Método fluorescente (diacetato de fluoresceína e brometo de etídeo) para o estudo da viabilidade de Cryptococcus neoformans em líquor

Padronizou-se método de fluorescência (solução de diacetato de fluoresceína DF e brometo de etídio BE) para análise de viabilidade de células fúngicas, em 40 amostras de liquor, provenientes de casos comprovados de neurocriptococose. A utilização de solução aquosa de saponina a 0,3% eliminou fluorescências interferentes emitidas por hemácias e leucócitos. Após o processamento dos materiais biológicos, foram retiradas alíquotas de 0,1 ml das supensões obtidas e misturadas a volumes iguais da solução DF-BE preparada pouco antes do uso. O tempo de coloração ideal foi de 30 minutos, resultando perfeita diferenciação entre microrganismos viáveis (fluorescência verde) e não viáveis (fluorescência vermelha).

Transporte do brometo de etídio através da parede celular de Mycrobacterium smegmatis: desenvolvimento e aplicação de metodologias de quantificação do transporte e correlação com a resistência aos antibióticos

O efluxo activo e a permeabilidade celular reduzida são consideradas as principais causas da resistência intrínseca micobacteriana a diversos antimicrobianos. Neste estudo comparámos a capacidade de extrusão do brometo de etídio (EtBr), um substrato universal de bombas de efluxo, pela estirpe selvagem M. smegmatis mc2155 com estirpes mutantes deletadas para LfrA e MspA, respectivamente a principal bomba de efluxo e porina de M. smegmatis, na presença e ausência dos inibidores de bombas de efluxo (IBEs) verapamil, tioridazina, clorpromazina e carbonil cianeto mclorofenilhidrazona (CCCP) e correlacionámos estes resultados com a capacidade destes IBEs em reduzir a susceptibilidade de M. smegmatis a diversos antibióticos. Os resultados obtidos mostram que, na ausência da principal porina de M. smegmatis, MspA, a acumulação de EtBr diminui e as células tornam-se igualmente mais resistentes a diversos antibióticos. Quando a principal bomba de efluxo de M. smegmatis, LfrA, é deletada a estirpe mutante apresenta um incremento na acumulação de EtBr e um aumento na susceptibilidade ao EtBr, isoniazida, rifampicina, etambutol e ciprofloxacina. Na presença dos IBEs testados, com excepção do CCCP, observa-se uma redução da concentração mínima inibitória para a estreptomicina, rifampicina, amicacina, ciprofloxacina, claritromicina e eritromicina. Estes resultados colocam em evidência que a porina MspA é um canal importante para a entrada de EtBr e antibióticos nas células e que o efluxo activo através da bomba de efluxo LfrA está envolvido na resistência de baixo nível a diversos antibióticos e EtBr em M. smegmatis. As metodologias desenvolvidas neste trabalho e os resultados obtidos proporcionaram um contributo para o melhor conhecimento dos dois principais mecanismos responsáveis pela resistência intrínseca micobacteriana a diversos antimicrobianos, considerada como um importante factor facilitador da aquisição e estabilização de um património genético que conduz ao desenvolvimento de resistência de alto nível em micobactérias.

Experimentos com os nematicidas D. D., E. D. B. e brometo de metilo no combate aos nematódeos causadores de galhas em raízes de plantas (Meloidogyne spp.)

Tomato roots heavily disfigured by root-knot nematodes were throughly mixed with soil. At various time intervals, samples were taken from the mixture and treated in closed containers by each of the folio wing nematicides: D.D., E.D.B. and M.B. The efficacy of the treatment was tested by setting indicator plants in the treated soil and by examining their roots for the presence of galls two months later. In other words, the ability of the three nematicides to penetrate nematode galls after various periods of rotting, which varied from 5 to 30 days was studied. The main conclusions drawn are as follows: a) no nematicide among the three listed above showed the ability for complete destruction of the nematodes protected inside the roots, for a number of small galls developed on the root system of the indicator plant in all treatments; b) smaller and less numerous galls were present on the roots of the indicator plants grown in soil treated after a rotting period of 30 days; c) however, the control obtained seems to be quite satisfactory economically, since the check plants grew poorly and have developed a very unhealthy root system. This is in accordance with STARK & LEAR (1947), LEAR (1951) and CICCARONE's (1951) statements. The results of the present experiments show again that awaiting for the rotting of galls of the root-knot nematodes is not indispensable for an economically convenient soil fumigation. Fields in which many fleshy infected roots from previous crops have been buried can be economically fumigated immediately, without any loss of time. Notwithstanding, when thick woody roots are present in the soil, the above statements may not hold true. This should constitute a new problem calling for further experiments. Another essay dealing with methyl bromide alone, consisted in treating cotton roots heavily disfigured by Meloidogyne incognita in a container (diameter = 28cm, height = 32 cm), which remained closed for five days. After the treatment, the roots were mixed with soil, in which tomato seedlings were planted. After a growing period of two months, the roots of the tomato plants were washed in running water and examined for the presence of galls. As an early infeccion was present in the root system of all plants, the inefficacy of the treatment has been proved.

Excitação eletrônica das moléculas de metacrilato de metila e estireno na região do ultravioleta de vácuo

Angle-resolved electron energy-loss spectra have been measured for the methyl methacrylate (MMA) and styrene molecules in the 0 - 50 eV energy range. The spectra have been obtained at 1 keV incident energy, with an energy resolution of 0.8 eV and covering an angular range of 2.0 to 7.0 degrees. Within our knowledge, this is the first gas-phase excitation spectrum for MMA and styrene in this energy range. The spectra of MMA at small scattering angles are dominated by an intense peak at 6.7 eV followed by a broad band centered at about 16 eV. In the case of styrene, six bands can be observed in the spectra. Based on the angular behaviour of the excitation spectra of these molecules, the low-lying peaks observed are considered to be associated predominantly with dipole-allowed processes. In both cases, new bands can be observed for excitation energies greater than 20 eV. This could be associated with dipole-forbidden transitions to shake-up and doubly-excited states.

Validação do método de determinação simultânea dos íons brometo e bromato por cromatografia iônica em águas de consumo humano

European Directive (98/83/CE), compulsory after 2008, states that bromate in drinking water must be controlled at levels below 10 mg L-1. Supporting implementation of the Directive, the European Comission has established project SMT4-CT96 2134, in collaboration with various european institutions, aiming at the identification of the interferents to the current analytical method (Ionic Chromatography with Conductimetric Detection - IC/CD), their removal and the automation of pre-treatment and injection steps, as well as the development of alternative methods. EPAL, responsible for the water supply to a great deal of Portuguese regions, has taken steps to meet these requirements. Although not part of such project, this work (the result of a project conducted under a protocol of collaboration between EPAL SA and FCUL - Faculty of Sciences, University of Lisbon), reports on studies of usefulness to laboratories planning to monitor bromate in ozone treated waters, in conditions different from those described in EPA 300.1. Simultaneous determination of bromide is justified by its role as bromate precursor.

Estudo da adsorção de brometo de etídeo em resina XAD-7

The adsorption of ethidium bromide on XAD-7 resin was studied. The Freundlich model was the most representative isotherm model to describe the sorption behavior. A solid-liquid equilibrium model was proposed to explain the resin mass influence on the sorption. The equilibrium constant value estimated was 2.31. The results showed an ethidium bromide ion-pair physical adsorption, with adsorption enthalpy equals to -19.33 kJ/mol. A pK2 value equals to 4.69 ± 0.01 was estimated by two distinct methods. The results will be applied to the ethidium bromide preconcentration aiming its decomposition.

Determinação de ácido valpróico em soro por cromatografia líquida de alta eficiência com detector de arranjo de diodos (CLAE-DAD), após derivatização com brometo de fenacila

Valproic acid (VA) is a drug used to control seizures in several epileptic conditions. In VA pharmacotherapy, therapeutic drug monitoring is recommended to obtain adequate seizure control and avoid toxicity. The aim of this study was to validate a method for the determination of valproic acid in serum, employing high performance liquid chromatography with diode array detection (HPLC-DAD), after derivatization with phenacyl bromide. The calibration curve (y=0.0133x-0.0025) presented good linearity with r²=0.9999. Accuracy (101-115%), intra-assay precision (4.53-8.15%) and inter-assay precision (3.15-6.77%) were acceptable. The quantification limit was 2.0 µg/mL. The method presented similar results to enzyme immunoassay.

Recentes aplicações sintéticas de compostos orgânicos tricloro(bromo)metila substituídos

This review presents the recent applications of some trihalomethyl carbinols, α,α,α-trihalogenated carbonyl compounds and trichloro(bromo)methyl-containing heterocycles, acting as high versatile synthetic precursors in organic chemistry, based on the last 10 years' literature.

Síntese e caracterização de argilas organofílicas contendo diferentes teores do surfactante catiônico brometo de hexadeciltrimetilamônio

The major applications of organoclays are in adsorption of organic polluents. The objective of this work was the synthesis and characterization of organoclays using differents amounts of cationic surfactant hexadecyltrimethylammonium bromide. The clays were characterized by low angle x-ray diffraction (XRD), scanning electron microscope (SEM), infrared with Fourier tranformation (FTIR), BET surface area, elemental analysis (CHN), Foster swell and adsorption of methylene blue. The surfactant can adsorb in differents forms in the interlamelar region changed the basal spacing. The presence of the surfactant adsorbed can be favorable or not in adsorption of the methylene blue due the different interactions dye-organoclays.

Hidroformilação do oleato de metila catalisada por complexos de ródio

In this work, we describe the hydroformylation of methyl oleate catalyzed by several rhodium complexes. Parameters including total pressure, phosphorous/rhodium and CO/H2 ratio, temperature and phosphorous ligands were scanned. Total conversion of the starting double bonds was achieved while maintaining excellent selectivity in aldehydes.

Quantificação de salicilato de metila em quatro gêneros de polygalaceae, por CLAE-DAD

Polygalaceae is represented in Brazil by ten genera and 191 species, of which the Polygala is the most representative, characterized by the occurrence of methyl salicylate. Seventeen species of Polygalaceae have been analyzed by HPLC-DAD and the technique proved to be selective, precise, accurate, and with low limits of quantification and detection. The analysis of plant material confirmed the presence of methyl salicylate, with concentration values ​​ranging from 14.1 a 126.9 µg/g.

Atividades biológicas dos óleos essenciais de Endlicheria citriodora, uma lauraceae rica em geranato de metila

The essential oils of branches and leaves of Endlicheria citiodora were obtained by hydrodistillation and analysed using GC-FID, GC-MS and both NMR 13C and ¹H, resulting in the identification of methyl geranate as major constituent (93%) in both oils. Cytotoxicity, tyrosinase-inhibition and antioxidant activities were studied and characterized. High antioxidant potential (15.52 and 13.53 µg/mL), low cytotoxicity and tyrosinase inhibition (53.85%) were observed. This is the first paper reporting the biological activities and composition of the essential oils of this species.

Desevolvimento de nanocompósitos de poli(metacrilato de metila)/argila processados por mistura no estado fundido

Poly(methyl methacrylate)/clay nanocomposites were prepared by melt mixing using a montmorillonite-rich clay (MMT). The clay in natura was treated with acrylic acid to facilitate the dispersion in the polymer matrix. A masterbatch of PMMA/clay was prepared and combined with the pure PMMA and then subjected to extrusion process using singlescrew and twin-screw extruders followed by injection. Nanocomposites were processed with clay contents of 1, 3, 5 and 8 wt.%. The effect of shear processing on the morphology of the nanocomposites was evaluated by XRD, SEM and TEM. Thermal and mechanical properties of the nanocomposites were investigated through TGA, DSC, HDT, VICAT, tensile and impact tests, to evaluate the effect of the addition of clay to the PMMA matrix. Flammability tests were also conducted to investigate the effect of the addition of clay on the flame retardation properties. SEM images of the nanocomposites indicated the presence of clay agglomerates, which resulted in the reduction of properties such as thermal stability, mechanical strength and impact resistance, and increased the rate of burning for materials processed by both extrusion routes

Estudo morfológico da blenda polimérica poli(metacrilato de metila)/poli(tereftalato de etileno) reciclado (PMMA/PET)

Among the options for plastics modification more convenient, both from a technical-scientific and economic, is the development of polymer blends by processing in the molten state. This work was divide into two stages, with the aim to study the phase morphology of binary blend PMMA / PET blend and this compatibilized by the addition of the poly(methyl methacrylate-co-glycidyl methacrylate-co-ethyl acrylate) copolymer (MMA-GMA-EA). In the first stage is analyzed the morphology of the blend at a preliminary stage where we used the bottle-grade PET in a Haake torque rheometer and the effect of compatibilizer in this blend was evaluated. In the second stage the blend was processed using the recycled PET in a single screw extruder and subsequently injection molding in the shape of specimens for mechanical tests. In both stages we used a transmission electron microscopy (TEM) to observe the morphologies of the samples and an image analyzer to characterize them. In the second stage, as well as analysis by TEM, tensile test, scanning electron microscopy (SEM) and atomic force microscopy (AFM) was performed to correlate the morphology with the mechanical properties. The samples used in morphological analyzes were sliced by cryo-ultramicrotomy technique for the analysis by TEM and the analysis by SEM and AFM, we used the flat face of the block after cut cryogenic. It was found that the size of the dispersed phase decreased with the addition of MMA-GMA-EA in blends prepared in a Haake. In the tensile test, the values of maximum tensile strength and modulus of elasticity is maintained in a range between the value of pure PMMA the pure PET, while the elongation at break was influenced by the composition by weight of the PMMA mixture. The coupling agent corroborated the results presented in the blend PMMA / PETrec / MMA-GMA-EA (80/15/5 %w/w), obtained by TEM, AFM and SEM. It was concluded that the techniques used had a good morphologic correlation, and can be confirmed for final analysis of the morphological characteristics of the blends PMMA / PET

Efeito da adição dos azocorantes DR73 e DB79 nas propriedades do poli(metacrilato de metila)

Currently new polymeric materials have been developed to replace other of traditionally materials classes. The use of dyes allows to expand and to diversify the applications in the polymeric materials development. In this work the behavior and ability of azo dyes Disperse Blue 79 (DB79) and Disperse Red 73 (DR73) on poly(methyl methacrylate) (PMMA) were studied. Two types of mixtures were used in the production of masterbatches: 1) rheometer 2) solution. Processing by extrusion-blow molding of PMMA was carried out in order to evaluate the applications of polymeric films. Thermal analysis were performed by thermogravimetry to evaluate polymer and azo dyes thermal stability. Colorimetric analysis were obtained through monitoring the spectral variations associated with sys/trans/anti azo dyes isomerization process Colorimetric data were treated and evaluated in accordance to the color system RGB and CIEL*ab, by monitoring the color change as function of time. Mechanical properties, characterized by tensile tests, were evaluated and correlated with the presence and content of azo dyes in the samples. Analyses by scanning electronic microscopy (SEM) were performed on the surfaces of samples to check the azo dye dispersion after the mixing process. It was concluded that the production of PMMA/azo dyes is possible and feasible, and the mixtures produced had synergy of properties for use in various applications