Simultaneous determination of food dyes by first derivative spectrophotometry with sorption onto polyurethane foam.

A simple spectrophotometric method is described for resolving binary mixtures of some food dyes: Amaranth, Brilliant Blue, Sunset Yellow and Tartrazine, using the first-derivative spectra with measurements at zero-crossing wavelengths. Analytical curves are linear up to 20 mg L(-1). Standard deviations of 1.30, 2.22, 1.93 and 0.81% were obtained for synthetic binary mixtures of 2 mg L(-1) of Amaranth, Brilliant Blue, Sunset Yellow and Tartrazine, respectively. Before the spectrophotometric measurements, the dyes were sorbed onto polyurethane foam and recovered in sodium dodecyl benzene sulfonate solution. Therefore, matrix complexity was eliminated and simple spectra were obtained. The method was very satisfactorily used for determining the colorants in synthetic mixtures, with recoveries in the 96 - 101% range. Detection limit values were dependent on the colorant combination investigated. Commercial products containing binary combinations of these dyes in different ratios (from 1:1 to 1:8) were analyzed. The results were compared with those obtained by HPLC; very similar values were found by the two methods.     

Differentiation of isomeric flavone/isoflavone aglycones by MS2 ion trap mass spectrometry and a double neutral loss of CO

The fragmentation behaviour of seven pairs of isomeric flavone/isoflavone aglycones (solely hydroxylated and/or methoxylated) was studied using ion trap mass spectrometry with atmospheric pressure ionisation (API, both electrospray and APCI) in the positive and negative ion modes. A major difference was found in the neutral loss of 56 u, which was a common feature of all isoflavones in API(+). It was identified as a double loss of CO by accurate mass tandem mass spectrometric (MS/MS) measurements using a hybrid quadrupole time-of-flight (Q-TOF) instrument. Fragmentation of daidzein with (13)C-isotope labelling of the carbon C2 showed that this double loss occurred from the central ring of the molecule. A mechanism for this selective fragmentation is given. Further isoflavone-specific fragmentations were used to develop a guideline for the identification of isoflavone structures. A software-based neutral loss scan of 56 u in the API(+)-MS(2) mode was applied to extracts of leaves of Lupinus albus and to soy flour. The structure elucidation guideline allowed identification of hydroxy and/or methoxy isoflavones. Structures could be confirmed for those available as reference compounds.     

METABOLISM OF tRNAs IN GROWING CELLS OF Escherichia coli ILLUMINATED WITH NEAR-ULTRAVIOLET LIGHT

Abstract— The tRNA metabolism which accompanies illumination of growing E. coli cells has been examined in conditions that led to growth delay. (i) The in vivo formation of the 8–13 link was followed by a fluorimetric procedure and revealed pseudo-first order kinetics very close to those obtained in vim under the same illumination conditions. The yield of 8–13 link appears to be quantitative (± 10%). Comparison of these kinetics with the radiochromatographic data of Blanchetot et al . (1984) suggests the transient formation during illumination of a new RNase-T,-resistant dinucleotide in tRNA distinct from the 8–13 link. (ii) Evidence is provided that under illumination some tRNA molecules lack one or more bases in a specific position in the sequence, thus yielding discrete fragments after aniline treatment. (iii) During the growth lag, uracil incorporation into nucleic acids occurs at an apparent rate between 4–8% of that normally observed during exponential growth. Evidence is provided however that the pyrimidine ribonucleoside triphosphate pools are strongly perturbed after illumination. Comparison of exogenous [³H]uracil incorporation into two strains proficient or deficient in uracil biosynthesis suggests a derepression of the endogenous path after light treatment. In addition, the UTP-to-CTP conversion is inhibited. In spite of preferential incorporation of exogenously labelled uracil in tRNA after illumination, a possible pyrimidine base turnover cannot be proved. These data are compatible with tRNA repair (Blanchetot et al ., 1984) involving a few tRNA species.     

Human BCL-2 Expression Delays Ultraviolet-Induced Apoptosis in Marsupial Cells

We have introduced the human bcl-2 gene under the control of the human metallothionein MTII_A promoter into the rat kangaroo PtK2 cell line. Two independent clones were obtained in which the levels of Bcl-2 protein expression can be controlled by the addition of metals in the culture medium. These cell lines were employed to investigate the effects of this protein in UV-induced apoptosis. Overexpression of Bcl-2 in PtK2 cells resulted in a delay in the appearance of apoptosis markers, such as chromatin condensation and internucleosomal DNA fragmentation. However, colony survival after UV was not affected, suggesting that Bcl-2 did not impose a definitive block for cell death. The elimination of cyclobutane py-rimidine dimers through photoreactivation 24 h after irradiation in cells overexpressing Bcl-2 did not affect apoptosis. This indicates that irreversible events in the signaling pathway of apoptosis occur in the period between irradiation and photoreactivation even in the presence of high levels of Bcl-2 in the cell. Therefore, although the human Bcl-2 protein can delay the onset of UV-induced apoptosis in these marsupial cells, early events triggered by the pyrimidine dimers, upstream from the Bcl-2 action, lead the cell to a state committed to die.     

Superparamagnetic poly(methyl methacrylate) nanoparticles surface modified with folic acid presenting cell uptake mediated by endocytosis

The encapsulation of superparamagnetic nanoparticles (MNPs) in polymeric nanoparticles (NPs) with modified surfaces can improve targeted delivery and induce cell death by hyperthermia. The goals of this study were to synthesize and characterize surface modified superparamagnetic poly(methyl methacrylate) with folic acid (FA) prepared by miniemulsion polymerization (MNPsPMMA-FA) and to evaluate their in vitro cytotoxicity and cellular uptake in non-tumor cells, murine fibroblast (L929) cells and tumor cells that overexpressed folate receptor (FR) β, and chronic myeloid leukemia cells in blast crisis (K562). Lastly, hemolysis assays were performed on human red blood cells. MNPsPMMA-FA presented an average mean diameter of 135 nm and a saturation magnetization (Ms) value of 37 emu/g of iron oxide, as well as superparamagnetic behavior. The MNPsPMMA-FA did not present cytotoxicity in L929 and K562 cells. Cellular uptake assays showed a higher uptake of MNPsPMMA-FA than MNPsPMMA in K562 cells when incubated at 37 °C. On the other hand, MNPsPMMA-FA showed a low uptake when endocytosis mechanisms were blocked at low temperature (4 °C), suggesting that the MNPsPMMA-FA uptake was mediated by endocytosis. High concentrations of MNPsPMMA-FA showed hemocompatibility when incubated for 24 h in human red blood cells. Therefore, our results suggest that these carrier systems can be an excellent alternative in targeted drug delivery via FR.     

A structure-activity study of antibacterial diterpenoids

An analysis of the antibacterial activities of 15 terpenoids, eleven of which were previously described by us and four were extracted from the literature, suggested two structural requirements for activity of these and related compounds: a hydrophobic moiety,consisting of a substituted decalin skeleton, and a hydrophilic region possessing one hydrogen-bond-donor group. These structural requirements are responsible for an optimal insertion of these and related compounds into cell membranes, as suggested by the results of docking some of these compounds into a model phospholipid bilayer.     

On-line redox speciation analysis of antimony using l-proline immobilized on controlled pore glass and hydride generation inductively coupled plasma optical emission spectrometry for detection

l-proline was immobilized on controlled pore glass to study the ability of this material for the separation and preconcentration of Sb(III) and Sb(V). The substrate was packed in a minicolumn and incorporated in a flow injection system. The effluents of the on-line solid phase extraction (before and after elution) were directly coupled to the hydride generation inductively coupled plasma optical emission spectrometry system. The effect of pH, sample (and eluent) volume, flow rates of sample loading and elution on separation of Sb(III) e Sb(V) were evaluated. Our experiments demonstrated that Sb(V) was not retained and it was selectively determined during the loading step, while retained Sb(III) was determined after elution. The proposed system was also used for the selective preconcentration of Sb(III). In this case, a preconcentration factor of 11 and a limit of detection of 90 ng L⁻¹ for Sb(III) were achieved when 8 mL of sample were loaded into the column. The speciation analysis of inorganic Sb in river water and effluent samples was performed using the proposed method. The values obtained for total Sb (obtained by sum of Sb(III) and Sb(V)) were in good agreement with expected values. Recoveries of Sb(III) and Sb(V) in the river water Standard Reference Material 1640 (from National Institute of Standard and Technology) and spiked river waters were between 83 and 111%.     

On an evolution equation with acoustic boundary conditions

In this paper, we analyze from the mathematical point of view a model for small vertical vibrations of an elastic string with weak internal damping and quadratic term, coupled with mixed boundary conditions of Dirichlet type and acoustic type. Our goal is to extend some of the results of Frota‐Goldstein work in the sense of considering a weaker internal damping and one more quadratic nonlinearity in the elastic string equation. Copyright © 2011 John Wiley & Sons, Ltd.     

Conley's spectral sequence via the sweeping algorithm

In this article we consider a spectral sequence (Er,dr) associated to a filtered Morse-Conley chain complex (C,Δ), where Δ is a connection matrix. The underlying motivation is to understand connection matrices under continuation. We show how the spectral sequence is completely determined by a family of connection matrices. This family is obtained by a sweeping algorithm for Δ over fields F as well as over Z. This algorithm constructs a sequence of similar matrices Δ⁰=Δ,Δ¹,… , where each matrix is related to the others via a change-of-basis matrix. Each matrix Δr over F (resp., over Z) determines the vector space (resp., Z-module) Er and the differential dr. We also prove the integrality of the final matrix ΔR produced by the sweeping algorithm over Z which is quite surprising, mainly because the intermediate matrices in the process may not have this property. Several other properties of the change-of-basis matrices as well as the intermediate matrices Δr are obtained.     

Two-part fractional regression model with conditional FDH responses: an application to Brazilian agriculture

We extend the notion of a two-part fractional regression model with conditional free disposal hull efficiency responses to accommodate two-stage regression analysis. The two-part regression model includes the binomial model with a nonlinear specification for the expected response in (0,1] and is a more general formulation in the context of fractional regressions. We use nonlinear least squares to assess the effect of covariates in the conditional efficiency response. The approach is applied to Brazilian agricultural county data, as reported in the Brazilian agricultural census of 2006. The efficiency measure is output oriented and assumes variable returns to scale. Output is rural gross income and inputs are land expenses, labor expenses and expenses on other technological inputs. The covariates affecting production are credit, technical assistance, a rural development index, income concentration, measured by the Gini index, and regional dummies. Overall Brazilian rural production performance responds positively to all covariates.