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Greek plant extracts exhibit selective estrogen receptor modulator (SERM)-like properties.

To prevent bone loss that occurs with increasing age, nutritional and pharmacological factors are needed. Traditional therapeutic agents (selective estrogen receptor modulators or SERMs, biphosphonates, calcitonin) may have serious side effects or contraindications. In an attempt to find food components potentially acting as SERMs, we submitted four plant aqueous extracts derived from Greek flora (Sideritis euboea, Sideritis clandestina, Marticaria chamomilla, and Pimpinella anisum) in a series of in vitro biological assays reflective of SERM profile. We examined their ability (a) to stimulate the differentiation and mineralization of osteoblastic cell culture by histochemical staining for alkaline phosphatase and Alizarin Red-S staining, (b) to induce, like antiestrogens, the insulin growth factor binding protein 3 (IGFBP3) in MCF-7 breast cancer cells, and (c) to proliferate cervical adenocarcinoma (HeLa) cells by use of MTT assay. Our data reveal that all the plant extracts studied at a concentration range 10-100 microg/mL stimulate osteoblastic cell differentiation and exhibit antiestrogenic effect on breast cancer cells without proliferative effects on cervical adenocarcinoma cells. The presence of estradiol inhibited the antiestrogenic effect induced by the extracts on MCF-7 cells, suggesting an estrogen receptor-related mechanism. In conclusion, the aqueous extracts derived from Sideritis euboea, Sideritis clandestina, Marticaria chamomilla, and Pimpinella anisum may form the basis to design "functional foods" for the prevention of osteoporosis.

Breast Neoplasms↗

Molecularly imprinted solid-phase extraction of naphthalene sulfonates from water.

A new polymeric sorbent synthesised by exploiting molecular imprinting technology has been used to selectively extract naphthalene sulfonates (NSs) directly from aqueous samples. In the non-covalent molecular imprinting approach used to prepare this polymer, 1-naphthalene sulfonic acid (1-NS) and 4-vinylpyridine (4-VP) were used as a template molecule and functional monomer, respectively, and both dissolved in a mixture of methanol/water (4:1) as porogen together with the cross-linker ethylene glycol dimethacrylate. The new non-covalent molecularly imprinted polymer (MIP) prepared in aqueous environment was used as a sorbent in solid-phase extraction (SPE) to selectively extract a group of naphthalene mono- and disulfonates. When one litre of a standard aqueous solution, which contained a mixture of eight NSs, was percolated through the SPE cartridge, all the NSs were retained on the MIP because of the cross-reactivity of the polymer. Recoveries were higher than 80% for all the compounds even after a clean-up step with methanol (MeOH). The MIP was also used to analyse water from the Ebro river.

Naphthalenesulfonates↗

Isolation of myelin basic protein from whole tissue extracts by selective pH-dependent solubilization.

A previous study of the selective solubility of myelin basic protein (MBP) of tissue extracts at pH 9.0 has raised issues of its quantitative recovery, and the differential solubility of its charge isomers. The pH-dependent solubility of proteins of acid extracts of delipidated tissue of bovine spinal cord was therefore reexamined. MBP of whole extracts was completely soluble up to pH 8.0 only, and less so by 25% at pH 9.0, and 43% at pH 10.0. The proteins other than MBP were virtually insoluble between pH 5.0 to 6.0, and 9.0 to 10.0. The solubility of the main charge isomers I to III of MBP of 18.5 kDa was found not to be affected by pH. Either pH 5.0 or 9.0 is therefore suitable for the selective isolation of MBP from whole tissue extracts, only pH 5.0 providing for the complete recovery of MBP. The pH-dependent solution behaviour was also examined following the separation of proteins of whole extracts by anion exchange chromatography at pH 10.4. Purified MBP and several related minor cationic components of lower molecular weight were soluble throughout. In contrast, the anionic proteins were only partly soluble between pH 4.0 to 10.0, i.e. by 4 to 20%. The results are consistent with specific protein-protein interactions of the proteins of whole extracts, either enhancing the solubility of non-MBP proteins, e.g. at pH 7.0, or impairing that of MBP between pH 8.0 to 10.0.

Animals↗

Effect of electron beam irradiation and storage at 5 degrees C on thiobarbituric acid reactive substances and carbonyl contents in chicken breast meat infused with antioxidants and selected plant extracts.

This study evaluated the effectiveness of synthetic and natural antioxidants, green tea, commercial grape seed extracts/combinations, and TBHQ, with varying concentrations of lipid oxidation of nonirradiated and irradiated chicken breast meats stored at 5 degrees C for 12 days. Fresh boneless and skinless chicken breast meats were vacuum-infused with varying concentrations of antioxidants: green tea, grape seed extracts alone/in combination, and TBHQ. The irradiation dosage was 3.0 kGy. Carbonyl values of raw chicken meat and thiobarbituric acid reactive substances (TBARS) values of raw and cooked chicken meat were determined for 0-12 days at 5 degrees C storage. TBARS values for 0-12 days of storage at 5 degrees C ranged from 1.21 to 7.3 and 1.22 to 8.51 mg malondialdehyde/100 g chicken for nonirradiated and irradiated raw chicken, respectively. TBARS values of cooked chicken ranged from 2.19 to 35.83 and 2.45 to 45.72 mg malondialdehyde/100 g chicken for nonirradiated and irradiated chicken, respectively. Irradiation increased TBARS values of both controls and plant extracts. The carbonyl content in meat lipid ranged from 1.7 to 2.9 and 1.7 to 4.41 micromol acetophenone/10 g of nonirradiated and irradiated chicken meat, respectively, and meat protein ranged from 1.4 to 2.07 and 1.41 to 2.72 micromol/10 g meat. Infusion of chicken meat with selected plant extracts is an effective method to minimize lipid oxidation and volatiles developments caused by irradiation.

Acetophenones↗

Mixed monolayer-protected gold nanoclusters as selective peptide extraction agents for MALDI-MS analysis.

Cationic and anionic nanoparticles selectively target peptides with low and high isoelectric points, respectively. Additionally, their high surface area-to-volume ratios make these nanoparticles (approximately 2-nm core diameter) very efficient extraction and concentration agents. Upon extraction, the peptide-bound nanoparticles can be analyzed by MALDI-MS to provide highly sensitive detection of the targeted peptides. We demonstrate that MALDI-MS can detect peptide concentrations as low as 500 pM from 250-microL solutions using these nanoparticle scaffolds as extraction and concentration agents.

Gold↗

Extraction of selected drugs from serum using microwave irradiation.

Microwave irradiation is used as an alternative heating method for extraction over more conventional hot plate methods. We describe a fast, efficient method for the determination of selected drugs in human blood/serum using microwave extraction. The microwave extraction of organic substances requires special instrumentation and the results have been compared with the results from classical liquid/liquid extraction. The present microwave extractions were performed in an 'atmospheric pressure' system. Before irradiation with microwaves, an appropriate solvent mixture was added to the buffered specimen. Lidocaine, methadone, diazepam, nordiazepam, propoxyphene and norpropoxyphene were tested as model substances. The quantitation was performed by GC/NPD. The procedure has been applied successfully to a number of forensic cases. The use of microwaves decreases the time of extraction and the solvent consumption.

Analgesics↗

Identification of carbohydrate structures in glycoprotein peptide maps by the use of LC/MS with selected ion extraction with special reference to tissue plasminogen activator and a glycosylation variant produced by site directed mutagenesis.

Electrospray ionization mass spectrometry utilizing a single quadrupole on line with reversed-phase HPLC (LC/MS) enables the characterization of glycoproteins in a relatively short period of time. In this approach the protein is digested with a suitable protease and the peptides are separated by reversed-phase HPLC and detected by electrospray ionization mass spectrometry. The glycopeptides are initially observed as a cluster of negatively sloping ions in a contour plot of data from the LC/MS run (m/z vs retention time) or as a characteristics series of masses at different elution times. The search for a particular glycopeptide is based on previously known carbohydrate structures and on consensus glycosylation sites. Further structural information is obtainable with glycosidase digestion and LC/MS analysis. The mass shifts following glycosidase digestion allow further confirmation of the structure. This approach identifies the site of attachment of two hybrid glycoforms to the T11 tryptic peptide in a reversed-phase tryptic map of recombinant tissue plasminogen activator (rt-PA). Use of selected ion extraction of the LC/MS data files allows one to graphically describe the elution order of closely related glycopeptides. The potential of LC/MS for the characterization of small amounts of unknown glycoproteins is shown by the study of an rt-PA mutant. A new potential site for glycosylation is created by site directed mutagenesis of wild type rt-PA with replacement of a threonine residue with asparagine at residue 103. An examination of a tryptic map shows that the mutant contains two new complex carbohydrate chains. The introduction of the new asparagine proximal to asparagine 117 changes this native high-mannose site in rt-PA to a complex-type glycosylation. This method allows rapid identification of carbohydrate containing peptides and yields useful structural information on microgram amounts of material.

Amino Acid Sequence↗

Molecularly imprinted polymers for the clean-up of a basic drug from environmental and biological samples.

A molecularly imprinted polymer (MIP) was synthesized and evaluated to selectively extract an alpha-blocker, i.e. alfuzosin, from human plasma. The synthesis of the MIP was performed in dichloromethane with methacrylic acid as monomer and the target drug as template. A first series of experiments was carried out in dichloromethane to estimate the potential of the MIP in its specific recognition medium, i.e. dichloromethane, by developing a selective procedure and by measuring the capacity of the sorbent. An optimized procedure was developed for the selective extraction of alfuzosin with a recovery close to 100% in this medium and a specific capacity of 1.3 micromol g(-1) of MIP was measured. A study in aqueous media was also carried out by a comprehensive approach of the retention mechanism in order to build a selective procedure of extraction. The effects of the amount and of the charge of cations were studied and an optimal pH value was defined to limit matrix effects. Then, the alfuzosin MIP was then directly used to selectively extract the target drug from human plasma with an extraction recovery of 60%. Lastly, a soil was extracted by a pressurized solvent and the resulting extract was cleaned up on the MIP, showing the possibility to use this selective sorbent for the sample treatment of various complex matrices.

Adrenergic alpha-Antagonists↗

High-yield selection and extraction of two promoter-defined phenotypes of neural stem cells from the fetal human brain.

Neural stem and precursor cells reside in the ventricular lining of the fetal forebrain, and may provide a cellular substrate for brain repair. To selectively identify and extract these cells, we infected dissociated fetal human brain cells with adenoviruses bearing the gene for green fluorescence protein (GFP), placed under the control of enhancer/promoters for two genes (nestin and musashi1) that are expressed in uncommitted neuroepithelial cells. The cells were then sorted by fluorescence-activated cell sorting (FACS) on the basis of E/nestin- or P/musashi1-driven GFP expression. Both P/musashi1:hGFP- and E/nestin:EGFP-sorted cells were multipotent: limiting dilution with clonal expansion as neurospheres, in tandem with retroviral lineage analysis and xenograft to E17 and P0-2 rat forebrain, revealed that each phenotype was able to both self-renew and co-generate neurons and glia. Thus, fluorescent genes placed under the control of early neural promoters allow neural stem cells to be specifically targeted, isolated, and substantially enriched from the fetal human brain.

Adenoviridae↗

Microwave-assisted extraction of embelin from Embelia ribes.

A rapid and efficient microwave-assisted extraction (MAE) process for the selective extraction of embelin from Embelia ribes was developed. Solvent selection, microwave energy input and solid loading were optimized. The rate of extraction and purity of embelin depended upon the solvent used and exposure time to microwaves. Maximum MAE was achieved in acetone with total yield of 92% (w/w) embelin with 90% (w/w) purity with 1% (w/v) raw material loading at 150 W power level in 80 s. Non-polar solvents, such as hexane and dichloromethane, were not effective for the selective extraction of embelin.

Benzoquinones↗

Antiviral activities of extracts and selected pure constituents of Ocimum basilicum.

1. Ocimum basilicum (OB), also known as sweet basil, is a well known medicinal herb in traditional Chinese medicine preparations. In the present study, extracts and purified components of OB were used to identify possible antiviral activities against DNA viruses (herpes viruses (HSV), adenoviruses (ADV) and hepatitis B virus) and RNA viruses (coxsackievirus B1 (CVB1) and enterovirus 71 (EV71)). 2. The results show that crude aqueous and ethanolic extracts of OB and selected purified components, namely apigenin, linalool and ursolic acid, exhibit a broad spectrum of antiviral activity. Of these compounds, ursolic acid showed the strongest activity against HSV-1 (EC50 = 6.6 mg/L; selectivity index (SI) = 15.2), ADV-8 (EC50 = 4.2 mg/L; SI = 23.8), CVB1 (EC50 = 0.4 mg/L; SI = 251.3) and EV71 (EC50 = 0.5 mg/L; SI = 201), whereas apigenin showed the highest activity against HSV-2 (EC50 = 9.7 mg/L; SI = 6.2), ADV-3 (EC50 = 11.1 mg/L; SI = 5.4), hepatitis B surface antigen (EC50 = 7.1 mg/L; SI = 2.3) and hepatitis B e antigen (EC50 = 12.8 mg/L; SI = 1.3) and linalool showed strongest activity against AVD-II (EC50 = 16.9 mg/L; SI = 10.5). 3. No activity was noted for carvone, cineole, beta-caryophyllene, farnesol, fenchone, geraniol, beta-myrcene and alpha-thujone. 4. The action of ursolic acid against CVB1 and EV71 was found to occur during the infection process and the replication phase. 5. With SI values greater than 200, the potential use of ursolic acid for treating infection with CVB1 and EV71 merits further investigation.

Acyclic Monoterpenes↗

Effect of redox potential on fixation of 137Cs in lake sediment.

Fixation of 137Cs was determined in lake sediment suspensions under controlled redox potentials in the laboratory. The activity of previously added 137Cs on various clay sites was determined by time-series selective extractions. Monovalent cations, particularly NH+4, were much more effective at displacing 137Cs than divalent cations or Na+. Ammonium ion (NH+4) and Na+ were used to extract 137Cs from selective and non-selective 137Cs-binding sites, respectively. The activity of water-soluble 137Cs and Na-extractable 137Cs was significantly higher under anaerobic redox conditions (-200 mV), when soluble NH+4 concentrations in the anaerobic suspensions were 1000 microM or higher. Activities of 137Cs were highest (initially 40-50% of the 137Cs added) on the NH+4-extractable site. Over the long term, activities of NH+4-extractable 137Cs decreased linearly to below 10% as 137Cs was fixed on inter-lattice sites. Water-soluble 137Cs was significantly correlated with Na-extractable 137Cs in short-term experiments, suggesting the existence of an equilibrium between the different clay sites. High concentrations of NH+4 under anaerobic redox conditions could shift this equilibrium, resulting in increases in water-soluble 137Cs and increases in the activities of 137Cs bound on non-selective clay sites. Additional studies are necessary to evaluate the interaction of 137Cs with sulfides, iron oxides, and other reactive chemical species which may attenuate 137Cs in sediment.

Cesium Radioisotopes↗

Pressurized liquid extraction of selected molecular biomarkers in deep sea sediments used as proxies in paleoceanography.

Pressurized liquid extraction has been performed on a suite of deep-sea sediments to assess its capability as an extraction technique in the analysis of molecular biomarkers used in paleoceanography. Specific compounds assessed comprise long-chain alkenones, n-alkanes, n-alcohols and, additionally, one diol and one keto-ol. These have been extracted by both pressurized liquid extraction and ultrasonication for comparison. One key result is that the U37(K') index (based on the degree of unsaturation of the alkenones and used as a paleothermometer in paleoceanography) remains intact after both extraction techniques. In terms of biomarker concentrations, which are often used to qualitatively assess changes in marine productivity and/or terrigenous inputs, pressurized liquid extraction is substantially more efficient than ultrasonication, providing higher amounts of extracted constituents, particularly for polar compounds.

Biomarkers↗

Antibacterial activity of coffee extracts and selected coffee chemical compounds against enterobacteria.

The in vitro antimicrobial activity of commercial coffee extracts and chemical compounds was investigated on nine strains of enterobacteria. The antimicrobial activity investigated by the disc diffusion method was observed in both the extracts and tested chemical compounds. Even though pH, color, and the contents of trigonelline, caffeine, and chlorogenic acids differed significantly among the coffee extracts, no significant differences were observed in their antimicrobial activity. Caffeic acid and trigonelline showed similar inhibitory effect against the growth of the microorganisms. Caffeine, chlorogenic acid, and protocatechuic acid showed particularly strong effect against Serratia marcescens and Enterobacter cloacae. The IC(50) and IC(90) for the compounds determined by the microtiter plate method indicated that trigonelline, caffeine, and protocatechuic acids are potential natural antimicrobial agents against Salmonella enterica. The concentrations of caffeine found in coffee extracts are enough to warrant 50% of the antimicrobial effect against S. enterica, which is relevant to human safety.

Alkaloids↗

Extraction solvent selection in environmental analysis.

A method for the prediction of a suitable solvent for the extraction of pesticides is outlined. The procedure is based on the Hildebrand solubility parameter, delta(t). The solubility parameter is broken down into three individual components, which are calculated by the addition of group contributions. To demonstrate the applicability of the approach pressurized fluid extraction was used to extract 4,4'-DDT [1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane] and its metabolites, 4,4'-DDD [1,1-dichloro-2,2-bis(p-chlorophenyl)ethane] and 4,4'-DDE [1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene], from an historically contaminated soil from the United States and pentachlorophenol from a certified reference material (CRM524) using various solvents. Visual representation of the individual parameters predicted the ideal extraction solvent to be DCM for 4,4'-DDT and its metabolites and a mixture of acetonitrile and dichloromethane (1:1, v/v) for PCP. These findings were confirmed by the experimental results.

Environmental Pollutants↗

Antibacterial activity of aqueous extracts of selected chewing sticks.

This aim of this study was to determine the antibacterial activity in extracts obtained from various Nigerian chewing sticks. Aqueous extracts from seventeen chewing sticks and the fruit of C. ferruginea, one fruit used in oral hygiene in Nigeria, were screened for antibacterial activity against type cultures of Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa. Eleven of the test extracts showed activity against at least two of these referenced organisms. Minimum inhibitory concentrations (MIC) of these eleven extracts against clinical isolates from orofacial infection were determined. All the extracts demonstrated activity against Staphylococcal and Streptococcal isolates. Over half of the extracts were active against Enterobacteriaceae and obligate anaerobic isolates, including Prevotella melaninogenica, Porphyromonas gigivalis, Fusobacterium nucleatum, and Peptostreptococcus prevotii. Extracts of the Vitellaria paradoxa root, Bridellia ferruginea stem and twigs, Garcinia cola stem, Terminalia glaucescens root, Morinda lucida root, and Cnestis ferruginea fruit showed appreciable activity against all classes of bacterial isolates. The extracts of these plants may serve as sources for chemotherapeutic agents for the management of orofacial infections.

Bacteria, Anaerobic↗

Electrospray characterization of selected medicinal plant extracts.

Extracts of selected medicinal plants were examined by electrospray mass spectrometry (ESI-MS). This technique allowed identification of the main components of each extract, thereby providing a typical finger-print of the examined plants. More specifically, anthocyanins (Vaccinium myrtillus), isoflavones (Glycine max, soybean), flavonol-glycosides and terpenes (Ginkgo biloba), triterpenes (Centella asiatica), caffeoyl-quinic acids (Cynara scolymus, artichoke), ginsenosides (Panax ginseng), catechins (Camellia sinensis, green tea) and flavones and flavanones (Propolis) were detected rapidly at levels in the range of 0.1-1 microg/ml, using 0.2-1 mg/ml of each medicinal plant extract.

Plant Extracts↗