Antiproliferative activity of extracts of Euphorbia tirucalli L (Euphorbiaceae) from three regions of Brazil

Purpose: To investigate Euphorbia tirucalli extract for probable geographic variations in its antiproliferative activity.Methods: The aerial parts of E. tirucalli were collected in the Brazilian states of Mato Grosso, Rio de Janeiro, Pará, Minas Gerais and Santa Catarina. The 70 % ethanol extract was obtained according to the procedure described in Brazilian Homeopathic Pharmacopeia. The antiproliferative activity of extracts, in concentrations of 62, 125, 250, and 500 μg mL-1, was tested against leukemia (HL-60), lymphoma (Daudi) and melanoma (B16F10) cell lines using methyl thiazol tetrazolium assay (MTT). Phytochemical analysis were carried out using High-performance liquid chromatography-diode array (HPLC-UV-DAD) and electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI(-) FT-ICR MS) assays.Results: There was significant regional variability in the cytotoxicity of E. tirucalli extracts in a dosedependent manner. The extracts had similar activity towards leukemia cell line HL-60, decreasing cell viability to about 60 – 70 %. The extract showed the presence of ellagitannins, flavonoids, veracylglucan, and acid triterpenes as the major compounds.Conclusion: While the results support the ethnopharmacological use of E. tirucalli throughout Brazil, regional quantitative differences found in some classes of secondary metabolites may explain the variations observed in antitumor activity.Keywords: Aveloz, Cancer, Cytotoxicity, Antiproliferative, Ethnopharmacological, Traditional medicin

Determining the distribution of triclosan and methyl triclosan in estuarine settings

We have developed a method for the analysis of two sewage-derived contaminants: triclosan (TCS), an antibacterial agent, and methyl triclosan (MTCS), a TCS metabolite. For solid samples (4 g), extraction and cleanup were integrated into the same step using pressurized liquid extraction (PLE) with in-cell-clean-up (1 g of florisil). The extraction was performed using dichloromethane at 100 °C, 1500 psi and 3 static extraction cycles of 5 min each. For water samples (100 mL), stir bar sorptive extraction–liquid desorption (SBSE–LD) was used. Bars were stirred for 10 h and analytes were later desorbed using acetonitrile. Finally, MTCS and a silylated derivative of TCS were determined by gas chromatography–mass spectrometry (GC–MS). Recovery experiments in water and sediments were performed and the results ranged from 67% to 78%. Limits of detection (LODs) were 5 ng L−1 for TCS and 1 ng L−1 for MTCS, in water samples, and 0.1 ng g−1 for TCS and MTCS in solid samples. The method was applied then to determine the levels of these compounds in the estuary of Guadalete River (SW Spain). TCS and MTCS concentrations up to 9.6 ng g−1 in sediments and 310 ng L−1 in water were measured. Their distribution was strongly influenced by the presence of wastewater sources, treated and untreated, along the sampling area, where maximum concentrations were detected. Highest values were reached in the water column during low tides as the water volume in the estuary becomes lower

Environmentally friendly analysis of emerging contaminants by pressurized hot water extraction-stir bar sorptive extraction-derivatization and gas chromatography-mass spectrometry

This work describes the development, optimiza- tion, and validation of a new method for the simultaneous determination of a wide range of pharmaceuticals (beta- blockers, lipid regulators ... ) and personal care products (fragrances, UV filters, phthalates ... ) in both aqueous and solid environmental matrices. Target compounds were extracted from sediments using pressurized hot water ex- traction followed by stir bar sorptive extraction. The first stage was performed at 1,500 psi during three static extrac- tion cycles of 5 min each after optimizing the extraction temperature (50 – 150 °C) and addition of organic modifiers (% methanol) to water, the extraction solvent. Next, aqueous extracts and water samples were processed using polydime- thylsiloxane bars. Several parameters were optimized for this technique, including extraction and desorption time, ionic strength, presence of organic modifiers, and pH. Fi- nally, analytes were extracted from the bars by ultrasonic irradiation using a reduced amount of solvent (0.2 mL) prior to derivatization and gas chromatography – mass spectrome- try analysis. The optimized protocol uses minimal amounts of organic solvents (<10 mL/sample) and time ( ≈ 8 h/sam- ple) compared to previous ex isting methodologies. Low standard deviation (usually below 10 %) and limits of de- tection (sub-ppb) vouch for the applicability of the method- ology for the analysis of target compounds at trace levels. Once developed, the method was applied to determin

Exogenous HIV-1 Nef Upsets the IFN-γ-Induced Impairment of Human Intestinal Epithelial Integrity

The mucosal tissues play a central role in the transmission of HIV-1 infection as well as in the pathogenesis of AIDS. Despite several clinical studies reported intestinal dysfunction during HIV infection, the mechanisms underlying HIV-induced impairments of mucosal epithelial barrier are still unclear. It has been postulated that HIV-1 alters enterocytic function and HIV-1 proteins have been detected in several cell types of the intestinal mucosa. In the present study, we analyzed the effect of the accessory HIV-1 Nef protein on human epithelial cell line.We used unstimulated or IFN-γ-stimulated Caco-2 cells, as a model for homeostatic and inflamed gastrointestinal tracts, respectively. We investigated the effect of exogenous recombinant Nef on monolayer integrity analyzing its uptake, transepithelial electrical resistance, permeability to FITC-dextran and the expression of tight junction proteins. Moreover, we measured the induction of proinflammatory mediators. Exogenous Nef was taken up by Caco-2 cells, increased intestinal epithelial permeability and upset the IFN-γ-induced reduction of transepithelial resistance, interfering with tight junction protein expression. Moreover, Nef inhibited IFN-γ-induced apoptosis and up-regulated TNF-α, IL-6 and MIP-3α production by Caco-2 cells while down-regulated IL-10 production. The simultaneous exposure of Caco-2 cells to Nef and IFN-γ did not affect cytokine secretion respect to untreated cells. Finally, we found that Nef counteracted the IFN-γ induced arachidonic acid cascade.Our findings suggest that exogenous Nef, perturbing the IFN-γ-induced impairment of intestinal epithelial cells, could prolong cell survival, thus allowing for accumulation of viral particles. Our results may improve the understanding of AIDS pathogenesis, supporting the discovery of new therapeutic interventions

Analysis and distribution of estrogens and progestogens in sewage sludge, soils and sediments

9 pages, 4 tables.This article focuses on solid samples and reviews the main findings so far concerning the source, the presence and the fate of estrogens and progestogens in the aquatic environment. We discuss the very few existing analytical methods for determination of estrogens and progestogens in environmental matrices (soils, sediments and sludge). Estrogens are continuously released in the aquatic environment mainly because treatment plants are unsuccessful in removing them. Studies show that estrogens and progestogens are easily distributed in the environment and are likely to accumulate in river sediments and in soils. However, it is not yet clear whether sorption or biodegradation processes play a major role in their elimination from the aquatic environment.This work was supported by the Energy, Environmental and Sustainable Development Program (Project ARTDEMO EVK1-CT2002-00114), and by the Spanish Ministry of Science and Technology (Projects BQU2002-10903-E and PPQ2001-1805-CO3-01).Peer reviewe

Analysis of pesticides in water by liquid chromatography-tandem mass spectrometric techniques

17 pages, 7 figures, 3 tables.-- PMID: 16705628 [PubMed].Pesticide residues continue to be the focus of many environmental studies, and the number of articles describing the development of more advanced, multiresidue analytical methodologies does not decline. The use of liquid chromatography-mass spectrometry based on single quadrupole or ion trap analyzers is consolidated for this purpose. The implementation, in the near future, of more sophisticated mass analyzers, such as triple quadrupole and hybrid quadrupole-time-of-flight is anticipated for routine analysis. This article reviews the various works published so far in the literature for the determination of pesticides and transformation products (TPs) in water by means of liquid chromatography (LC) coupled to tandem mass spectrometry. It discusses the various ionization sources and analyzers used for this purpose, as well as the extraction procedures employed for previous sample preconcentration. Because of the widespread use of triple quadrupole analyzers for the generation of pesticides levels in water using tandem mass spectrometry, a table compiling the transitions monitored for ca. 70 compounds is also included.This work has been supported by the EU through the projects ARTDEMO (EVK1-CT2002-00114) and MODELKEY (SSPICT-2003-511237-2), and by the Ministerio de Ciencia y Tecnología through the project EVITA (CTM2004-06265-C03-01/TECNO). Marina Kuster gratefully acknowledges the I3P Program (Itinerario integrado de inserción profesional; co-financed by CSIC and European Social Funds) for a predoctoral grant.Peer reviewe

Squaraine Planar-Heterojunction Solar Cells

The photovoltaic performance of squaraine-based organic solar cells is investigated. Two squaraine derivatives with extraordinarily high extinction coefficients are used as electron donors in bilayer heterojunctions with fullerene as electron acceptor. Due to the very strong squaraine absorption band in the red spectral domain, antibatic behavior due to light filtering is observed in the photocurrent spectrum for film thicknesses of 35 nm to 40 nm. At reduced film thicknesses of 20 nm, this filtering effect at maximum absorption can be alleviated and power conversion efficiencies under simulated AM 1.5 full sun irradiation of 0.59% and 1.01% are obtained for the two squaraine derivatives, respectively. The photovoltaic properties of these cells are investigated with respect to electrode materials and chemical doping.ISSN:1110-662XISSN:1687-529

Comparative study of an estradiol enzyme-linked immunosorbent assay kit, liquid chromatography-tandem mass spectrometry, and ultra performance liquid chromatography-quadrupole time of flight mass spectrometry for part-per-trillion analysis of estrogens in water samples

10 pages, 5 tables.-- PMID: 17540393 [PubMed].-- Printed version published on Aug 10, 2207.-- Issue title: 23rd Montreux Symposium on Liquid Chromatography-Mass Spectrometry, Supercritical Fluid Chromatography-Mass Spectrometry, Capillary Electrophoresis-Mass Spectrometry and Tandem Mass Spectrometry (Montreaux, Switzerland, Nov 8-10, 2006).Presented at the 2nd International Workshop on Liquid-Chromatography-Tandem Mass Spectrometry for Screening and Trace Level Quantitation in Environmental and Food Samples, Barcelona, Spain, September 18–19, 2006.àáEstrogens have been often identified as the major contributors to the endocrine-disrupting activity observed in environmental waters. However, their analysis in these, sometimes very complex, matrices is still challenging due to the very low detection limits and the selectivity required for their reliable determination at the very low concentrations at which they are physiologically active. In this work, a polyclonal enzyme-linked immunosorbent assay (ELISA) kit for 17-prueba-estradiol analysis, high-performance liquid chromatography-tandem mass spectrometry (HPLC–MS/MS) based on triple-quadrupole analyzer (QqQ), and a newly developed method based on ultra performance liquid chromatography–quadrupole time of flight mass spectrometry (UPLC–Q-TOF-MS) have been evaluated in terms of performance for the rapid screening, quantitative analysis, and unequivocal identification of some selected, environmentally relevant estrogens in different water matrices, including urban wastewater, river water, and ground water, after solid phase extraction. Compounds quantified and/or identified included the estrogens 17-β-estradiol, estrone, 17-α-ethynyl estradiol and estriol, and the isoflavones daidzein, genistein, and biochanin A. Except for a moderate overestimation using the ELISA kit, especially in the analysis of complex wastewater samples, results obtained by all the investigated techniques were in very good, general agreement. The instrumental sensitivity achieved increased in the order: UPLC–Q-TOF-MS < polyclonal ELISA kit < HPLC–MS/MS (QqQ). Direct analysis of water samples by using the ELISA kit permitted to reach a limit of detection of 2.5 ng L(−1). However, using an appropriated sample pretreatment method detection limits at nanogram to picogram per liter levels can be obtained with all techniques and the risk for matrix effects is minimized. In terms of selectivity, both HPLC–MS/MS (QqQ) and UPLC–Q-TOF-MS show outstanding performance, but the latter allows, in addition, shorter analysis times (16 min vs. 45 min) and the identification of non-target, unknown compounds. The identification of unknown compounds is based on the accurate mass measurements for the precursor and product ions, that permit the elemental compositions calculation and the chemical structures to be identified searching against different databases.This work has been supported by the EU through the projects SWIFT-WFD (SSPI-CT-2003-502492), and MODELKEY (SSPI-CT-2003-511237-2), and the financial support from the Spanish Ministry of Education and Science, through the projects CTM2005-24255-E and EVITA(CTM2004-06265-C03-01/TECNO). Marinella Farré thanks the support from the Spanish Ministry of Education and Science through the Juan de la Cierva program.Peer reviewe

Evaluation of two aquatic passive sampling configurations for their suitability in the analysis of estrogens in water

The presence of estrogens in the aquatic environment has been the target of several studies in the last decade. Newly developed passive sampling techniques for polar organic chemicals show great promise for the assessment of long-time exposure of aquatic organisms to emerging contaminants. In the present work, two configurations of the Chemcatcher® passive sampler have been tested for their applicability to the analysis of seven estrogens in water. Accumulation experiments in the laboratory, to calculate the uptake rates, and a field trial show that the polar configuration of this device may be used for the efficient sampling and determination of estrogens in water. Time weighted average concentrations were determined in the field trial and compared with spot sampling concentrations. The detection of estriol using passive sampling, although not found with spot sampling, clearly demonstrates the value of this technique in assessing relevant concentrations of estrogens in the aquatic media.This work has been financially supported by the European Union under the STAMPS (EVK1-CT2002-00119) project and by the Spanish Ministry of Science and Innovation through the projects CEMAGUA (CGL2007-64551/HID), SCARCE (Consolider-Ingenio 2010 CSD2009-00065) and SOSTAQUA (led by Aguas de Barcelona and founded by CDTI in the framework of the Ingenio 2010 Programme under the CENIT call), and reflects the author's view. The EU is not liable for any use that may be made of the information contained in it. Many thanks to Arturo Faura, Lluis Tirapu and Antoni Ginebreda from the Catalonian Water Agency for helping with the choice of the sampling sites and for the use of their facilities, as well as for providing the water parameters monitored with SAIH and XACQA. Marina Kuster gratefully acknowledges the I3P Program (Itinerario integrado de inserción profesional; co-financed by CSIC and European Social Funds) for a predoctoral grant. Merck (Darmstadt, Germany) is acknowledged for the gift of SPE and LC columns.Peer reviewe