RESUMO
MAIN CONCLUSION: A stable genetic transformation system for Erigeron breviscapus was developed. We cloned the EbYUC2 gene and genetically transformed it into Arabidopsis thaliana and E. breviscapus. The leaf number, YUC2 gene expression, and the endogenous auxin content in transgenic plants were significantly increased. Erigeron breviscapus is a prescription drug for the clinical treatment of cardiovascular and cerebrovascular diseases. The rosette leaves have the highest content of the major active compound scutellarin and are an important component in the yield of E. breviscapus. However, little is known about the genes related to the leaf number and flowering time of E. breviscapus. In our previous study, we identified three candidate genes related to the leaf number and flowering of E. breviscapus by combining resequencing data and genome-wide association study (GWAS). However, their specific functions remain to be characterized. In this study, we cloned and transformed the previously identified full-length EbYUC2 gene into Arabidopsis thaliana, developed the first stable genetic transformation system for E. breviscapus, and obtained the transgenic plants overexpressing EbYUC2. Compared with wild-type plants, the transgenic plants showed a significant increase in the number of leaves, which was correlated with the increased expression of EbYUC2. Consistently, the endogenous auxin content, particularly indole-3-acetic acid, in transgenic plants was also significantly increased. These results suggest that EbYUC2 may control the leaf number by regulating auxin biosynthesis, thereby laying a foundation for revealing the molecular mechanism governing the leaf number and flowering time of E. breviscapus.
Assuntos
Arabidopsis , Erigeron , Erigeron/genética , Arabidopsis/genética , Estudo de Associação Genômica Ampla , Ácidos Indolacéticos , Folhas de Planta/genética , Plantas Geneticamente Modificadas , Transformação GenéticaRESUMO
In this study, a photoelectrochemical (PEC) sensor based on perylene diimide derivatives (PDIs) was developed for the ultrasensitive quantification of dopamine (DA). PDIs were able to form self-assembled semiconductor nanostructures by strong π-π stacking, suitable for photoactive substances. Moreover, the shape of the PDI significantly affected the PEC properties of these nanostructures. The results showed that amino PDI with two-dimensional (2D) wrinkled layered nanostructures exhibited superior PEC properties relative to one-dimensional (1D) nanorods and fiber-based nanostructures (methyl and carboxyl PDIs). Based on these results, a mechanism for PEC sensor action was then proposed. The presence of 2D amino-PDI resulted in accelerated charge separation and transport. Furthermore, dopamine acted as effective electron donor to cause an increase in photocurrent. The as-obtained sensor was then used to detect small molecules like DA. A blue light optimized sensor at an applied potential of 0.7 V showed a detection limit of 1.67 nM with a wide linear range of 5 nM to 10 µM. On the other hand, the sensor presented acceptable reliability in determining DA in real samples. A recovery rate between 97.99 and 101.0% was obtained. Overall, controlling the morphology of semiconductors can influence PEC performance, which is a useful finding for the future development of PEC sensors.
Assuntos
Técnicas Biossensoriais , Nanoestruturas , Perileno , Dopamina , Perileno/química , Reprodutibilidade dos Testes , Técnicas Eletroquímicas/métodos , Técnicas Biossensoriais/métodos , Limite de DetecçãoRESUMO
Electrochemical nanotags with strong signal input are necessary for a ratiometric electrochemical sensor to overcome the drawbacks of inaccurate detection results. In this paper, the metal-organic framework (MOF) UiO-66 was utilized as an electrochemical signal tag. A stable and strong current response at +0.9 V can be detected in neutral conditions. MicroRNA (miRNA) was employed as the model analyte. Herein, an enzyme-free DNA-walker-based ultrasensitive ratiometric electrochemical biosensor in combination with Zr MOF (UiO-66) signal tags to detect miRNA was demonstrated. In the presence of miRNA, the autonomous walker movement can be initiated by miRNA, leading to the release of biotin-modified fragments. Thus, streptavidin-labeled UiO-66 nanomaterials were not bound to the electrode, generating a low signal response of UiO-66 at +0.9 V. However, the current signal of electrolyte solution as reference at +0.2 V was increased due to the enhancement of electrode conductivity. This ratiometic sensor demonstrated high sensitivity, selectivity, and reproducibility. It can eliminate the disturbance of environmental factors and basic electrode characteristics, providing more accurate signals. A limit of detection (LOD) of 0.17 fM was achieved. Moreover, the method was also used to detect miRNA-21 spiked in real serum samples.
Assuntos
Técnicas Biossensoriais , Estruturas Metalorgânicas , MicroRNAs , Técnicas Biossensoriais/métodos , Biotina , DNA/genética , Técnicas Eletroquímicas/métodos , Eletrólitos , Limite de Detecção , Ácidos Ftálicos , Reprodutibilidade dos Testes , EstreptavidinaRESUMO
Ochratoxin A (OTA) is a highly toxic food contaminant and is harmful to human beings. Herein, a ratiometric electrochemical aptasensor based on a DNA tetrahedral nanomaterial (NTH) was developed in combination with the signal tag of a zirconium metal-organic framework (UiO-66) for the detection of OTA. In the sensor, UiO-66 and a [Fe(CN)6]3-/4- electrolyte solution were used as the signal probe and the internal reference probe, respectively. In the presence of OTA, the OTA aptamer was released from the electrode due to the specific binding of OTA. Thus, signal probe P1 labeled-UiO-66 was captured on the electrode surface by hybridization with DNA NTH. Since signal probe P1 labeled-UiO-66 was close to the electrode, it leads to an increased signal current of UiO-66 at +0.9 V. As the conductivity of the modified electrode decreased, the current signal of [Fe(CN)6]3-/4- at +0.2 V also decreased. The proposed ratiometric electrochemical aptasensor could effectively eliminate external environmental influences and could avoid electrochemical background signals. The aptasensor demonstrated high specificity for OTA, and achieved a good linear range of 1 pg mL-1-100 ng mL-1 with a detection limit of 330 fg mL-1. The developed electrochemical aptamer biosensor effectively detected OTA in corn kernel samples, verifying its practical application for the determination of OTA in actual samples.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Estruturas Metalorgânicas , Nanoestruturas , Ocratoxinas , DNA , Técnicas Eletroquímicas , Eletrólitos , Ouro , Humanos , Limite de Detecção , Ocratoxinas/análise , Ácidos Ftálicos , Zea mays , ZircônioRESUMO
The rapid, sensitive, and selective detection of ascorbic acid (AA) is of significance in medical assays and diagnostics. In this work, a new aminoperylenediimide (APDI) derived ratiometric fluorescent probe based on the specific redox reaction of cobalt oxyhydroxide (CoOOH) and AA was constructed. APDI exhibited dual fluorescence emission peaks at 549 and 596 nm with an excitation wavelength of 494 nm. In the presence of CoOOH, the dual fluorescence could be quenched. The dominant fluorescence quenching mechanism was caused by the inner filter effect. Using the red emission as a reference, the fluorescence intensity ratio (F549 /F596 ) was linearly correlated with the concentration of AA over a range of 0.05 to 1 µM. The limit of detection for AA was found to be 17 nM. Importantly, the probe was successfully used to detect AA in living cells. Therefore, this high sensitivity and selectivity strategy could directly survey the AA levels in real samples.
Assuntos
Ácido Ascórbico , Pontos Quânticos , Cobalto , Fluorescência , Corantes Fluorescentes , Imidas , Limite de Detecção , Óxidos , Perileno/análogos & derivadosRESUMO
An ultrasensitive electrochemical sensor has been constructed for the detection of single nucleotide polymorphisms (SNPs) based on DNA-functionalized Cd-MOFs-74 as cascade signal amplification probe under enzyme-free conditions. Interestingly, the introduction of an auxiliary probe did not disturb the detection of SNP targets, but could bind more Cd-MOFs-74 signal elements to enhance the different pulse voltammetry electrochemical signal 2~3 times as compared to sensing system without auxiliary probe, which obviously improves the sensitivity of the proposed sensor. Experimental results taking p53 tumor suppressor gene as SNP model demonstrated that the proposed method can be employed to sensitively and selectively detect target p53 gene fragment with a linear response ranging from 0.01 to 30 pmol/L (detection limit of 6.3 fmol/L) under enzyme-free conditions. Utilizing this strategy, the ultrasensitive SNP electrochemical sensor is a promising tool for the determination of SNPs in biomedicine. Graphical Abstract.
Assuntos
Cádmio/química , DNA/química , Ouro/química , Grafite/química , Nanopartículas Metálicas/química , Estruturas Metalorgânicas/química , Sequência de Bases , Quitosana/química , Técnicas Eletroquímicas , Genes p53 , Limite de Detecção , Técnicas de Amplificação de Ácido Nucleico , Polimorfismo de Nucleotídeo Único , Sensibilidade e EspecificidadeRESUMO
An ultrasensitive and nonenzymatic electrochemical sandwich-type immunoassay using covalent organic framework (COF-LZU1) material applied as a fixed matrix was developed for the determination of C-reactive protein (CRP). COFs with large specific surface area, good conductivity and stability were employed for functionalisation of the surface. Au nanoparticles were loaded on COF-LZUl to immobilise the CRP antibody (anti-CRP) on the surface of a glassy carbon electrode. Microwave method was employed for the synthesis of the Pt/Ru/C nanoparticles to imitate the protein enzyme with high catalytic activity. The as-synthesised activated carbon-supported bimetallic Pt/Ru/C nanoparticle composite was used to label secondary CRP antibody because it exhibited excellent catalytic behaviour toward hydrogen peroxide. After incubation of CRP, Pt/Ru/C-labelled anti-CRP was combined with CRP through specific antibody-antigen recognition process. The reduction current of H202 at - 0.2 V catalysing by tag Pt/Ru/C as measured by a chronoamperometric method is proportional to the concentration of CRP. Under optimal experimental conditions, employing chronoamperometry to investigate the CRP, the obtained linear range was 0.2 to 20 ng/mL with a detection limit of 0.1 ng/mL. This immunosensor provides an attractive platform for the applicability of COF-LZU1 materials and Pt/Ru/C nanoparticles in electrochemical assays. Graphical abstract An ultrasensitive and nonenzymatic electrochemical immunoassay using covalent organic frameworks (COF-LZU1) material as the fixed matrix was developed for the detection of C-reactive protein (CRP). Microwave method was employed to synthesis the bimetallic metal composites Pt/Ru/C nanoparticles, which exhibited excellent catalytic behavior toward small molecules H2O2. COFs with large specific surface area, good conductivity and stability were employed for surface functionalization. Our proposed biosensor is highly sensitive, with the detection limit of 0.1 ng/mL.
Assuntos
Proteína C-Reativa/análise , Técnicas Eletroquímicas/métodos , Imunoensaio/métodos , Nanopartículas Metálicas/química , Estruturas Metalorgânicas/química , Anticorpos Imobilizados/imunologia , Anticorpos Monoclonais/imunologia , Proteína C-Reativa/imunologia , Carbono/química , Limite de Detecção , Platina/química , Rutênio/químicaRESUMO
A novel and simple electrochemical immunoassay for C-reactive protein was developed using metal-organic frameworks (Au-MOFs) as signal unit. In this study, we found MOFs could be used as signal probe. And this new class of signal probe differs from traditional probe. The signal of the copper ions (Cu2+) from MOFs could be directly detected without acid dissolution and preconcentration, which would greatly simplify the detection steps and reduce the detection time. Moreover, MOFs contain large amounts of Cu2+ ions, providing high electrochemical signals. Our report represents the first example of using MOFs themselves as electrochemical signal probe for biosensors. Platinum nanoparticle modified covalent organic frameworks (Pt-COFs) with high electronic conductivity was employed as the substrate, which is the first time demonstrating the use of Pt-COFs for electrochemical immunoassay. Under the optimized experimental conditions, the proposed sensing strategy provides a linear dynamic ranging from 1 to 400 ng/mL. A detection limit of 0.2 ng/mL was obtained, indicating an improved analytical performance. With these merits, this stable, simple, low-cost, sensitive and selective electrochemical immunoassay shows promise for applications in the point-of-care diagnostics of dieses and environmental monitoring.
Assuntos
Proteína C-Reativa/análise , Técnicas Eletroquímicas/métodos , Imunoensaio/métodos , Estruturas Metalorgânicas/química , Nanoestruturas/química , Técnicas Biossensoriais/métodos , Cobre/química , Transporte de Elétrons , Ouro/química , Humanos , Limite de Detecção , Nanoestruturas/ultraestrutura , Platina/químicaRESUMO
The monitoring of acetylcholinesterase (AChE) activity and the screening of its inhibitors are significance of the diagnosis and drug therapy of nervous diseases. A metal ions-mediated signal amplification strategy was developed for the highly sensitive and multicolor assay of AChE activity and visually screening its drug inhibitors. After the specific reaction between AChE and acetylthiocholine (ATCh), the hydrolysis product thiocholine (TCh) can directly and decompose the α-FeOOH nanorods (NRs) to release amounts of Fe2+, which was regarded as Fenton reagent to efficiently catalyze H2O2 to produce ·OH. Then, the as-formed ·OH can further largely shorten the gold nanobipyramids (Au NBPs), generating a series of palpable color variations. The linear range for AChE activity was 0.01-500.0 U/L with the limit of detection as low as 0.0074 U/L. The vivid visual effects could be easily distinguished for the multicolor assay of AChE activity by naked eye in visible light. To achieve the point-of-care testing, Au NBPs were further assembled on polymeric electrospun nanofibrous films (ENFs) surface as test strips for the easy-to-use test of AChE activity by RGB values with a smartphone. Fascinatingly, this proposed strategy can be used for the visual screening AChE inhibitors or non-inhibitors. Comparing with the clinical drugs (rivastigmine tartrate, and donepezil), some natural alkaloids such as evodiamine, caffeine, camptothecin, and berberine hydrochloride were selected as inhibitor modes to confirm the drug screening capability of this method. This proposed strategy may have great potential in the other disease-related enzymatic biomarkers assay and the rapid screening of drug therapy.
Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Acetilcolinesterase , Peróxido de Hidrogênio , Avaliação Pré-Clínica de Medicamentos/métodos , Técnicas Biossensoriais/métodos , Inibidores da Colinesterase/farmacologia , Inibidores da Colinesterase/análise , Testes ImediatosRESUMO
The construction of efficient methods for highly sensitive and rapid detection of disease markers is essential for the early diagnosis of serious diseases. In this paper, taking advantage of the UiO-66-NH2 signal molecule in combination with a waste-free entropy-driven DNA machine, a novel homogeneous electrochemical ratiometric platform is developed to detect MircoRNA (miRNA). Metal-organic framework materials (UiO-66-NH2 MOF) and ferrocene were utilized as electrochemical signal tags and reference probes, respectively. The target-initiated waste-free three-dimensional (3D) entropy-driven DNA nanomachine is activated in the presence of miRNA, resulting in DNA-labeled-UiO-66-NH2 falling off from the electrode, leading to a decrease in the signal of UiO-66-NH2 at 0.83V. Our strategy can mitigate false positive responses induced by the DNA probes immobilized on electrodes in traditional distance-dependent signal adjustment ratiometric strategies. The proposed ratiometric platform demonstrates superior sensitivity (a detection limit of 9.8 fM), simplified operation, high selectivity, and high repeatability. The ratiometric biosensor is also applied to detect miRNA content in spiked serum samples.
Assuntos
Técnicas Biossensoriais , Técnicas Eletroquímicas , Entropia , Estruturas Metalorgânicas , MicroRNAs , MicroRNAs/sangue , MicroRNAs/análise , Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , Humanos , Estruturas Metalorgânicas/química , DNA/química , Limite de Detecção , Eletrodos , Sondas de DNA/química , Sondas de DNA/genética , Compostos Ferrosos/química , Metalocenos/químicaRESUMO
Brain ageing leads to atrophy and degeneration of the cholinergic nervous system, resulting in profound neurobehavioral and cognitive dysfunction from decreased acetylcholine biosynthesis and reduced secretion of growth and neurotrophic factors. Human adipose tissue-derived mesenchymal stem cells (ADMSCs) were intravenously (1 × 10(6) cells) or intracerebroventricularly (4 × 10(5) cells) transplanted into the brains of 18-month-old mice once or four times at 2-week intervals. Transplantation of ADMSCs improved both locomotor activity and cognitive function in the aged animals, in parallel with recovery of acetylcholine levels in brain tissues. Transplanted cells differentiated into neurons and, in part, into astrocytes and produced choline acetyltransferase proteins. Transplantation of ADMSCs restored microtubule-associated protein 2 in brain tissue and enhanced Trk B expression and the concentrations of brain-derived neurotrophic factor and nerve growth factor. These results indicate that human ADMSCs differentiate into neural cells in the brain microenvironment and can restore physical and cognitive functions of aged mice not only by increasing acetylcholine synthesis but also by restoring neuronal integrity that may be mediated by growth/neurotrophic factors. © 2013 Wiley Periodicals, Inc.
Assuntos
Tecido Adiposo/citologia , Envelhecimento/fisiologia , Transtornos Cognitivos/cirurgia , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/fisiologia , Atividade Motora/fisiologia , Acetilcolina/metabolismo , Animais , Aprendizagem da Esquiva/fisiologia , Encéfalo/metabolismo , Encéfalo/patologia , Contagem de Células , Diferenciação Celular/fisiologia , Colina O-Acetiltransferase/metabolismo , Transtornos Cognitivos/etiologia , Transtornos Cognitivos/fisiopatologia , Modelos Animais de Doenças , Regulação da Expressão Gênica/fisiologia , Humanos , Masculino , Aprendizagem em Labirinto/fisiologia , Camundongos , Camundongos Endogâmicos ICR , Proteínas do Tecido Nervoso/metabolismo , Fatores de TempoRESUMO
Ochratoxin A (OTA) is a naturally occurring mycotoxin that poses serious threats, such as kidney damage, to human health. Therefore, we developed a DNA walker-based dual-signal electrochemical ratiometric platform for OTA detection, which could overlook the variations in environmental and instrumental factors and DNA load densities. Cobalt metal-organic frameworks (Co-MOFs) and toluidine blue were used as the electrochemical signal tag and internal reference probe, respectively. In the presence of OTA, this developed machine resulted in the DNA labelled-Co-MOFs far away from the electrode. Thus, Co-MOFs signal at -1.18 V decreased, while toluidine blue at -0.28 V increased. This proposed strategy has displayed superior sensitivity (limit of detection = 0.31 fg/mL, linear range = 1-50 ng/mL) and high reproducibility. The sensor was also applied for determining OTA content in red wine samples and the results were comparable to those of commercial enzyme-linked immunoassay kits with satisfactory results.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Estruturas Metalorgânicas , Ocratoxinas , Humanos , Aptâmeros de Nucleotídeos/genética , Aptâmeros de Nucleotídeos/química , Técnicas Eletroquímicas/métodos , Técnicas Biossensoriais/métodos , Cobalto , Cloreto de Tolônio , Reprodutibilidade dos Testes , Ocratoxinas/análise , DNA/química , Limite de Detecção , Ouro/químicaRESUMO
Precise and sensitive microRNA (miRNA) analysis is very significant for early disease diagnosis. In this work, a dual DNA nanomachines-based homogeneous electrochemical biosensor was constructed for the sensitively ratiometric detection of miRNA by a nicking enzyme (Nt.AlwI)-assisted cycling signal amplification strategy. The Co-based metal organic frameworks (Co-MOFs) and toluidine blue (TB) were employed as signal probes and internal reference probes, respectively. The introduction of internal reference probes can actually calibrate the interferent factors of the analytical system to improve the stability in detection procedure. In addition, with the help of the magnetic separation technique, the homogeneous electrochemical biosensor provides a more simpler way for the development of immobilization-free electrochemical miRNA biosensors, avoiding the complex modification procedure of traditional electrochemical biosensing interfaces. Consequently, taking advantages of this proposed dual DNA nanomachines-based homogeneous electrochemical biosensor, the highly sensitive and selective detection of miRNA-141 as model could be accomplished in ranging from 1 fM to 10 nM with detection limit of 0.46 fM. This strategy exhited good sensitivity and stability to integrate the nicking enzyme-powered dual DNA nanomachines with the ratiometric electrochemical output modes, which open new opportunities for the sensitive and reliable diagnosis of miRNA-related diseases.
Assuntos
Técnicas Biossensoriais , Estruturas Metalorgânicas , MicroRNAs , MicroRNAs/análise , Técnicas Eletroquímicas/métodos , DNA/genética , Técnicas Biossensoriais/métodos , Limite de DetecçãoRESUMO
PURPOSE: The objective was to confirm the anti-obesity activity of a silk peptide (SP) and a silkworm pupa peptide (SPP) in rats fed a high-fat diet (HFD) and to elucidate their action mechanism(s) in a preadipocyte culture system. METHODS: In an in vitro mechanistic study, the differentiation and maturation of 3T3-L1 preadipocytes were stimulated with insulin (5 µg/mL), and effects of SP and SPP on the adipogenesis of mature adipocytes were assessed. In an in vivo anti-obesity study, male C57BL/6 mice were fed an HFD containing SP or SPP (0.3, 1.0, or 3.0%) for 8 weeks, and blood and tissue parameters of obesity were analyzed. RESULTS: Hormonal stimulation of preadipocytes led to a 50-70% increase in adipogenesis. Polymerase chain reaction and Western blot analyses revealed increases in adipogenesis-specific genes (leptin and Acrp30) and proteins (peroxisome proliferator-activated receptor-γ and Acrp30). The hormone-induced adipogenesis and activated gene expression was substantially inhibited by treatment with SP and SPP (1-50 µg/mL). The HFD markedly increased body weight gain by increasing the weight of epididymal and mesenteric fat. Body and fat weights were significantly reduced by SP and SPP, in which decreases in the area of abdominal adipose tissue and the size of epididymal adipocytes were confirmed by magnetic resonance imaging and microscopic examination, respectively. Long-term HFD caused hepatic lipid accumulation and increased blood triglycerides and cholesterol, in addition to their regulatory factors Acrp30 and leptin. However, SP and SPP recovered the concentrations of Acrp30 and leptin, and attenuated steatosis. CONCLUSIONS: SP and SPP inhibit the differentiation of preadipocytes and adipogenesis by modulating signal transduction pathways and improve HFD-induced obesity by reducing lipid accumulation and the size of adipocytes.
Assuntos
Adipócitos/efeitos dos fármacos , Adipogenia/efeitos dos fármacos , Bombyx/química , Proteínas de Insetos/farmacologia , Peptídeos/farmacologia , Seda/química , Células 3T3-L1 , Adipócitos/metabolismo , Adipogenia/genética , Adiponectina/genética , Adiponectina/metabolismo , Animais , Fármacos Antiobesidade/farmacologia , Peso Corporal/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Dieta Hiperlipídica , Insulina/sangue , Leptina/genética , Leptina/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Obesidade/prevenção & controle , PPAR gama/genética , PPAR gama/metabolismo , Pupa/químicaRESUMO
Intelligently walking DNA nanomachines have sprung up an upsurge in various nucleic acid testing, but the rapid and sensitive test methods toward disease biomarker proteins based on the signal amplification strategy of DNA nanomachines were still ongoing development. In this work, an electrochemical aptasensor coupling the magnetic separation technique with the nuclease-powered walking DNA nanomachine was established for Mucin 1 (MUC1) detection. The magnetic beads (MBs) were modified by MUC1 aptamer hybridized with blocker DNA probe (BDP). After reacting with MUC1 proteins, the BDP was released from MBs to trigger the opening of capture hairpin DNA on Au nanoparticle (Au NPs)/MXene-modified electrode surface. In the presence of exonuclease III (Exo III), the BDP as a DNA walker is activated to autonomously move on the electrode. Then, lots of residual DNA fragments can still stay on electrode, further hybridizing with hairpin DNA, which can capture more UiO-66-NH2 metal-organic frameworks (MOFs). The amounts of ligands in MOFs can generate enhanced differential pulse voltammetry (DPV) signal probes. Furthermore, the concentrations of MUC1 can convert into the amplified DPV signals by introducing the signal amplification between the BDP as DNA walkers and Exo III as driven forces. This proposed electrochemical aptasensor achieved MUC1 detection ranging from 5 pg/mL to 50 ng/mL with detection limit of 0.72 pg/mL. Consequently, the designed and nuclease-powered walking DNA nanomachine provided an efficient strategy for the quantitative analysis of proteins by the interconversion between protein and BDP as a walker, which exhibited practical applicability of MUC1 detection in human serum.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Nanopartículas Metálicas , Estruturas Metalorgânicas , Aptâmeros de Nucleotídeos/metabolismo , Técnicas Biossensoriais/métodos , DNA , Sondas de DNA/genética , Técnicas Eletroquímicas/métodos , Endonucleases , Ouro , Humanos , Mucina-1/análise , Ácidos FtálicosRESUMO
A visual colorimetric aptasensor combining platinum nanoparticles (Pt NPs) loaded on Fe-based metal organic gel (Fe-MOG) as peroxidase mimics with magnetic separation technique was designed for the detection of fumonisin B1 (FB1). Interestingly, based on the inherent properties of Fe-MOG such as porous nanostructures, abundant Fe2+/Fe3+ active metal centers and synergetic effect between Pt NPs and Fe-MOG, Pt NPs/Fe-MOG composites can be employed as excellent peroxidase mimetic enzyme to oxidize 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of H2O2. This process was accompanied by a color variation of the solution from colorless to blue. Kinetic analysis showed that the Pt NPs/Fe-MOG composites make an effective peroxidase mimic with low Michaelis constant (Km), high substrate binding affinity and rapid catalytic velocity (Vm). Furthermore, the Pt NPs/Fe-MOG based colorimetric aptasensor was used to detect FB1. The prepared Pt NPs/Fe-MOG achieved lower detection limit (2.7 pg/mL, 3σ/k) with a wide linear range of 0.01-2000.0 ng/mL. Finally, as a proof of concept of the rapidity and the simplicity of this colorimetric aptasensor, Pt NPs/Fe-MOG was used for the detection of FB1 in real corn samples.
Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Colorimetria/métodos , Fumonisinas , Géis , Peróxido de Hidrogênio/química , Ferro/química , Cinética , Nanopartículas Metálicas/química , Oxirredutases/metabolismo , Peroxidase/química , Peroxidases/metabolismo , Platina/química , Zea maysRESUMO
The concentration of tumor biomarker Mucin 1 (MUC 1) is highly related with many diseases, which can be employed for the early diagnosis of cancer. In this paper, an electrochemical ratiometric aptasensor with intrinsic self-calibration property for the detection of MUC 1 is presented. In this paper, Co-MOFs themselves were employed as signal substances. This strategy was fabricated by using gold nanoparticles@black phosphorus (BP) as the substrate on the electrode, followed by modification of DNA nanotetrahedrons (DTN) via Au-S bond. The terminal of DTN contains MUC 1 aptamer. In the presence of MUC 1, the signal of DNA-labeled Co-MOFs can be detected. The current signal of Co-MOFs increased and that of thionine (as reference) was unchanged upon the addition of MUC 1. Thus, an intrinsic self-calibration aptasensor was achieved. In order to simplify the modification procedure, the electrolyte solution thionine was employed as an inner reference probe. Moreover, coupling of the hybridization chain reaction (HCR) with these MOFs signal tags presents an enzyme-free method for signal amplification, endowing the proposed ratiometric biosensor detection with high reproducibility and high sensitivity. The current ratio (IIR/ISP) remained stable over 30 individual measurements performed on ten different working electrodes. Even ten repeated scans performed on a single electrode exhibited a constant current ratio. The electrochemical ratiometric aptasensor is highly sensitivity for MUC 1 with the detection limit of 1.34 fM. Our proposed ratiometric sensor has great potential for the detection of cancer-related biomarkers.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Nanopartículas Metálicas , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Calibragem , DNA/química , Técnicas Eletroquímicas/métodos , Ouro/química , Limite de Detecção , Nanopartículas Metálicas/química , Mucina-1 , Reprodutibilidade dos TestesRESUMO
BACKGROUND: Since cyclophosphamide is metabolically activated to teratogenic acrolein and cytotoxic phosphoramide mustard by cytochrome P-450 type 2B (CYP2B), we assessed the effects of licorice, a CYP2B inducer, on the fetal defects induced by cyclophosphamide. METHODS: Pregnant Sprague-Dawley rats were daily administered with licorice (100 mg/kg) by gavage for 7 days, from the 6th to 12th day of gestation, and intraperitoneally administered with cyclophosphamide (11 mg/kg) 1 hr after the final licorice treatment. On the 20th day of gestation, maternal and fetal abnormalities were determined by Cesarian section. RESULTS: Cyclophosphamide was found to reduce fetal and placental weights without increasing resorption or death. In addition, it induced malformations in live fetuses; 93.8, 41.1, and 100% of the external (skull and limb defects), visceral (cleft palate and ureteric dilatation), and skeletal (acrania, vertebral/costal malformations, and delayed ossification) abnormalities, respectively. When pre-treated with licorice, cyclophosphamide-induced body weight loss and abnormalities of fetuses were remarkably aggravated. Moreover, repeated treatment with licorice greatly increased mRNA expression and activity of hepatic CYP2B. CONCLUSIONS: The results indicate that repeated intake of licorice may aggravate cyclophosphamide-induced body weight loss and malformations of fetuses by upregulating CYP2B.
Assuntos
Antineoplásicos Alquilantes/toxicidade , Ciclofosfamida/toxicidade , Citocromo P-450 CYP2B1/genética , Glycyrrhiza/química , Extratos Vegetais/toxicidade , RNA Mensageiro/efeitos dos fármacos , Teratogênicos/toxicidade , Anormalidades Induzidas por Medicamentos , Animais , Peso Corporal/efeitos dos fármacos , Citocromo P-450 CYP2B1/metabolismo , Feminino , Desenvolvimento Fetal/efeitos dos fármacos , Reação em Cadeia da Polimerase , Gravidez , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , Regulação para CimaRESUMO
The analytical methods based on colorimetric detection of various analytes have attracted intensive interest. However, most of them display relatively low sensitivity. Herein, a novel colorimetric strategy based on iodide/metal-organic frameworks (MOF)-mediated amplification was developed for low-cost, naked-eye detection and quantification of H2O2,Cr2O72-, and H2S. Cu-MOFs could catalyze the oxidation of the colorless peroxidase substrate TMB to produce a blue product. The published researches mainly focused on the immobilization or integration of a macromolecule, such as natural enzymes, to enhance MOFs catalytic abilities. The use of small molecules to improve the catalytic performance of MOFs has rarely reported. Due to the negligible steric hindrance, iodide could easily be adsorbed in the framework pore of MOFs to conduct the synergic catalytic effect, and shows a high catalytic effect. As a result, the catalytic activity of Cu-MOFs was dramatically enhanced, and thus, the nanocatalyst could act as an amplifier system for target detection. The detection limits obtained by the amplified method are 25, 30, and 0.2 nM, respectively, which are about 200-fold lower than that of the unamplified colorimetric assays. The colorimetric strategy developed herein provides a novel system for the detection of low concentrations of analytes in complex biological samples.
Assuntos
Colorimetria , Estruturas Metalorgânicas , Peróxido de Hidrogênio , Iodetos , PeroxidasesRESUMO
In this paper, a simple, rapid, and low-cost colorimetric method was designed based on Co/Fe-MOFs-iodide composite for the quantitative detection of H2S. It is know that iodide can improve the catalytic activity of bimetallic porous material Co/Fe-MOFs via adsorption into the framework of MOFs. Herein, we demonstrate a novel strategy to enhance the peroxidase-like activity of MOFs. Compared to horseradish peroxidase (HRP), the kinetic measurement results show that Co/Fe-MOFs-iodide exhibits excellent affinity to substrates, promoting electron transfer. Due to the synergetic effect of Co/Fe-MOFs and iodide, and rapid electron transfer process, Co/Fe-MOFs-iodide demonstrates improved peroxidase-like activity. As a proof-of-concept application, a novel, highly sensitive H2S colorimetric method is established with a detection limit (LOD) of 0.33 nM. In the absence of iodide, LOD is approx. 200-fold higher than that of the amplified colorimetric assay. The proposed method can also accurately detect traces of H2S in serum samples.