Comprehensive Analysis of the Chemical and Bioactivity Profiles of Endemic Crataegus turcicus Dönmez in Comparison with Other Crataegus Species.

Crataegus turcicus is a plant endemic to Türkiye. For the first time, this study aimed to comparatively assess its flower-bearing branches, leaves, and fruits with other well-known Crataegus species (C. monogyna, C. pentagyna, and C. orientalis) in terms of chemical composition and bioactivity studies to evaluate its potential use as a food supplement. Firstly, the contents of total phenolics (TPC), flavonoids (TFC), proanthocyanidin (TPAC), and anthocyanin (TAC) in different plant parts of Crataegus species were evaluated. The highest TPAC was found in the hydroalcoholic extract of C. turcicus flower-bearing branches. Moreover, all plant parts had comparatively higher amounts of TPC, TFC, and TAC compared to other Crataegus species. The chemical screening by high-performance thin-layer chromatography (HPTLC) resulted that C. turcicus parts were rich with chlorogenic acid, neochlorogenic acid, quercetin and vitexin derivatives, epicatechin, procyanidin, etc., and their quantities were evaluated by high-performance liquid chromatography (HPLC). In terms of several in vitro antioxidant activity outcomes, the flower-bearing branches of C. turcicus showed the highest antioxidant activity by a 2,2-diphenyl-1-picrylhydrazyl (DPPH) test among the assessed antioxidant assays. Additionally, hydroalcoholic extracts of C. turcicus significantly decreased LPS-induced nitric oxide, tumor necrosis factor-alpha, and interleukin-6 production more potently than indomethacin (positive control). In addition to its remarkable anti-inflammatory activity, C. turcicus showed analgesic activity by reducing prostaglandin E2 levels.

Comprehensive estrogenic/anti-estrogenic, anticancer, mutagenic/anti-mutagenic, and genotoxic/anti-genotoxic activity studies on chemically characterized black poplar and Eurasian aspen propolis types.

Propolis is mainly composed of plant resins, and its type is named according to the primary plant origin in its composition. Identification of propolis botanical origin is essential for predicting and repeating its pharmacological activity because of the variations in chemical composition. This study aimed to compare chemical composition of black poplar (Populus nigra L.) type-propolis (PR1 and PR2) and Eurasian aspen (P. tremula L.)-type propolis (PR3) by liquid chromatography-tandem mass spectrometry (LC-MS/MS) technique and to evaluate their biological activity profiles. According to LC-MS/MS results, in addition to marked caffeic acid phenethyl ester content in PR1 and PR2, flavonoid aglycones such as pinocembrin, chrysin, pinobanksin, and galangin were found to be dominant in these samples. On the other hand, PR3 contained relatively high concentrations of phenolic acids such as ferulic acid, p-coumaric acid, and trans-cinnamic acid. The anti-estrogenic activity test showed that PR2 exerted the highest anti-estrogenic activity by inhibiting cell proliferation by 44.6%. All propolis extracts showed anticancer activity, which was justified by decreasing activity on the 3D spheroid size in a concentration-dependent manner. Besides, all extracts showed moderate or potent antimutagenic activity in Salmonella typhimurium TA98 and TA100 strains with and without metabolic activation, respectively. In addition, the Comet assay results revealed that propolis extracts have a geno-protective effect against H2O2-induced DNA damage in CHO-K1 cells at 0.625 and 1.25 µg/mL concentrations. Overall, the result of this study may help in preparing standardized propolis extracts and developing products with defined pharmacological benefits in the food supplements industry.

Secondary metabolites from the underground parts of Valeriana sisymbriifolia Vahl. and their in vitro cytotoxic activities.

Cytotoxic activity-guided isolation studies on the underground parts of Valeriana sisymbriifolia Vahl. led to the isolation of 12 secondary metabolites including two undescribed iridoids, sisymbriifolivaltrate and sisymbriifolioside, and two unreported sesquiterpene lactones, sisymbriifolins A and B. Chemical structures of the isolates were established by extensive 1D and 2D NMR analyses as well as HR-ESI-MS. The in vitro cytotoxic activities of the extract, sub-fractions and isolates on lung (A549), breast (MCF7), gastric (HGC27) and prostate (PC3) cancer cell lines were evaluated by MTS assay. Sisymbriifolivaltrate, didrovaltrate, valtrate, 7-homovaltrate and 1-α-acevaltrate exhibited promising cytotoxic activity on MCF7 cell line with IC50 values ranging from 2.5 to 12.3 µM, while valtrate demonstrated the best cytotoxicity against A549 cells with the IC50 value of 7.5 µM. Valtrate and 7-homovaltrate were found to exert noteworthy cytotoxicity towards HGC27 cell line (IC50 values: 2.3 and 3.7 µM, respectively), whereas valtrate, 7-homovaltrate and 1-α-acevaltrate (IC50 values: 2.3-9.7 µM) were found to be potent cytotoxic against PC3 cells. Among the tested compounds, particularly valepotriate-type iridoids were found to be the main cytotoxic principles of V. sisymbriifolia.

Activity-Guided Isolation of Cytotoxic Non-Glycosidic Ester Iridoids from Valeriana alliariifolia Adams and Unravelling Their Cell Death Mechanisms.

The aim of this study was to isolate the cytotoxic compounds from V. alliariifolia via activity-guided isolation and to determine the mechanism of actions of the most potent ones. The crude EtOH extract as well as CHCl3 and AcOEt subextracts demonstrated remarkable cytotoxic activities against A549, MCF7, HGC27 and PC3 cancer cells. Sequential chromatographic separations on active subextracts yielded 14 secondary metabolites, including 11 iridoids (1-11) most of which belong to non-glycosidic ester iridoids, two phenylpropanoids (12 and 13) and one lignan (14). The chemical structures of purified compounds were elucidated by NMR and MS analysis. Among the isolates, 7-deisovaleroylvaltrate (3) was isolated for the first time as a natural product. According to the cytotoxic assay compounds, 2, 4-6 and 8 were found to be the potent cytotoxic compounds (IC50 <10 µM) against at least one of the tested cancer cell lines. Thus, 2, 4-6 and 8 were investigated for their effects on apoptotic, necrotic and autophagic pathways as well as cell cycle progression. They exerted anticancer activities by inducing different cell death mechanisms depending on the cancer cells. The results demonstrated that 2, 4-6 and 8 could be potential anticancer drug leads that deserve further in vivo and clinical studies on the way to discover novel natural compounds with anticancer properties.

A comprehensive study to evaluate the wound healing potential of okra (Abelmoschus esculentus) fruit.

ETHNOPHARMACOLOGICAL RELEVANCE: Okra fruit (Abelmoschus esculentus (L.) Moench) has been extensively used for the treatment of skin damage and subcutaneous tissue abscess for many years in Turkish folk medicine. AIM OF STUDY: In this study, we aimed to investigate the wound healing potential of okra fruit by in vitro and in vivo experimental models in detail. Furthermore, based on the results of experiments, a wound healing formulation was developed and its activity profile was studied. MATERIALS AND METHODS: For this purpose, the phenolic, flavonoid and proanthocyanidin contents and chemical profile of aqueous and ethanolic extracts prepared from okra fruits cultivated in two different locations of Turkey, i.e. Aegean and Kilis regions, were comparatively determined and the tryptophan levels, which is known to be an influential factor in wound healing, were measured. Antioxidant activity of the okra fruit extracts was determined by DPPH test, ABTS radical scavenger activity, iron-binding capacity, total antioxidant capacity and copper reduction capacity assays. Moreover, antibacterial activity potentials of the aqueous and ethanolic extracts of okra fruits were determined. The protective effect of the extracts against H2O2-induced oxidative stress and anti-inflammatory activity were assessed in HDF (human dermal fibroblast) cells and in RAW 264.7 murine macrophages, respectively. The biocompatibility of the gel formulations prepared with the best performing extract were evaluated by human Epiderm™ reconstituted skin irritation test model. Wound-healing activity was investigated in rats by in vivo excision model and, histopathological examination of tissues and gene expression levels of inflammation markers were also determined. RESULTS: According to our findings, the aqueous and ethanolic extracts of okra fruits were found to possess a rich in phenolic content. Besides, isoquercitrin was found to be a marker component in ethanolic extracts of okra fruits. Both extracts exhibited antioxidant activity with significant protective effect against H2O2-induced damage in HDF cells by diminishing the MDA level. Also, the highest dose of ethanolic extracts has displayed a potent anti-inflammatory activity on LPS-induced RAW264.7 cells. Besides, both water and ethanolic extracts were shown to possess antimicrobial activity. On the other hand, the formulations prepared from the extracts were found non-irritant on in vitro Epiderm™-SIT. In vivo excision assay showed that tissue TGF-ß and IL-1ß levels were significantly decreased by the 5% okra ethanolic gel formulation. The histopathological analysis also demonstrated that collagenisation and granulation tissue maturation were found higher in 5% (w/v) okra ethanolic extract-treated group. CONCLUSION: 5% of okra ethanolic extract might be suggested as a potent wound healing agent based on the antimicrobial, antioxidant and anti-inflammatory tests. The proposed activity was also confirmed by the histopathological findings and gene expression analysis.

Influence of In Vitro Human Digestion Simulation on the Phenolics Contents and Biological Activities of the Aqueous Extracts from Turkish Cistus Species.

Oxidative stress is one of the significant precursors of various metabolic diseases such as diabetes, Parkinson's disease, cardiovascular diseases, cancer, etc. Various scientific reports have indicated that secondary plant metabolites play an important role in preventing oxidative stress and its harmful effects. In this respect, this study was planned to investigate the phenolic profile and antioxidant and antidiabetic potentials of the aqueous extracts from Turkish Cistus species by employing in vitro methods. In vitro digestion simulation procedure was applied to all extracts to estimate the bioavailability of their phenolic contents. Total phenolic, flavonoid, phenolic acid and proanthocyanidin contents were determined for all phases of digestion. In addition, changes in the quantity of the assigned marker flavonoids (tiliroside, hyperoside and quercitrin) were monitored by High-Performance Thin Layer Chromatography (HPTLC) analysis. The antioxidant activity potentials of the extracts were studied by various methods to reveal their detailed activity profiles. On the other hand, in vitro α-amylase and α-glucosidase enzymes and advanced-glycation end product (AGE) inhibitory activities of the extracts were determined to evaluate the antidiabetic potentials of extracts. The results showed that aqueous extracts obtained from the aerial parts of Turkish Cistus species have rich phenolic contents and potential antioxidant and antidiabetic activities; however, their bioactivity profiles and marker flavonoid concentrations might significantly be affected by human digestion. The results exhibited that total phenolic contents, antioxidant activities and diabetes-related enzyme inhibitions of the bioavailable samples were lower than non-digested samples in all extracts.

The Protective Effect of Nigella sativa Oil against Experimentally Induced Cisplatin Ototoxicity: An Animal Study.

OBJECTIVES: The objective of this study was to investigate the potential protective effect of Nigella sativa oil (NSO) against cis-diamminedichloroplatinum or cisplatin (CDDP)-induced ototoxicity. MATERIALS AND METHODS: Twenty-four Wistar albino rats were randomly and equally divided into four groups. Groups 1 and 2 were given a total of 15 mg/kg CDDP intraperitoneally, which was divided equally into three doses on days 1, 3, and 5. Group 2 was treated via gavage feeding with 15 ml NSO that was divided into five doses on days 1, 3, 5, 7, and 9. Groups 3 and 4 received only 15 ml of NSO and 15 ml of 0.9% saline solution, respectively, which were orally administered and divided into five doses on days 1, 3, 5, 7, and 9. Baseline high-frequency (8, 12, 16, and 32 kHz) auditory brainstem response (ABR) measurements were collected in all the groups before the medical administrations and were repeated on the 14th day before sacrifice. Afterward, a histopathological evaluation of the cochlea was performed. RESULTS: There was a significant difference in the histopathological changes between group 1 and the other groups (p<0.01). Changes in the spiral ganglion cells, the stria vascularis, and the external ciliated cells were significantly different between groups 1 and 2 (p=0.019, 0.039, and 0.045, respectively). The ABR results revealed significant differences in the 16 and 32 kHz measurements between groups 1 and 2 (p=0.013 and p<0.01, respectively). CONCLUSION: According to the results, NSO may have a protective effect on cochlear function against the disruptive effects of CDDP in rats.

Investigation of the Protective Effect of Nigella Sativa Oil in Cisplatin Induced Oral Mucositis: An Experimental Study.

OBJECTIVE: The aim of this experimental study was to investigate the histopathologic effect of Nigella Sativa oil (NSO) on cisplatin (Cis) induced oral mucositis (OM) in rats. METHODS: Twenty-four rats were divided into four equal groups. The animals in Group 1 and Group 2 were given 5 mg/kg intraperitoneal (ip) Cis systemically on the 1st, 3rd and 5th days of the study. Additionally, 15 mL NSO were given to the rats in Group 2, with gavage feeding on days 1, 3, 5, 7, and 9. The animals in Group 3 were given per oral 15 ml NSO on days 1, 3, 5, 7 and 9. As the control group, Group 4 received a total of 15 mL 0.9% saline solution divided into 5 doses on days 1, 3, 5, 7 and 9 by oral gavage. On the 14th day, animals were euthanized and buccal mucosa from both sides, including submucosal tissues, were excised and taken to histopathological examination. RESULTS: The mean mucosal thicknesses of the groups were 224.58 µm, 276.1 µm, 323.33 µm, and 331.33 µm, respectively for Groups 1, 2, 3, and 4 (p<0.05). When the degree of mucosal inflammation was examined, the most intense inflammation was detected in Group 1 and the least intense inflammation was in Group 4 (p<0.01). The degree of inflammation in Group 2 and Group 3 were similar to Group 4 (p>0.05). CONCLUSION: According to the results of this study, NSO, for which anti-inflammatory and antioxidant properties have been shown in previous studies, may also be effective in preventing Cis-induced OM.

Phenolic compounds from the aerial parts of Clematis viticella L. and their in vitro anti-inflammatory activities.

Phytochemical investigations on the EtOH extract of Clematis viticella led to the isolation of six flavonoid glycosides, isoorientin (1), isoorientin 3'-O-methyl ether (2), quercetin 7-O-α-L-rhamnopyranoside (3), quercetin 3,7-di-O-α-L-rhamnopyranoside (4), manghaslin (5) and chrysoeriol 7-O-ß-D-glucopyranoside (6), one phenylethanol derivative, hydroxytyrosol (7), along with three phenolic acids, caffeic acid (8), (E)-p-coumaric acid (9) and p-hydroxybenzoic acid (10). The structures of the isolates were elucidated on the basis of NMR and HR-MS data. All compounds were isolated from C. viticella for the first time. Compounds 7 and 8 showed significant anti-inflammatory activity at 100 µM by reducing the release of NO in LPS-stimulated macrophages comparable to positive control indomethacin. Compounds 3 and 7 exhibited anti-inflammatory activity through lowering the levels of TNF-α while 1, 3 and 5 decreased the levels of neopterin better than the positive controls.

Sambulin A and B, non-glycosidic iridoids from Sambucus ebulus, exert significant in vitro anti-inflammatory activity in LPS-induced RAW 264.7 macrophages via inhibition of MAPKs's phosphorylation.

ETHNOPHARMACOLOGICAL RELEVANCE: The leaves of Sambucus ebulus L. (Adoxaceae) are widely used in Turkish folk medicine particularly against inflammatory disorders. The fresh leaves after wilted over fire or the poultices prepared are directly applied externally to heal burns, edema, eczema, urticarial and abscess. Two iridoids were recently isolated (sambulin A, sambulin B) from the leaves of S. ebulus. AIM OF THE STUDY: This study aims to investigate the in vitro anti-inflammatory activities of these iridoids on LPS-induced RAW 264.7 macrophages. MATERIALS AND METHODS: Raw 264.7 macrophages were treated with 12.5, 25 and 50µg/ml Sambulin A and 6.25, 12.5 and 25µg/ml Sambulin B and induced with 1µg/ml lipopolysaccaharides (LPS). Effect of the compounds on nitric oxide (NO) production and cytokines (TNFα, IL-6) were determined by Griess and ELISA assays respectively. iNOS and the phosphorylation levels of MAPKs (ERK, JNK) were examined by Western Blot. RESULTS: Sambulin A and sambulin B inhibited 52.82% and 72.88% of NO production at 50 and 25µg/ml concentrations respectively. The levels of iNOS were significantly decreased by both molecules, sambulin B at 25µg/ml almost completely decreased iNOS levels (97.53%). Both molecules significantly inhibited TNFα productions. However, only sambulin B inhibited IL-6 production. Consequently, it was shown that sambulin B exerted its effect through the inhibition of ERK and JNK phosphorylations. CONCLUSION: The prominent bioactivities exerted by two iridoids will contribute to explanation of the usage of S. ebulus in traditional medicine against rheumatoid diseases.

Caspase-mediated Apoptotic Effects of Ebenus boissieri Barbey Extracts on Human Cervical Cancer Cell Line HeLa.

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).

The effect of Nigella sativa oil on prevention of myringosclerosis in a Guinea pig model.

OBJECTIVES: In this study, our aim was to identify the possible effects of Nigella sativa L. (NS) [blackcumin] seed oil on the prevention of experimentally induced myringosclerosis (MS). MATERIALS AND METHODS: Fourteen Guinea pigs were used and they were divided into three groups. Tympanic membranes (TM) of all animals were perforated and then group I was treated with saline soaked gel foams as a control group, group II was treated with 0.5 ml NS oil soaked gel foams at 0, 24 and 48 h and group III was treated with 5 ml NS oil orally at 0, 24, 48, 72 and 120 h. After 15 days, all animals were euthanized. Tympanic membranes were evaluated macroscopically and histopathologically. RESULTS: Groups I showed extensive myringosclerosis in contrast to those of Groups II and III which had significantly less changes (p < 0.05). The fibrosis and inflammation in the lamina propria of the tympanic membranes of Groups I was found to be significantly more pronounced (p < 0.05). The tympanic membranes were found to be significantly thinner in Groups II and III when compared with Groups I (p < 0.05). CONCLUSIONS: The results of this study suggested that topical or oral administration of NS oil supressed the inflammation and fibroblastic activity in the lamina propria of the myringotomized TMs of the Guinea pigs. For providing further evidence to use plant extracts as antioxidant and antiinflammatory therapy after myringotomy or ventilation tube insertion, further clinical studies with larger population will be essential.

A mixture of St. John's wort and sea buckthorn oils regresses endometriotic implants and affects the levels of inflammatory mediators in peritoneal fluid of the rat: A surgically induced endometriosis model.

OBJECTIVE: Sea buckthorn (Hippophae rhamnoides L.) and St. John's wort (Hypericum perforatum L.) are used as an emmenagog and for the treatment of other gynecological disorders including uterus inflammation and endometriosis. The aim of the present study is to investigate the potential of a mixture of sea buckthorn and St. John's wort oils (HrHp oil) in the treatment of endometriosis. MATERIALS AND METHODS: The activity was assessed in surgically induced endometriosis in rats. A 15-mm piece of endometrium was sutured into the abdominal wall. Twenty-eight days later, a second laparotomy was performed to calculate the endometrial foci areas and to score intra-abdominal adhesions. The rats were treated with either vehicle, HrHp oil formulation, or the reference (buserelin acetate). At the end of the experiment all rats were sacrificed and endometriotic foci areas and intra-abdominal adhesions were re-evaluated. The tissue sections were analyzed histopathologically. Peritoneal fluids of the experimental animals were collected in order to detect the levels of tumor necrosis factor-α, vascular endothelial growth factor, and interleukin-6, which might be involved in the etiology of endometriosis. RESULTS: In the HrHp oil-treated group, the volumes of endometriotic implants were found to be significantly decreased (from 50.8 mm3 to 18.6 mm3, p<0.001) without any adhesion (0.0±0.0, p<0.001) when compared to the control group (3.1±0.9). The levels of tumor necrosis factor-α decreased from 7.02±1.33 pg/mL to 4.78±1.02 pg/mL (p<0.01); vascular endothelial growth factor from 17.39±8.52 pg/mL to 9.67±5.04 pg/mL (p<0.01); and interleukin-6 from 50.95±22.84 pg/mL to 29.11±7.45 pg/mL (p<0.01), respectively, after HrHp oil treatment. CONCLUSION: HrHp oil may be a promising alternative for the treatment of endometriosis.

A Guidance Manual for the Toxicity Assessment of Traditional Herbal Medicines.

Herbal remedies have been used for thousands of years in worldwide traditional medicines for their potential health benefits. Although they are generally presumed safe unless a significant risk has been identified in humans, increasing number of case reports notify acute or chronic intoxications resulting from their use. This study aims to produce a scientific guide for the evaluation of traditional herbal medicines (THMs) in terms of their toxicity risks based on the published regulatory documents. For this purpose recommended in vitro and in vivo toxicity tests on medicinal products for human use issued by the international regulatory bodies are overviewed and they are then adopted to be used for the toxicity assessment of THMs. Accordingly, based on compilation of these issued regulations, the following tests are recommended for the toxicity assessment of THMs; in vitrocytotoxicity, genotoxicity, acute and repeated dose toxicity, carcinogenicity, reproductive and developmental toxicity, local tolerance tests, toxicokinetic studies, and additional toxicity tests including safety pharmacology, immunotoxicity and antigenicity, endocrine system toxicity, gastro-intestinal toxicity, renal and hepatotoxicity, and drug interaction studies. This study describes and discusses the applicability of these tests for the risk assessment in THMs.

Cytotoxic and apoptotic effects of Ebenus boissieri Barbey on human lung cancer cell line A549.

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 fruit extract of Berberis crataegina DC: exerts potent antioxidant activity and protects DNA integrity.

BACKGROUND: Dried fruits of Berberis crataegina (Berberidaceae) have been frequently consumed as food garniture in Turkish cuisine, while its fruit paste has been used to increase stamina and in particular to prevent from cardiovascular dysfunctions in Northeastern Black Sea region of Turkey. This study investigated this folkloric information in order to explain the claimed healing effects as well as to evaluate possible risks. METHODS: Total phenolic, flavonoid and proanthocyanidin contents and antioxidant capacity of the methanolic fruit extract were evaluated through several in vitro assays. The cytotoxic and genotoxic effects of B. crataegina fruit extract were also assessed in both cervical cancer cell line (HeLa) and human peripheral blood lymphocytes. RESULTS: The extract showed protective effects against ferric-induced oxidative stress and had a relatively good antioxidant activity. It also ameliorated the H2O2 mediated DNA damage in lymphocytes, suggesting the protective effect against oxidative DNA damage. CONCLUSION: The methanolic extract of B. crataegina fruits may be a potential antioxidant nutrient and also may exert a protective role against lipid peroxidation as well as oxidative DNA damage.

Effect of in vitro gastrointestinal digestion on the bioavailability of phenolic components and the antioxidant potentials of some Turkish fruit wines.

This study was designed to evaluate the effects of in vitro gastrointestinal simulation method on the antioxidant potentials and phenolic profile of some Turkish fruit wines and to compare the results with a Turkish red wine prepared from native grape varieties (Papazkarasi). For this purpose, blueberry, black mulberry and cherry wines were studied since they are widely consumed in Turkey. Papazkarasi wine was chosen due to the lack of studies regarding this type of wine. Antioxidant potentials of samples were measured with four different methods: DPPH radical-scavenging activity, H2O2-scavenging activity, cupric reducing capacity and total antioxidant capacity assays. The phenolic profiles of samples were evaluated by the determination of total phenolic content and HPLC-DAD analysis of seven different molecules. The results of this study provided information not only the effect of gastrointestinal digestion on parameters mentioned above, but also the bioaccessibility about the phenolic compounds found in these four different wine samples.

Appraisal of in vitro and in vivo antioxidant activity potential of cornelian cherry leaves.

The antioxidant activity of the 80% methanolic extract of Cornus mas L. leaves (CMM) was evaluated by various methods both in vitro and in vivo. In vitro screening tests indicated that CMM had high antioxidant activity in terms of free radical scavenging and metal reducing activity. In vivo antioxidant activity studies in normal healthy rats demonstrated that the total antioxidant capacity of liver homogenates were increased, although no changes were observed in the activities of antioxidant enzymes superoxide dismutase, catalase and glutathione peroxidase or in the level of lipid peroxidation. Studies on CCl4-treated rats also showed that CMM restored the activities of antioxidant enzymes, lowered the level of lipid peroxidation and elevated the total antioxidant capacities of both the total blood and liver homogenates of the animals. Further activity-guided fractionation studies led to the isolation of gallic acid, a well-known antioxidant, as one of the active components.

A comparative study on the in vitro antioxidant potentials of three edible fruits: cornelian cherry, Japanese persimmon and cherry laurel.

This study was designed in order to investigate in vitro antioxidant potentials of 80% methanolic extracts prepared from three edible fruits, Cornus mas L., Diospyros kaki L., Laurocerasus officinalis Roem. For this purpose, 8 different tests were performed including 2,2-diphenyl-1-picrylhydrazyl (DPPH) and superoxide radical scavenging tests, ferric reducing antioxidant power (FRAP), cupric reducing antioxidant capacity (CUPRAC), metal-chelating capacity, determination of total antioxidant capacity, ß-carotene bleaching test in a linoleic acid emulsion system and trolox equivalent antioxidant capacity. In addition, for evaluating the phenolic profile, total phenolic, flavonoid and proanthocyanidin contents were measured spectrophotometrically. Among the three fruits analyzed, Diospyros kaki L. showed the highest activity in all tests, except ß-carotene bleaching test. Whereas, neither of three fruits showed metal-chelating activity. Also, a good correlation was found between the phenolic content and antioxidant parameters.

Efficacy of Daphne oleoides subsp. kurdica used for wound healing: identification of active compounds through bioassay guided isolation technique.

ETHNOPHARMACOLOGICAL RELEVANCE: In Turkish traditional medicine, the aerial parts of Daphne oleoides Schreber subsp. kurdica (DOK) have been used to treat malaria, rheumatism and for wound healing. The aim was to evaluate the ethnopharmacological usage of the plant using in vivo and in vitro pharmacological experimental models, and to perform bioassay-guided fractionation of the 85% methanolic extract of DOK for the isolation and identification of active wound-healing component(s) and to elucidate possible mechanism of the wound-healing activity. MATERIALS AND METHODS: In vivo wound-healing activity was evaluated by the linear incision and the circular excision wound models. Anti-inflammatory and antioxidant activities, which are known to support the wound healing process, were also assessed by the Whittle method and the DPPH (2,2-diphenyl-1-picrylhydrazyl) radical-scavenging assays, respectively. The total phenolic content of the extract and subextracts was estimated to establish any correlation between the phenolic content and the antioxidant activity. The methanolic extract of DOK was subjected to various chromatographic separation techniques leading to the isolation and identification of the active component(s). Furthermore, in vitro hyaluronidase, collagenase and elastase enzymes inhibitory activity assays were conducted on the active components to explore the activity pathways of the remedy. RESULTS: After confirmation of the wound-healing activity, the methanolic extract was subjected to successive solvent partitioning using solvents of increasing polarity creating five subextracts. Each subextract was tested on the same biological activity model and the ethyl acetate (EtOAc) subextract had the highest activity. The EtOAc subextract was subjected to further chromatographic separation for the isolation of components 1, 2 and 3. The structures of these compounds were elucidated as daphnetin (1), demethyldaphnoretin 7-O-glucoside (2) and luteolin-7-O-glucoside (3). Further in vivo testing revealed that luteolin-7-O-glucoside was responsible for the wound-healing activity of the aerial parts. It was also found to exert significant anti-inflammatory, antioxidant, anti-hyaluronidase and anti-collagenase activities. CONCLUSION: The present study explored the wound-healing potential of Daphne oleoides subsp. kurdica. Through bioassay-guided fractionation and isolation techniques, luteolin-7-O-glucoside was determined as the main active component of the aerial parts. This compound exerts its activity through inhibition of hyaluronidase and collagenase enzymes activity as well as interfering with the inflammatory stage.