RESUMO
More sustainable materials have been becoming an important concern of worldwide scientists, and cellulosic materials are one alternative in water decontamination. An efficient strategy to improve removal capacity is functionalizing or incorporating nanomaterials in cellulose-based materials. The new hybrid cDAC/ZnONPs was produced by green synthesis of zinc oxide nanoparticles (ZnONPs), promoting the in situ reduction and immobilization on the cationic dialdehyde cellulose microfibers (cDAC) surface to remove Congo red dye from water. cDAC/ZnONPs was characterized by scanning electron microscopy (SEM-EDS) and infrared spectroscopy (FTIR), which showed efficient nanoparticles reduction. Adsorption efficiency on cationic cellulose surface was investigated by pH, contact time, initial concentration, and dye selectivity tests. The material followed the H isotherm model, which resulted in a maximum adsorption capacity of 1091.16 mg/g. Herein, was developed an efficient and ecologically correct new adsorbent, highly effective in Congo red dye adsorption even at high concentrations, suitable for the remediation of contaminated industrial effluents.
Assuntos
Vermelho Congo , Poluentes Químicos da Água , Purificação da Água , Óxido de Zinco , Óxido de Zinco/química , Vermelho Congo/química , Vermelho Congo/isolamento & purificação , Poluentes Químicos da Água/química , Poluentes Químicos da Água/isolamento & purificação , Adsorção , Purificação da Água/métodos , Química Verde , Concentração de Íons de Hidrogênio , Celulose/química , Celulose/análogos & derivados , Corantes/química , Nanopartículas/química , Cinética , Nanopartículas Metálicas/química , Celulose Oxidada/química , Cátions/químicaRESUMO
This work aimed to describe the adsorption behavior of Congo red (CR) onto activated biochar material prepared from Haematoxylum campechianum waste (ABHC). The carbon precursor was soaked with phosphoric acid, followed by pyrolysis to convert the precursor into activated biochar. The surface morphology of the adsorbent (before and after dye adsorption) was characterized by scanning electron microscopy (SEM/EDS), BET method, X-ray powder diffraction (XRD), and Fourier-transform infrared spectroscopy (FTIR) and, lastly, pHpzc was also determined. Batch studies were carried out in the following intervals of pH = 4-10, temperature = 300.15-330.15 K, the dose of adsorbent = 1-10 g/L, and isotherms evaluated the adsorption process to determine the maximum adsorption capacity (Qmax, mg/g). Kinetic studies were performed starting from two different initial concentrations (25 and 50 mg/L) and at a maximum contact time of 48 h. The reusability potential of activated biochar was evaluated by adsorption-desorption cycles. The maximum adsorption capacity obtained with the Langmuir adsorption isotherm model was 114.8 mg/g at 300.15 K, pH = 5.4, and a dose of activated biochar of 1.0 g/L. This study also highlights the application of advanced machine learning techniques to optimize a chemical removal process. Leveraging a comprehensive dataset, a Gradient Boosting regression model was developed and fine-tuned using Bayesian optimization within a Python programming environment. The optimization algorithm efficiently navigated the input space to maximize the removal percentage, resulting in a predicted efficiency of approximately 90.47% under optimal conditions. These findings offer promising insights for enhancing efficiency in similar removal processes, showcasing the potential of machine learning in process optimization and environmental remediation.
Assuntos
Teorema de Bayes , Carvão Vegetal , Vermelho Congo , Aprendizado de Máquina , Carvão Vegetal/química , Adsorção , Vermelho Congo/química , Cinética , Poluentes Químicos da Água/química , Concentração de Íons de Hidrogênio , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
Laccase is a superfamily of ligninolytic enzymes known to degrade a wide variety of xenobiotics, including synthetic dyes. Congo Red (CR) has a diazo dye function, carcinogenic and mutagenic potential, and is currently applied in clinical analysis. The objective of this work was to produce and characterize the crude extract of Lentinus sp. in semi-solid fermentation (FSS) and perform in vitro and in silico studies to assess the potential of the crude extract to discolor the CR dye. Laccase activity was determined using ABTS as substrate and characterized. The in vitro discoloration was carried out using experimental design 22 at room temperature and monitored at 340 nm for 24h. Molecular docking and molecular dynamics simulations were performed between laccase and CR. The maximum laccase activity production was 29.63 U L-1 with six days of FSS. The optimal temperature and pH were 50 °C and 3.0, respectively. Discoloration of the CR dye was obtained only in tests containing CuSO4. Laccase formed stable complexes with the dye, presenting negative binding energy values ranging from -70.94 to -63.16 kcal mol-1 and the occurrence of seven hydrogen bonds. Molecular dynamics results showed the stability of the system (RMSD ranging from 1.0 to 2.5 Ä) and protein-ligand interaction along simulation. RMSF values pointed residues at the end of chains A (residues 300 to 305, 480 to 500) and B (residues 650 to 655 and 950 to 1000) as the most flexible regions of the laccase. This study highlighted the enzymatic action in the bioremediation of CR in vitro in agreement with the in silico simulations that demonstrate the enzyme potential.Communicated by Ramaswamy H. Sarma.
Assuntos
Vermelho Congo , Lentinula , Vermelho Congo/química , Corantes/química , Lacase/química , Lacase/metabolismo , Simulação de Acoplamento Molecular , Lentinula/metabolismo , Projetos de Pesquisa , Misturas ComplexasRESUMO
The removal of dyes from effluents of textile industries represents a technological challenge, due to their significant environmental impact. The application of halloysite (Hal) and palygorskite (Pal) clay minerals as adsorbents for the removal of Congo red (CR) and methylene blue (MB) was evaluated in this work. The materials were applied both in natural and acid-treated forms, and characterized by XRD, XPS, SEM-EDS, FTIR, and N2 adsorption-desorption isotherm techniques to identify their properties and main active sites. The adsorbents showed potential to remove CR (> 98%) and MB (> 85%) within 180 min, using 0.3 g adsorbent and initial dye concentration of 250 mg L-1. Semi-empirical quantum mechanical calculations (SQM) confirmed the interaction mechanism between dyes and the adsorbents via chemisorption (- 69.0 kcal mol-1 < Eads < - 28.8 kcal mol-1), which was further observed experimentally due to the high fit of adsorption data to pseudo-second order kinetic model (R2 > 0.99) and Langmuir isotherm (R2 > 0.98). The use of Pal and Hal to remove dyes was proven to be economically and environmentally viable for industrial application.
Assuntos
Argila , Corantes , Compostos de Silício , Poluentes Químicos da Água , Adsorção , Argila/química , Corantes/química , Poluentes Químicos da Água/química , Compostos de Silício/química , Minerais/química , Azul de Metileno/química , Silicatos de Alumínio/química , Compostos de Magnésio/química , Cinética , Vermelho Congo/químicaRESUMO
In this article, we propose a simple photochemical method to synthesize pure La2Ti2O7 films and La2Ti2O7 films doped with silver at 1.0, 3.0, and 5.0 mol%. After annealing the photo-deposited films at 900 °C, XRD, SEM, and XPS analyses showed the formation of a monoclinic La2Ti2O7 phase and the presence of Ag and AgO in doped samples. Photocatalytic tests for Congo red degradation demonstrated that pure La2Ti2O7 achieved 25.4% degradation, while doped samples reached a maximum of 92.7% degradation. Moreover, increasing silver doping on La2Ti2O7 films significantly reduced the growth of Staphylococcus aureus, indicating potential antibacterial properties. The enhanced photoactivity was attributed to the formation of a type I heterojunction between La2Ti2O7 and AgO, and a degradation mechanism was proposed based on Congo red degradation.
Assuntos
Vermelho Congo , Staphylococcus aureus , Vermelho Congo/química , Prata/farmacologia , Prata/química , Titânio/química , Antibacterianos/farmacologia , Antibacterianos/químicaRESUMO
The thermodynamics and kinetics of arachin-Congo red (ARA-CR) and conarachin-Congo red (CON-CR) interactions were studied using surface plasmon resonance. KCl led to a reduction of up to 55% in the values of the associated kinetic constants, but it had less influence on the dissociation rates (less than 12%). The change in ionic strength had little effect on the thermodynamic stability of the complexes, but it did reduce their affinities ( [Formula: see text] from 3.52 to 2.44 × 103 M-1 and [Formula: see text] from 15.1 to 12.5 × 103 M-1). The shielding of the electrical double layer favored ARA-CR hydrophilic interactions ( [Formula: see text] decreased from -30.60 to -42.98 kJ mol-1). On the other hand, hydrophobic interactions came to dominate during the formation of [CON-CR]0 ( [Formula: see text] increased from -11.21 to 28.34 kJ mol-1 and [Formula: see text] increased from 12.64 to 51.73 kJ.mol-1). The data presented here improve our understanding of plant-based protein nanocarriers of small bioactive molecules.
Assuntos
Vermelho Congo , Sondas Moleculares , Vermelho Congo/química , Cinética , Proteínas de Plantas , TermodinâmicaRESUMO
A sensitive and selective strategy to identify insulin fibrils remains a challenge for researchers in amyloid protein research. Thus, it is critical to detect, in vitro, the species generated during amyloid aggregation, particularly the fibrillar species. Here we demonstrate that the luminescent complex cis-[Ru(phen)2(3,4Apy)2]2+ (RuApy; phen = 1,10-phenanthroline; 3,4Apy = 3,4-diaminopyridine) is a rapid, low-cost alternative to in vitro detection of fibrillar insulin, using conventional optical techniques. The RuApy complex displays emission intensity enhancement at 655 nm when associated with insulin, which enables imaging of the conformational changes of the protein's self-aggregation. The complex shows high sensitivity to fibrillar insulin with a limit of detection of 0.85 µM and binding affinity of 12.40 ± 1.84 µM which is comparable to those of Thioflavin T and Congo red, with the advantage of minimizing background fluorescence, absorption of light by biomolecules, and light scattering from physiologic salts in the medium.
Assuntos
Amiloide/análise , Corantes Fluorescentes/química , Insulina/análise , Rutênio/química , Amiloide/metabolismo , Peptídeos beta-Amiloides/metabolismo , Proteínas Amiloidogênicas/metabolismo , Animais , Benzotiazóis/química , Linhagem Celular , Vermelho Congo/química , Fluorescência , Insulina/metabolismo , Luminescência , Camundongos , Fenantrolinas/química , Agregados Proteicos , RatosRESUMO
GRASP55, one of the two human GRASP proteins, has been implicated in the organization of Golgi stacks and in unconventional protein secretion. However, the detailed molecular mechanisms supporting GRASP55 participation in those processes remain mostly unclear. We have shown that GRASP55 exists as monomers in solution, which transitions to amorphous aggregates with increasing temperatures. Here, we further investigated the formation of higher order structures of GRASP55 by exploring its amyloid fibrillation at 37 °C. Sequence-based AGGRESCAN analysis revealed that GRASP55 has ten aggregation "hot spots", preferentially concentrated in its N-terminal half. Congo Red, ThT, and circular dichroism assays suggested GRASP55 formed amyloid-like fibrils in a time-dependent manner at 37 °C. Dynamic light scattering showed the mean hydrodynamic radius of GRASP55 amyloid-like fibrils increased with increasing incubation times at 37 °C. Transmission electron microscopy and intrinsic fluorescence lifetime imaging showed that, upon increasing incubation time at 37 °C, GRASP55 yielded amyloid-like fibrils in a nucleation-dependent process via a sequence of events: lag-phase (monomers to oligomers), growth phase (oligomers to organized protofibrils), and plateau phase (protofibrils to amyloid-like fibrils). The insights gained herein may help in better understanding the mechanisms of GRASP55 amyloid fibrillation in vivo and its potential association with neurological disorders.
Assuntos
Amiloide/química , Complexo de Golgi/fisiologia , Proteínas da Matriz do Complexo de Golgi/química , Benzotiazóis/química , Dicroísmo Circular , Biologia Computacional , Vermelho Congo/química , Humanos , Hidrodinâmica , Cinética , Luz , Microscopia Eletrônica de Transmissão , Doenças do Sistema Nervoso/fisiopatologia , Conformação Proteica , Domínios Proteicos , Proteínas Recombinantes/química , Espalhamento de Radiação , TemperaturaRESUMO
AIM AND OBJECTIVE: A common method used for massive detection of cellulolytic microorganisms is based on the formation of halos on solid medium. However, this is a subjective method and real-time monitoring is not possible. The objective of this work was to develop a method of computational analysis of the visual patterns created by cellulolytic activity through artificial neural networks description. MATERIALS AND METHODS: Our method learns by an adaptive prediction model and automatically determines when enzymatic activity on a chromogenic indicator such as the hydrolysis halo occurs. To achieve this goal, we generated a data library with absorbance readings and RGB values of enzymatic hydrolysis, obtained by spectrophotometry and a prototype camera-based equipment (Enzyme Vision), respectively. We used the first part of the library to generate a linear regression model, which was able to predict theoretical absorbances using the RGB color patterns, which agreed with values obtained by spectrophotometry. The second part was used to train, validate, and test the neural network model in order to predict cellulolytic activity based on color patterns. RESULTS: As a result of our model, we were able to establish six new descriptors useful for the prediction of the temporal changes in the enzymatic activity. Finally, our model was evaluated on one halo from cellulolytic microorganisms, achieving the regional classification of the generated halo in three of the six classes learned by our model. CONCLUSION: We assume that our approach can be a viable alternative for high throughput screening of enzymatic activity in real time.
Assuntos
Celulose/química , Biblioteca Gênica , Redes Neurais de Computação , Reconhecimento Automatizado de Padrão , Celulose/metabolismo , Cor , Corantes/química , Vermelho Congo/química , Hidrólise , Aprendizado de Máquina , Modelos MolecularesRESUMO
GRASPs are proteins involved in cell processes that seem paradoxical: responsible for shaping the Golgi cisternae and involved in unconventional secretion mechanisms that bypass the Golgi. Despite its physiological relevance, there is still a considerable lack of studies on full-length GRASPs. Our group has previously reported an unexpected behavior of the full-length GRASP from the fungus C. neoformans: its intrinsically-disordered characteristic. Here, we generalize this finding by showing that it is also observed in the GRASP from S. cerevisae (Grh1), which strongly suggests it might be a general property within the GRASP family. Furthermore, Grh1 is also able to form amyloid-like fibrils either upon heating or when submitted to changes in the dielectric constant of its surroundings, a condition that is experienced by the protein when in close contact with membranes of cell compartments, such as the Golgi apparatus. Intrinsic disorder and fibril formation can thus be two structural properties exploited by GRASP during its functional cycle.
Assuntos
Proteínas Amiloidogênicas/química , Proteínas Intrinsicamente Desordenadas/química , Peptídeos/química , Proteínas de Saccharomyces cerevisiae/química , Proteínas de Transporte Vesicular/química , Sequência de Aminoácidos , Amiloide/química , Benzotiazóis/química , Dicroísmo Circular , Vermelho Congo/química , Complexo de Golgi/metabolismo , Proteínas da Matriz do Complexo de Golgi/química , Imagem Óptica , Conformação Proteica em Folha beta , Desnaturação Proteica , Saccharomyces cerevisiae/metabolismo , Solventes/química , Espectrometria de Fluorescência , Temperatura , Rede trans-Golgi/metabolismoRESUMO
In this paper, we report the combination of two metal oxides (TiO2ZnO) that allows mixed density of states to reduce band gap energy, facilitating the photo-oxidation of Congo red dye under visible light. For the oxidation, a possible mechanism is proposed after analyzing the intermediates by GC-MS, and it is consistent with Density Functional Theory (DFT). The nanohybrids were characterized comprehensibly by several analytical techniques such as X-Ray diffraction (XRD), Transmission Electron Microscopy (TEM), Atomic Force Microscopy (AFM), and X-ray Photoelectron Spectroscopy (XPS). For the addition of ZnO to TiO2, a dominance of anatase phase was found rather than other phases (rutile or brookite). A broad band (â¼550â¯nm) is observed in UV-Visible spectra for TiO2ZnO/Ag NPs nm because of Surface Plasmon properties of Ag NPs. The band gap energy was calculated for TiO2ZnO/Ag system, and then it has been further studied by DFT in order to show why the convergence of two semiconductors allows a mixed density of states, facilitating the reduction of the energy gap between occupied and unoccupied bands; ultimately, it improves the performance of catalysts under visible light. Significantly, the interaction of crystal planes (0 0 I) of TiO2 anatase and (0 0 1) of ZnO crucially plays as an important role for the reduction of energy band-gap. Additionally, TiO2ZnOAg NPs were used recognize Saccharomyces cerevisiae cells by con-focal fluorescence microscope, showing that it develops bright bio-images for the cells; while for TiO2 or ZnO or TiO2ZnO NPs, no fluorescent response was seen within the cells.
Assuntos
Vermelho Congo/química , Luz , Fotólise , Titânio/química , Catálise , Vermelho Congo/efeitos da radiação , Microscopia , Nanopartículas/química , Semicondutores , Análise Espectral , Óxido de Zinco/químicaRESUMO
Mannheimia haemolytica causes respiratory disease in cattle. Amyloid proteins are a major component of biofilms; they aid in adhesion and confer resistance against several environmental insults. The amyloid protein curli is highly resistant to protease digestion and physical and chemical denaturation and binds Congo red (CR) dye. The purpose of this study was to characterize an approximately 50-kDa CR-binding amyloid-like protein (ALP) expressed by M. haemolytica. This protein resisted boiling and formic acid digestion and was recognized by a polyclonal anti-Escherichia coli curli serum, suggesting its relationship with amyloid proteins. Immunolabeling and transmission electron microscopy showed that antibodies bound long, thin fibers attached to the bacterial surface. Mass spectrometry analysis indicated that these fibers are M. haemolytica OmpP2-like proteins. The purified protein formed filaments in vitro, and antiserum against it reacted positively with biofilms. An in silico analysis of its amino acid sequence indicated it has auto-aggregation properties and eight amyloid peptides. Rabbit polyclonal antibodies generated against this ALP diminished the adhesion of ATCC 31612 and BA1 M. haemolytica strains to A549 human epithelial cells, indicating its participation in cell adhesion. ALP expressed by M. haemolytica may be important in its pathogenicity and ability to form biofilms.
Assuntos
Proteínas Amiloidogênicas/química , Proteínas da Membrana Bacteriana Externa/química , Biofilmes/crescimento & desenvolvimento , Mannheimia haemolytica/química , Células A549 , Sequência de Aminoácidos , Proteínas Amiloidogênicas/biossíntese , Proteínas Amiloidogênicas/genética , Proteínas Amiloidogênicas/isolamento & purificação , Animais , Aderência Bacteriana , Proteínas da Membrana Bacteriana Externa/biossíntese , Proteínas da Membrana Bacteriana Externa/genética , Proteínas da Membrana Bacteriana Externa/isolamento & purificação , Bovinos , Vermelho Congo/química , Expressão Gênica , Humanos , Soros Imunes/química , Soros Imunes/isolamento & purificação , Mannheimia haemolytica/genética , Mannheimia haemolytica/isolamento & purificação , Mannheimia haemolytica/metabolismo , Modelos Moleculares , Peso Molecular , Pasteurelose Pneumônica , Ligação Proteica , Estrutura Secundária de Proteína , Coelhos , Alinhamento de Sequência , OvinosRESUMO
To optimize the therapeutic applications of Congo red (CR), a potential inhibitor of protein aggregation, the kinetics and thermodynamics of the interactions between CR and a model protein need to be understood. We used surface plasmon resonance (SPR) and fluorescence techniques to determine the dynamics and thermodynamic parameters for the formation of complexes between CR and bovine serum albumin (BSA). CR interacts with BSA through a transition complex; the activation energy for association (Eact(a)) was determined to be 35.88kJmol-1, while the activation enthalpy (ΔH), entropy (ΔS), and Gibbs free energy (ΔG) are 33.41kJmol-1, 0.18Jmol-1K-1, and 33.35kJmol-1, respectively. When this intermediate transforms into the final CR-BSA complex, the entropy of the system increases and part of the absorbed energy is released; this process is associated with a reverse activation energy (Eact(d)) of 20.17kJmol-1, and values of ΔH, ΔS, and ΔG of 17.69kJmol-1, -162.86Jmol-1K-1, and 66.25kJmol-1, respectively. A comparison of the SPR and fluorescence results suggests that there is more than one site where BSA interacts with CR.
Assuntos
Vermelho Congo/química , Soroalbumina Bovina/química , Animais , Cinética , Ressonância de Plasmônio de Superfície , TermodinâmicaRESUMO
Congo red (CR), one of the most commonly used dyes for the identification of amyloid fibril aggregates, is also a ligand of native bovine serum albumin (BSA). Induced circular dichroism (ICD) is a phenomenon observed when a chiral compound induces chirality in an achiral one. Here, we study the spectral properties and analytical applications of ICD in Congo red provoked by its interaction with BSA. The complex BSA:CR displays a strong ICD spectrum with a positive band at 412 nm and two negative bands at 356 and 490 nm. The use of site I and site II albumin ligands as warfarin and ibuprofen, respectively, provoked different alterations in the Congo red ICD spectrum. The BSA binding sites were modified by oxidation and the ICD signal was sensitive to this alteration. The thermal treatment of the BSA:CR complex (30-90 °C) was monitored by ICD at 490 nm and showed a sigmoidal pattern typical of phase transition in proteins. The altered ICD spectrum is consistent with the formation of amyloid-like fibril aggregates in BSA, which was confirmed by thioflavin T and Rayleigh scattering assays. In conclusion, the ICD provoked by the binding of Congo red to albumin may represent a new spectroscopic technique for studying alterations in the structure of albumin regarding its binding sites and the formation of amyloid aggregates.
Assuntos
Amiloide/química , Dicroísmo Circular , Vermelho Congo/química , Agregados Proteicos , Soroalbumina Bovina/química , Animais , Sítios de Ligação , Bovinos , Vermelho Congo/metabolismo , Ligantes , Oxirredução , Ligação Proteica , Soroalbumina Bovina/metabolismoRESUMO
Poly(ethylene glycol) (PEG)-decorated polystyrene (PS) nanoparticles with mean hydrodynamic diameter (D) and zeta-potential (ζ) of (286 ± 15) nm and (-50 ± 5) mV, respectively, were modified by the adsorption of Congo red (CR). The PS/PEG/CR particles presented D and ζ values of (290 ± 19) nm and (-36 ± 5) mV, respectively. The adsorption of lipase onto PS/PEG or PS/PEG/CR particles at (24 ± 1) °C and pH 7 changed the mean D value to (380 ± 20) and (405 ± 11) nm, respectively, and ζ value to (-32 ± 4) mV and (-25 ± 2) mV, respectively. The kinetic parameters of the hydrolysis of p-nitrophenyl butyrate were determined for free lipase, lipase immobilized onto PS/PEG and PS/PEG/CR particles. Lipase on PS/PEG/CR presented the largest Michaelis-Menten constant (KM), but also the highest Vmax and kcat values. Moreover, it could be recycled seven times, losing a maximum 10% or 30% of the original enzymatic activity at 40 °C or 25 °C, respectively. Although lipases immobilized onto PS/PEG particles presented the smallest KM values, the reactions were comparatively the slowest and recycling was not possible. Hydrolysis reactions performed in the temperature range of 25 °C to 60 °C with free lipases and lipases immobilized onto PS/PEG/CR particles presented an optimal temperature at 40 °C. At 60 °C free lipases and lipases immobilized onto PS/PEG/CR presented ~80% and ~50% of the activity measured at 40 °C, indicating good thermal stability. Bioconjugation effects between CR and lipase were evidenced by circular dichroism spectroscopy and spectrophotometry. CR molecules mediate the open state conformation of the lipase lid and favor the substrate approaching.
Assuntos
Butiratos/metabolismo , Enzimas Imobilizadas/farmacocinética , Lipase/metabolismo , Lipase/farmacocinética , Nanopartículas/metabolismo , Adsorção , Butiratos/química , Candida/enzimologia , Domínio Catalítico , Dicroísmo Circular , Vermelho Congo/química , Enzimas Imobilizadas/química , Interações Hidrofóbicas e Hidrofílicas , Lipase/química , Nanopartículas/química , Polietilenoglicóis/química , Poliestirenos/química , EspectrofotometriaRESUMO
Poly(ethylene glycol), PEG, decorated polystyrene (PS) nanoparticles were synthesized and characterized by means of dynamic light scattering (DLS), zeta (ζ) potential measurements, Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). The adsorption of Congo red (CR) onto PS/PEG particles was evidenced by the decrease of ζ potential values and increase in the particles mean diameter in comparison to bare particles. Cholesterol oxidase (ChOx), the main enzyme in the oxidation of cholesterol, adsorbed onto PS/PEG and PS/PEG/CR particles, as revealed by the increase in the particles mean size and spectrophotometry. The enzymatic activity of free and immobilized ChOx was determined as a function of time by means of a coupled reaction with horseradish peroxidase. The activity of free ChOx decreased with time, while the activity of immobilized ChOx increased with time; after 1h reaction the latter was half of the former. Freeze-drying the ChOx covered PS/PEG/CR particles allowed their storage for at least one month under room conditions without loss of enzymatic activity. Conjugation effects between CR and ChOx or cholesterol evidenced by circular dichroism and spectrophotometry rendered a conformational state of ChOx, such that the enzymatic action was favored. ChOx adsorbed onto PS/PEG presents no enzymatic activity, probably due to ChOx denaturation or unfavorable orientation. Freeze-dried and freshly prepared dispersions of ChOx immobilized onto PS/PEG/CR particles yielded linear response in the cholesterol concentration range of 100mgdL(-1) (lowest limit of normal blood concentration) to 300mgdL(-1) (high risk level).
Assuntos
Colesterol Oxidase/metabolismo , Vermelho Congo/metabolismo , Enzimas Imobilizadas/metabolismo , Nanopartículas/metabolismo , Polímeros/metabolismo , Adsorção , Colesterol Oxidase/química , Vermelho Congo/química , Ativação Enzimática , Enzimas Imobilizadas/química , Estrutura Molecular , Nanopartículas/química , Tamanho da Partícula , Polímeros/síntese química , Polímeros/química , Propriedades de SuperfícieRESUMO
This work assessed the application of one- and two-stage mesophilic anaerobic systems to colour removal of sulphonated mono and diazo dyes with ethanol as electron donor. The dyes Congo Red (CR), Reactive Black 5 (RB5) and Reactive Red 2 (RR2) were selected as model compounds and tested separately in seven different periods. The one-stage system (R(1)) consisted of a single up-flow anaerobic sludge blanket (UASB) reactor, whereas the two-stage system (R(2)) consisted of an acidogenic UASB reactor (R(A)), a settler and a methanogenic UASB reactor (R(M)). For CR and RB5, no remarkable difference was observed between the colour removal performance of both anaerobic systems R(1) and R(2). The experiments with RR2 revealed that R(2) was more efficient on colour removal than R(1), showing efficiencies almost 2-fold (period VI) and 2.5-fold (period VII) higher than those found by R(1). Additionally, R(2) showed a higher stability, giving a good prospect for application to textile wastewaters. Finally, the acidogenic reactor (R(A)) had an important role in the overall decolourisation achieved by R(2) during the experiments with CR and RB5 (>78 %), whereas for RR2, a more recalcitrant dye, R(A) was responsible for up to 38 % of the total colour removal.
Assuntos
Corantes/química , Vermelho Congo/química , Elétrons , Naftalenossulfonatos/química , Esgotos/química , Triazinas/química , Anaerobiose , Reatores Biológicos , Cor , Concentração de Íons de Hidrogênio , Oxirredução , Têxteis , Eliminação de Resíduos Líquidos , Purificação da Água/métodosRESUMO
Staphylococcus epidermidis in immunocompromised patients can cause bacteremia related to the use of catheter due to biofilm production. There are different phenotypic methods to detect biofilm formation. One method is based on culture in brain heart infusion agar (BHIA) containing sucrose and red Congo dye (original Congo red agar). Our group created a new CRA formula and we have confirmed its capacity to detect biofilm production in 210 S. epidermidis strains, including 76 (36.2%) icaAB gene-positive strains. Other parameters were also evaluated. The new CRA formula that gave the best results was BHIA with sucrose (5%), Congo red (0.08%), NaCl (1.5%), glucose (2%), and vancomycin (0.5 mg/mL) (vancomycin-modified CRA-CRAmod). The CRAmod plus vancomycin may be a promising tool and can help to determine the real participation of S. epidermidis in the infectious process.
Assuntos
Técnicas Bacteriológicas/métodos , Biofilmes , Vermelho Congo/química , Meios de Cultura/química , Staphylococcus epidermidis/isolamento & purificação , Genes Bacterianos , Glucose/química , Humanos , Reação em Cadeia da Polimerase Multiplex , Fenótipo , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Cloreto de Sódio/química , Infecções Estafilocócicas/sangue , Infecções Estafilocócicas/microbiologia , Staphylococcus epidermidis/química , Staphylococcus epidermidis/efeitos dos fármacos , Staphylococcus epidermidis/genética , Sacarose/química , Vancomicina/farmacologiaRESUMO
In this study, we assessed the catalytic effect of anthraquinone-2,6-disulfonate (AQDS) to enhance the reductive decolourisation of the azo dyes Reactive Red 2 and Congo Red in batch and continuous-flow experiments. While testing the anaerobic sludge 1 in assays free of AQDS, the highest values for the first-order kinetic constant (k1) were found with co-substrates formate and glucose. In the assays that contained 50 microM of AQDS, the k1 values increased with all co-substrates tested, increasing by 3.5-fold when ethanol was the electron donor. The upflow anaerobic sludge blanket (UASB) reactors R1 (AQDS-free) and R2 (AQDS-supplemented) reached excellent decolourisation efficiencies (higher than 90%) even for the high Congo Red concentration tested (1.2 mM). However, electron donor depletion in the influent drastically decreased the colour removal capacity in both bioreactors. Reactor R2 presented higher stability and decolourisation efficiency compared to R1, indicating that the addition of a redox mediator can be valuable for treating dye-coloured wastewaters.
Assuntos
Antraquinonas/química , Bactérias Anaeróbias/metabolismo , Cor , Vermelho Congo/metabolismo , Naftalenossulfonatos/metabolismo , Esgotos/química , Esgotos/microbiologia , Triazinas/metabolismo , Biodegradação Ambiental , Catálise , Vermelho Congo/química , Naftalenossulfonatos/química , Triazinas/química , Poluentes Químicos da Água/metabolismo , Purificação da Água/métodosRESUMO
Lung involvement in Sjögren's syndrome (SS) can affect trachea, bronchus, small airways, pleura and may cause interstitial lung injury. It may also be associated with malignancies, particularly non-Hodgkin's lymphoma, which is a well-recognized complication of this disease. Here we describe the occurrence of localized amyloidosis presenting as pulmonary nodules in a patient with newly diagnosed SS. We highlight this rare occurrence as a diagnostic possibility that should be considered in the evaluation of pulmonary involvement in this disease.