Effects of polyphenolic grape extract on the oxidative status of muscle and endothelial cells.

A grape pomace extract enhanced antioxidant mechanisms in muscle and endothelial cells both in the absence and in the presence of oxidative stress-induced agent tert-butyl hydroperoxide (tBHP). In particular, muscle (C2C12) and endothelial (EA.hy926) cells were treated with the extract at noncytotoxic concentrations for 24 h, and the oxidative stress markers, total reactive oxygen species (ROS), glutathione (GSH), thiobarbituric reactive substances (TBARS), and protein carbonyl levels were assessed. The results showed that the grape extract treatment reduced significantly ROS, TBARS, and protein carbonyl levels and increased GSH in C2C12 cells, while it increased GSH and decreased protein carbonyl levels in EA.hy926 cells. In the presence of tBHP, the grape extract treatment in C2C12 cells reduced significantly ROS, TBARS, and protein carbonyls and increased GSH compared with tBHP alone treatment, while, in EA.hy926 cells, the extract decreased significantly TBARS and protein carbonyls but increased GSH. The antioxidant potency of the extract was different between muscle and endothelial cells suggesting that the antioxidant activity depends on cell type. Moreover, the antioxidant activity of the grape extract, in both cell lines, exerted, at least in part, through increase in GSH levels. The present work is the first to report the effects of grape extract shown for skeletal muscle cells.

Ursolic acid triggers apoptosis and Bcl-2 downregulation in MCF-7 breast cancer cells.

In this report we determine the ability of ursolic acid (UA) to induce apoptosis and to modulate glucocorticoid receptor (GR) and Activator Protein-1 (AP-1) in MCF-7 cells. The UA-induced apoptosis (53 microM), the PARP cleavage, and the decrease in Bcl-2 protein (53 microM) support the notion that UA induces apoptosis through the intrinsic mitochondrial pathway. UA binds GR (relative binding affinity: 2.57) and translocates GR into nucleus, suggesting its potential as a GR modulator. UA had no effect on GRE- or TRE-driven gene expression. In summary, UA is a GR modulator and may be considered as a potential anticancer agent in breast cancer.

Assessment of antioxidant activity of extracts from unique Greek varieties of Leguminosae plants using in vitro assays.

BACKGROUND: It is believed that legumes are a very good source of micronutrients and phytochemicals that present chemopreventive activity against diseases such as diabetes, coronary heart disease and colon cancer. Methanolic and aqueous extracts from 11 unique varieties of Leguminosae family plants cultured in Greece were tested using three different in vitro assays in order to investigate the mechanisms by which phytochemicals present in these legumes exert their chemoprevention. MATERIALS AND METHODS: The extracts were tested by the 1, -diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, the hydroxyl radical- and the peroxyl radical-induced DNA strand scission assays. Hydroxyl (OH*) and peroxyl (ROO*) radicals were generated from ultraviolet (UV) photolysis of hydrogen peroxide (H2O2) and thermal decomposition of 2,2'-azobis-(2-amidinopropane hydrochloride) (AAPH) respectively. RESULTS: In the DPPH assay, all the tested extracts displayed potent radical scavenging efficiency. Furthermore, most of the Leguminosae family plant extracts exerted significant protective activity against DNA damage induced by both reactive oxygen species, although they were more effective in inhibiting ROO*-induced rather than OH*-induced DNA strand scission. CONCLUSION: The results suggest that the free radical scavenging activity of Leguminosae plants may be one of the mechanisms accounting for their chemoprevention.

Ursolic acid, a naturally occurring triterpenoid, demonstrates anticancer activity on human prostate cancer cells.

PURPOSE: Glucocorticoids are widely used as adjuvant therapy in hormonal refractory prostate cancer; their therapeutic role, however, remains unclear. Ursolic acid, a natural triterpene, structurally similar to dexamethasone, exhibits antitumor effects in various cell types. Our main objective was to investigate the effects of ursolic acid on cell viability, apoptosis and bcl-2 protein, in human hormone refractory and androgen-sensitive prostate cancer cells. METHODS: The ursolic acid-induced changes in cell viability, apoptosis and bcl-2 protein were examined in human hormone refractory prostate cancer PC-3 cells and androgen-sensitive LNCaP cells, by MTT assay, flow cytometry and western blot analysis, respectively. RESULTS: Ursolic acid inhibited significantly the cell viability and induced apoptosis in PC-3 cells at 55 microM and in LNCaP cells at 45 microM associated with a downregulation of bcl-2 protein. CONCLUSIONS: The antiproliferative and apoptotic effects of ursolic acid in PC-3 and LNCaP cells implicate its potential therapeutic use for the treatment of hormone refractory and androgen-sensitive prostate cancer. The downregulation of bcl-2 may be one of the molecular mechanisms via which it induces apoptosis in PC-3 and LNCaP cells.

Acretoside, a new sucrose ester from Aristolochia cretica.

A new sucrose ester, acretoside, has been isolated from the roots of the Greek endemic species Aristolochia cretica and identified as 6-O-p-coumaroyl-beta-D-fructofuranosyl-(2 --> 1)-alpha-D-glucopyranoside (1). In addition, a known sucrose ester, identified as arillatose B, two phenylpropanoid glucose esters, and five derivatives of aristolochic acids have been isolated. Their structures have been elucidated on the basis of MS and NMR data.

Composition of the essential oil of two Nepeta species and in vitro evaluation of their activity against Helicobacter pylori.

The chemical composition of the essential oils obtained from the aerial parts of Nepeta camphorata and Nepeta argolica ssp. dirphya were analysed by GC-MS. A total of 52 components were identified and significant differences (qualitative and quantitative) were observed between the two samples. 1,8-Cineol and two nepetalactones were found to be the major components of the oil of N. camphorata and N. argolica ssp. dirphya respectively. The in vitro activity, of the two oils and the three above mentioned isolated compounds, against 25 clinically isolated and commercial strains of Helicobacter pylori was investigated and some activity was found.

Composition and antimicrobial activity of the essential oils of two Origanum species.

The essential oils obtained from the aerial parts of Origanum scabrum and Origaum microphyllum, both endemic species in Greece, were analyzed by means of GC and GC-MS. Forty-eight constituents were identified, representing 98.59 and 98.66% of the oils, respectively. Carvacrol, terpinen-4-ol, linalool, sabinene, alpha-terpinene, and gamma-terpinene were found as the major components. Furthermore, both samples exhibited a very interesting antimicrobial profile after they were tested against six Gram-negative and -positive bacteria and three pathogenic fungi.

Samioside, a new phenylethanoid glycoside with free-radical scavenging and antimicrobial activities from Phlomis samia.

A new phenylethanoid glycoside, samioside, was isolated from the aerial parts of Phlomis samia and identified as 1-O-3,4-(dihydroxyphenyl)ethyl beta-D-apiofuranosyl-(1-->4)-alpha-L-rhamnopyranosyl-(1-->3)-4-O-caffeoyl-beta-D-glucopyranoside (1). In addition, one known phenylethanoid glycoside and three known flavonoids were identified as acteoside (2), apigenin, chrysoeriol, and ermanin, respectively. The structure of 1 was elucidated on the basis of its spectroscopic data. Samioside (1) demonstrated scavenging properties toward the DPPH radical and antimicrobial activity against Gram-positive and -negative bacteria.

Flavonoids as cycline-dependent kinase inhibitors: inhibition of cdc 25 phosphatase activity by flavonoids belonging to the quercetin and kaempferol series.

In an effort to detect potential inhibitors of cdc25 phosphatase, nineteen flavonoids belonging to the quercetin and kaempferol series have been evaluated, using a colorimetric assay of recombinant human cdc25A tyrosine phosphatase as a cell cycle-specific target. Compounds bearing two benzyl or methyl groups in positions 7 and 4' and acetyl on the hydroxy groups of the sugar moiety showed the maximal activity.

New hemisynthetic manoyl oxide derivatives with antimicrobial activity.

The synthesis and antimicrobial activity of ten labdane-type diterpenes derived from ent-3-beta-hydroxy-13-epi-manoyl oxide (ribenol) is reported. The chloroethyl carbamidic ester 9 showed the strongest antimicrobial activity against all the tested gram (+), gram (-) bacteria and pathogenic fungi. Moreover, the glycoside 11 exhibited an interesting activity against the three tested fungi.

Furomegistines I and II, two furanopyridine alkaloids from the bark of Sarcomelicope megistophylla.

Two alkaloids, furomegistine I (1) and furomegistine II (2), were isolated from the bark of Sarcomelicope megistophylla. Their structures have been elucidated on the basis of MS and NMR data. Both belong to the category of furanopyridine alkaloids and should be considered as oxidation products of a furo[2,3-b]quinoline precursor. The two alkaloids exhibited moderate cytotoxic activity.

New semisynthetic antimicrobial labdane-type diterpenoids derived from the resin "ladano" of Cistus creticus.

The antimicrobial activity of fifteen semisynthetic labdane-type diterpenes derived from the two major natural compounds 3 and 4 of the resin "ladano" of Cistus creticus is reported. The chloroethyl carbamidic esters 15 and 20 showed the strongest antimicrobial activity against Gram(+), Gram(-) bacteria and pathogenic fungi.

Composition and antimicrobial activity of the essential oils of five taxa of Sideritis from Greece.

The chemical compositions of the essential oils obtained from the aerial parts of five taxa of Sideritis were analyzed using various GC-MS techniques. A total of 99 different compounds was identified, and significant differences (qualitative and quantitative) were observed between the samples. The in vitro antimicrobial activity of the essential oils against six bacteria and three fungi is also reported.

Chemistry of plants from Crete: stachyspinoside, a new flavonoid glycoside and iridoids from Stachys spinosa.

The new flavonoid glycoside stachyspinoside (1), and the three iridoids, 7-O-acetyl-8-epi-loganic acid (2), ajugol (3) and harpagide (4) were isolated from Stachys spinosa. The structures of these compounds were established on the basis of mass spectrometry (ESMS and tandem MS), one- and two-dimensional nuclear magnetic resonance experiments (COSY, COSY LR, HMQC, TOCSY and HMBC) as well as simple chemical derivatization.

Antileukemic activity of synthetic daunomycinone derivatives bearing modifications in the glycosidic moiety.

The antileukemic activities of the daunomycinone glycosides synthesized in our laboratories (compounds 4 and 7, code names S12 and S13, respectively) were characterized in L1210 cells in vitro. S13 inhibits tumor cell proliferation and viability at day 4 (IC50: 150-200 nM) more effectively than S12 (IC50: 250-450 nM), suggesting that the 4'-trifluoracetamido substitution of the glycosidic moiety of these 3'-halo daunonycinone derivatives has greater antitumor potential than the 4'-azido substitution. Since S12 and S13 do not increase but rather decrease the mitotic index of L1210 cells at 24 hours, they are not antitubulin drugs but might arrest the early stages of cell cycle progression. Pretreatments for 1.5-3 hours with S12 and S13 are sufficient to partially inhibit the rates of DNA and RNA syntheses (IC50: 4-10 microM) determined over 30- to 60-minute periods of pulse-labeling in L 1210 cells in vitro, but these daunomycinone glycosides alter neither the cellular transport of purine and pyrimidine nucleosides nor the rate of protein synthesis. After 24 hours, the concentration-dependent induction of DNA cleavage by S13 reaches a plateau at 10 microM but the weaker S12 requires 48 hours to maximally stimulate DNA cleavage like S13. The mechanism by which S13 induces DNA fragmentation is inhibited by actinomycin D, cycloheximide, benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone, benzyloxycarbonyl-Ile-Glu-Thr-Asp-fluoromethyl ketone, N-tosyl-L-phenylalanine chloromethyl ketone and ZnSO4, suggesting that S13 triggers apoptosis by caspase and endonuclease activation. Since microM concentrations of S12 and S13 are cytostatic and cytotoxic, but do not sufficiently inhibit RNA and protein syntheses to block their own ability to sustain the active process of apoptosis and DNA fragmentation, such 3'-halo daunomycinone glycosides might be valuable to develop new means of polychemotherapy.

Design, synthesis and biological activity of 7-O-(4-O-acetyl-3-iodo-2,3,6-trideoxy-alpha-L-arabino-hexopyranosyl) daunomycinone and 7-O-(3-chloro-2,3,6-trideoxy-4-O-propanoyl-alpha-L-lyxo- hexopyranosyl)daunomycinone.

The synthesis and pharmacological evaluation of 7-O-(4-O-acetyl-3-iodo-2,3,6-trideoxy-alpha-L-arabino-hexopyranosyl) daunomycinone and 7-O-(3-chloro-2,3,6-trideoxy-4-O-propanoyl-alpha-L-lyxo-hexopyrano syl) daunomycinone are described. Their cytotoxic activity was evaluated against normal and resistant cell lines. Both compounds exhibited activity against the adriamycin resistant cell line KB-A1. These results support the hypothesis that the increased lipophilicity of the sugar part of anthracyclines is associated with their ability to overcome multidrug resistance (MDR).

Three new arylobenzofurans from Onobrychis ebenoides and evaluation of their binding affinity for the estrogen receptor.

Three new 2-phenyl-benzofurans, ebenfuran I, ebenfuran II, and ebenfuran III, were isolated from Onobrychis ebenoides. Their structures were elucidated on the basis of chemical and spectral data as 2-(2,4-dihydroxyphenyl)-5-hydroxy-6-methoxy-benzofuran (1), 2-(2,4-dihydroxyphenyl)-3-formyl-4-hydroxy-6-methoxy-benzofuran (2), and 2-(2, 4-dihydroxyphenyl)-3-formyl-4-hydroxy-6-methoxy-5-(3-methyl-buten- 2-y l)-benzofuran (3). The affinity of these compounds for the estrogen receptor was studied using a receptor-binding assay.

Verbaspinoside, a new iridoid glycoside from Verbascum spinosum.

A new iridoid glycoside, verbaspinoside (1), was isolated from the aerial parts of Verbascum spinosum. Its structure was elucidated on the basis of chemical and spectral data as 6-O-[(2' '-O-trans-cinnamoyl)-alpha-L-rhamnopyranosyl]-catalpol. Additionally, three known iridoids (aucubin, catalpol, and ajugol) and three phenylpropanoid glycosides [acteoside, angoroside A (2), and angoroside C (3)] were isolated and identified.