RESUMO
A low cost and versatile colorimetric platform is developed for selective detections of various enzymes. Similar to peroxidases, free copper ion catalyzes the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of H2O2 and turns TMB into a blue product. Bindings from ligands toward copper ions inhibit this catalysis. Enzymes catalyze the reactions of related substrates with generation or consumption the ligands for the binding and thus in turn alter the color changes as responses toward the enzymes. With suitable substrates, exemplary enzymes, including trypsin, acid phosphatase, and tyrosinase, can be sensitively measured, with limits of detection of 0.003 µg/mL, 0.004 U/L, and 0.02 U/mL, respectively. This platform is built with directly available reagents, and the signals can be obtained with inexpensive photometers or visual observations. The low cost and convenience make it suitable for cases where complicated instrumentations are not available, such as point-of-care testing.
Assuntos
Colorimetria , Cobre , Benzidinas , Catálise , Ensaios Enzimáticos , Peróxido de Hidrogênio , Limite de Detecção , OxirreduçãoRESUMO
Silver ion detection with ultra-high sensitivity was established. We synthesized copper nanoclusters (CuNCs) with blue fluorescence through a one-pot process. Instead of a direct quencher toward the CuNCs, silver ions activated the strong oxidation from persulfate and subsequently converted divalent manganese ion into manganese dioxide (MnO2). The surface charges of MnO2 and the CuNCs brought them together and quenched the fluorescence from the latter. Due to silver ions' role as the catalyst in the process, it cycled and even a small amount leads to a significant fluorescence change. This signaling provided the determination of silver ions in the range 5 pM~1 nM, with a detection limit of 1.2 pM. The method is selective, and its applicability was validated through practical water sample analyses.
Assuntos
Corantes Fluorescentes/química , Nanopartículas Metálicas/química , Povidona/química , Prata/análise , Cobre/química , Água Potável/análise , Fluorescência , Lagos/análise , Limite de Detecção , Manganês/química , Compostos de Potássio/química , Espectrometria de Fluorescência/métodos , Sulfatos/química , Poluentes Químicos da Água/análiseRESUMO
A method for sensitive detection of nitrite is presented. It is found that the red fluorescence of gold nanoclusters (with excitation/emission maxima at 365/635 nm) is quenched by traces of iodine via etching. Free iodide is formed by oxidation of iodide by bromate anion under the catalytic effect of nitrite. This catalytic process provides a sensitive means for nitrite detection. Under the optimal conditions, fluorescence linearly dropos in the 10 nM to 0.8 µM nitrite concentration range. The limit of detection is 1.1 nM. This is a few orders of magnitude lower than the maximum concentration allowed by authorities. Graphical abstract Schematic representation of a method for detection of nitrite via a redox reaction. Iodine was produced in the reaction and subsequently quenched the fluorescence from gold nanoclusters by etching their metallic cores, and a sensitive assay for nitrite down to 1.1 nM was developed.
RESUMO
A fluorimetric method is described for the determination of alkaline phosphatase (ALP) activity. It is based on the use of polyethyleneimine-coated silver nanoclusters (AgNCs), which display an intense blue fluorescence peaking at 450 nm (under 375 nm excitation). ALP catalyzes the dephosphorylation of the thiophosphate amifostine to generate a thiol that binds to the AgNCs and causes its fluorescence to be quenched. Under the optimal experimental conditions, fluorescence linearly drops in the 0.08-2.0 U L-1 ALP activity range, and the limit of detection is 0.02 U L-1. The method was successfully applied to the determination of ALP activity in spiked human serum samples. Graphical abstract Alkaline phosphatase (ALP) catalyzes the degradation of amifostine with a generation a thiol product. The thiol quenches the fluorescence of silver nanoclusters, and a method for the detection of ALP down to 0.02 U L-1 was developed.
Assuntos
Fosfatase Alcalina/sangue , Ensaios Enzimáticos/métodos , Fluorometria/métodos , Nanopartículas Metálicas/química , Prata/química , Fosfatase Alcalina/química , Amifostina/química , Fluorescência , Humanos , Limite de Detecção , Nanopartículas Metálicas/efeitos da radiação , Polietilenoimina/química , Compostos de Sulfidrila/síntese química , Raios UltravioletaRESUMO
The occurrence rate of senile dementia among diabetes patients is high, but there is still no effective therapeutic method. This study aimed to explore curative effect of exosomes deriving from MSCs treating diabetes-induced cognition impairment. After BM-MSCs culture was purified, Western Blot and electron microscope were used to identify exosomes which were injected into diabetes mouse through intracranial injection. Purified exosomes in Western blot analysis and electron microscope observation were verified. After therapy with exosomes deriving BM-MSC, mouse cognition impairment and histologic abnormity were recovered. Exosomes deriving from BM-MSCs can be a new preparation treating cognition with application prospect.
Assuntos
Transtornos Cognitivos/etiologia , Transtornos Cognitivos/terapia , Diabetes Mellitus/fisiopatologia , Exossomos/fisiologia , Células-Tronco Mesenquimais/citologia , Animais , Cognição/fisiologia , Modelos Animais de Doenças , Camundongos , Camundongos Endogâmicos C57BLRESUMO
A method is described for ratiometric fluorometric assays of H2O2 by using two probes that have distinct response profiles. Under the catalytic action of ferrous ion, the 615 nm emission of protein-stabilized gold nanoclusters (under 365 nm photoexcitation) is quenched by H2O2, while an increased signal is generated with a peak at 450 nm by oxidizing coumarin with the H2O2/Fe(II) system to form a blue emitting fluorophore. These decrease/increase responses give a ratiometric signal. The ratio of the fluorescences at the two peaks are linearly related to the concentration of H2O2 in the range from 0.05 to 10 µM, with a 7.7 nM limit of detection. The detection scheme was further coupled to the urate oxidase catalyzed oxidation of uric acid which proceeds under the formation of H2O2. This method provides an simple and effective means for the construction of ratiometric fluorometric (enzymatic) assays that involve the detection of H2O2. Graphical abstract Under catalysis by ferrous ion, hydrogen peroxide quenches the luminescence of gold nanoclusters (AuNCs) and oxidizes coumarin into a fluorescent derivative, which rendered fluorescence ON and OFF at two distinct wavelengths for ratiometric measurements.
Assuntos
Técnicas Biossensoriais/métodos , Corantes Fluorescentes/química , Ouro/química , Peróxido de Hidrogênio/química , Nanopartículas Metálicas/química , Urato Oxidase/metabolismo , Ácido Úrico/análise , Biocatálise , Humanos , Limite de Detecção , Ácido Úrico/sangue , Ácido Úrico/químicaRESUMO
Acetylcholinesterase (AChE) is an important enzyme in the central and peripheral nervous system that regulates the balance of the neurotransmitter acetylcholine. In this work, a simple, selective and sensitive fluorescence assay was developed toward AChE activity. A conventional AChE substrate acetylthiocholine iodide (ATCI) was applied. Instead directly rendering a signaling, it was found that free iodide ions was released during the enzymatic hydrolysis of ATCI. These ions further catalyzed the oxidation of non-emissive o-phenylenediamine (OPD) into a fluorescent product. This gave a response differed from frequently-adopted sulfhydryl- -based signals and thus minimized related interferences. All materials included in this process were directly available and no additional syntheses were required. Due to the extra iodide-based catalysis included, this scheme was capable of providing a sensitive response toward AChE in the range of 0.01-8 U/L, with a limit of detection at 0.006 U/L. This method was further extended onto chlorpyrifos as an exemplary AChE inhibitor, with a detection down to 3 pM.
Assuntos
Acetilcolinesterase , Acetiltiocolina/análogos & derivados , Iodetos , Peroxidase , Fluorescência , Catálise , Corantes , PeroxidasesRESUMO
Coronary heart disease (CHD) poses an important threat to human health, and its pathogenesis is the formation of atheromatous plaques in coronary ventricles. Compared to other biomarkers, lipoprotein-associated phospholipase A2 (Lp-PLA2), which is involved in multiple processes of atherosclerosis, is a noticeable inflammatory biomarker related to CHD. Herein, using a multifunctional nanocomposite containing a CoFe Prussian blue analogue (PBA) and Au nanoparticles (AuNPs) (AuNPs@CoFe PBA) as a sensing substrate, an electrochemiluminescent (ECL) immunosensor was developed for the highly sensitive detection of Lp-PLA2. Benefiting from the synergistic effect of the PBA and AuNPs, the nanocomposite exhibits excellent peroxidase-like activity and can catalyze the luminol-ECL reaction, amplifying the ECL signal by â¼29-fold. Meanwhile, the enlarged specific surface area of the nanocomposite and the presence of abundant AuNPs allow the immobilization of more antibody proteins, thereby improving the sensing response of the immunosensor. When the target Lp-PLA2 is captured by the antibody on the sensor surface, the sensor emits a reduced ECL signal because of the increased mass and electron transfer resistance due to the formation of the immune complex. Under optimized conditions, the constructed ECL immunosensor exhibits a broad linear range from 1 to 2200 ng/mL and a low detection limit of 0.21 ng/mL. Additionally, the ECL immunosensor exhibits high specificity, stability, and reproducibility. This work provides a new approach to diagnose CHD and broadened the application of the PBA in the field of ECL sensors.
Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Humanos , Ouro , 1-Alquil-2-acetilglicerofosfocolina Esterase , Reprodutibilidade dos Testes , Limite de Detecção , Imunoensaio , AnticorposRESUMO
Trypsin is a serine protease playing a key role in regulating pancreatic exocrine function and can be applied as a marker for the diagnosis of pancreatitis. In this work, a convenient and sensitive fluorescent assay was developed toward trypsin. Hydrogen peroxide slowly oxidized a non-fluorescent o-phenylenediamine (OPD) into a fluorescent product 2,3-diaminophenothiazine (DAP) under the catalytic from copper ions. After the introduction of bovine serum albumin (BSA), the combination of BSA with copper ions formed a peroxidase mimic and significantly accelerated the reaction rate. As an efficient protease, trypsin cleaved the lysine and arginine residues in BSA. This destroyed the binding between Cu2+ and BSA, and brought in a reduction of the catalytic effect. The accompanying decrease in fluorescence provided a response to trypsin in the range of 0.01-600 ng/mL, with a detection limit of 0.007 ng/mL. The scheme had a good selectivity and was successfully applied to the detection of real samples.
Assuntos
Cobre , Soroalbumina Bovina , Tripsina/química , Soroalbumina Bovina/química , Fluorescência , Cobre/química , Peroxidase , Peroxidases , Corantes Fluorescentes/química , Íons , Espectrometria de FluorescênciaRESUMO
With TiO(2) nanoparticles as carrier, a supported nano-material of Au atomic cluster/TiO(2) nano-hybrid was synthesized. It was then modified onto the surface of indium tin oxide (ITO) by Nafion to act as a working electrode for exciting the electrochemiluminescence (ECL) of luminol. The properties of the nano-hybrid and the modified electrode were characterized by XRD, XPS, electronic microscopy, electrochemistry and spectroscopy. The experimental results demonstrated that the modification of this nano-hybrid onto the ITO electrode efficiently intensified the ECL of luminol. It was also revealed that the ECL intensity of luminol on this modified electrode showed very sensitive responses to oxygen and hydrogen peroxide. The detection limits for dissolved oxygen and hydrogen peroxide were 2 µg L(-1) and 5.5 × 10(-12) M, respectively. Besides the discussion of the intensifying mechanism of this nano-hybrid for ECL of luminol, the developed method was also applied for monitoring dissolved oxygen and evaluating the scavenging efficiency of reactive oxygen species of the Ganoderma lucidum spore.
RESUMO
A method for a sensitive fluorescence detection of glutathione was established. Glutathione-stabilized copper nanoclusters (CuNCs) were synthesized via a facile process. These CuNCs showed blue fluorescence with a peak around 450 nm. In the presence of p-benzoquinone (PBQ), the electron transfer from the copper nanoclusters to PBQ quenched the fluorescence of the CuNCs. Glutathione (GSH), as a reducing agent, formed a complex with PBQ. This formation inhibited the quenching from PBQ, and a restored fluorescence was obtained. This interaction provided a fluorescence enhancement for the measurement of GSH. Under the optimal condition, linear responses were obtained toward GSH in the ranges of 0.06-6.0 µM, with a limit of detection at 20 nM. This developed assay was easy in operation with high sensitivity and selectivity. The applicability was approved with successful glutathione measurements in real samples.
Assuntos
Cobre , Nanopartículas Metálicas , Bioensaio , Glutationa , Limite de Detecção , Espectrometria de FluorescênciaRESUMO
A simple colorimetric method was developed for sensitive and selective detections of I- and Hg2+. Histidine stabilized gold nanoclusters (His-AuNCs) were synthesized and catalyzed the oxidation of colorless 3,3',5,5'-tetramethylbenzidine (TMB) to a blue product. As a strong ligand toward gold, iodide (I-) attached to the surface of the His-AuNCs and significantly enhanced the oxidase-like activity of the His-AuNCs. Based on this enhancement, a sensitive colorimetric response toward I- was obtained. Furthermore, the strong interaction between Hg2+ and I- was adopted for an indirect Hg2+ detection. Under the optimal conditions, the platform presented high selectivity for the determinations of I- and Hg2+ in the ranges 0.02-1 µM and 0.05-0.8 µM, with detection limits as 3.3 nM and 8 nM respectively. This colorimetric assay was successfully applied for analysis of real samples.
Assuntos
Mercúrio , Nanopartículas Metálicas , Colorimetria/métodos , Ouro , Iodetos , Mercúrio/análiseRESUMO
We have developed a photoresponsive DNA-cross-linked hydrogel that can be photoregulated by two wavelengths with a reversible sol-gel conversion. This photoinduced conversion can be further utilized for precisely controllable encapsulation and release of multiple loads. Specifically, photosensitive azobenzene moieties are incorporated into DNA strands as cross-linkers, such that their hybridization to complementary DNAs (cDNAs) responds differently to different wavelengths of light. On the basis of the rheology variation of hydrogels, it is possible to utilize this material for storing and releasing molecules and nanoparticles. To prove the concept, three different materials--fluorescein, horseradish peroxidase, and gold nanoparticles--were encapsulated inside the gel at 450 nm and then released by photons at 350 nm. Further experiments were carried out to deliver the chemotherapy drug doxorubicin in a similar manner in vitro. Our results show a net release rate of 65% within 10 min, and the released drug maintained its therapeutic effect. This hydrogel system provides a promising platform for drug delivery in targeted therapy and in biotechnological applications.
Assuntos
Reagentes de Ligações Cruzadas/química , DNA/química , Portadores de Fármacos/química , Hidrogéis/química , Luz , Neoplasias/tratamento farmacológico , Compostos Azo/química , Linhagem Celular Tumoral , DNA Complementar/química , Preparações de Ação Retardada , Doxorrubicina/química , Doxorrubicina/uso terapêutico , Portadores de Fármacos/toxicidade , Fluoresceína/química , Ouro/química , Peroxidase do Rábano Silvestre/química , Humanos , Teste de Materiais , Nanopartículas Metálicas/química , Modelos Moleculares , Hibridização de Ácido Nucleico , Transição de Fase , Conformação Proteica , Estereoisomerismo , TermodinâmicaRESUMO
A disulfide linked naphthalimide dimer probe was designed for mercury ion (Hg(2+)) recognition in this work. The recognition was based on the strong affinity of mercury for sulfur. The experimental results revealed that the probe exhibited high selectivity and sensitivity toward Hg(2+) in comparison to other metal ions via a turn-on and reversible response to Hg(2+) in neutral buffer solution. More importantly, the probe demonstrated a linear response for Hg(2+) over a concentration range from 0 to 150 µM with a detection limit of 0.38 µM, which is just the limit of the safe concentration for humans. Upon addition of 150 µM Hg(2+), the enhancement of fluorescence reached a maximum (â¼7-fold). The performances of the probe indicated that it could meet the selectivity requirements for biomedical and environmental application and also was sensitive enough to detect Hg(2+) in environmental and biological samples.
Assuntos
Dissulfetos/química , Corantes Fluorescentes/química , Mercúrio/análise , Naftalimidas/química , Dimerização , Corantes Fluorescentes/síntese química , Concentração de Íons de Hidrogênio , Estrutura Molecular , Naftalimidas/síntese química , Soluções , Espectrometria de Fluorescência , Água/químicaRESUMO
Dopamine (DA) is one of the most important neurotransmitters in human bodies and its sensitive detection remains a challenge. Herein, protein stabilized gold-silver nanoclusters (Au-AgNCs) were synthesized at first. It was found that the introduction of dopamine lead to a significant enhancement of the fluorescence from the nanoclusters, together with a red-shift of the peak. Through related spectroscopic and electrochemical studies, the fluorescence enhancement was attributed to the reduction of the nanoclusters by dopamine. This enhancement was then adopted for quantitative measurements, and linear responses toward dopamine in the ranges 0.01-1.7 µM and 1.7-10 µM were constructed. A limit of detection was obtained at 6.9 nM. The present study provided a facile and efficient method for the determination of dopamine, and the method was successfully applied for related measurements in serum samples.
Assuntos
Ouro , Nanopartículas Metálicas , Dopamina , Fluorescência , Humanos , Limite de Detecção , Prata , Espectrometria de FluorescênciaRESUMO
A ï¬uorescence method was developed for a turn-on measurement of acid phosphatase (ACP) activity. It was found that cerous ion (Ce3+) could lead to an enhancement of glutathione protected gold nanocluster fluorescence through an aggregation-induced-emission (AIE) process, while its higher valent counterpart ceric ion (Ce4+) could not. In a weakly acidic environment, ACP catalyzed the dephosphorylation of a phosphate ester of ascorbic acid, with a generation of ascorbic acid (AA). AA reduced Ce4+ into Ce3+, which subsequently enhanced the nanocluster fluorescence. This kind of turn-on fluorescence linearly related to the ACP activity in the range of 0.005-2.4 U/L, with a limit of detection as 0.001 U/L. Human serum samples were measured after a trichloroacetic acid treatment and a simple dilution. The whole analyses were accomplished in 1.5 h with results in good accordance with a reference method.
Assuntos
Ouro , Nanopartículas Metálicas , Fosfatase Ácida , Glutationa , Humanos , Espectrometria de FluorescênciaRESUMO
OBJECTIVE: To assess the clinical efficacy of Chinese herbal medicine for removing blood stasis combined with acupuncture in the treatment of sequelae of cerebral infarction. METHODS: Ninety patients with cerebral infarction admitted to our hospital from April 2018 to April 2020 were enrolled and equally allocated to an experimental group and a control group. The control group was treated with aspirin, and the experimental group was treated with Chinese herbal medicine for removing blood stasis combined with acupuncture. The recovery of the ability of daily living (ADL), recovery of hemiplegic limb function, blood viscosity, total cholesterol (TC), triglyceride (TG), and quality of life were evaluated. RESULTS: After treatment, the ADL of patients in the two groups witnessed a remarkable recovery, with superior results in the experimental group than the control group (P < 0.05). The hemiplegic limb recovery of the experimental group was observed to be significantly improved when compared with the control group (P < 0.05). Remarkably lower blood viscosity-related indexes of TC and TG of the experimental group compared to the control group were identified (P < 0.05). As to the total remission rate (TRR), the experimental group demonstrated a higher level than the control group (P < 0.05). The scores of quality of life of patients in the experimental group after treatment were evidently higher than those of the control group (P < 0.05). CONCLUSION: Chinese herbal medicine for removing blood stasis combined with acupuncture treatment can better improve the hemiplegic limb function and the quality of life, and reduce blood viscosity of patients with sequelae of cerebral infarction.
RESUMO
Detection of hydrogen peroxide is of significant importance for biological assays, and fluorescence methods are intensively reported for this purpose. Due to the highly oxidative property of this species, usually fluorescence quenching is obtained during the interactions and decreased signals are rendered. In this report, this oxidative property was adopted for an increased fluorescence signaling. Photoluminescent silver nanoclusters (AgNCs) were synthesized with polyethyleneimine as the stabilizer. This fluorescence from these nanoclusters could be quenched by reduced glutathione (GSH) through an interaction from its thiol group. As an oxidant, hydrogen peroxide converted GSH into an oxidized form (GSSG) with an elimination of the free thiols, and inhibited the quenching. This interaction presented an increased response toward hydrogen peroxide in the range of 0.1-20 µM with a detection limit of 35 nM. The scheme was further coupled with glucose oxidase for a glucose analysis down to 0.11 µM. This method was selective and was successfully applied for glucose measurement in human serum samples.
Assuntos
Nanopartículas Metálicas , Prata , Fluorescência , Glucose , Humanos , Peróxido de Hidrogênio , Limite de Detecção , Polietilenoimina , Espectrometria de FluorescênciaRESUMO
Automatic, inexpensive, simple and sensitive methods for DNA sensing and quantification are highly desirable for biomedical research. The rapid development of both the fundamentals and applications of electrochemiluminescence (ECL) over the past years has demonstrated its potential for analytical and bio-analytical chemistry. This paper reports the quenching effect of DNA on the ECL of luminol and the further development of a DNA sensing device. With the pre-functionalization by a composite of carbon nano-tubes (CNTs) and Au nanoparticles (AuNPs), the sensor provides a novel and valuable label-free approach for DNA sensing. Here the ECL intensity was remarkably decreased when more than 1.0 × 10(-12) molar of DNA were adsorbed on the sensor. Linearity of the DNA amount with the reciprocal of ECL intensity was observed. A saturated sensor caused a 92.8% quenching effect. The research also proposes the mechanism for the quenching effect which could be attributed to the interaction between luminol and DNA and the elimination of reactive oxygen species (ROSs) by DNA.
Assuntos
DNA/análise , Eletroquímica/instrumentação , Medições Luminescentes/instrumentação , Luminol/química , Ouro/química , Nanopartículas Metálicas/química , Nanotubos de Carbono/químicaRESUMO
Molybdenum (Mo) is an essential nutrient for the proper functioning of some enzymes in living organisms as human beings. Conventional methods for its detection require complicated instrumentations as atomic absorption or mass spectrometers. In this work, a sensitive kinetic fluorescence was developed as an alternative. Gold nanoclusters (AuNCs) with red fluorescence emission were synthesized, and this fluorescence was effectively quenched by iodine through an etching process. It was found that the presence of Mo significantly speeded up a reaction for the generation of iodine, and thus enhanced the quenching. This effect was then adopted for the development of a sensitive fluorescent measurement toward Mo. The method was capable of detecting Mo down to 0.2 nM and was successfully applied for the analyses of mung bean and tea leaf samples.