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1.
J Hazard Mater ; 474: 134742, 2024 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-38810577

RESUMO

Developing a multifunctional material that can detect and remove carcinogens in water environments, simultaneously monitor their toxic metabolites in living organisms is significant for environmental remediation and human health. However, most research only focused on detection or adsorption carcinogens due to the difficulty of integrating multiple functions into one material, let alone monitoring their toxic metabolites. Here, a multifunctional Tb/Eu@TATB-HOF (1) is first developed to monitor two carcinogens, malachite green (MG) and its metabolites leucomalachite green (LMG), and simultaneously remove MG from the contaminated water. 1, as the dual-emission fluorescence sensor, can achieve ultrasensitive and highly visualized sensing for MG and LMG with different response modes. Even in actual samples, 1 still exhibits satisfactory sensing performances. As the adsorbent, 1 displays good recyclability and high adsorption capacity for MG. The sensing and adsorption mechanisms are explored through experiments and theoretical calculations. This work not only provides a novel insight for environmental remediation and human health through detection and removal of carcinogens, simultaneously monitoring their toxic metabolites, but first reveals the enormous potential of HOFs as multifunctional materials simultaneously for fluorescence sensing and adsorption.


Assuntos
Carcinógenos , Corantes de Rosanilina , Poluentes Químicos da Água , Adsorção , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/química , Carcinógenos/análise , Corantes de Rosanilina/química , Corantes de Rosanilina/análise , Purificação da Água/métodos , Corantes Fluorescentes/química
2.
Food Chem ; 452: 139543, 2024 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-38735107

RESUMO

Malachite green (MG), a widely used antiparasitic agent, poses health risks to human due to its genotoxic and carcinogenic properties. Herein, a stable dual-emission fluoroprobe of carbon dots/copper nanoclusters is prepared for highly selective detection of MG based on the inner filter effect. This probe exhibits characteristic emission bands at 435 and 625 nm when excited at 376 nm. After adding MG, the both emission signals were significantly quenched, and the ratio of fluorescence intensity (F435/F625) was linearly related to the concentration of MG in the range of 0.05-40 µmol L-1 with a limit of detection of 18.2 nmol L-1. Meanwhile, the two signals exhibit linear relationships with the concentration of MG, respectively, and the corresponding detection results were consistent. The fluoroprobe was successfully used for the detection of MG in fish samples with the recoveries ranging from 96.0% to 103.8% and a relative standard deviation of <3.3%.


Assuntos
Carbono , Cobre , Peixes , Nanocompostos , Pontos Quânticos , Corantes de Rosanilina , Corantes de Rosanilina/química , Corantes de Rosanilina/análise , Cobre/química , Cobre/análise , Animais , Pontos Quânticos/química , Carbono/química , Nanocompostos/química , Espectrometria de Fluorescência/métodos , Contaminação de Alimentos/análise , Limite de Detecção , Fluorescência , Corantes Fluorescentes/química
3.
Food Chem ; 451: 139399, 2024 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-38663240

RESUMO

Malachite green (MG) has been illicitly employed in aquaculture as a parasiticide, however, its teratogenic and carcinogenic effects pose a significant human health threat. Herein, a dual-mode colorimetric and electrochemical aptasensor was fabricated for MG detection, capitalizing on the robust catalytic and peroxidase-like activity of P-CeO2NR@Mxene and good capture efficiency of a tetrahedral DNA nanostructure (TDN) designed with multiple aptamers (m-TDN). P-CeO2NR@Mxene-modified complementary DNA (cDNA) served as both colorimetric and electrochemical probe. m-TDN was attached to AuE to capture MG and P-CeO2NR@Mxene/cDNA. The superior aptamer and MG binding to cDNA regulated signals and enabled precise MG quantification. The further introduced Exo I enabled aptamer hydrolysis, releasing MG for further binding rounds, allowing target recycling amplification. Under the optimal conditions, the aptasensor reached an impressively low detection limit 95.4 pM in colorimetric mode and 83.6 fM in electrochemical mode. We believe this dual-mode approach holds promise for veterinary drug residue detection.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Colorimetria , Técnicas Eletroquímicas , Corantes de Rosanilina , Aptâmeros de Nucleotídeos/química , Corantes de Rosanilina/química , Corantes de Rosanilina/análise , Técnicas Biossensoriais/instrumentação , Exodesoxirribonucleases/química , Exodesoxirribonucleases/metabolismo , Limite de Detecção , Contaminação de Alimentos/análise
4.
Food Chem ; 394: 133517, 2022 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-35749877

RESUMO

Malachite green (MG) is an organic dye compound that is frequently used as a fungicide and antiseptic in aquaculture. However, human or animal exposure to MG causes carcinogenic, teratogenic and mutagenic effects. Herein, a novel fluorescent assay was designed for the detection of MG using manganese dioxide nanosheets (MnO2 NS) as an energy acceptor to quench the fluorescence of branched poly(ethylenimine) carbon dots (BPEI-CDs) via Förster resonance energy transfer. When butyrylcholinesterase is introduced to form thiocholine in the presence of S-butyrylthiocholine iodide, MnO2 NS can be recovered by thiocholine to Mn2+, resulting in restoration of the fluorescence of BPEI-CDs. Exploiting these changes in fluorescence intensity in the above system, a fluorescence probe was successfully developed for the quantitative detection of MG. Besides, this assay was applied to fish samples, verifying the high potential for practical application of the proposed sensor for the monitoring of MG in aquatic products.


Assuntos
Medições Luminescentes , Pontos Quânticos , Animais , Aquicultura , Butirilcolinesterase , Carbono/química , Peixes , Corantes Fluorescentes/química , Limite de Detecção , Medições Luminescentes/métodos , Compostos de Manganês/química , Nanoestruturas/química , Óxidos , Polietilenoimina , Pontos Quânticos/química , Corantes de Rosanilina/análise , Corantes de Rosanilina/química , Tiocolina
5.
Int J Biol Macromol ; 194: 790-799, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34838577

RESUMO

Different groups of synthetic dyes might lead to environmental pollution. The binding affinity among hazardous materials with biomolecules necessitates a detailed understanding of their binding properties. Malachite Green might induce a change in the iron transfer by Apo-transferrin. Spectroscopic studies showed malachite green oxalate (MGO) could form the apo-transferrin-MGO complex and change the Accessible Surface Area (ASA) of the key amino acids for iron transfer. According to the ASA results the accessible surface area of Tyrosine, Aspartate, and Histidine of apo-transferrin significantly were changed, which can be considered as a convincing reason for changing the iron transfer. Moreover, based on the fluorescence data MGO could quench the fluorescence intensity of apo-transferrin in a static quenching mechanism. The experimental and Molecular Dynamic simulation results represented that the binding process led to micro environmental changes, around tryptophan residues and altered the tertiary structure of apo-transferrin. The Circular Dichroism (CD) spectra result represented a decrease in the amount of the α-Helix, as well as, increase in the ß-sheet volumes of the apo-transferrin structure. Moreover, FTIR spectroscopy results showed a hypochromic shift in the peaks of amide I and II. Molecular docking and MD simulation confirmed all the computational findings.


Assuntos
Substâncias Perigosas/química , Ferro/química , Corantes de Rosanilina/química , Transferrina/química , Transporte Biológico , Humanos , Modelos Químicos , Modelos Moleculares , Conformação Molecular , Estrutura Molecular , Ligação Proteica , Análise Espectral , Relação Estrutura-Atividade
6.
Int J Biol Macromol ; 191: 385-395, 2021 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-34537301

RESUMO

Recently, removal of synthetic dyes, especially cationic dye of malachite green (MG), and inhibition of the growth of pathogenic microorganism from drinking water have gained much interest due to their high toxic potency for aquatic biosystems. Herein, a new dye adsorbent with outstanding antibacterial activity was fabricated based on xanthan gum (XG) and SiO2 nanoparticles through ultrasonication followed by the crosslinking polymerization with vinyl imidazole monomer. The nano adsorbents were characterized with various techniques such as FTIR, XRD, SEM, EDX, and TEM. The nanocomposites were applied as a filter for discarding MG dye and killing the growth of bacterial strains such as E.coli and S.aureus which are considered as the common impurities for drinking water. The data revealed that a maximum adsorption capacity was recorded as 99.5% (Qmax = 588.2 mg/g) at optimum conditions including 10 mg nanocomposite, 10 mL of MG dye (450 ppm), pH = 7, the temperature of 30 °C, and the adsorption time was adjusted within 6 h. The process of dye adsorption was applied to the common isotherm models of Langmuir, Temkin, and Freundlich, and the findings showed that the adsorption behavior was well fitted with the Langmuir one (R2 = 0.9983). Moreover, different adsorption kinetic models such as pseudo-first order, pseudo-second order, and intra-particle diffusion were studied for understanding the mechanism of MG adsorption onto nanocomposite surface. It was found that both intraparticle diffusion and pseudo-first-order have participated evenly in the adsorption mechanism of MG dye. Ultimately, the as-prepared nanocomposites were tested against the growth of S. aureus, and E.coli manifesting a superior inhibition diameter as 23.5 ± 0.50, and 25.33 ± 0.47 mm against E.coli, and S. aureus, respectively. Therefore, our new XG-g-PVI/SiO2 adsorbent is a very promising adsorbent for the fast and efficient capture of dyes from aqueous solutions.


Assuntos
Anti-Infecciosos/química , Corantes/química , Nanocompostos/química , Polissacarídeos Bacterianos/química , Corantes de Rosanilina/química , Dióxido de Silício/química , Adsorção , Anti-Infecciosos/farmacologia , Staphylococcus aureus/efeitos dos fármacos
7.
Food Chem ; 352: 129415, 2021 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-33711728

RESUMO

Furazolidone (FZD) and its metabolite called 3-amino-2-oxazolidinone (AOZ) would induce carcinogenic and mutagenic effects to human. In this work, to develop a novel, stable, and simple point of care testing (POCT) with a potential to social applied for FZD detection, we utilized the aspect of protein staining of coomassie brilliant blue (CBB) to exploit a new CBB-LFIA strategy free of NPs. Only one mixing step is needed during the probe manufacturing process, which requires just 2 h and is a great time saving strategy compared with other methods (requiring 4-33 h for probe preparation). Besides, the cost of CBB-LFIA is 300 times lesser than other LFIA with respect to obtaining the label. The developed CBB-LFIA was successfully applied to detect AOZ with a detection limit of 2 ng mL-1, without any influence from other potential interfering compounds. The proposed CBB-LFIA exhibited prominent practical application, and possesses considerable utilization potential in the related field.


Assuntos
Custos e Análise de Custo , Furazolidona/análise , Furazolidona/química , Imunoensaio/economia , Imunoensaio/métodos , Sistemas Automatizados de Assistência Junto ao Leito/economia , Corantes de Rosanilina/química , Humanos , Limite de Detecção , Fatores de Tempo
8.
Mikrochim Acta ; 187(11): 597, 2020 10 09.
Artigo em Inglês | MEDLINE | ID: mdl-33033874

RESUMO

A fiber mat was developed to visually determine the oxidation of edible oils, based on the colorimetric reaction of Schiff's reagent and aldehydes - the major volatile formed during lipid oxidation. The mixtures of polyvinyl alcohol (PVA) and Schiff's reagent containing various amounts of glycerol were electrospun to form the fiber mats. The response of the PVA/Schiff's reagent fiber mats to gaseous hexanal (model aldehyde) was investigated. Oxidized soybean oils were used to evaluate the effectiveness of the PVA/Schiff's reagent fiber mat for indicating oxidation of the oils. The results showed that the fiber mats obtained had average fiber diameters of less than 100 nm. Upon hexanal exposure, the fiber mats turned from white to purple. Higher amount of glycerol led to larger color change of the fiber mats and shorter response time to hexanal. A linear relationship (R2 = 0.96) was observed between the color change of the mat and hexanal concentration (15-117 µmol L-1). The visual determination limit of the mat for hexanal was 29 µmol L-1. The color change of the PVA/Schiff's reagent fiber mat was increased with an increase of soybean oil oxidation. Out of the seven soybean oils tested, the PVA/Schiff's reagent fiber mat was able to correctly indicate the oxidation states of six oils. The result suggested that the visual determination method developed is a promising method to indicate the oxidation of edible oils, which can be performed easily by non-experts.Graphical abstract.


Assuntos
Análise de Alimentos/instrumentação , Nanofibras , Álcool de Polivinil/química , Óleo de Soja/química , Análise de Alimentos/métodos , Oxirredução , Corantes de Rosanilina/química
9.
Mikrochim Acta ; 187(11): 604, 2020 10 10.
Artigo em Inglês | MEDLINE | ID: mdl-33037925

RESUMO

A novel ultrasensitive surface-enhanced Raman spectroscopy (SERS)-based method was developed for the determination of hyaluronidase (HAase), which was based on hyaluronic acid-coated gold nanoparticles (HA-AuNPs) as a substrate, via a facile one-pot method. The detection mechanism is based on HAase which can hydrolyze HA on HA-AuNPs into hyaluronic acid oligomers, causing the originally uniformly dispersed HA-AuNPs to be disintegrated into many smaller HA-AuNPs. These oligomers in turn increase the surface shielding of AuNPs, resulting in high aggregation tendencies. As a result, the original SERS substrate was disassembled, leading to a weakening of the SERS signal at 1173 cm-1. Malachite green was also used as a Raman probe to detect the change of SERS peak intensity and to quantify HAase. Compared with other methods for the determination of HAase, this method is more convenient and efficient; its determination limit was 0.4 mU mL-1. The recoveries of HAase spiked into human urine samples ranged from 97.2 to 103.9%.


Assuntos
Ácido Hialurônico/química , Hialuronoglucosaminidase/urina , Nanopartículas Metálicas/química , Análise Espectral Raman/métodos , Corantes/química , Ouro/química , Humanos , Hialuronoglucosaminidase/química , Limite de Detecção , Corantes de Rosanilina/química
10.
J Mater Chem B ; 8(36): 8242-8248, 2020 09 23.
Artigo em Inglês | MEDLINE | ID: mdl-32794526

RESUMO

Three different malachite green leuco derivatives (MG-Xs) are incorporated in liposomes. In all three cases, a substituent (X) is covalently linked to the central carbon atom, abbreviated as MG-OH, MG-OCH3, and MG-CN. The three MG-X compounds are solubilized separately in liposome membranes and become cationic (MG+) and water soluble under acidic conditions. MG+ is consequently released from the liposome to the aqueous exterior. Their release behavior corresponds to their ionization ability: MG-OH > MG-OCH3 > MG-CN. The cellular uptake of the liposomes, the cytotoxic effect, and the location of MG+ in cancer cells are investigated using murine cells derived from colon cancer (Colon 26 cells) and human embryonic kidney cells (HEK 293 cells). The toxic effect on cancer cells is correlated to the ionization ability of MG-Xs. The liposomes effectively deliver MG+via the endocytic pathway, resulting in the cytotoxicity of liposomes containing MG-OH which is higher than that of free MG-OH and MG+. The difference in the phospholipids constituting the liposome membranes barely had an effect on the ionization ratio and the cytotoxicity of MG-OH. Confocal fluorescence microscopic observations revealed that MG+ is ultimately transported into the nuclei after being released in acidic cellular compartments.


Assuntos
Antineoplásicos/farmacologia , Portadores de Fármacos/química , Lipossomos/química , Corantes de Rosanilina/farmacologia , Animais , Antineoplásicos/química , Antineoplásicos/toxicidade , Linhagem Celular Tumoral , Liberação Controlada de Fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Células HEK293 , Humanos , Concentração de Íons de Hidrogênio , Camundongos , Corantes de Rosanilina/química , Corantes de Rosanilina/toxicidade , Solubilidade
11.
Food Chem Toxicol ; 143: 111526, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32603830

RESUMO

Malachite Green (MG) has a worldwide application in aquaculture as a therapeutic agent; however, its use in food producing animals is illegal, due to potential carcinogenicity and persistence of residues. This pilot study, the first conducted in Armenia, aimed to determine the concentration of MG residues in flesh of fish grown in artificial ponds of Armenia and conduct dietary exposure assessment to characterize possible health risks to consumers. Detection of MG residues, including the major metabolite leucomalachite, was carried out in 29 fish composite samples by ELISA. The results were confirmed by LC-MS/MS. To determine fish consumption values, a food frequency questionnaire was used. Possible health risks were evaluated by calculating the Margin of Exposure (MOE) based on BMDL of 13 (neoplastic effects) and 6 (non-neoplastic effects) mg/kg bw. In 34.5% of the investigated fish samples MG residues exceeded the minimum required performance limit. For BMDL10 and BMDL0.5, the MOEs ranged 3.36E+06-3.37E+07 and 1.55E+06-1.55E+07, respectively. The MOE for neoplastic effects was more than 10,000 and for non-neoplastic effects was more than 100. The results do not indicate public health concerns. However, the results highlight issues concerning the illegal use of MG in Armenian aquaculture, which deserves further attention.


Assuntos
Resíduos de Drogas/química , Resíduos de Drogas/toxicidade , Peixes , Corantes de Rosanilina/química , Corantes de Rosanilina/toxicidade , Animais , Anti-Infecciosos Locais/química , Anti-Infecciosos Locais/toxicidade , Aquicultura , Armênia , Pesqueiros , Contaminação de Alimentos/análise , Água Doce
12.
Int J Biol Macromol ; 161: 457-469, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-32526305

RESUMO

The article is related to sunlight and UV-visible mineralization of harmful magenta-O (FB) dye. The nanocomposite used is a cross linked network of acrylic acid synthesized inside poly(acrylamide) grafted Guggul gum in the presence of UV-visible respondent bismuth ferrite nanoparticles. The synthesis of poly(acrylamide) grafted Guggul gum (Sample I) and synthesizing a crosslinked network inside it (Sample II) involved a two-step synthesis for optimizing various reaction parameters. The maximum % water uptake obtained for polymeric samples I and II was calculated as 1227.78% and 387.97%, respectively. Average particle size of bismuth ferrite nanoparticles was 47.34 nm. The nanocomposite could maximum uptake-mineralize FB dye as 97.3% and 98.8% under sunlight and photochemical reactor, respectively for 500 mg nanocomposite dose in 10 mg/L concentrated FB solution. Dye uptake occurs through ionic interactions. However, mineralization is a consequence of advanced oxidation process involving free radical species (OH and O2-.). The overall process of uptake-mineralization resembled second order kinetics and Langmuir theorem (monolayer adsorption). Intraparticle diffusion model gave an idea about the multistep (three steps) process of adsorption. Physico-chemical properties of FB dye got changed after mineralization except for the pH. The maximum uptake-mineralization was observed to be 76.2% after consecutive reuse of the nanocomposite hydrogel for five cycles.


Assuntos
Corantes/química , Nanocompostos/química , Nanogéis/química , Extratos Vegetais/química , Gomas Vegetais/química , Corantes de Rosanilina/química , Resinas Acrílicas/química , Adsorção , Commiphora , Compostos Férricos/química , Cinética , Polímeros/química , Água/química , Poluentes Químicos da Água/química , Purificação da Água/métodos
13.
Int J Nanomedicine ; 15: 1823-1835, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32214814

RESUMO

BACKGROUND: Biodegradation of toxic organic dye using nanomaterial-based microbial biocatalyst is an ecofriendly and promising technique. MATERIALS AND METHODS: Here, we have investigated the novel properties of functionalized Au-Ag bimetallic nanoparticles using extremophilic Deinococcus radiodurans proteins (Drp-Au-AgNPs) and their degradation efficiency on the toxic triphenylmethane dye malachite green (MG). RESULTS AND DISCUSSION: The prepared Drp-Au-AgNPs with an average particle size of 149.8 nm were capped by proteins through groups including hydroxyl and amide. Drp-Au-AgNPs demonstrated greater degradation ability (83.68%) of MG than D. radiodurans cells and monometallic AuNPs. The major degradation product was identified as 4-(dimethylamino) benzophenone, which is less toxic than MG. The degradation of MG was mainly attributed to the capping proteins on Drp-Au-AgNPs. The bimetallic NPs could be reused and maintained MG degradation ability (>64%) after 2 cycles. CONCLUSION: These results suggest that the easily prepared Drp-Au-AgNPs have potential applications as novel nanomedicine for MG detoxification, and nanomaterial for biotreatment of a toxic polyphenyl dye-containing wastewater.


Assuntos
Proteínas de Bactérias/metabolismo , Deinococcus/química , Nanopartículas Metálicas/química , Corantes de Rosanilina/metabolismo , Proteínas de Bactérias/química , Corantes/química , Corantes/metabolismo , Difusão Dinâmica da Luz , Cromatografia Gasosa-Espectrometria de Massas , Ouro/química , Ouro/metabolismo , Tamanho da Partícula , Corantes de Rosanilina/química , Prata/química , Prata/metabolismo , Espectrofotometria Ultravioleta , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios X
14.
Anal Chim Acta ; 1102: 84-90, 2020 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-32043999

RESUMO

Sensitive imaging of intracellular microRNAs (miRNAs) in cells is of great significance in clinical diagnoses and disease treatments, and it remains a major challenge to achieve this goal. Herein, we report a new in situ rolling circle transcription synchronization machinery (RCTsm) of lighting-up RNA aptamer strategy for highly sensitive imaging and selective differentiation of miRNA expression levels in cells. Such a RCTsm approach utilizes a DNA promoter to recycle the target miRNAs to trigger the initiation of multiple RCT process for the yield of many lighting-up RNA aptamers. The malachite green dye further binds these aptamers to show significantly enhanced fluorescence for completely label-free detection of the target miRNAs with a high sensitivity in vitro with a low femtomolar detection limit. More importantly, sensitive detection of under-expressed miRNAs in cells and distinct differentiation of the miRNA expression variations in different cells can also be realized with this RCTsm approach in a washing-free format, making it a versatile and useful tool for imaging trace miRNAs in single cells with the great potential for early cancer diagnosis as well as biomedical research.


Assuntos
Aptâmeros de Nucleotídeos/química , MicroRNAs/análise , Sequência de Bases , Linhagem Celular Tumoral , DNA/química , DNA/genética , Sondas de DNA/química , Sondas de DNA/genética , Corantes Fluorescentes/química , Humanos , Limite de Detecção , MicroRNAs/genética , Microscopia Confocal/métodos , Microscopia de Fluorescência/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Hibridização de Ácido Nucleico , Corantes de Rosanilina/química
15.
Chemosphere ; 247: 125803, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-31972482

RESUMO

The present study explored removal of carcinogenic cationic and anionic dyes from aqueous medium using green fabricated zinc oxide nanoparticles (ZnO-NPs). The ZnO-NPs were synthesized employing biogenic green reduction and precipitation approach. The characterization of ZnO NPs was done using various techniques such as FESEM, XRD, BET, TGA, HRTEM, EDX, and FTIR. All experiments were conducted in batch mode. Maximum removal was achieved at pH 6.0 and pH 8.0 for Congo Red (CR) and Malachite Green (MG) dyes respectively. Dye adsorption process showed better fit with Langmuir and Temkin isotherm models for CR dye and MG dye respectively. Maximum adsorption capacity of ZnO NPs was 48.3 mg/g for CR dye and 169.5 mg/g for MG dye. The dye adsorption followed pseudo-second order model and values of thermodynamic parameters confirmed that the adsorption process was spontaneous and favourable. Reusability efficiency of the nanoparticle was explored using ethanol and water and based on results it was inferred that ZnO-NPs can be reused for dye removal. Effect of salinity on the removal of CR and MG dyes was also explored and found that presence of salinity in aqueous medium have adverse impact on the dye removal efficiency of ZnO-NPs.


Assuntos
Corantes/química , Eucalyptus/química , Nanopartículas Metálicas/química , Folhas de Planta/química , Águas Residuárias/química , Purificação da Água/métodos , Óxido de Zinco/química , Adsorção , Vermelho Congo/química , Concentração de Íons de Hidrogênio , Cinética , Corantes de Rosanilina/química , Termodinâmica , Poluentes Químicos da Água/química
16.
Biotechnol Prog ; 36(2): e2916, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31587519

RESUMO

Laccases are multi copper oxidases that can oxidize both phenolic and nonphenolic lignin related compounds. Consequently, there has been continuous demand for laccases for the oxidative degradation of phenolic dyes in effluents. In view of this, the present work was focused on laccase production by solid substrate fermentation using a newly isolated fungus Perenniporia tephropora-L168. To intensify the laccase production, the process parameters pH, nitrogen, inducer, and substrate: water ratio were optimized by using statistical model. A set of optimal conditions noted were pH 3, nitrogen 0.001 g/L; inducer 0.5% and substrate: water ratio (1:10), which yielded laccase 1,160 U/g. The crude laccase exhibited noteworthy potential to degrade a triaryl-methane dye especially Malachite green. Also, during bioremediation studies, the statistical process optimization could achieve 81% decolourization within 180 min. The laccase treatment brought chemical transformation in malachite green as evident from UV-Visible spectra, FTIR, HPLC while toxicity against bacteria and fungi was also reduced. During phytotoxicity study, effect of treated and untreated dye on germination of seed was analyzed. Interestingly, the germination index for Vigna aconitifolia and Vigna radiata was increased by two and fourfold, respectively. Overall, this work demonstrates optimized production of laccase using Perenniporia tephropora-L168 and its efficient bioremediation potential for triaryl-methane dye.


Assuntos
Corantes/metabolismo , Fermentação , Lacase/biossíntese , Polyporaceae/metabolismo , Corantes de Rosanilina/metabolismo , Corantes/química , Concentração de Íons de Hidrogênio , Lacase/química , Polyporaceae/isolamento & purificação , Corantes de Rosanilina/química
17.
Methods Mol Biol ; 2091: 59-71, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31773570

RESUMO

The yeast Saccharomyces cerevisiae has given us much information on the metabolism and function of inositol polyphosphates and inorganic polyphosphate. To expand our knowledge of the metabolic as well as functional connections between inositol polyphosphates and inorganic polyphosphate, we have refined and developed techniques to extract and analyze these molecules in a second eukaryotic experimental model, the amoeba Dictyostelium discoideum. This amoeba, possessing a well-defined developmental program, is ideal to study physiological changes in the levels of inositol polyphosphates and inorganic polyphosphate, since levels of both molecules increase at late stages of development. We detail here the methods used to extract inositol polyphosphates using perchloric acid and inorganic polyphosphate using acidic phenol. We also present the postextraction procedures to visualize and quantify these molecules by polyacrylamide gel electrophoresis and by malachite green assay.


Assuntos
Dictyostelium/crescimento & desenvolvimento , Polifosfatos/análise , Fracionamento Químico , Dictyostelium/química , Eletroforese em Gel de Poliacrilamida , Modelos Biológicos , Percloratos/química , Fenóis/química , Corantes de Rosanilina/química
18.
Methods Enzymol ; 629: 269-289, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31727245

RESUMO

CD73 is a membrane-anchored ectoenzyme that degrades extracellular AMP into adenosine, a potent immunosuppressive factor. In physiological conditions, induction of the CD73-adenosine pathway acts as natural feedback mechanism to prevent excessive immune reactions and subsequent tissue damage. In the past few years, the CD73-adenosine pathway has emerged as a major immunosuppressive mechanism by which multiple types of cancer evade anti-tumor immunity. Research from our group and others have established that blocking the CD73-adenosine pathway represents a promising approach to improve cancer immunotherapy. In this context, an increasing number of research laboratories are becoming interested in CD73 biology and in the development/characterization of CD73 inhibitors. Implementation of simple, rapid and HTS-compatible assays to evaluate CD73 enzymatic active is a critical step for any laboratory willing to study the CD73-adenosine pathway. Over the years, we developed, optimized or adapted various methodologies to assess CD73 enzymatic activity using in vitro assays. In this chapter, we describe two different in vitro assays adapted to the measurement of CD73 enzymatic activity. Both assays are simple, robust, HTS-compatible and can be used in a cell-based fashion.


Assuntos
5'-Nucleotidase/análise , Ensaios Enzimáticos/métodos , Ensaios de Triagem em Larga Escala/métodos , Neoplasias/tratamento farmacológico , 5'-Nucleotidase/antagonistas & inibidores , 5'-Nucleotidase/metabolismo , Adenosina/imunologia , Adenosina/metabolismo , Antineoplásicos Imunológicos/farmacologia , Antineoplásicos Imunológicos/uso terapêutico , Linhagem Celular Tumoral , Colorimetria/instrumentação , Colorimetria/métodos , Corantes/química , Ensaios Enzimáticos/instrumentação , Estudos de Viabilidade , Proteínas Ligadas por GPI/análise , Proteínas Ligadas por GPI/antagonistas & inibidores , Proteínas Ligadas por GPI/metabolismo , Ensaios de Triagem em Larga Escala/instrumentação , Humanos , Luciferases/química , Luciferases/metabolismo , Luminescência , Neoplasias/imunologia , Neoplasias/patologia , Receptores Purinérgicos P1/imunologia , Receptores Purinérgicos P1/metabolismo , Proteínas Recombinantes/análise , Proteínas Recombinantes/metabolismo , Corantes de Rosanilina/química , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/imunologia
19.
Mol Pharmacol ; 96(3): 355-363, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31263019

RESUMO

The P2X7 receptor is a trimeric ligand-gated ion channel activated by ATP. It is implicated in the cellular response to trauma/disease and considered to have significant therapeutic potential. Using chimeras and point mutants we have mapped the binding site of the P2X7R-selective antagonist AZ11645373 to the known allosteric binding pocket at the interface between two subunits, in proximity to, but separated from the ATP binding site. Our structural model of AZ11645373 binding is consistent with effects of mutations on antagonist sensitivity, and the proposed binding mode explains variation in antagonist sensitivity between the human and rat P2X7 receptors. We have also determined the site of action for the P2X7R-selective antagonists ZINC58368839, brilliant blue G, KN-62, and calmidazolium. The effect of intersubunit allosteric pocket "signature mutants" F88A, T90V, D92A, F103A, and V312A on antagonist sensitivity suggests that ZINC58368839 comprises a binding mode similar to AZ11645373 and other previously characterized antagonists. For the larger antagonists, brilliant blue G, KN-62, and calmidazolium, our data imply an overlapping but distinct binding mode involving the central upper vestibule of the receptor in addition to the intersubunit allosteric pocket. Our work explains the site of action for a series of P2X7R antagonists and establishes "signature mutants" for P2X7R binding-mode characterization.


Assuntos
Mutação Puntual , Antagonistas do Receptor Purinérgico P2X/farmacologia , Receptores Purinérgicos P2X7/química , Receptores Purinérgicos P2X7/metabolismo , 1-(5-Isoquinolinasulfonil)-2-Metilpiperazina/análogos & derivados , 1-(5-Isoquinolinasulfonil)-2-Metilpiperazina/química , 1-(5-Isoquinolinasulfonil)-2-Metilpiperazina/farmacologia , Trifosfato de Adenosina/metabolismo , Sítio Alostérico , Amidas/química , Amidas/farmacologia , Sítios de Ligação , Humanos , Imidazóis/química , Imidazóis/farmacologia , Indóis/química , Indóis/farmacologia , Modelos Moleculares , Simulação de Acoplamento Molecular , Antagonistas do Receptor Purinérgico P2X/química , Receptores Purinérgicos P2X7/genética , Corantes de Rosanilina/química , Corantes de Rosanilina/farmacologia , Tiazóis/química , Tiazóis/farmacologia
20.
Indian J Cancer ; 56(2): 114-118, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31062728

RESUMO

INTRODUCTION: The recommended technique of sentinel lymph node biopsy (SLNB) in breast cancer is a combination of blue dye and radiotracer. In the Indian scenario, SLNB is still not routinely practiced due to lack of nuclear medicine facilities and unavailability of isosulfan blue or patent blue violet (PBV). This study was conducted for optimizing SLN identification techniques by comparing the identification rate using PBV and methylene blue (MB) in combination with radiotracer. MATERIALS AND METHODS: Single-blinded two-arm parallel design randomized control trial was conducted at an apex teaching and research medical institute in India. Patients with axillary LN-negative breast cancer were included. Blue dye and radio tracer were injected preoperatively, and SLNB was performed using a combination technique. Frozen section was performed. Demographic, clinical, radiological, operative, and histopathological data were recorded. Descriptive statistics were used to represent patient characteristics. Baseline characteristics for entire cohort and between groups were compared using Student's t-test for quantitative variables and Chi-square test for qualitative variables. RESULTS: A total of 119 patients were randomized for mapping with MB and 118 patients with PBV between 2011 and 2015. SLN was identified in 116 patients with MB and 115 with PBV. SLN identification proportions were 97.4% (MB) and 96.6% (PBV). In patients undergoing axillary lymph node dissection, concordance with SLNB was 98.5% and 96.61% in MB and PBV, respectively. False-negative proportion for MB was 2.56% and 7.69% for PBV, respectively. The cost of MB is about INR 15 per ~10-mL vial. The cost of PBV is approximately ~$91 per ampoule (equivalent to approximately INR 8190). CONCLUSION: SLNB using MB can be recommended as the technique of choice in low-resource settings.


Assuntos
Neoplasias da Mama/diagnóstico , Metástase Linfática/diagnóstico , Biópsia de Linfonodo Sentinela/métodos , Linfonodo Sentinela/diagnóstico por imagem , Adulto , Idoso , Idoso de 80 Anos ou mais , Axila/diagnóstico por imagem , Axila/patologia , Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/patologia , Estudos de Coortes , Feminino , Humanos , Índia , Excisão de Linfonodo/métodos , Metástase Linfática/patologia , Azul de Metileno/química , Pessoa de Meia-Idade , Corantes de Rosanilina/química , Linfonodo Sentinela/patologia
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