RESUMO
CLEC12A, a member of the C-type lectin receptor family involved in immune homeostasis, recognizes MSU crystals released from dying cells. However, the molecular mechanism underlying the CLEC12A-mediated recognition of MSU crystals remains unclear. Herein, we reported the crystal structure of the human CLEC12A-C-type lectin-like domain (CTLD) and identified a unique "basic patch" site on CLEC12A-CTLD that is necessary for the binding of MSU crystals. Meanwhile, we determined the interaction strength between CLEC12A-CTLD and MSU crystals using single-molecule force spectroscopy. Furthermore, we found that CLEC12A clusters at the cell membrane and seems to serve as an internalizing receptor of MSU crystals. Altogether, these findings provide mechanistic insights for understanding the molecular mechanisms underlying the interplay between CLEC12A and MSU crystals.
Assuntos
Lectinas Tipo C , Receptores Mitogênicos , Ácido Úrico , Humanos , Gota/metabolismo , Lectinas Tipo C/química , Lectinas Tipo C/imunologia , Receptores Mitogênicos/química , Receptores Mitogênicos/imunologia , Ácido Úrico/química , Ácido Úrico/imunologia , Domínios Proteicos , Cristalografia por Raios X , Imagem Individual de Molécula , Linhagem CelularRESUMO
C-type lectin receptors (CLRs) are a family of pattern recognition receptors, which detect a broad spectrum of ligands via small carbohydrate-recognition domains (CRDs). CLEC12A is an inhibitory CLR that recognizes crystalline structures such as monosodium urate crystals. CLEC12A also recognizes mycolic acid, a major component of mycobacterial cell walls, and suppresses host immune responses. Although CLEC12A could be a therapeutic target for mycobacterial infection, structural information on CLEC12A was not available. We report here the crystal structures of human CLEC12A (hCLEC12A) in ligand-free form and in complex with 50C1, its inhibitory antibody. 50C1 recognizes human-specific residues on the top face of hCLEC12A CRD. A comprehensive alanine scan demonstrated that the ligand-binding sites of mycolic acid and monosodium urate crystals may overlap with each other, suggesting that CLEC12A utilizes a common interface to recognize different types of ligands. Our results provide atomic insights into the blocking and ligand-recognition mechanisms of CLEC12A and leads to the design of CLR-specific inhibitors.
Assuntos
Lectinas Tipo C , Receptores Mitogênicos , Lectinas Tipo C/imunologia , Lectinas Tipo C/química , Lectinas Tipo C/metabolismo , Humanos , Receptores Mitogênicos/química , Receptores Mitogênicos/imunologia , Receptores Mitogênicos/metabolismo , Cristalografia por Raios X , Ligantes , Ligação Proteica , Sítios de Ligação , Modelos Moleculares , Ácido Úrico/química , Ácido Úrico/metabolismo , Ácido Úrico/imunologiaRESUMO
Uric acid is typically measured through blood tests, which can be inconvenient and uncomfortable for patients. Herein, we propose a wearable surface-enhanced Raman scattering (SERS) chip, incorporating a hydrogel membrane with integrated plasmonic trimers, for noninvasive monitoring of uric acid in sweat. The plasmonic trimers feature sub 5 nm nanogaps, generating strong electromagnetic fields to boost the Raman signal of surrounding molecules. Simultaneously, the hydrogel membrane pumps sweat through these gaps, efficiently capturing sweat biomarkers for SERS detection. The chip can achieve saturation adsorption of sweat within 5 min, eliminating variations in individual sweat production rates. Dynamic SERS tracking of uric acid and lactic acid levels during anaerobic exercise reveals a temporary suppression of uric acid metabolism, likely due to metabolic competition with lactic acid. Furthermore, long-term monitoring correlates well with blood test results, confirming that regular exercise helps reduce serum uric acid levels and supporting its potential in managing hyperuricemia.
Assuntos
Hidrogéis , Análise Espectral Raman , Suor , Ácido Úrico , Dispositivos Eletrônicos Vestíveis , Ácido Úrico/análise , Ácido Úrico/sangue , Ácido Úrico/química , Humanos , Suor/química , Análise Espectral Raman/métodos , Hidrogéis/química , Ácido Láctico/análise , Ácido Láctico/sangueRESUMO
We introduce the FlexoSERS sensor, which is notable for its high stretchability, sensitivity, and patternability. Featuring a hierarchically oriented jellyfish-like architecture constructed from stretchable gold nanowires, this sensor provides an ultrasensitive SERS signal even under 50% strain, with an enhancement factor (EF) of 3.3 × 1010. Impressively, this heightened performance remains consistently robust across 2,500 stretch-release cycles. The integration of nanowires with 3D-printed hydrogel enables a customizable FlexoSERS sensor, facilitating localized sweat collection and detection. The FlexoSERS sensor successfully detects and quantifies uric acid (UA) in both artificial and human sweat and functions as a pH sensor with repeatability and sensitivity across a pH range of 4.2-7.8, enabling real-time sweat monitoring during exercise. In summary, the rational architectural design, scalable fabrication process, and hydrogel integration collectively position this nanowire-based FlexoSERS sensor as a highly promising platform for customizable wearable sweat diagnostics.
Assuntos
Ouro , Nanofios , Suor , Dispositivos Eletrônicos Vestíveis , Ouro/química , Nanofios/química , Suor/química , Humanos , Técnicas Biossensoriais/instrumentação , Ácido Úrico/análise , Ácido Úrico/química , Hidrogéis/química , Concentração de Íons de HidrogênioRESUMO
The effect of strong metal-support interaction (SMSI) has never been systematically studied in the field of nanozyme-based catalysis before. Herein, by coupling two different Pd crystal facets with MnO2, i.e., (100) by Pd cube (Pdc) and (111) by Pd icosahedron (Pdi), we observed the reconstruction of Pd atomic structure within the Pd-MnO2 interface, with the reconstructed Pdc (100) facet more disordered than Pdi (111), verifying the existence of SMSI in such coupled system. The rearranged Pd atoms in the interface resulted in enhanced uricase-like catalytic activity, with Pdc@MnO2 demonstrating the best catalytic performance. Theoretical calculations suggested that a more disordered Pd interface led to stronger interactions with intermediates during the uricolytic process. In vitro cell experiments and in vivo therapy results demonstrated excellent biocompatibility, therapeutic effect, and biosafety for their potential hyperuricemia treatment. Our work provides a brand-new perspective for the design of highly efficient uricase-mimic catalysts.
Assuntos
Hiperuricemia , Compostos de Manganês , Óxidos , Urato Oxidase , Hiperuricemia/tratamento farmacológico , Urato Oxidase/química , Urato Oxidase/uso terapêutico , Urato Oxidase/metabolismo , Óxidos/química , Compostos de Manganês/química , Compostos de Manganês/farmacologia , Humanos , Paládio/química , Paládio/farmacologia , Animais , Catálise , Ácido Úrico/química , CamundongosRESUMO
Development of reliable methods for the detection of potential biomarkers is of the utmost importance for an early diagnosis of critical diseases and disorders. In this study, a novel lanthanide-functionalized carbon dot-based fluorescent probe Zn-CD@Eu is reported for the ratiometric detection of dipicolinic acid (DPA) and uric acid (UA). The Zn-CD@Eu nanoprobe was obtained from a simple room-temperature reaction of zinc-doped carbon dots (Zn-CD) and the EDTA-Eu lanthanide complex. Under optimal conditions, a good linear response was obtained for DPA in two concentration ranges of 0-55 and 55-100 µM with a limit of detection of 0.53 and 2.2 µM respectively, which is significantly below the infectious dosage of anthrax (â¼55 µM). Furthermore, the Zn-CD@Eu/DPA system was employed for the detection of UA with a detection limit of 0.36 µM in the linear range of 0-100 µM. The fluorescent probe was successfully implemented for determining DPA and UA in human blood serum, sweat, and natural water bodies with considerable recovery rates. In addition, the potential of the nanoprobe for ex vivo visualization of UA was demonstrated in fruit fly (Drosophila melanogaster) as a model organism.
Assuntos
Corantes Fluorescentes , Ácidos Picolínicos , Ácido Úrico , Zinco , Corantes Fluorescentes/química , Ácidos Picolínicos/análise , Ácidos Picolínicos/química , Ácido Úrico/análise , Ácido Úrico/química , Humanos , Zinco/química , Zinco/análise , Animais , Európio/química , Pontos Quânticos/química , Cádmio/análise , Cádmio/química , Carbono/química , Limite de Detecção , Imagem Óptica , Drosophila melanogasterRESUMO
Gout is a common inflammatory arthritis caused by precipitation of monosodium urate (MSU) crystals in individuals with hyperuricemia. Acute flares are accompanied by secretion of proinflammatory cytokines, including interleukin-1ß (IL-1ß). Clonal hematopoiesis of indeterminate potential (CHIP) is an age-related condition predisposing to hematologic cancers and cardiovascular disease. CHIP is associated with elevated IL-1ß, thus we investigated CHIP as a risk factor for gout. To test the clinical association between CHIP and gout, we analyzed whole exome sequencing data from 177 824 individuals in the MGB Biobank (MGBB) and UK Biobank (UKB). In both cohorts, the frequency of gout was higher among individuals with CHIP than without CHIP (MGBB, CHIP with variant allele fraction [VAF] ≥2%: odds ratio [OR], 1.69; 95% CI, 1.09-2.61; P = .0189; UKB, CHIP with VAF ≥10%: OR, 1.25; 95% CI, 1.05-1.50; P = .0133). Moreover, individuals with CHIP and a VAF ≥10% had an increased risk of incident gout (UKB: hazard ratio [HR], 1.28; 95% CI, 1.06-1.55; P = .0107). In murine models of gout pathogenesis, animals with Tet2 knockout hematopoietic cells had exaggerated IL-1ß secretion and paw edema upon administration of MSU crystals. Tet2 knockout macrophages elaborated higher levels of IL-1ß in response to MSU crystals in vitro, which was ameliorated through genetic and pharmacologic Nlrp3 inflammasome inhibition. These studies show that TET2-mutant CHIP is associated with an increased risk of gout in humans and that MSU crystals lead to elevated IL-1ß levels in Tet2 knockout murine models. We identify CHIP as an amplifier of NLRP3-dependent inflammatory responses to MSU crystals in patients with gout.
Assuntos
Dioxigenases , Gota , Animais , Hematopoiese Clonal , Proteínas de Ligação a DNA/genética , Dioxigenases/genética , Gota/genética , Humanos , Inflamassomos/genética , Interleucina-1beta/genética , Camundongos , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Ácido Úrico/química , Ácido Úrico/farmacologiaRESUMO
The exact prognostic role of cardiovascular (CV) risk factors in patients with BCR::ABL1 negative chronic myeloproliferative neoplasms (MPNs) remains unknown as it is often masked by other MPN-related features that bear strong prognostic impact on thrombotic risk. Therefore, current MPN treatment is not primarily guided by presence of CV risk factors. Treatment of CV risk factors in MPN patients usually mirrors that from the general population, despite the fact that CV risk factors in MPNs have their own specificities. Moreover, the optimal target levels for different metabolic deflections in MPNs (i.e., low-density lipoprotein, serum uric acid, or glycated hemoglobin levels) have not been defined. In the current review, we separately discuss the most important aspects of every individual CV risk factor (arterial hypertension, hyperlipidemia, chronic kidney disease, smoking, diabetes mellitus, hyperuricemia, and obesity and cachexia) in MPNs, summarize recent advances in the field, and propose future directions and research areas which may be needed to appropriately manage CV risk factors in MPNs.
Assuntos
Neoplasias da Medula Óssea , Doenças Cardiovasculares , Transtornos Mieloproliferativos , Humanos , Neoplasias da Medula Óssea/genética , Neoplasias da Medula Óssea/terapia , Doenças Cardiovasculares/epidemiologia , Doenças Cardiovasculares/etiologia , Transtornos Mieloproliferativos/genética , Transtornos Mieloproliferativos/terapia , Fatores de Risco , Ácido Úrico/sangue , Ácido Úrico/química , Proteínas de Fusão bcr-ablRESUMO
Herein, the electrochemical sensing efficacy of carboxylic acid functionalized multiwalled carbon nanotubes (C-MWCNT) intertwined with coexisting phases of gadolinium monosulfide (GdS) and gadolinium oxide (Gd2O3) nanosheets is explored for the first time. The nanocomposite demonstrated splendid specificity for nonenzymatic electrochemical detection of uric acid (UA) in biological samples. It was synthesized using the coprecipitation method and thoroughly characterized. The presence of functional groups and disorder in the as-synthesized nanocomposite are confirmed using Fourier transform infrared spectroscopy and Raman spectroscopy. Furthermore, field emission scanning electron microscopy, high-resolution transmission electron microscope, X-ray powder diffraction, and X-ray photoelectron spectroscopy provides a clear understanding of the morphology, coexisting phases, and elemental composition of the as-synthesized nanocomposites. The differential pulse voltammetry technique was utilized to elaborate the electrochemical sensing of UA using a GdS-Gd2O3/C-MWCNT modified glassy carbon electrode (GCE), The sensor showed an enhanced current response by more than 2-fold compared to bare GCE. Also, the sensor's performance was further improved by dispersing the nanocomposite in an ionic liquid with the exceptional reproducibility (SD = 0.0025, n = 3). The fabricated UA sensor GdS-Gd2O3/C-MWCNT/IL/GCE demonstrated a wide linear detection range from 0.5-30 µM and 30-2000 µM, effectively covering the entire physiological range of UA in biological fluids with a limit of detection (LOD) of 0.380 µM (+3SD of blank) and a sensitivity of 356.125 µA mM-1 cm-2. Moreover, the electrodes exhibited storage stability for 2 weeks with decrease in zero-day current by only 4.5%. The sensor was validated by quantifying UA in 12 unprocessed clinical human urine and serum samples, and its comparison with the gold standard test yielded remarkable results (p < 0.05). Hence, the proposed nonenzymatic electrochemical UA sensor is selective, sensitive, reproducible, and stable, making it reliable for point-of-care diagnostics.
Assuntos
Técnicas Eletroquímicas , Gadolínio , Nanotubos de Carbono , Ácido Úrico , Humanos , Nanotubos de Carbono/química , Ácido Úrico/sangue , Ácido Úrico/urina , Ácido Úrico/química , Técnicas Eletroquímicas/métodos , Técnicas Eletroquímicas/instrumentação , Gadolínio/química , Eletrodos , Nanocompostos/química , Limite de Detecção , Ácidos Carboxílicos/químicaRESUMO
This work presents the synthesis and characterization of an innovative F,S-doped carbon dots/CuONPs hybrid nanostructure obtained by a direct mixture between F,S-doped carbon dots obtained electrochemically and copper nitrate alcoholic solution. The hybrid nanostructures synthesized were characterized by absorption spectroscopy in the Ultraviolet region (UV-vis), high-resolution transmission electron microscopy (HRTEM), X-ray photoelectron spectroscopy (XPS), and different electrochemical techniques. The fluoride and sulfur-doped carbon dots/CuONPs nanostructures were used to prepare a non-enzymatic biosensor on a printed carbon electrode, exhibiting excellent electrocatalytic activity for the simultaneous determination of NADH, dopamine, and uric acid in the presence of ascorbic acid with a detection limit of 20, 80, and 400 nmol L-1, respectively. The non-enzymatic biosensors were also used to determine NADH, dopamine, and uric acid in plasma, and they did not suffer significant interference from each other.
Assuntos
Técnicas Biossensoriais , Carbono , Cobre , Dopamina , Técnicas Eletroquímicas , Limite de Detecção , NAD , Ácido Úrico , Ácido Úrico/sangue , Ácido Úrico/química , Técnicas Biossensoriais/métodos , Dopamina/sangue , Dopamina/análise , Carbono/química , NAD/química , NAD/sangue , Cobre/química , Técnicas Eletroquímicas/métodos , Humanos , Enxofre/química , Fluoretos/química , Pontos Quânticos/química , Nanoestruturas/química , EletrodosRESUMO
Metal nanozymes have offered attractive opportunities for biocatalysis and biomedicine. However, fabricating nanozymes simultaneously possessing highly catalytic selectivity and activity remains a great challenge due to the lack of three-dimensional (3D) architecture of the catalytic pocket in natural enzymes. Here, we integrate rhodium nanocluster (RhNC), reduced graphene oxide (rGO), and protamine (PRTM, a typical arginine-rich peptide) into a composite facilely based on the single peptide. Remarkably, the PRTM-RhNC@rGO composite displays outstanding selectivity, activity, and stability for the catalytic degradation of uric acid. The reaction rate constant of the uric acid oxidation catalyzed by the PRTM-RhNC@rGO composite is about 1.88 × 10-3 s-1 (4 µg/mL), which is 37.6 times higher than that of reported RhNP (k = 5 × 10-5 s-1, 20 µg/mL). Enzyme kinetic studies reveal that the PRTM-RhNC@rGO composite exhibits a similar affinity for uric acid as natural uricase. Furthermore, the uricase-like activity of PRTM-RhNC@rGO nanozymes remains in the presence of sulfur substances and halide ions, displaying incredibly well antipoisoning abilities. The analysis of the structure-function relationship indicates the PRTM-RhNC@rGO composite features the substrate binding site near the catalytic site in a confined space contributed by 2D rGO and PRTM, resulting in the high-performance of the composite nanozyme. Based on the outstanding uricase-like activity and the interaction of PRTM and uric acid, the PRTM-RhNC@rGO composite can retard the urate crystallization significantly. The present work provides new insights into the design of metal nanozymes with suitable binding sites near catalytic sites by mimicking pocket-like structures in natural enzymes based on simple peptides, conducing to broadening the practical application of high-performance nanozymes in biomedical fields.
Assuntos
Grafite , Ródio , Ácido Úrico , Grafite/química , Ácido Úrico/química , Ácido Úrico/metabolismo , Ródio/química , Urato Oxidase/química , Urato Oxidase/metabolismo , Peptídeos/química , Peptídeos/farmacologia , Oxirredução , Arginina/química , Nanopartículas Metálicas/químicaRESUMO
MoO3-x NPs was rapidly synthesized at room temperature by an easy stirring method. It was interesting to find that MoO3-x NPs induce OH- to generate active free radicals (ROS), which is a highly promising property in chemiluminescence (CL). Benefiting from the abundant oxygen vacancy, MoO3-x NPs adsorbs H2O2 and turn it into ·OH. The oxidase activity of fluorescein under visible light had already been reported, which catalyzes dissolved oxygen to become O2-· and continue to convert to H2O2. By creating the synergy effect with fluorescein, MoO3-x NPs strengthen the CL intensity of K3[Fe(CN)6]-fluorescein system significantly. Utilizing the quench effect of uric acid for the CL intensity, we developed a rapid, simple, and highly sensitive CL platform for uric acid detection. The linear range was 5-80 µM and the detection limit (LOD) for uric acid was 3.11 µM (S/N = 3). This work expanded the application of MoO3-x NPs in the CL field and developed a simple and highly sensitive CL sensing system to detect UA in human saliva.
Assuntos
Fluoresceína , Limite de Detecção , Molibdênio , Óxidos , Saliva , Ácido Úrico , Ácido Úrico/análise , Ácido Úrico/química , Saliva/química , Humanos , Fluoresceína/química , Óxidos/química , Molibdênio/química , Medições Luminescentes/métodos , Peróxido de Hidrogênio/químicaRESUMO
The synthesis of cobalt nanocrystal-graphene quantum dot-Ti3C2TX monolithic film electrode (Co-GQD-Ti3C2TX) is reported via self-assembly of Ti3C2TX nanosheets induced by protonated arginine-functionalized graphene quantum dot and subsequent reduction of cobalt (III). The resulting Co-GQD-Ti3C2TX shows good monolithic architecture, mechanical property, dispersibility and conductivity. The structure achieves excellent supercapacitor and sensing behavior. The self-charging supercapacitor produced by printing viscous Co-GQD-Ti3C2TX hydrogel on the back of flexible solar cell surface provides high specific capacitance (296 F g-1 at 1 A g-1), high-rate capacity (153 F g-1 at 20 A g-1), capacity retention (98.1% over 10,000-cycle) and energy density (29.6 W h kg-1 at 299.9 W kg-1). The electrochemical chip produced by printing Co-GQD-Ti3C2TX hydrogel on paper exhibits sensitive electrochemical response towards uric acid. The increase of uric acid between 0.01 and 800 µM causes a linear increase in differential pulse voltammetry signal with a detection limit of 0.0032 µM. The self-powered sensing platform integrating self-charging supercapacitor, electrochemical chip and micro electrochemical workstation was contentedly applied to monitoring uric acid in sweats and shows one broad application prospect in wearable electronic health monitoring device.
Assuntos
Cobalto , Capacitância Elétrica , Técnicas Eletroquímicas , Eletrodos , Grafite , Limite de Detecção , Pontos Quânticos , Suor , Titânio , Ácido Úrico , Cobalto/química , Pontos Quânticos/química , Grafite/química , Suor/química , Humanos , Técnicas Eletroquímicas/métodos , Técnicas Eletroquímicas/instrumentação , Titânio/química , Ácido Úrico/análise , Ácido Úrico/química , Técnicas Biossensoriais/métodos , Nanopartículas/químicaRESUMO
Copper-cobalt bimetallic nitrogen-doped carbon-based nanoenzymatic materials (CuCo@NC) were synthesized using a one-step pyrolysis process. A three-channel colorimetric sensor array was constructed for the detection of seven antioxidants, including cysteine (Cys), uric acid (UA), tea polyphenols (TP), lysine (Lys), ascorbic acid (AA), glutathione (GSH), and dopamine (DA). CuCo@NC with peroxidase activity was used to catalyze the oxidation of TMB by H2O2 at three different ratios of metal sites. The ability of various antioxidants to reduce the oxidation products of TMB (ox TMB) varied, leading to distinct absorbance changes. Linear discriminant analysis (LDA) results showed that the sensor array was capable of detecting seven antioxidants in buffer and serum samples. It could successfully discriminate antioxidants with a minimum concentration of 10 nM. Thus, multifunctional sensor arrays based on CuCo@NC bimetallic nanoenzymes not only offer a promising strategy for identifying various antioxidants but also expand their applications in medical diagnostics and environmental analysis of food.
Assuntos
Antioxidantes , Carbono , Colorimetria , Cobre , Nitrogênio , Nitrogênio/química , Colorimetria/métodos , Carbono/química , Antioxidantes/química , Antioxidantes/análise , Cobre/química , Cobalto/química , Peróxido de Hidrogênio/química , Humanos , Catálise , Limite de Detecção , Glutationa/química , Glutationa/sangue , Dopamina/sangue , Dopamina/análise , Dopamina/química , Benzidinas/química , Polifenóis/química , Polifenóis/análise , Ácido Ascórbico/química , Ácido Ascórbico/sangue , Ácido Ascórbico/análise , Oxirredução , Ácido Úrico/sangue , Ácido Úrico/química , Ácido Úrico/análise , Cisteína/química , Cisteína/sangueRESUMO
Nanorod structures exhibit a high surface-to-volume ratio, enhancing the accessibility of electrolyte ions to the electrode surface and providing an abundance of active sites for improved electrochemical sensing performance. In this study, tetragonal α-MnO2 with a large K+-embedded tunnel structure, directly grown on microfibrous carbon paper to form densely packed nanorod arrays, is investigated as an electrocatalytic material for non-enzymatic glucose sensing. The MnO2 nanorods electrode demonstrates outstanding catalytic activity for glucose oxidation, showcasing a high sensitivity of 143.82 µA cm-2 mM-1 within the linear range from 0.01 to 15 mM, with a limit of detection (LOD) of 0.282 mM specifically for glucose molecules. Importantly, the MnO2 nanorods electrode exhibits excellent selectivity towards glucose over ascorbic acid and uric acid, which is crucial for accurate glucose detection in complex samples. For comparison, a gold electrode shows a lower sensitivity of 52.48 µA cm-2 mM-1 within a linear range from 1 to 10 mM. These findings underscore the superior performance of the MnO2 nanorods electrode in both sensitivity and selectivity, offering significant potential for advancing electrochemical sensors and bioanalytical techniques for glucose monitoring in physiological and clinical settings.
Assuntos
Técnicas Biossensoriais , Técnicas Eletroquímicas , Eletrodos , Glucose , Compostos de Manganês , Nanotubos , Óxidos , Compostos de Manganês/química , Óxidos/química , Nanotubos/química , Glucose/análise , Glucose/química , Técnicas Eletroquímicas/métodos , Técnicas Biossensoriais/métodos , Técnicas Biossensoriais/instrumentação , Limite de Detecção , Carbono/química , Oxirredução , Papel , Ácido Úrico/análise , Ácido Úrico/química , Catálise , Ácido Ascórbico/química , Ácido Ascórbico/análiseRESUMO
Two-dimensional MXenes have become an important material for electrochemical sensing of biomolecules due to their excellent electric properties, large surface area and hydrophilicity. However, the simultaneous detection of multiple biomolecules using MXene-based electrodes is still a challenge. Here, a simple solvothermal process was used to synthesis the Ti3C2Tx coated with TiO2 nanosheets (Ti3C2Tx@TiO2 NSs). The surface modification of TiO2 NSs on Ti3C2Tx can effectively reduce the self-accumulation of Ti3C2Tx and improve stability. Glassy carbon electrode was modified by Ti3C2Tx@TiO2 NSs (Ti3C2Tx@TiO2 NSs/GCE) and was able simultaneously to detect dopamine (DA), ascorbic acid (AA) and uric acid (UA). Under concentrations ranging from 200 to 1000 µM, 40 to 300 µM and 50 to 400 µM, the limit of detection (LOD) is 2.91 µM, 0.19 µM and 0.25 µM for AA, DA and UA, respectively. Furthermore, Ti3C2Tx@TiO2 NSs/GCE demonstrated remarkable stability and reliable reproducibility for the detection of AA/DA/UA.
Assuntos
Ácido Ascórbico , Dopamina , Nanoestruturas , Titânio , Ácido Úrico , Titânio/química , Ácido Úrico/análise , Ácido Úrico/química , Dopamina/análise , Ácido Ascórbico/análise , Ácido Ascórbico/química , Nanoestruturas/química , Limite de Detecção , Técnicas Eletroquímicas/métodos , Eletrodos , Reprodutibilidade dos Testes , Técnicas Biossensoriais/métodosRESUMO
Five mini proteins mimicking uricase comprising 20, 40, 60, 80, and 100 amino acids were designed based on the conserved active site residues within the same dimer, using the crystal structure of tetrameric uricase from Arthrobacter globiformis (PDB ID: 2yzb) as a template. Based on molecular docking analysis, the smallest mini protein, mp20, shared similar residues to that of native uricase that formed hydrogen bonds with uric acid and was chosen for further studies. Although purified recombinant mp20 did not exhibit uricase activity, it showed specific binding towards uric acid and evinced excellent thermotolerance and structural stability at temperatures ranging from 10°C to 100°C, emulating its natural origin. To explore the potential of mp20 as a bioreceptor in uric acid sensing, mp20 was encapsulated within zeolitic imidazolate framework-8 (mp20@ZIF-8) followed by the modification on rGO-screen printed electrode (rGO/SPCE) to maintain the structural stability. An irreversible anodic peak and increased semicircular arcs of the Nyquist plot with an increase of the analyte concentrations were observed by utilizing cyclic voltammetry and electrochemical impedance spectroscopy (EIS), suggesting the detection of uric acid occurred, which is based on substrate-mp20 interaction.
Assuntos
Grafite , Ácido Úrico , Ácido Úrico/análise , Ácido Úrico/química , Urato Oxidase/genética , Urato Oxidase/química , Urato Oxidase/metabolismo , Simulação de Acoplamento MolecularRESUMO
One of the major challenges for in vivo electrochemical measurements of dopamine (DA) is to achieve selectivity in the presence of interferents, such as ascorbic acid (AA) and uric acid (UA). Complicated multimaterial structures and ill-defined pretreatments have been frequently utilized to enhance selectivity. The lack of control over the realized structures has prevented establishing associations between the achieved selectivity and the electrode structure. Owing to their easily tailorable structure, carbon nanofiber (CNF) electrodes have become promising materials for neurobiological applications. Here, a novel yet simple strategy to control the sensitivity and selectivity of CNF electrodes toward DA is reported. It consists of adjusting the lengths of CNF by modulating the growth phase during the fabrication process while keeping the surface chemistries similar. It was observed that the sensitivity of the CNF electrodes toward DA was enhanced with the increase in the fiber lengths. More importantly, the increase in the fiber length induced (i) an anodic shift in the DA oxidation peak and (ii) a cathodic shift in the AA oxidation peak. As the UA oxidation peak remained unaffected at high anodic potentials, the electrodes with long CNFs showed excellent selectivity. Electrodes without proper fibers showed only a single broad peak in the solution of AA, DA, and UA, completely lacking the ability to discriminate DA. Hence, the simple strategy of controlling CNF length without the need to carry out any complex chemical treatments provides us a feasible and robust route to fabricate electrode materials for neurotransmitter detection with excellent sensitivity and selectivity.
Assuntos
Dopamina , Nanofibras , Dopamina/química , Carbono/química , Técnicas Eletroquímicas , Eletrodos , Ácido Ascórbico/química , Ácido Úrico/química , OxirreduçãoRESUMO
Although many studies have focused on the role of individual biomolecules or metal ions in the crystallization behavior of sodium urate, the regulatory effects of multiple molecular species still remain mysterious. The synergistic cooperation of biomolecules and metal ions may contribute to unprecedented regulatory effects. Here, the cooperative effect of arginine-rich peptides (APs) and copper ions on the phase behavior, crystallization kinetics, and size/morphology of urate crystals was first investigated. Compared with the individual copper ion and AP, the nucleation induction time of sodium urate is prolonged dramatically (about 48 h), and the nucleation rate of sodium urate is reduced efficiently in a saturated solution due to the synergistic effect of Cu2+ and AP in stabilizing amorphous sodium urate (ASU). The length of sodium urate monohydrate crystals decreases obviously under the synergistic effect of Cu2+ and AP. The comparative experiments of common transition metal cations show that only copper ions can cooperate with AP, which may be due to the strong coordination effect between copper ions with urate and AP. Further studies show that the synergistic effect of copper ions and APs with different chain lengths on the crystallization behavior of sodium urate is significantly different. Both the guanidine functional groups and the length of peptide chains simultaneously determine the synergistic inhibition effect of polypeptides and Cu2+. This work highlights the synergistic inhibition effect of metal ions and cationic peptides on the crystallization of sodium urate, which enriches the understanding of the regulating mechanism of biological mineral crystallization using the synergy of multispecies and offers a new strategy for designing efficient inhibitors for sodium urate crystallization in gout stone diseases.
Assuntos
Gota , Ácido Úrico , Humanos , Ácido Úrico/química , Cobre , Cristalização , Peptídeos , ÍonsRESUMO
Pseudogout is crystalline arthritis. It has a similar clinical picture to that of gout, and it is difficult to distinguish the two diseases using conventional analysis methods. However, it is important to identify the different crystals responsible for these two cases because the treatment strategies are different. In a previous study, we reported magnetic orientation of monosodium urate (MSU) crystals, which are the causative agent of gout, at the permanent magnet level. In this study, we investigated the effect of an applied magnetic field on calcium pyrophosphate (CPP) crystals, which are the causative agent of pseudogout, and the difference in the magnetic responses of CPP and MSU crystals. We found that the CPP crystals were oriented in a magnetic field on milli-Tesla order because of the anisotropy of the diamagnetic susceptibility. In addition, the CPP crystals exhibited different anisotropic magnetic properties from those of MSU crystals, which led to a characteristic difference between the orientations of the two crystals. That is, we found that the causative agents of gout and pseudogout responded differently to a magnetic field. This report suggests that the discrimination between CPP and MSU by optical measurements is possible by application of magnetic fields appropriately. © 2023 Bioelectromagnetics Society.