RESUMEN
Antibiotic residues in the environment pose a serious threat to ecosystems and human health. Therefore, it is important to develop sensitive and rapid in situ detection methods. In this work, the designed nanozymes, with excellent four enzyme activities, were proved to be constituted of unique hollow nanocage structures (CoZnSe@CN HCs). Based on the peroxidase-like enzymes, a portable colorimetric sensor was constructed for the on-site determination of tetracycline (TC) in real samples. The linear range of TC detection was 0.1-100 µM, and the detection limit was 0.02 µM. At the same time, colorimetric detection and smartphones have also been combined for on-site colorimetric detection of TC. In-depth exploration of the detection mechanism showed that TC could be bound with the material, inhibiting the production of oxidized 3,3',5,5'-tetramethylbenzidine. The sensor was also used for the detection of TC in environmental soil and water samples. This study can provide an intelligent detection method for environmental monitoring.
Asunto(s)
Ecosistema , Realidad Virtual , Humanos , Teléfono Inteligente , Tetraciclina , AntibacterianosRESUMEN
Since the catalyst's surface was the major active location, the inner structure's contribution to catalytic activity was typically overlooked. Here, ZnO-Co3O4-v nanozymes with several surfaces and bulk oxygen vacancies were created. The O atoms of H2O2 moved inward to preferentially fill the oxygen vacancies in the interior and form new "lattice oxygen" by the X-ray photoelectron spectroscopy depth analysis and X-ray absorption fine structure. The internal Co2+ continually transferred electrons to the surface for a continuous catalytic reaction, which generated a significant amount of reactive oxygen species. Inner and outer double-layer electron cycles accompanied this process. A three-dimensional model of ZnO-Co3O4-v was constructed using virtual reality interactive modelling technology to illustrate nanozyme catalysis. Moreover, the bactericidal rate of ZnO-Co3O4-v for Methionine-resistant Staphylococcus aureus and Multiple drug resistant Escherichia coli was as high as 99%. ZnO-Co3O4-v was biocompatible and might be utilized to heal wounds following Methionine-resistant Staphylococcus aureus infection. This work offered a new idea for nanozymes to replace of conventional antibacterial medications.
Asunto(s)
Staphylococcus aureus Resistente a Meticilina , Realidad Virtual , Óxido de Zinc , Oxígeno/química , Óxido de Zinc/farmacología , Óxido de Zinc/química , Electrones , Peróxido de Hidrógeno , Escherichia coli , Cicatrización de Heridas , MetioninaRESUMEN
Although the application of nanozymes has been widely studied, it is still a huge challenge to develop highly active and multifunctional nanozyme catalysts with a wider application prospect. Co3O4/CoFe2O4 hollow nanocubes (HNCs) with oxygen vacancies were proposed in this study, which had a porous oxide heterostructure with CoFe2O4 as the core and Co3O4 as the shell. The Co3O4/CoFe2O4 HNCs had three enzyme activities: peroxidase-like, oxidase-like, and catalase-like. Combining XPS depth profiling with density functional theory (DFT), the catalytic mechanism of peroxidase-like activity was explored in depth, which was mainly originated from ·OH produced by the synergistic effect between the outer oxygen and inner oxygen and electron transfer between Co and Fe. A colorimetry/smartphone dual sensing platform was designed based on the peroxidase-like activity. Especially, a multifunctional intelligent sensing platform based on deep learning-YOLO v3 algorithm-assisted smartphone was constructed to realize real-time and rapid in situ detection of l-cysteine, norfloxacin, and zearalenone. Surprisingly, the detection limit of norfloxacin was low at 0.015 µM, which was better than that of the newly published detection method in the field of nanozymes. Meanwhile, the detection mechanism of l-cysteine and norfloxacin was successfully investigated by in situ FTIR. In fact, it also showed outstanding applications in detecting l-cysteine in the food environment and norfloxacin in drugs. Furthermore, Co3O4/CoFe2O4 HNCs also could degrade 99.24% of rhodamine B, along with good reusability even after 10-cycle runs. Therefore, this work provided an in-depth understanding of the synergistic effect between the outer and inner oxygen in the reaction mechanism and an efficient method for establishing a deep-learning-assisted intelligent detection platform. In addition, this research also offered a good guideline for the further development and construction of nanozyme catalysts with multienzyme activities and multifunctional applications.
Asunto(s)
Aprendizaje Profundo , Oxígeno , Oxidación-Reducción , Oxígeno/química , Cisteína , Norfloxacino , Teléfono Inteligente , Peroxidasa/química , Colorimetría/métodos , Peróxido de HidrógenoRESUMEN
Voltage-gated calcium channels containing alpha1 subunits encoded by Ca(v)2 family genes are critical in regulating release of neurotransmitter at chemical synapses. In Drosophila, cac is the only Ca(v)2-type gene. Cacophony (CAC) channels are localized in motor neuron terminals where they have been shown to mediate evoked, but not AP-independent, release of glutamate at the larval neuromuscular junction (NMJ). Cultured embryonic neurons also express CAC channels, but there is no information about the properties of CAC-mediated currents in adult brain nor how these channels regulate transmission in central neural circuits where fast excitatory synaptic transmission is predominantly cholinergic. Here we report that wild-type neurons cultured from late stage pupal brains and antennal lobe projection neurons (PNs) examined in adult brains, express calcium currents with two components: a slow-inactivating current sensitive to the spider toxin Plectreurys toxin II (PLTXII) and a fast-inactivating PLTXII-resistant component. CAC channels are the major contributors to the slow-inactivating PLTXII-sensitive current based on selective reduction of this component in hypomorphic cac mutants (NT27 and TS3). Another characteristic of cac mutant neurons both in culture and in whole brain recordings is a reduced cholinergic miniature excitatory postsynaptic current frequency that is mimicked in wild-type neurons by acute application of PLTXII. These data demonstrate that cac encoded Ca(v)2-type calcium channels regulate action potential (AP)-independent release of neurotransmitter at excitatory cholinergic synapses in the adult brain, a function not predicted from studies at the larval NMJ.
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Potenciales de Acción/fisiología , Encéfalo/fisiología , Canales de Calcio Tipo N/genética , Canales de Calcio Tipo N/fisiología , Canales de Calcio/genética , Canales de Calcio/fisiología , Proteínas de Drosophila/genética , Proteínas de Drosophila/fisiología , Neurotransmisores/metabolismo , Sistema Nervioso Parasimpático/metabolismo , Sinapsis/metabolismo , Animales , Encéfalo/citología , Encéfalo/efectos de los fármacos , Células Cultivadas , Drosophila , Electrofisiología , Potenciales Postsinápticos Excitadores/fisiología , Proteínas Fluorescentes Verdes , Inmunohistoquímica , Microscopía Electrónica , Actividad Motora/fisiología , Neuronas/metabolismo , Neurotoxinas/farmacología , Sistema Nervioso Parasimpático/efectos de los fármacos , Técnicas de Placa-Clamp , Órganos de los Sentidos/fisiología , Sinapsis/efectos de los fármacosRESUMEN
In Drosophila, nicotinic acetylcholine receptors (nAChRs) mediate fast excitatory synaptic transmission in mushroom body Kenyon cells, a neuronal population involved in generation of complex behaviors, including responses to drugs of abuse. To determine whether activation of nAChRs can induce cellular changes that contribute to functional plasticity in these neurons, we examined nicotine-evoked responses in cells cultured from brains of late stage OK107-GAL4 pupae. Kenyon cells can be identified by expression of green fluorescent protein (GFP+). Nicotine activates alpha-bungarotoxin-sensitive nAChRs, causing a rapid increase in intracellular calcium levels in over 95% of the Kenyon cells. The nicotine-evoked calcium increase has a voltage-gated calcium channel (VGCC) dependent component and a VGCC-independent component that involves calcium influx directly through nAChRs. Thapsigargin treatment reduces the nicotine response consistent with amplification by calcium release from intracellular stores. The response to nicotine is experience-dependent: a short conditioning pulse of nicotine causes a transient 50% reduction in the magnitude of the response to a test pulse of nicotine when the interpulse interval is 4 h. This cellular plasticity is dependent on activation of the VGCC-component of the nicotine response and on cAMP-signaling, but not on protein synthesis. These data demonstrate that activation of nAChRs induces a calcium-dependent plasticity in Kenyon cells that could contribute to adult behaviors involving information processing in the mushroom bodies including responses to nicotine.
Asunto(s)
Señalización del Calcio/fisiología , Drosophila/metabolismo , Cuerpos Pedunculados/metabolismo , Plasticidad Neuronal/fisiología , Neuronas/metabolismo , Receptores Nicotínicos/metabolismo , Animales , Bungarotoxinas/farmacología , Calcio/metabolismo , Canales de Calcio/efectos de los fármacos , Canales de Calcio/metabolismo , Señalización del Calcio/efectos de los fármacos , Células Cultivadas , Inhibidores Enzimáticos/farmacología , Femenino , Masculino , Cuerpos Pedunculados/citología , Cuerpos Pedunculados/efectos de los fármacos , Vías Nerviosas/efectos de los fármacos , Vías Nerviosas/metabolismo , Plasticidad Neuronal/efectos de los fármacos , Neuronas/efectos de los fármacos , Agonistas Nicotínicos/farmacología , Receptores Nicotínicos/efectos de los fármacos , Transmisión Sináptica/efectos de los fármacos , Transmisión Sináptica/fisiología , Tapsigargina/farmacologíaRESUMEN
Spontaneous calcium oscillations in mushroom bodies of late stage pupal and adult Drosophila brains have been implicated in memory consolidation during olfactory associative learning. This study explores the cellular mechanisms regulating calcium dynamics in Kenyon cells, principal neurons in mushroom bodies. Fura-2 imaging shows that Kenyon cells cultured from late stage Drosophila pupae generate spontaneous calcium transients in a cell autonomous fashion, at a frequency similar to calcium oscillations in vivo (10-20/h). The expression of calcium transients is up regulated during pupal development. Although the ability to generate transients is a property intrinsic to Kenyon cells, transients can be modulated by bath application of nicotine and GABA. Calcium transients are blocked, and baseline calcium levels reduced, by removal of external calcium, addition of cobalt, or addition of Plectreurys toxin (PLTX), an insect-specific calcium channel antagonist. Transients do not require calcium release from intracellular stores. Whole cell recordings reveal that the majority of voltage-gated calcium channels in Kenyon cells are PLTX-sensitive. Together these data show that influx of calcium through PLTX-sensitive voltage-gated calcium channels mediates spontaneous calcium transients and regulates basal calcium levels in cultured Kenyon cells. The data also suggest that these calcium transients represent cellular events underlying calcium oscillations in the intact mushroom bodies. However, spontaneous calcium transients are not unique to Kenyon cells as they are present in approximately 60% of all cultured central brain neurons. This suggests the calcium transients play a more general role in maturation or function of adult brain neurons.