Solubilization of proteins from human lymph node tissue and two-dimensional gel storage.

In the present study, we compared six different solubilization buffers and optimized two-dimensional electrophoresis (2-DE) conditions for human lymph node proteins. In addition, we developed a simple protocol for 2-D gel storage. Efficient solubilization was obtained with lysis buffers containing (a) 8 M urea, 4% CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate), 40 mM Tris base, 65 mM DTT (dithiothreitol) and 0.2% carrier ampholytes; (b) 5 M urea, 2 M thiourea, 2% CHAPS, 2% SB 3-10 (N-decyl-N,N-dimethyl-3-ammonio-1-propanesulfonate), 40 mM Tris base, 65 mM DTT and 0.2% carrier ampholytes or (c) 7 M urea, 2 M thiourea, 4% CHAPS, 65 mM DTT and 0.2% carrier ampholytes. The optimal protocol for isoelectric focusing (IEF) was accumulated voltage of 16,500 Vh and 0.6% DTT in the rehydration solution. In the experiments conducted for the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), best results were obtained with a doubled concentration (50 mM Tris, 384 mM glycine, 0.2% SDS) of the SDS electrophoresis buffer in the cathodic reservoir as compared to the concentration in the anodic reservoir (25 mM Tris, 192 mM glycine, 0.1% SDS). Among the five protocols tested for gel storing, success was attained when the gels were stored in plastic bags with 50% glycerol. This is the first report describing the successful solubilization and 2D-electrophoresis of proteins from human lymph node tissue and a 2-D gel storage protocol for easy gel handling before mass spectrometry (MS) analysis.

Electrophysiological responses of Atta sexdens rubropilosa workers to essential oils of Eucalyptus and its chemical composition

The leaf-cutting ant Atta sexdens rubropilosa Forel, 1908 is the most harmful of the Eucalyptus pests, causing severe losses in wood production through defoliation. Various strategies have been tried and effort spent on the development of methods to control this pest, however no practical and environmentally acceptable one currently exists. In this work the chemical composition of the essential oil of seven Eucalyptus species was identified and the selectivity and sensitivity of antennal receptors of A. sexdens rubropilosa workers to the volatile compounds were determined using the electroantennographic technique (EAG and GC-EAD). Analysis by GC-EAD showed in E. cloesiana and E. maculata, respectively, seventeen and sixteen terpenes that elicited responses in ant workers' antennae, indicating the potential role of the essential oils as allelochemicals that determine the choice of the foraging material. © 2006 Verlag der Zeitschrift für Naturforschung.

Yield and phytochemical characterization of essential oil from Ocimum selloi B. obtained by hydrodistillation and supercritical fluid extraction

The yield and chemical composition of essential oils from leaves of Ocimum selloi B. submitted to organic and mineral fertilization, obtained by hydrodistillation and supercritical fluid extraction (SFE) were compared. Essential oil was extracted in a Clevenger-type apparatus for 2 h 30 min and analyzed by GC-MS (Shimadzu, QP 5050-DB-5 capillary column - 30 m × 0.25 mm × 0.25 μm). Carrier gas was helium (1.7 ml/min); split ratio: 1:30. Temperature program: 50°C, rising to 180°C at 5°C/min, 180°C, rising to 280°C at 10°C/min. Injector temperature: 240°C and detector temperature: 230°C. Identifications of chemical compounds were made by matching their mass spectra and Kovat's indices (IK) values with known compounds reported in the literature. An Applied Separations-apparatus (Speed SFE, model 7071, Allentown, PA, EUA) was used for SFE extractions. They were conducted at pressure 200 bar and temperature 30°C (20 min in static mode and 40 min in dynamic mode). The supercritical CO2 flow rate was (6.8±0.7)×10-5 kg-CO2/s. The essential oil collected was immersed in ethylene glycol bath (5°C). The yield of essential oils obtained by SFE was larger than hydrodistillation in both fertilization treatments (279 and 333% for organic and mineral fertilizations, respectively). There were no differences between the fertilization treatments. The amount of the volatile components showed by GC-MS chromatogram was highest in the essential oil obtained by hydrodistillation than SFE. The main volatile constituents of the essential oils were trans-anethole (Hydrodistillation: organic - 52.4%; mineral - 55.0%/ SFE: Hydrodistillation - 62.8%; mineral - 66.8%) and methyl-chavicol (Hydrodistillation: organic - 37.3%; mineral - 38.3%/ SFE: organic - 8.4%; mineral - 4.3%). A reduction of methyl-chavicol relative proportion of essential oil obtained by SFE was observed. Cys-anethole, α-copaene, trans-cariofilene, germacrene-D, β-selinene, biciclogermacrene and spathulenol were expressed only in hydrodistillation. The extraction of essential oil by SFE presented larger yield of essential oil than hydrodistillation technique, presenting, however, these essential oils, different phytochemical profiles.

Optimal single-embryo mass spectrometry fingerprinting

In pre-implantation embryos, lipids play key roles in determining viability, cryopreservation and implantation properties, but often their analysis is analytically challenging because of the few picograms of analytes present in each of them. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) allows obtaining individual phospholipid profiles of these microscopic organisms. This technique is sensitive enough to enable analysis of individual intact embryos and monitoring the changes in membrane lipid composition in the early stages of development serving as screening method for studies of biology and biotechnologies of reproduction. This article introduces an improved, more comprehensive MALDI-MS lipid fingerprinting approach that considerably increases the lipid information obtained from a single embryo. Using bovine embryos as a biological model, we have also tested optimal sample storage and handling conditions before the MALDI-MS analysis. Improved information at the molecular level is provided by the use of a binary matrix that enables phosphatidylcholines, sphingomyelins, phosphatidylserines, phosphatidylinositols and phosphoethanolamines to be detected via MALDI(±)-MS in both the positive and negative ion modes. An optimal MALDI-MS protocol for lipidomic monitoring of a single intact embryo is therefore reported with potential applications in human and animal reproduction, cell development and stem cell research. Copyright © 2013 John Wiley & Sons, Ltd. Copyright © 2013 John Wiley & Sons, Ltd.

Atividade antibacteriana de óleos essenciais e avaliação de potencial sinergético

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Caracterização da composição química dos óleos essenciais de Lychnophora pinaster Mart. em função da sazonalidade

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Substâncias húmicas aquáticas: caracterização e interação com íons crômio e proposta de um indicador de matéria orgânica lábil

Pós-graduação em Química - IBILCE

Métodos de obtenção de lignanas de Holostylis reniformis provenientes de duas regiões brasileiras

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Padronização de extratos vegetais: Astronium urundeuva (Anacardiaceae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo metalômico do mercúrio em leite materno coletado da população ribeirinha da área de influência do Ahe Jirau-bacia do rio Madeira

Pós-graduação em Biologia Geral e Aplicada - IBB

Aplicação de β-glicosidases produzidas pelos microorganismos Aureobasidium pullulans e Thermoascus aurantiacus ao processo fermentativo da aguardente de cana com foco na produção de terpenos

Pós-graduação em Alimentos e Nutrição - FCFAR

Estudo dos fotoiniciadores utilizados em resinas compostas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Remoção pela chuva de diferentes formulações de flutriafol aplicada em soja, com e sem a adição de óleo mineral na calda

Pós-graduação em Agronomia (Energia na Agricultura) - FCA

Determination of myricetin derivatives in Chrysobalanus icaco L. (Chrysobalanaceae)

A fração flavonoídica do extrato hidroalcoólico de folhas de Chrysobalanus icaco L. (Chrysobalanaceae), que são largamente utilizadas na medicina tradicional do Norte do Brasil para controlar a glicemia de pacientes diabéticos, foi caracterizada. Miricetina-3-O-glucuronídeo (miricitrina) e quercitrina, entre outros derivados de miricetina minoritários, foram evidenciados por análises com sistemas CLAE/DAD e CLAE/EM

Biotransformation of chalcones by the endophytic fungus Aspergillus flavus isolated from Paspalum maritimum trin

O fungo Aspergillus flavus isolado como endofítico da planta Paspalum maritimum Trin. foi avaliado quanto ao seu potencial de aplicação em reações de biotransformações. Os compostos chalcona (1), 3,4,5-trimetoxichalcona (2) e 2,3,4,4'-tetrametoxichalcona (3) foram biotransformados, respectivamente, na diidrochalcona (4), 3,4,5-trimetoxidiidrochalcona (5) e 2,3,4,4'-tetrametoxidiidrochalcona (6). As estruturas destes compostos foram determinadas por análises de RMN uni e bidimensionais e por espectrometria de massas. As diidrochalconas 5 e 6 são substâncias inéditas.