves were extracted in 70% methanol at 40 °C under reflux. To obtain extracts of different polarities, the crude extract was fractionated with n-hexane, dichloromethane, and n-butanol. Phenolic content was determined using liquid chromatography-high resolution mass spectrometry (LC-HRMS). 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and antityrosinase activities of all extracts were investigated using spectrophotometric methods. Furthermore, the DNA-damage protective properties of extracts were investigated using electrophoretic methods. The n-butanol extract was found to have the highest total phenolic content, with 72.58 ± 4.52 mg GAE/g dry weight. According to LC-HRMS analysis, rutin (40.13 g/kg) was the most abundant compound in the n-butanol extract. The n-butanol extract, which was found to have the highest tyrosinase inhibitory effects among the extracts, demonstrated radical scavenging activity of 37.01 ± 1.15% and 82.57 ± 0.88% at 80 and 200 µg/mL, respectively. The n-butanol extract had the highest protective effects against Fenton's reagent, UV radiation, and singlet oxygen. Given these findings, it is possible to argue that F. carica leaves can be evaluated for developing products that could be used to treat various diseases.

Dihydroisocoumarins and Phenylglycosides from Scorzonera longiana, Their Antimicrobial Activities and Molecular Docking Studies.

Five new phenyl dihydroisocoumarin glycosides (1-5) and two known compounds (6-7) were identified from the butanol fraction of Scorzonera longiana. The structures of 1-7 were elucidated based on spectroscopic methods. Antimicrobial, antitubercular, and antifungal evaluation of compounds 1-7 were carried out using the microdilution method against nine microorganisms. Compound 1 was active only against Mycobacterium smegmatis (Ms) with a MIC value of 14.84 µg/mL. All tested compounds (1-7) were active against Ms but only compounds 3-7 were active against fungi (C. albicans, S. cerevisiae) with MIC values of 25.0-125 µg/mL. In addition, molecular docking studies were conducted against Ms DprE1 (PDB ID: 4F4Q), Mycobacterium tuberculosis (Mbt) DprE1 (PDB ID: 6HEZ), and arabinosyltransferase C (EmbC, PDB ID: 7BVE) enzymes. Compounds 2, 5, and 7 are the most effective Ms 4F4Q inhibitors. Compound 4 was the most promising inhibitory activity on Mbt DprE with the lowest binding energy of -9,9 kcal/mol.

Two new dihydroisocoumarins and terpenoids from Scorzonera longiana Sümbül an endemic species to Turkey and their antimicrobial activity.

Two new dihydroisocoumarins (scorzolongin I (1), and scorzolongin II (2)) and nine known compounds (3',5'-dimethoxy hydrangenol (scorzolongin III, 3), cladantholide (4), dammar-24-ene-3ß-ol (5), taraxasterol (6), ß-sitosterol (7), mangifgerursanone (8), and a mixture of α-amyrenone (9a), ß-amyrenone (9b), and dammar-24-ene-3-one (9c) in about 1:1:2 ratio) were identified from the dichloromethane fraction of Scorzonera longiana. The structure of all compounds (1-9a-c) were elucidated by extensive 1D and 2D NMR (1H, 13C/APT, COSY, HMBC, HSQC, and NOESY) spectroscopy, UV, FT-IR, and LC-QTOF-MS data and by comparison of their NMR data with the literature. These compounds have been isolated from S. longiana for the first time. An antimicrobial assay against eight microorganisms was applied to isolated compounds 1-3. Scorzolongin I, and scorzolongin II, and scorzolongin III showed notable activity against gram (-) (Escherichia coli and Yersinia pseudotuberculosis) and fungi (Candida albicans, Saccharomyces cerevisiae) with 20 mm inhibition zone each. Scorzolongin II (2) exhibited strong activity against E. coli, Y. pseudotuberculosis, Mycobacterium smegmatis C. albicans, S. cerevisiae with MIC value of 33.8 µg/mL.

Wound healing acceleration and anti-inflammatory potential of Prunella vulgaris L.: From conventional use to preclinical scientific verification.

ETHNOPHARMACOLOGICAL RELEVANCE: The genus Prunella L. (Lamiaceae) is represented by nine species in the world and four species in Turkey. The infusion prepared from the aerial parts of Prunella vulgaris L. is used internally for abdominal pain and as an expectorant, the decoction prepared from all parts is used internally or externally as a wound healing. AIM OF THE STUDY: This study aims to investigate the wound healing potential of Prunella vulgaris L. on the scientific platform. MATERIAL AND METHODS: The aerial parts of the plant were extracted with 80% methanol. The resulting aqueous methanol extract was partitioned with n-hexane and ethyl acetate, and sub-extracts were obtained. The wound healing effects of the methanol extract and sub-extracts were studied in mice and rats using linear incision and circular excision wound models, and the anti-inflammatory effect was investigated using acetic acid-induced capillary permeability test. Isolation studies were performed using the ethyl acetate sub-extract, which exhibited the highest activity. RESULTS: Using various chromatographic methods, 6 compounds were isolated from the ethyl acetate sub-extract. The structures of the compounds were identified as methyl arginolate, ursolic acid, chlorogenic acid, rosmarinic acid, methyl 3-epimaclinate, and ethyl rosmarinate by spectroscopic techniques (UV, IR, 13C-NMR, 1H-NMR, 2D-NMR, MS). The wound healing mechanisms of the pure compounds were investigated by performing assays to inhibit the enzymes hyaluronidase, collagenase, and elastase. Ursolic acid, chlorogenic acid, and rosmarinic acid were found to be responsible for the anti-inflammatory and wound healing effects. CONCLUSION: The experimental study revealed that Prunella vulgaris showed significant wound healing and anti-inflammatory activities.

Immunomodulatory properties of triterpenes.

The immune system is one of the main defence mechanisms of the human body. Inadequacy of this system or immunodeficiency results in increased risk of infections and tumours, whereas over-activation of the immune system causes allergic or autoimmune disorders. A well-balanced immune system is important for protection and for alleviation of these diseases. There is a growing interest to maintain a well-balanced immune system, especially after the Covid-19 pandemic. Many biological extracts, as well as natural products, have become popular due to their wide array of immunomodulatory effects and influence on the immune system. Triterpenes, one of the secondary metabolite groups of medicinal plants, exhibit immunomodulatory properties by various mechanisms. Different triterpenes, including components of commonly consumed plants, can promote some protection and alleviation of disease symptoms linked with immune responses and thus enhance overall well-being. This review aims to highlight the efficacy of triterpenes in light of the available literature evidence regarding the immunomodulatory properties of triterpenes. We have reviewed widely investigated immunomodulatory triterpenes; oleanolic acid, glycyrrhizin, glycyrrhetinic acid, pristimerin, ursolic acid, boswellic acid, celastrol, lupeol, betulin, betulinic acid, ganoderic acid, cucumarioside, and astragalosides which have important immunoregulatory properties. In spite of many preclinical and clinical trials were conducted on triterpenes related to their immunoregulatory actions, current studies have several limitations. Therefore, especially more clinical studies with optimal design is essential.

Anti-inflammatory secondary metabolites from Scrophularia kotschyana.

The species belonging to Scrophularia genus grow mainly in Irano-Turanian and Mediterranean regions and have been used as folk remedy for inflammatory-related diseases since ancient times. The present study was aimed to evaluate the anti-inflammatory activity of the extracts of Scrophularia kotschyana as well as the isolated compounds. The aerial parts and the roots of the plant were separately extracted with methanol. Anti-inflammatory activities of both extracts were evaluated with formalin test in mice. As the methanolic extract of the aerial parts significantly (p < .05) inhibited inflammation, it was then submitted to successive solvent extractions with n-hexane, dichloromethane, ethyl acetate and n-butanol to yield subextracts. Anti-inflammatory activities of the subextracts were evaluated within the same test system. Among the subextracts tested, the n-butanol subextract produced a significant (p < .05) anti-inflammatory activity at all doses (5, 10, and 30 mg/kg, ip.). Sequential chromatographic separation of the n-butanol subextract yielded 8-O-acetyl-4'-O-(E)-p-coumaroylharpagide, 8-O-acetyl-4'-O-(Z)-p-coumaroylharpagide, ß-sitosterol 3-O-ß-glucopyranoside, apigenin 7-O-ß-glucopyranoside, apigenin 7-O-rutinoside, luteolin 7-O-ß-glucopyranoside and luteolin 7-O-rutinoside. The anti-inflammatory activities of the isolates were evaluated at 5 mg/kg dose. Luteolin 7-O-ß-glucopyranoside and apigenin 7-O-rutinoside caused a significant (p < .05) inhibition of oedema formation.

Analysis of plant secondary metabolism using stable isotope-labelled precursors.

INTRODUCTION: Analysis of biochemical pathways typically involves feeding a labelled precursor to an organism, and then monitoring the metabolic fate of the label. Initial studies used radioisotopes as a label and then monitored radioactivity in the metabolic products. As analytical equipment improved and became more widely available, preference shifted the use stable 'heavy' isotopes like deuterium (2 H)-, carbon-13 (13 C)- and nitrogen-15 (15 N)-atoms as labels. Incorporation of the labels could be monitored by mass spectrometry (MS), as part of a hyphenated tool kits, e.g. Liquid chromatography (LC)-MS, gas chromatography (GC)-MS, LC-MS/MS. MS offers great sensitivity but the exact location of an isotope label in a given metabolite cannot always be unambiguously established. Nuclear magnetic resonance (NMR) can also be used to pick up signals of stable isotopes, and can give information on the precise location of incorporated label in the metabolites. However, the detection limit for NMR is quite a bit higher than that for MS. OBJECTIVES: A number of experiments involving feeding stable isotope-labelled precursors followed by NMR analysis of the metabolites is presented. The aim is to highlight the use of NMR analysis in identifying the precise fate of isotope labels after precursor feeding experiments. As more powerful NMR equipment becomes available, applications as described in this review may become more commonplace in pathway analysis. CONCLUSION AND PROSPECTS: NMR is a widely accepted tool for chemical structure elucidation and is now increasingly used in metabolomic studies. In addition, NMR, combined with stable isotope feeding, should be considered as a tool for metabolic flux analyses.

The Volatile Chemical Compositions of the Essential Oil/SPME and Enzyme Inhibitory and Radical Scavenging Activities of Solvent Extracts and the Essential oils from Coronilla orientalis Miller and C. varia L. grows in Turkey.

The volatile organic compounds (VOCs) of two Coronilla species (Coronilla orientalis Miller and Coronilla varia L.) obtained by hydrodistillation (HD) and solid phase microextraction (SPME) techniques were identified by GC-FID/MS. The major compounds identified in the SPME extracts were limonene (43.4%) in Coronilla orientalis (C. orientalis), (Z)-ß-ocimene and (E)-ß-ocimene (34.3% and 32.4%) in Coronilla varia (C. varia), whereas, the essential oils of C. orientalis and C. varia were rich with γ-terpinene (22.4%) and phytol (30.7%), respectively. In addition, acetylcholinesterase (AChE), butyrylcholinesterase (BuChE), tyrosinase, α-glucosidase enzyme inhibitory, and radical scavenging activities (DPPH) of chloroform, ethyl acetate, methanol, and water extracts, and also essential oils obtained from C. orientalis and C. varia were investigated. The tyrosinase activity was studied at the doses of 25 µg/mL, 50 µg/mL and 100 µg/mL. Tyrosinase inhibition percentage was observed to increase by dose and methanol extracts of the both species were found to have the highest activity. Essential oils of the both species were found to have significant acetylcholinesterase and butyrylcholinesterase inhibition activities. α-Glucosidase enzyme inhibition of the ethyl acetate and water extracts of C. orientalis was determined as 80.11 ± 4.07% and 80.32 ± 3.47% at the 100 µg/mL concentration, respectively. Essential oils, chloroform, ethyl acetate, methanol, and water extracts were determined to have moderate DPPH radical scavenging activities.

α-Glucosidase inhibitory effects of polyphenols from Geranium asphodeloides: Inhibition kinetics and mechanistic insights through in vitro and in silico studies.

Some Geranium species have been used to treat diabetes. To evaluate the scientific basis of this ethnopharmacological use, we aimed to isolate potent α-glucosidase inhibitory metabolites of Geranium asphodeloides Burm. through in vitro bioactivity-guided fractionation. All the tested extracts showed high α-glucosidase inhibitory effect compared to acarbose. Among the tested extracts, the ethyl acetate subextract showed the highest activity with an IC50 value of 0.85 ±â€¯0.01 µM. A hydrolysable tannin, 1,2,4-tri-O-galloyl-ß-d-glucopyranose (1), and five flavonoid glycosides, kaempferol-3-O-α-rhamnopyranoside (2), kaempferol-3-O-α-arabinofuranoside (3), quercetin-3-O-ß-glucopyranoside (4), quercetin-3-O-α-rhamnopyranoside (5), and quercetin-3-O-α-rhamnofuranoside (6), were isolated from the ethyl acetate subextract. Their structures were identified by 1D- and 2D-NMR experiments. 1 exhibited the highest α-glucosidase inhibitory effect, approximately 61 times more potent than positive control, acarbose, with an IC50 value of 0.95 ±â€¯0.07 µM. Also, 2 was more potent than acarbose. An enzyme kinetics analysis revealed that compounds 2, 3 and 4 were competitive, whereas 1 and 6 uncompetitive inhibitors. Molecular docking studies were performed to get insights into inhibition mechanisms of the isolated compounds in the light of the enzyme kinetic studies using various binding sites of the enzyme model.

Evaluation of in vivo analgesic activity of Scrophularia kotscyhana and isolation of bioactive compounds through activity-guided fractionation.

The present study was undertaken to evaluate the in vivo analgesic activities of the extracts prepared from the aerial parts and roots of Scrophularia kotscyhana and to isolate the bioactive metabolites from the most active extract. Analgesic activities of all extracts and subextracts at the doses of 5, 10 and 30 mg/kg (i.p.) were examined using hot plate test in mice. Among the tested extracts, MeOH extract prepared from the aerial parts and the n-butanol subextract prepared thereof displayed the best analgesic activity at all doses. Phytochemical studies on n-butanol subextract led to the isolation of two new iridoid glycosides as an inseparable mixture, 8-O-acetyl-4'-O-(E)-(p-coumaroyl)-harpagide (1) and 8-O-acetyl-4'-O-(Z)-(p-coumaroyl)-harpagide (2) along with five known secondary metabolites, ß-sitosterol 3-O-ß-glucopyranoside (3), apigenin 7-O-ß-glucopyranoside (4), apigenin 7-O-rutinoside (5), luteolin 7-O-ß-glucopyranoside (6) and luteolin 7-O-rutinoside (7). The iridoid mixture (1 and 2), 3 and 4 elicited significant inhibition of pain at 5 mg/kg dose.

In Vitro Protection by Crataegus microphylla Extracts Against Oxidative Damage and Enzyme Inhibition Effects.

OBJECTIVES: Crataegus species have been used as food and also in folk medicine for the treatment of various diseases. The present study aimed to make investigations on the biologic properties of different extracts prepared from Crataegus microphylla C. Koch, which was collected from Turkey. MATERIALS AND METHODS: Dried leaf, stem bark, and fresh fruit samples of C. microphylla were separated and ethanol extract, acidified (0.5% HCl, pH: 2.5) ethanol extract, ethanol:water (1:1) extract, methanol extract, acidified (0.5% HCl, pH: 2.5) methanol extract, methanol:water (1:1) extract, water extract, and acidified (0.5% HCl, pH: 2.5) water extract were prepared for each. Various biologic effects such as the prevention of oxidative DNA damage, acetylcholinesterase, tyrosinase, α-glucosidase inhibition, and antioxidant effects with 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, PRAP, and FRAP assays of these extracts at different concentrations were studied. RESULTS: Acidified methanol extract of stem barks exhibited the highest acetylcholinesterase and tyrosinase inhibitions among the other extracts with IC50 values of 204.02±0.95 µg/mL and 37.30±0.27 µg/mL, respectively. Acidified ethanol extract of leaves was the most efficient extract against α-glucosidase, giving an IC50 of 15.78±0.14 µg/mL. The IC50 value of the acidified ethanol extract for DPPH was 9.89±0.09 µg/mL. Methanol extracts of leaves and stem barks at the dose of 125 µg/mL exhibited significant protective activity against DNA strand scission by hydroxyl radicals (.OH) on supercoiled pBR322 DNA. CONCLUSION: Acidified methanol or ethanol extracts prepared with stem bark and leaf from C. microphylla have potential antioxidant, hypoglycemic, and neuroprotective effects.

Carbonic Anhydrase and Urease Inhibitory Potential of Various Plant Phenolics Using in vitro and in silico Methods.

Plant phenolics are known to display many pharmacological activities. In the current study, eight phenolic compounds, e.g., luteolin 5-O-ß-glucoside (1), methyl rosmarinate (2), apigenin (3), vicenin 2 (4), lithospermic acid (5), soyasaponin II (6), rubiadin 3-O-ß-primeveroside (7), and 4-(ß-d-glucopyranosyloxy)benzyl 3,4-dihydroxybenzoate (8), isolated from various plant species were tested at 0.2 mm against carbonic anhydrase-II (CA-II) and urease using microtiter assays. Urease inhibition rate for compounds 1 - 8 ranged between 5.0 - 41.7%, while only compounds 1, 2, and 4 showed a considerable inhibition over 50% against CA-II with the IC50 values of 73.5 ± 1.05, 39.5 ± 1.14, and 104.5 ± 2.50 µm, respectively, where IC50 of the reference (acetazolamide) was 21.0 ± 0.12 µm. In silico experiments were also performed through two docking softwares (Autodock Vina and i-GEMDOCK) in order to find out interactions between the compounds and CA-II. Actually, compounds 6 (30.0%) and 7 (42.0%) possessed a better binding capability toward the active site of CA-II. According to our results obtained in this study, among the phenolic compounds screened, particularly 1, 2, and 4 appear to be the promising inhibitors of CA-II and may be further investigated as possible leads for diuretic, anti-glaucoma, and antiepileptic agents.

Phytochemical Content, Antioxidant and Cytotoxic Activities of Sedum spurium.

Sedum L. species are used for their hemostatic, antidiarrheal, antifungal, diuretic and wound healing properties, and there is growing interest in these species because of their usage in folk medicine. DPPH, SO, NO, and ABTS radical scavenging activities and protective effects against H(2)(02) induced cytotoxicity, as well as cytotoxic activities against the Hep-2 cell line of various extracts from Sedum spurium Bieb. were investigated. Besides, the total phenol, flavonoid, and flavonol contents of the extracts were determined to clarify their biological and phytochemical properties. Chromatographic studies on the most active extract led to the isolation of the major compound, identified as 2-methyl-erythritol by (1)H and (13)C NMR techniques. The EtOAc extract is found to be the most active extract in all tests. However, major compound of EtOAc extract did not possess tested activities. The EtOAC extract of S. spurium could be effective to improve antihemolytic defences of erythrocytes, and radical scavenging potential of the antioxidant mechanism. The extracts should be investigated in detail for their cytotoxic activities because of their possible pro-oxidant effects at high concentrations.

Comparative assessment of dermal wound healing potentials of various Trifolium L. extracts and determination of their isoflavone contents as potential active ingredients.

ETHNOPHARMACOLOGICAL RELEVANCE: Trifolium species are used in Turkish folk medicine as a wound healing agent, expectorant, antiseptic, sedative and to alleviate pain in rheumatism. In the present study, the aqueous methanolic extracts (80%) of 13 Trifolium species (Trifolium ambigum, Trifolium arvense var. arvense, Trifolium campestre, Trifolium canescens, Trifolium hybridum var. anatolicum, Trifolium hybridum var. hybridum, Trifolium pannonicum, Trifolium pratense var. pratense, Trifolium purpureum var. purpureum, Trifolium repens var. repens, Trifolium resupinatum var. microcephalum, Trifolium spadiceum and Trifolium trichocephalum) collected from different regions of Anatolia were evaluated for their in vivo wound healing effects. MATERIALS AND METHODS: In vivo wound healing activities of the plant aqueous methanolic extracts were evaluated by linear incision and circular excision wound models subsequent to histopathological analysis. Active constituents were determined by a validated high performance liquid chromatographic method. Precision of the method was performed by the evaluation of intra-day and inter-day variations of the each standard at limits of quantification (LOQ) levels. RESULTS: The aqueous methanolic extracts of Trifolium canescens and Trifolium pretense var. pratense possessed better wound healing activity compared to the other extracts and control groups. The animal groups treated with the Trifolium canescens extract demonstrated increased contraction (48.96%) on excision and a significant increase in wound tensile strength (35.6%) on incision models. The main compounds were detected as genistein and biochanin A for Trifolium canescens. CONCLUSION: The results of the present study revealed the wound healing potential of Trifolium canescens. This might be due to the combined effect of the isoflavones genistein, formononetin, daidzein, and biochanin A present in the extract.