Anthraquinone profile, antioxidant and antimicrobial activity of bark extracts of Rhamnus alaternus, R. fallax, R. intermedia and R. pumila.

The quantity of phenols, as well as antioxidant and antimicrobial activities, were investigated in bark of Rhamnus alaternus L., R. fallax Boiss., R. intermedia Steud. et Hochst., and R. pumila Turra from natural stands in Croatia. The most abundant anthraquinones in the investigated extracts were chrysophanol in R. alaternus (3.14 mg/g), emodin in R. pumila (0.339 mg/g), and physcion in R. fallax (2.70 mg/g) and R. intermedia (0.285 mg/g). The species exhibiting the highest antioxidant activity were R. fallax and R. pumila. A positive correlation was observed between total phenolic and flavonoid levels of the extracts and antioxidant activity in some of the assays. All species showed antimicrobial activity against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Candida albicans, Aspergillus niger and Microsporum gypseum with minimal inhibitory concentrations equal to or below 2.500 mg/mL. The results indicate that the investigated Rhamnus species are a source of anthraquinones and other phenols, which act as multifunctional antioxidants with antimicrobial activity.

Licorice and its potential beneficial effects in common oro-dental diseases.

Licorice, the name given to the roots and stolons of Glycyrrhiza species, has been used since ancient times as a traditional herbal remedy. Licorice contains several classes of secondary metabolites with which numerous human health benefits have been associated. Recent research suggests that licorice and its bioactive ingredients such as glycyrrhizin, glabridin, licochalcone A, licoricidin, and licorisoflavan A possess potential beneficial effects in oral diseases. This paper reviews the effects of licorice and licorice constituents on both the oral microbial pathogens and the host immune response involved in common ora-dental diseases (dental caries, periodontitis, candidiasis, and recurrent aphthous ulcers). It also summarizes results of clinical trials that investigated the potential beneficial effects of licorice and its constituents for preventing/treating oro-dental diseases.

In vitro effects of natural prenyloxycinnamic acids on human cytochrome P450 isozyme activity and expression.

Previous studies demonstrated that natural prenyloxyphenylpropanoid derivatives have potent biological properties like anti-cancer effects in vitro and in vivo. Additionally they are extremely safe and associated with low toxicity, making them excellent candidates as chemopreventive agents. However, so far only little is known about possible interactions with isoforms of cytochrome P450 (CYPs) being involved in the metabolism of xenobiotics and representing a major site for drug-drug interactions. The aim of this study was to evaluate the effects of selected natural prenyloxyphenylpropanoids (prenyloxycinnamic acids) on expression and activity of some major CYPs and on the activity of the major drug efflux transporter P-glycoprotein (P-gp). Inhibition of CYP3A4, CYP2C19, and CYP2D6 was quantified using commercially available kits. P-gp inhibtion was quantified by calcein assay. Induction of CYP mRNA (CYP3A4, CYP2C19, CYP2C9, and CYP2B6) was measured in LS180 cells by quantitative real-time reverse transcriptase polymerase chain reaction using the LightCycler technology. Only boropinic acid revealed substantial inhibition of CYPs, especially of CYP2C19 (IC50 = 31±5µM). This compound also had the most pronounced effect on CYP mRNA expression among the prenyloxycinnamic acids tested. However all but 4'-isopentenyloxy-p-coumaric acid revealed inducing effects on CYPs with different induction profiles. P-gp was only significantly inhibited by 4'-geranyloxyferulic acid. This was the first study demonstrating modulating effects of prenyloxycinnamic acids on CYP activity and expression and on P-gp activity. The results suggest that boropinic acid is most prone to drug-drug interactions at the level of CYPs, whereas 4'-isopentenyloxy-p-coumaric acid does not modulate CYP activity and expression.

Antiproliferative, protective and antioxidant effects of artichoke, dandelion, turmeric and rosemary extracts and their formulation.

Artichoke, dandelion, turmeric extracts and rosemary essential oil are commonly used as ingredients in many herbal preparations to treat hepatic and gallbladder disorders. In the present work we compare the activity of each single extract with a commercial mixture for antiproliferative, antiradical and protective effects against induced oxidant stress effect. In ABTS and DPPH tests, turmeric extract is the most active, followed by artichoke and dandelion. All samples exhibited antiproliferative activity in a dose-dependent manner against HepG2 cells. In the same cell lines, the protective effect of pre-treatment with the extracts were detected by evaluating the prostaglandin E2 release, a marker of oxidative stress induced by hydrogen peroxide. The treatments with the extracts were efficient in reducing the release of PGE2 induced by oxidative stimulus. The positive results of the cell viability test, together with the protective and antiradical activity confirm the rationale for the use of these ingredients in commercial formulations as a health aid tool in modern phytotherapy.

Quantification of 4'-geranyloxyferulic acid, a new natural colon cancer chemopreventive agent, by HPLC-DAD in grapefruit skin extract.

Oxyprenylated natural products (isopentenyloxy-, geranyloxy- and the less spread farnesyloxy-compounds and their biosynthetic derivatives) represent a family of secondary metabolites that have been consider for years merely as biosynthetic intermediates of the most abundant C-prenylated derivatives. Many of the isolated oxyprenylated natural products were shown to exert in vitro and in vivo remarkable anti-cancer and anti-inflammatory effects. 4'-Geranyloxyferulic acid [3-(4'-geranyloxy-3'-methoxyphenyl)-2-trans-propenoic] has been discovered as a valuable chemopreventive agent of several types of cancer. After development of a high yield and "eco-friendly" synthetic scheme of this secondary metabolite, starting from cheap and non-toxic reagents and substrates, we developed a new HPLC-DAD method for its quantification in grapefruit skin extract. A preliminary study on C18 column showed the separation between GOFA and boropinic acid (having the same core but with an isopentenyloxy side chain), used as internal standard. The tested column were thermostated at 28+/-1 degrees C and the separation was achieved in gradient condition at a flow rate of 1 mL/min with a starting mobile phase of H(2)O:methanol (40:60, v/v, 1% formic acid). The limit of detection (LOD, S/N=3) was 0.5 microg/mL and the limit of quantification (LOQ, S/N=10) was 1 microg/mL. Matrix-matched standard curves showed linearity up to 75 microg/mL. In the analytical range the precision (RSD%) values were

Comparison of three different extraction methods and HPLC determination of the anthraquinones aloe-emodine, emodine, rheine, chrysophanol and physcione in the bark of Rhamnus alpinus L. (Rhamnaceae).

INTRODUCTION: Rhamnus alpinus L. (Rhamnaceae), a traditional plants in the flora of the Abruzzo region, is known to contain active anthraquinone secondary metabolites. However, the content of anthraquinones varies among R. alpinus samples depending on collection season and site. Thus, using simple, reliable and accurate analytical methods for the determination of anthraquinones in R. alpinus extracts allows comparative study of different methods of extraction. OBJECTIVE: After a partial validation of an HPLC method for the simultaneous determination of five anthraquinones, aloe-emodine, rheine, emodine, chrysophanol and physcione, in the bark of R. alpinus, we compared three different methods of extraction. METHODOLOGY: Anthraquinones were extracted from the bark of R. alpinus using different techniques (methanol maceration, ultrasonic and supercritical CO(2) extraction). Separation and quantification of anthraquinones were accomplished using a reversed-phase C(18) column with the mobile phase of H(2)O-methanol (40 : 60, v/v, 1% formic acid) at a wavelength of 254 nm. The qualitative analyses were also achieved at wavelength of 435 nm. RESULTS: All calibration curves were linear over the concentration range tested (10-200 mM) with the determination coefficients >or=0.991. The detection limits (S/N = 3) were 5 mM for each analytes. All five anthraquinones were found in the samples tested at concentrations reported in experimental data. CONCLUSION: The described HPLC method and optimised extraction procedure are simple, accurate and selective for separation and quantification of anthraquinones in the bark of R. alpinus and allow evaluation of the best extraction procedure between the tested assays.

Naringenin has anti-inflammatory properties in macrophage and ex vivo human whole-blood models.

BACKGROUND AND OBJECTIVE: Periodontitis is a chronic inflammatory disease of bacterial etiology, affecting tooth-supporting tissues. The host inflammatory response to periodontopathogens, notably the high and continuous production of cytokines, is considered a major factor causing the local tissue destruction observed in periodontitis. The aim of the present study was to investigate the effect of naringenin, a major flavanone in grapefruits and tomatoes, on the lipopolysaccharide-induced pro-inflammatory cytokine production by host cells, using two different models. MATERIAL AND METHODS: The effect of naringenin was characterized using macrophages stimulated with the lipopolysaccharide of either Aggregatibacter actinomycetemcomitans or Escherichia coli and using whole blood stimulated with A. actinomycetemcomitans lipopolysaccharide, in the presence or absence of naringenin. Lipopolysaccharide-induced interleukin-1 beta, interleukin-6, interleukin-8 and tumor necrosis factor-alpha production by macrophages and whole-blood samples treated with naringenin were evaluated using an enzyme-linked immunosorbent assay. Changes in the phosphorylation states of macrophage kinases induced by A. actinomycetemcomitans lipopolysaccharide and naringenin were characterized by immunoblot screening. RESULTS: Our results clearly indicated that naringenin is a potent inhibitor of the pro-inflammatory cytokine response induced by lipopolysaccharide in both macrophages and in whole blood. Naringenin markedly inhibited the phosphorylation on serines 63 and 73 of Jun proto-oncogene-encoded AP-1 transcription factor in lipopolysaccharide-stimulated macrophages. CONCLUSION: The results from the present study suggest that naringenin holds promise as a therapeutic agent for treating inflammatory diseases such as periodontitis.

Antifungal activity of some Cuban Zanthoxylum species.

Ethanolic extracts of the trunk bark of Zanthoxylum fagara, Z. elephantiasis and Z. martinicense showed activity against different species of fungi. No antibacterial activity was detected.