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A novel molecularly imprinted nanomaterial (Eu (BTC)-MPS@MIP) was synthesized on the surface of silanized europium-based metal-organic frameworks (Eu (BTC)-MPS) using 1, 3, 5-benzotrioic acid (H3BTC) as a ligand. The resulting Eu (BTC)-MPS@MIP was applied to constructing a smartphone sensing platform for the sensitive and selective detection of clothianidin (CLT) in vegetables. The synthesized Eu (BTC)-MPS@MIP demonstrated the successful formation of a typical core-shell structure featuring a shell thickness of approximately 70 - 80 nm. The developed sensing platform based on Eu (BTC)-MPS@MIP exhibited sensitivity in CLT detection with a detection limit of 4 µg/L and a linear response in the range 0.01 - 10 mg/L at excitation and emission wavelengths of 365 nm and 617 nm, respectively. The fluorescence sensing platform displayed excellent specificity for CLT detection, as evidenced by a high imprinting factor of 3.1. This specificity is primarily attributed to the recognition sites in the molecularly imprinted polymer (MIP) layer. When applied to spiked vegetable samples, the recovery of CLT ranged from 78.9 to 102.0%, with relative standard deviation (RSD) values falling between 2.2 and 6.2%. The quenching mechanism of Eu (BTC)-MPS@MIP toward CLT can be attributed to the inner filter effect (IFE), resulting from the optimal spectral overlap between the absorption spectrum of CLT and the excitation spectra of Eu (BTC)-MPS@MIP. The proposed method has the potential for extension to the detection of other pesticides by replacing the MIP recognition probes.
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Lead contamination is a major concern in food safety and, as such, many lead detection methods have been developed, especially aptamer-based biosensors. However, the sensitivity and environmental tolerance of these sensors require improvement. A combination of different types of recognition elements is an effective way to improve the detection sensitivity and environmental tolerance of biosensors. Here, we provide a novel recognition element, an aptamer-peptide conjugate (APC), to achieve enhanced affinity of Pb2+. The APC was synthesized from Pb2+ aptamers and peptides through clicking chemistry. The binding performance and environmental tolerance of APC with Pb2+ was studied through isothermal titration calorimetry (ITC); the binding constant (Ka) was 1.76*106 M-1, indicating that the APC's affinity was increased by 62.96% and 802.56% compared with the aptamers and peptides, respectively. Besides, APC demonstrated better anti-interference (K+) than aptamer and peptide. Through the molecular dynamics (MD) simulation, we found that more binding sites and stronger binding energy between APC with Pb2+are the reasons for higher affinity between APC with Pb2+. Finally, a carboxyfluorescein (FAM)-labeled APC fluorescent probe was synthesized and a fluorescent detection method for Pb2+ was established. The limit of detection of the FAM-APC probe was calculated to be 12.45 nM. This detection method was also applied to the swimming crab and showed great potential in real food matrix detection.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Chumbo , Aptâmeros de Nucleotídeos/química , Limite de Detecção , Corantes Fluorescentes/química , Técnicas Biossensoriais/métodosRESUMO
A turn-on fluorescent aptasensor based on a paper-based microfluidic chip was developed to detect arsenite via aptamer competition strategy and smartphone imaging. The chip was prepared by wax-printing hydrophilic channels on filter paper. It is portable, low-cost, and environmentally friendly. Double-stranded DNA consisting of aptamer and fluorescence-labeled complementary strands was immobilized on the reaction zone of the paper chip. Due to the specific strong binding between aptamer and arsenite, the fluorescent complementary strand was squeezed out and driven by capillary force to the detection area of the paper chip, so that the fluorescent signal arose in the detection area under the excitation wavelength of 488 nm. Arsenite can be quantified by using smartphone imaging and RGB image analysis. Under the optimal conditions, the paper-based microfluidic aptasensor exhibited excellent linear response over a wide range of 1 to 1000 nM, with a detection limit as low as 0.96 nM (3σ).
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Arsenitos , Smartphone , Oligonucleotídeos , Corantes , Dispositivos Lab-On-A-ChipRESUMO
Due to the threat to health of heavy metal contamination, simple and rapid detection methods for heavy metals are an urgent needed in environment protection and food safety. In this work, we have developed a fluorescent aptasensor for the 'turn-off' model detection of Pb2+ . The key feature of the aptasensor is that the dye-labelled nucleic acid strand can be folded into a G-quadruplex structure in the presence of Pb2+ . This conformational change induces photoinduced electron transfer (PET) between a G-quadruplex-hemin complex and 6-carboxyrhodamine X (ROX), which results in fluorescence quenching of ROX. We systematically investigated the interaction mechanism between Pb2+ and the aptasensor and the effects of several environmental parameters were also studied. Under the optimum conditions, the proposed method exhibited a good liner relationship between the concentration of Pb2+ and fluorescence quenching efficiency in the range 25-500 nM and the limit of detection was 1.02 nM. In addition, this method also manifested good performance in spiked lettuce samples with satisfactory recoveries of 87.10-109.6%. This target-induced PET platform can be further expanded to other heavy metal analysis.
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Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Quadruplex G , Elétrons , Hemina , Chumbo , Limite de DetecçãoRESUMO
The improper conformation of oligonucleotides on gold nanoparticle surfaces is caused by unintended base adsorption, which hinders DNA hybridization and lowers colloidal stability. In this work, we treated spherical nucleic acids with Br- , which serves as an efficient backfilling agent, to adjust the DNA conformation by displacing bases from the gold surface. To investigate the effect of DNA conformation on interfacial recognition, a kanamycin fluorescent aptasensor was constructed with bromide backfilled-treated spherical nucleic acids. In the presence of kanamycin, the anchored aptamer binds with the target and the partially complementary reporter strand is dissociated from the surface of the gold nanoparticles, resulting in the fluorescence recovery of labelled fluorophore on the reporter strand. Under optimum conditions, the apparent binding affinity of the aptasensor with bromide backfilling was 2.2-fold that without backfilled one. The proposed aptasensor exhibited a good liner relationship between the concentration of kanamycin and fluorescence intensity change in the range 200 nM to 10 µM and the limit of detection was calculated to be 71.53 nM. Moreover, this aptasensor was also successfully applied in a spiked milk sample assay and the satisfactory recoveries were obtained in the range 96.94-101.57%, which demonstrated its potential in practical applications.
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Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Nanopartículas Metálicas , Ácidos Nucleicos , Animais , Canamicina/análise , Canamicina/química , Ouro/química , Brometos , Ácidos Nucleicos/análise , Nanopartículas Metálicas/química , Aptâmeros de Nucleotídeos/química , Leite/química , Conformação de Ácido Nucleico , Técnicas Biossensoriais/métodos , Limite de DetecçãoRESUMO
Kanamycin fluorescence aptasensors were created using a series of di-block oligonucleotide modified gold nanoparticles with various lengths of poly-adenine. In the presence of kanamycin, the double strand structure of the aptamer-reporter strand complex is disrupted, and the dye-labelled reporter strand detaches from the surface of gold nanoparticles, resulting in fluorescence recovery (Ex/Em = 485/520 nm). By adjusting the number of consecutive adenines, the programable aptamer density can be implemented on the gold nanoparticle surface, and the conformation of nucleic acid changed from lying-down to up-right. The apparent binding constant, binding kinetics, and limit of detection of the prepared aptasensors were carefully examined to explore the influence of surface density. Under the optimum condition, the aptasensor had a tenfold lower limit of detection than the thiolated aptamer modified one, as low as 23.6 nM, when a di-block oligonucleotide with twenty consecutive adenines tailed. In addition, satisfactory recoveries ranging from 96.33 to 99.47% were achieved in spiked milk samples with relative standard deviation of 1.2-6.9% (n = 3). This surface density regulation strategy holds great promise in other aptamer-based interfacial recognition and sensing. Schematic presentation of di-block oligonucleotide modified gold nanoparticle with different surface densities and its kanamycin sensing application.
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Nanopartículas Metálicas , Ácidos Nucleicos , Animais , Ouro/química , Canamicina/análise , Nanopartículas Metálicas/química , Leite/química , Ácidos Nucleicos/análise , Oligonucleotídeos/análise , Poli ARESUMO
Lead contamination in aquatic products is one of the main hazard factors. The aptasensor is a promising detection method for lead ion (Pb(II)) because of its selectivity, but it is easily affected by pH. The combination of ion-imprinted polymers(IIP) with aptamers may improve their stability in different pH conditions. This paper developed a novel electrochemical biosensor for Pb(II) detection by using aptamer-imprinted polymer as a recognition element. The glassy carbon electrode was modified with gold nanoparticles and aptamers. After the aptamer was induced by Pb(II) to form a G-quadruplex conformation, a chitosan-graphene oxide was electrodeposited and cross-linked with glutaraldehyde to form an imprint layer, improving the stability of the biosensor. Under the optimal experimental conditions, the current signal change (∆I) showed a linear correlation of the content of Pb(II) in the range of 0.1-2.0 µg/mL with a detection limit of 0.0796 µg/mL (S/N = 3). The biosensor also exhibited high selectivity for the determination of Pb(II) in the presence of other interfering metal ion. At the same time, the stability of the imprinted layer made the sensor applicable to the detection environment with a pH of 6.4-8.0. Moreover, the sensor was successfully applied to the detection of Pb(II) in mantis shrimp.
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Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Grafite , Nanopartículas Metálicas , Polímeros/química , Ouro/química , Chumbo , Grafite/química , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , Limite de Detecção , EletrodosRESUMO
A novel visual detection mode is proposed to improve the detection sensitivity for the determination of ochratoxin A (OTA). The mode is based on aptamer recognition and the signal amplification of rolling circle amplification (RCA) products self-assembled DNA hydrogel. Moreover, gold nanoparticles (AuNPs) were directly assembled inside the DNA hydrogel by adjusting the padlock probe sequences to achieve a stronger binding force between the DNA hydrogel and AuNPs; this avoids the need for modification of AuNPs with DNA sequences. In the presence of OTA, DNA hydrogel is formed. With higher concentrations of OTA, a larger amount of DNA hydrogel is formed. When AuNPs are added to the DNA hydrogel, AuNPs can be enclosed inside the DNA hydrogel. With more DNA hydrogel, there is less AuNPs in the supernatant. Thus, the absorbance of the supernatant is anti-correlated with the concentration of OTA. After optimization of the experimental conditions, the change in the absorbance of the supernatant was linearly correlated with the concentration of OTA, in the range 0.05 to 10 ng/mL; the limit of detection was 0.005 ng/mL. The good specificity of the developed biosensor was confirmed in the presence of other mycotoxins that are coexistent with or analogues of OTA. By comparing the developed method with the ELISA method, the accuracy and stability of this new method were also verified, with good performance obtained in real samples. Diagram of the principle of the colorimetric aptasensor for OTA detection based on rolling circle amplification product self-assembled DNA hydrogel.
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Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , DNA/química , Hidrogéis/química , Ocratoxinas/análise , Cerveja/análise , Colorimetria/métodos , Contaminação de Alimentos/análise , Ouro/química , Limite de Detecção , Nanopartículas Metálicas/química , Técnicas de Amplificação de Ácido Nucleico , Ocratoxinas/químicaRESUMO
Saxitoxin (STX) is a major marine toxin from shellfish, and it is responsible for paralytic shellfish poisoning (PSP). In this study, a highly sensitive and rapid aptamer assay was developed for STX detection by combining fluorescence resonance energy transfer (FRET) and nuclease-assisted target recycling signal amplification. The aptamer STX-41 conjugated with graphene quantum dots (GQDs) was adsorbed on magnetic reduced graphene oxide (MRGO) to establish a fluorescence quenching system. Then, the binding between STX and aptamer induced the desorption of GQD-aptamer from MRGO and the restoring of fluorescence for the fluorescent determination of STX. The digestion of the target bound aptamer by DNase I could release the target for recycling thus achieving signal amplification. Under the optimized conditions, the aptamer assay showed a wide detection range (0.1-100 ng·mL-1), low detection limit (LOD of 0.035 ng·mL-1), high specificity, good recovery (86.75-94.08% in STX-spiked clam samples) and repeatability (RSD of 4.27-7.34%). Combined with fluorescent detection technology, signal amplification technology, and magnetic separation technology, the proposed method can be used to detect STX in seafood products successfully.
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Técnicas Biossensoriais/métodos , Corantes Fluorescentes/química , Grafite/química , Pontos Quânticos/química , Saxitoxina/análise , Animais , Aptâmeros de Nucleotídeos/química , Bivalves/química , Endodesoxirribonucleases/química , Transferência Ressonante de Energia de Fluorescência/métodos , Contaminação de Alimentos/análise , Limite de Detecção , Fenômenos Magnéticos , Saxitoxina/químicaRESUMO
Four new indolyl diketopiperazines, aspamides A-E (1-4) and two new diketopiperazines, aspamides F-G (5-6), along with 11 known diketopiperazines and intermediates were isolated from the solid culture of Aspergillus versicolor, which is an endophyte with the sea crab (Chiromantes haematocheir). Further chiral high-performance liquid chromatography resolution gave enantiomers (+)- and (-)-4, respectively. The structures and absolute configurations of compounds 1-6 were determined by the comprehensive analyses of nuclear magnetic resonance (NMR), high-resolution mass spectrometry (HR-MS), and electronic circular dichroism (ECD) calculation. All isolated compounds were selected for the virtual screening on the coronavirus 3-chymoretpsin-like protease (Mpro) of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), and the docking scores of compounds 1-2, 5, 6, 8 and 17 were top among all screened molecules, may be helpful in fighting with Corona Virus Disease-19 (COVID-19) after further studies.
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Antivirais , Organismos Aquáticos/química , Aspergillus/química , Cisteína Endopeptidases/metabolismo , Dicetopiperazinas/química , Dicetopiperazinas/metabolismo , Proteínas não Estruturais Virais/metabolismo , Antivirais/química , Antivirais/metabolismo , Betacoronavirus/metabolismo , Cromatografia Líquida de Alta Pressão , Proteases 3C de Coronavírus , Cisteína Endopeptidases/química , Avaliação Pré-Clínica de Medicamentos , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Simulação de Acoplamento Molecular , SARS-CoV-2 , Estereoisomerismo , Interface Usuário-Computador , Proteínas não Estruturais Virais/químicaRESUMO
The physiological processes and mechanisms of an arterial system are complex and subtle. Physics-based models have been proven to be a very useful tool to simulate actual physiological behavior of the arteries. The current physics-based models include high-dimensional models (2D and 3D models) and low-dimensional models (0D, 1D and tube-load models). High-dimensional models can describe the local hemodynamic information of arteries in detail. With regard to an exact model of the whole arterial system, a high-dimensional model is computationally impracticable since the complex geometry, viscosity or elastic properties and complex vectorial output need to be provided. For low-dimensional models, the structure, centerline and viscosity or elastic properties only need to be provided. Therefore, low-dimensional modeling with lower computational costs might be a more applicable approach to represent hemodynamic properties of the entire arterial system and these three types of low-dimensional models have been extensively used in the study of cardiovascular dynamics. In recent decades, application of physics-based models to estimate central aortic pressure has attracted increasing interest. However, to our best knowledge, there has been few review paper about reconstruction of central aortic pressure using these physics-based models. In this paper, three types of low-dimensional physical models (0D, 1D and tube-load models) of systemic arteries are reviewed, the application of three types of models on estimation of central aortic pressure is taken as an example to discuss their advantages and disadvantages, and the proper choice of models for specific researches and applications are advised.
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Aorta/fisiologia , Pressão Arterial , Fenômenos Biofísicos , Modelos Biológicos , HumanosRESUMO
Six new depsidones, curdepsidones B-G (1-6), were obtained from the marine-derived fungus Curvularia sp. IFB-Z10. Their planar structures were determined by comprehensive analysis of HRESIMS and 1D/2D-NMR data. The absolute configuration of curdepsidones B-C (1-2) were established by synergistic use of DFT/NMR (density functional theory/nuclear magnetic resonance) and TDDFT/ECD (time-dependent density functional theory/electronic circular dichroism) calculations. Partial isolated compounds were tested for their anti-inflammatory activities in Propionibacterium acnes-induced THP-1 cells. Curdepsidone C (2) displayed significant anti-inflammatory properties with an IC50 value of 7.47 ± 0.35 µM, and reduced the P. acnes-induced phosphorylation levels of JNK and ERK in a dose-dependent mechanism. The possible anti-inflammatory mechanism of 2 was also investigated by molecular docking.
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Anti-Inflamatórios/farmacologia , Depsídeos/química , Depsídeos/farmacologia , Lactonas/química , Lactonas/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Depsídeos/isolamento & purificação , Fungos , Humanos , Concentração Inibidora 50 , Lactonas/isolamento & purificação , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Simulação de Acoplamento Molecular , Estrutura Molecular , Propionibacterium acnes , Células THP-1/efeitos dos fármacosRESUMO
Food safety is a global health objective, and foodborne diseases represent a major crisis in health. Techniques that are simple and suitable for fast screening to detect and identify pathogenic factors in the food chain are vital to ensure food safety. At present, a variety of analytical methods have been reported for the detection of pathogenic agents. Whereas the sensitivity of detection and quantification are still important challenges, we expect major advances from new assay formats and synthetic bio-recognition elements, such as aptamers. Owing to the specific folding capability of aptamers in the presence of an analyte, aptasensors have substantially and successfully been exploited for the detection of a wide range of small and large molecules (e.g., toxins, antibiotics, heavy metals, bacteria, viruses) at very low concentrations. Here, we review the use of aptasensors for the development of highly sensitive and affordable detection tools for food analysis.
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Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Contaminação de Alimentos/análise , Inocuidade dos AlimentosRESUMO
A novel chemiluminescent aptasensor for the highly sensitive detection of chloramphenicol (CAP) in milk was successfully developed using biotinylated CAP aptamer-functionalized magnetic nanoparticles (MNPs) as capture probes and thiolated hybridized complementary strand-modified N-(4-aminobutyl)-N-ethylisoluminol (ABEI)-functionalized flower-like gold nanostructures (AuNFs) as signal probes. P-iodophenol (PIP) was also added to form an ABEI-H2O2-PIP steady-state chemiluminescence (CL) system. Based on a competitive format, the CL intensity was negatively correlated with the concentration of CAP in the range of 0.01-0.20 ng/mL and the detection limit was 0.01 ng/mL in buffer and 1 ng/mL in milk. The proposed method was successfully applied to measure CAP in milk samples and compared to a commercial ELISA method. The high sensitivity of AuNFs, excellent selectivity and stability of aptamers, and good overall stability of the chemiluminescent bioassay with magnetic separation make them a promising approach for the detection of small molecular illegal additives. Additionally, the high sensitivity, easy operation, and good reproducibility exhibited by the stable chemiluminescent bioassay demonstrate its applicability for the trace detection of CAP in applications, such as animal husbandry.
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Antibacterianos/análise , Aptâmeros de Nucleotídeos/química , Cloranfenicol/análise , Ouro/química , Luminol/análogos & derivados , Leite/química , Nanoestruturas/química , Animais , Técnicas Biossensoriais/métodos , Peróxido de Hidrogênio/química , Limite de Detecção , Medições Luminescentes/métodos , Luminol/química , Nanopartículas de Magnetita/química , Nanoestruturas/ultraestruturaRESUMO
The existing aptamers for cadmium (Cd2+), the common toxic heavy metal contaminant in food, cannot meet the requirements for detecting Cd2+ in rapid detection methods. In previous work, we found that coupling aptamer-peptide conjugates (APCs) with peptides and aptamers can provide a less disruptive method with a significantly improved affinity. Moreover, we found that the spatial conformation of aptamers and peptides is crucial for obtaining proper affinity in APC. Therefore, we describe a simple design strategy to obtain a series of APCs with different affinities by designing peptide orientations (N-terminal, C-terminal). The best affinity was found for APC(C1-N) with a binding constant (Ka) of 2.23 × 106 M-1, indicating that the APC(C1-N) affinity was significantly increased by 829.17% over aptamer. Finally, a rolling-circle amplification (RCA)-coupled ratio fluorescence-based biosensor for Cd2+ detection was established with a detection limit of 0.0036 nM, which has great potential for practical aquatic product detection.
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Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Corantes Fluorescentes/química , Cádmio , Aptâmeros de Nucleotídeos/química , Peptídeos , Técnicas Biossensoriais/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Limite de DetecçãoRESUMO
BACKGROUND AND OBJECTIVE: Diabetic foot (DF) complications often lead to severe vascular issues. This study investigated the effectiveness of enhanced external counterpulsation (EECP) and its derived innovative compression strategies in addressing poor perfusion in DF. Although developing non-invasive and efficient treatment methods for DF is critical, the hemodynamic alterations during EECP remain underexplored despite promising outcomes in microcirculation. This research sought to address this gap by developing a patient-specific 0D-1D model based on clinical ultrasound data to identify potentially superior compression strategies that could substantially enhance blood flow in patients with DF complications. METHODS: Data were gathered from 10 patients with DF utilizing ultrasound for blood flow rate and computed tomography angiography (CTA) to identify lower limb conditions. Clinical measurements during standard EECP, with varying cuff pressures, facilitated the creation of a patient-specific 0D-1D model through a two-step parameter estimation process. The accuracy of this model was verified via comparison with the clinical measurements. Four compression strategies were proposed and rigorously evaluated using this model: EECP-Simp-I (removing hip cuffs), EECP-Simp-II (further removing the cuffs around the lower leg), EECP-Impr-I (removing all cuffs around the affected side), and EECP-Impr-II (building a loop circulation from the healthy side to the affected side). RESULTS: The predicted results under the rest and standard EECP states were generally closely aligned with clinical measurements. The patient-specific 0D-1D model demonstrated that EECP-Simp-I and EECP-Impr-I contributed similar enhancement to perfusion in the dorsal artery (DA) and were comparable to standard EECP, while EECP-Simp-II had the least effect and EECP-Impr-II displayed the most significant enhancement. Pressure at the aortic root (AO) remained consistent across strategies. CONCLUSIONS: EECP-Simp-I is recommended for patients with DF, emphasizing device simplification. However, EECP-Simp-II is discouraged as it significantly diminished blood perfusion in this study, except in cases of limb fragility. EECP-Impr-II showed superior enhancement of blood perfusion in DA to all other strategies but required a more complex EECP device. Despite increased AO pressure in all the proposed compression strategies, safety could be guaranteed as the pressue remained within a safe range.
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Due to the threat of lead pollution to health, environmental and food safety, developing simple and fast detection methods is highly required. Whereas, traditional single-mode probe suffers from limited application scenario. In this study, a colorimetric and fluorometric dual-mode probe for Pb2+ determination was constructed by using bifunctional G-quadruplex-hemin complex. In this dual-mode probe, enzyme strand and substrate strand of 8-17 DNAzyme are labeled with G-quadruplex-hemin complex and fluorophore, respectively. In the absence of Pb2+, the self-assembly of enzyme strand and substrate strand inhibits intrinsic mimic peroxidase of G-quadruplex-hemin complex by base-pairing, which also quench the fluorescence via in proximity effect. When the DNAzyme is activated by Pb2+, the quenched fluorescence is restored as well as the inherent peroxidase mimetic activity, leading to dual signal output. Under optimal conditions, this dual-mode probe exhibit a good linear relationship between logarithm of Pb2+ concentration and signal difference within the range from 1.5 nM to 20 nM and 0.5 nM to 10 nM for colorimetric and fluorescence mode, respectively. The detection limits for the corresponding mode were estimated to be 1.29 nM and 0.16 nM, respectively. This dual-mode probe also successfully applied for the spiked river water assay with satisfactory recovery in the range of 93.2 %-115.3 %. This work paves a new way for DNAzyme based dual-mode probe construction.
Assuntos
Técnicas Biossensoriais , DNA Catalítico , Quadruplex G , Hemina , Chumbo , Colorimetria/métodos , Corantes , Peroxidases , Técnicas Biossensoriais/métodosRESUMO
BACKGROUND: Detection methods based on aptamer probes have great potential and progress in the field of rapid detection of heavy metal ions. However, the unstable conformation of aptamers often results in poor sensitivity due to the dissociation of aptamer-target complex in real environments. RESULTS: In this study, we developed a locking aptamer probe and combined it with AgInZnS quantum dots for the first time to detect cadmium ions. When cadmium ions are combined with the probe, the cadmium ions are fixed in the core-locking position, forming a stable cavity structure. The limit of detection (LOD) was achieved at a concentration of 6.9 nmol L-1, with a broad detection range from 10 nmol L-1 to 1000 µmol L-1, and good recovery rates (92.93%-102.8 %) were achieved in aquatic product testing. The locking aptamer probe with stable conformation effectively enhances the stability of the aptamer-target complex and remains good stability in four buffer environments as well as a 600 mmol L-1 salt solution; it also exhibits good stability at pH 6.5-7.5 and temperatures ranging from 25 °C to 35 °C. SIGNIFICANCE: Overall, our study presented a general, simple, and cost-effective strategy for stabilizing aptamer conformations, and used for highly sensitive detection of cadmium ions.
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Enhanced external counterpulsation (EECP) is a treatment and rehabilitation approach for ischemic diseases, including coronary artery disease. Its therapeutic benefits are primarily attributed to the improved blood circulation achieved through sequential mechanical compression of the lower extremities. However, despite the crucial role that hemodynamic effects in the lower extremity arteries play in determining the effectiveness of EECP treatment, most studies have focused on the diastole phase and ignored the systolic phase. In the present study, a novel siphon model (SM) was developed to investigate the interdependence of several hemodynamic parameters, including pulse wave velocity, femoral flow rate, the operation pressure of cuffs, and the mean blood flow changes in the femoral artery throughout EECP therapy. To verify the accuracy of the SM, we coupled the predicted afterload in the lower extremity arteries during deflation using SM with the 0D-1D patient-specific model. Finally, the simulation results were compared with clinical measurements obtained during EECP therapy to verify the applicability and accuracy of the SM, as well as the coupling method. The precision and reliability of the previously developed personalized approach were further affirmed in this study. The average waveform similarity coefficient between the simulation results and the clinical measurements during the rest state exceeded 90%. This work has the potential to enhance our understanding of the hemodynamic mechanisms involved in EECP treatment and provide valuable insights for clinical decision-making.
Assuntos
Contrapulsação , Análise de Onda de Pulso , Humanos , Reprodutibilidade dos Testes , Hemodinâmica , Extremidade Inferior , Contrapulsação/métodosRESUMO
Owing to advantage in high sensitivity and fast response, aptamer based electrochemical biosensors have attracted much more attention. However, inappropriate interfacial engineering strategy leads to poor recognition performance, which ascribe to the following factors of immobilized oligonucleotide strand including steric hindrance, interchain entanglement, and unfavorable conformation. In this work, we proposed a DNA tetrahedron based diblock aptamer immobilized strategy for the construction of label-free electrochemical biosensor. The diblock aptamer sequence is composite of T-rich anchor domain and recognition domain, where T-rich domain enabling anchored on the edge of DNA tetrahedron via Hoogsteen hydrogen bond at neutral condition. The DNA tetrahedron scaffold offers an appropriate lateral space for target recognition of diblock aptamer. More importantly, this trivalent aptamer recognition interface can be regenerated by simply adjusting the pH environment to alkaline, resulting in the dissociation of diblock aptamer. Under the optimum condition, proposed electrochemical aptasensor manifested a satisfied sensitivity for aminoglycosides antibiotic, kanamycin with a limit of detection of 0.69 nM, which is 45-fold lower than traditional Au-S immobilization strategy. Moreover, the proposed aptasensor had also successfully been extended to ampicillin detection by changing the sequence of recognition domain in diblock aptamer. This work paves a new way for the rational design of aptamer-based electrochemical sensor.