A high throughput colorimetric assay of beta-1,3-D-glucans by Congo red dye

Mushroom strains contain complex nutritional biomolecules with a wide spectrum of therapeutic and prophylactic properties. Among these compounds, β-d-glucans play an important role in immuno-modulating and anti-tumor activities. The present work involves a novel colorimetric assay method for β-1,3-d-glucans with a triple helix tertiary structure by using Congo red. The specific interaction that occurs between Congo red and β-1,3-d-glucan was detected by bathochromic shift from 488 to 516 nm (> 20 nm) in UV–Vis spectrophotometer. A micro- and high throughput method based on a 96-well microtiter plate was devised which presents several advantages over the published methods since it requires only 1.51 μg of polysaccharides in samples, greater sensitivity, speed, assay of many samples and very cheap. β-d-Glucans of several mushrooms (i.e., Coriolus versicolor, Ganoderma lucidum, Pleurotus ostreatus, Ganoderma carnosum, Hericium erinaceus, Lentinula edodes, Inonotus obliquus, Auricularia auricular, Polyporus umbellatus, Cordyseps sinensis, Agaricus blazei, Poria cocos) were isolated by using a sequence of several extractions with cold and boiling water, acidic and alkaline conditions and quantified by this microtiter plate method. FTIR spectroscopy was used to study the structural features of β-1,3-d-glucans in these mushroom samples as well as the specific interaction of these polysaccharides with Congo red. The effect of NaOH on triple helix conformation of β-1,3-d-glucans was investigated in several mushroom species.

Layer by layer TiO2 thin films and photodegradation of Congo red

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Albumin-induced circular dichroism in Congo red: applications for studies of amyloid-like fibril aggregates and binding sites

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

Seeing red: the use of Congo Red dye to identify cooked and damaged starch grains in archaeological residues

Starchy plant foods are significant in the diet of almost all peoples. Archaeologically, however, preservation of such plants is limited, and direct evidence of plant use by past people is also rare. Although starch grains can be preserved on artefacts used to process starchy plants, it is very difficult to identify grains damaged by processing methods such as milling or cooking. We present a method for identifying such damaged starch grains using Congo Red staining to identify cooking or milling activities in the past subsistence behaviour of Aboriginal people of southeast Queensland, Australia. (c) 2005 Elsevier Ltd. All rights reserved.

Cobalt promoted TiO2/GO for the photocatalytic degradation of oxytetracycline and Congo Red

A family of titania derived nanocomposites synthesized via sol-gel and hydrothermal routes exhibit excellent performance for the photocatalytic degradation of two important exemplar water pollutants, oxytetracycline and Congo Red. Low loadings of Co3O4 nanoparticles dispersed over the surfaces of anatase TiO2 confer visible light photoactivity for the aqueous phase decomposition of organics through the resulting heterojunction and reduced band gap. Subsequent modification of these Co3O4/TiO2 composites by trace amounts of graphene oxide nanosheets in the presence of a diamine linker further promotes both oxytetracycline and Congo Red photodegradation under simulated solar and visible irradiation, through a combination of enhanced photoresponse and consequent radical generation. Radical quenching and fluorescence experiments implicate holes and hydroxyl radicals as the respective primary and secondary active species responsible for oxidative photodegradation of pollutants.

Evaluation of phenotypic markers associated with pathogenicity in the genus Listeria

A total of 130 Listeria strains were tested in order to evaluate lecithinase production and capacity for Congo red adsorption as markers of pathogenicity. The strains were identified according to acid production from sugars and by the CAMP test and the data were correlated with the ability to produce keratoconjunctivitis in guinea pigs. L. monocytogenes cultures presented 51.8% and 88.8% positivity rates for Congo red adsorption and lecithinase production, respectively, whereas 80.8% and 100% for L. innocua cultures were negative for the two test, respectively

Adsorção do corante vermelho congo por derivados da o-carboximetilquitosana hidrofobicamente modificados

Hydrophobically modified O-carboxymethylchitosan derivatives were synthesized through a reaction with lauroyl chloride and applied for adsorption of congo red dye. The Langmuir-Freundlich isotherm model was found to be the most suitable one for the VC adsorption and maximum adsorption capacity obtained was 281.97 mg g-1 at a pH value of 7.0 for HL 1.0. The adsorption process follows the pseudo-second-order kinetics and the corresponding rate constants were obtained. The thermodynamic parameters showed that adsorption process is spontaneous (positive ∆Hº) and favorable (negative ∆Gº). The hydrophobic derivatives are able to adsorb the dye even in high pH values.

Macauba palm (acrocomia aculeata) cake from biodiesel processing: an efficient and low cost substrate for the adsorption of dyes

The potential of natural and heat-treated Macauba palm cake as adsorbent for the removal of Methylene Blue (MB) and Congo Red (CR) from solution has been investigated. Equilibrium adsorption was attained in <7 h and the process was favored at pH 5.0 for MB and pH 6.5 for CR with an adsorbent (g):adsorbate (mL) ratio of 1:200 and an initial concentration of adsorbate of 25 mg L−1. The maximum adsorption capacities of the natural and heat-treated materials were, respectively, 25.80 and 32.30 mg g−1 for MB, and 32.00 and 20.30 mg g−1 for CR. The isotherm model proposed by Sips represented most adequately the adsorption of MB and CR. The adsorptions of the dyes were best described in terms of a pseudo second-order reaction. Thermodynamic parameters such as ΔHo, ΔSo and ΔGo were calculated. The adsorption process was found to be endothermic and spontaneous. Macauba palm cake is adequate for the removal of waste dye from industrial effluents by virtue of its abundance, low cost and efficiency of adsorption.

Electrochemical Control of the Time-Dependent Intensity Fluctuations in Surface-Enhanced Raman Scattering (SERS)

Time-dependent fluctuations in surface-enhanced Raman scattering (SERS) intensities were recorded from a roughened silver electrode immersed in diluted solutions of rhodamine 6G (R6G) and congo red (CR). These fluctuations were attributed to a small number of SERS-active molecules probing regions of extremely high electromagnetic field (hot spots) at the nanostructured surface. The time-dependent distribution of SERS intensities followed a tailed statistics at certain applied potentials, which has been linked to single-molecule dynamics. The shape of the distribution was reversibly tuned by the applied voltage. Mixtures of both dyes, R6G and CR, at low concentrations were also investigated. Since R6G is a cationic dye and CR is an anionic dye, the statistics of the SERS intensity distribution of either dye in a mixture were independently controlled by adjusting the applied potential. The potential-controlled distribution of SERS intensities was interpreted by considering the modulation of the surface coverage of the adsorbed dye by the interfacial electric field. This interpretation was supported by a two-dimensional Monte Carlo simulation that took into account the time evolution of the surface configuration of the adsorbed species and their probability to populate a hypothetical hot spot. The potential-controlled SERS dynamics reported here is a first step toward the spectroelectrochemical investigation of redox processes at the single-molecule level by SERS.

Removal of dyes by lignocellulose adsorbents originating from biodiesel production

In this study, use was made of tucumã cake, in natura (TCN) and thermally treated (TCT), as potential alternative adsorbents for the adsorption of cationic and anionic dyes. The effects of the parameters: contact time, adsorbent: adsorbate mass ratio, and initial concentration of dye were analyzed. The adsorption isotherms were established from optimized adsorption parameters. The best conditions for adsorption were: equilibrium time of 7 h, concentration of 25 mg L 1 and ratio of 1:200 for the methylene blue dye; and pH 6.5, concentration of 25 mg L 1 and ratio of 1:200 for the congo red dye. The adsorption process was best represented by the Dubinin–Radushkevich and Sips isotherms. The kinetics of adsorption of the dyes were best described by the pseudo-second-order kinetic and Elovich models. TCT showed the best maximum adsorption capacity (Qm) for the methylene blue dye (63.92 mg g 1 ).

Isolation and characterization of Flavobacterium columnare (Bernardet et al. 2002) from four tropical fish species in Brazil

Flavobacterium columnare is the causative agent of columnaris disease in freshwater fish, implicated in skin and gill disease, often causing high mortality. The aim of this study was the isolation and characterization of Flavobacterium columnare in tropical fish in Brazil. Piracanjuba (Brycon orbignyanus), pacu (Piaractus mesopotamicus), tambaqui (Colossoma macropomum) and cascudo (Hypostomus plecostomus) were examined for external lesions showing signs of colunmaris disease such as greyish white spots, especially on the head, dorsal part and caudal fin of the fish. The sampling comprised 50 samples representing four different fish species selected for study. Samples for culture were obtained by skin and kidney scrapes with a sterile cotton swabs of columnaris disease fish and streaked onto Carlson and Pacha (1968) artificial culture medium (broth and solid) which were used for isolation. The strains in the liquid medium were Gram negative, long, filamentous, exhibited flexing movements (gliding motility), contained a large number of long slender bacteria and gathered into ‘columns'. Strains on the agar produced yellow-pale colonies, rather small, flat that had rhizoid edges. A total of four Flavobacterium columnare were isolated: 01 Brycon orbignyanus strain, 01 Piaractus mesopotamicus strain, 01 Colossoma macropomum strain, and 01 Hypostomus plecostomus strain. Biochemical characterization, with its absorption of Congo red dye, production of flexirubin-type pigments, H2S production and reduction of nitrates proved that the isolate could be classified as Flavobacterium columnare.

Superior oblique muscle amyloidosis mimicking myositis

A 47-year-old man presented with complaints of progressive diplopia in downgaze and a painful firm mass on the left medial superior canthus. On examination, there was marked hyperemia of the superior bulbar conjunctiva of the left eye. Systemic examination revealed erythematous papules on his trunk and pulmonary infiltrates. CT of the orbits revealed a fusiform enlargement of the left superior oblique muscle and diffuse infiltration of the left temporal region. Biopsy of the left superior oblique muscle and temporal muscle disclosed Congo red deposits that show apple-green birefringence under polarized light. A comprehensive systemic investigation failed to show any disease that could explain the amyloid deposits. The patient was then diagnosed as having primary systemic amyloidosis. We think that this case highlights the necessity of a biopsy in any atypical extraocular muscle enlargement before a diagnosis of myositis.

Molecular imaging agents for detection of β-amyloid plaques in Alzheimer’s disease

The formation of amyloid structures is a neuropathological feature that characterizes several neurodegenerative disorders, such as Alzheimer´s and Parkinson´s disease. Up to now, the definitive diagnosis of these diseases can only be accomplished by immunostaining of post mortem brain tissues with dyes such Thioflavin T and congo red. Aiming at early in vivo diagnosis of Alzheimer´s disease (AD), several amyloid-avid radioprobes have been developed for b-amyloid imaging by positron emission tomography (PET) and single-photon emission computed tomography (SPECT). The aim of this paper is to present a perspective of the available amyloid imaging agents, special those that have been selected for clinical trials and are at the different stages of the US Food and Drugs Administration (FDA) approval.

A novel colorimetric assay of beta-D-glucans in basidiomycete strains by alcian blue dye in a 96-well microtiter plate

Basidiomycete strains synthesize several types of beta-D-glucans, which play a major role in the medicinal properties of mushrooms. Therefore, the specific quantification of these beta-D-glucans in mushroom strains is of great biochemical importance. Because published assay methods for these beta-D-glucans present some disadvantages, a novel colorimetric assay method for beta-D-glucan with alcian blue dye was developed. The complex formation was detected by following the decrease in absorbance in the range of 620 nm and by hypsochromic shift from 620 to 606 nm (similar to 14 nm) in UV-Vis spectrophotometer. Analysis of variance was used for optimization of the slope of the calibration curve by using the assay mixture containing 0.017% (w/v) alcian blue in 2% (v/v) acetic acid at pH 3.0. The high-throughput colorimetric assay method on microtiter plates was used for quantification of beta-D-glucans in the range of 0-0.8 mu g, with a slope of 44.15 x 10(-2) and a limit of detection of 0.017 mu g/well. Recovery experiments were carried out by using a sample of Hericium erinaceus, which exhibited a recovery of 95.8% for beta-1,3-D-glucan. The present assay method exhibited a 10-fold higher sensitivity and a 59-fold lower limit of detection compared with the published method with congo red beta-D-glucans of several mushrooms strains were isolated from fruiting bodies and mycelia, and they were quantified by this assay method. This assay method is fast, specific, simple, and it can be used to quantify beta-D-glucans from other biological sources. (C) 2015 American Institute of Chemical Engineers

Biological and genetic characteristics of uropathogenic Escherichia coli strains

The aim of the present study was to determine biological characteristics such as expression of fimbriae, Congo red binding, production of hemolysin and aerobactin, adhesion to HeLa and uroepithelial cells and invasion of HeLa cells by Escherichia coli isolates obtained from patients showing clinical signs of urinary tract infection (UTI). Also, the presence of genes (apa, afa, spa) for fimbria expression and cytotoxic necrotizing factors (CNF1, CNF2) was assayed using specific primers in PCR. The data obtained were compared with the clonal relationships obtained by analysis of multilocus enzyme electrophoresis (MLEE), restriction fragment length polymorphism (RFLP) of the rDNA (ribotyping) and enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR). All isolates but one presented a combination of at least two of the characteristics studied, a fact suggesting the presence of pathogenicity islands (PAIs). Diffuse adherence type to HeLa cells was observed to occur in most of the strains, but adhesion to uroepithelial cells seems to be a more reliable test to verify pathogenicity. Although four strains seemed to be able to invade HeLa cells when assayed by light microscopy, electron microscopy studies demonstrated that these strains were not invasive. MLEE, RFLP and ERIC-PCR were able to group the isolates differently into main clusters that were not correlated with the presence of pathogenic traits.