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1.
iScience ; 27(9): 110648, 2024 Sep 20.
Artículo en Inglés | MEDLINE | ID: mdl-39286487

RESUMEN

The traditional parameter adjustment design makes it difficult to effectively regulate the acoustic insulation performance of periodic sandwich structures while meeting the lightweight and mechanical stiffness requirements. A dynamic three-field floating projection topology optimization (FPTO) method for periodic structures is proposed to meet the optimization requirements of low-noise and high-stiffness performance of lightweight periodic sandwich structures. The sound transmission loss is taken as the optimization objective, and the lightweight volume and mechanical stiffness performance are taken as the multiple constraints. The results show that a smooth topology configuration with superior sound insulation performance, high stiffness, and a freely customizable number of periodic cores can be obtained via the proposed method. The accuracy and effectiveness of the presented method are verified via 3D printing technology and impedance tube sound insulation experiments, providing an important reference for the optimal design of lightweight composite structures for vibration and noise reduction in transportation equipment.

2.
Front Plant Sci ; 15: 1459013, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-39290736

RESUMEN

Introduction: Fruit color significantly influences the quality of horticultural crops, which affects phytochemical diversity and consumer preferences. Despite its importance, the genetic basis of the white-colored fruit in tomatoes remains poorly understood. Methods: In this study, we demonstrate that white-fleshed tomato varieties accumulate fewer carotenoids than yellow-fleshed varieties. We developed various segregating populations by hybridizing red, yellow, and white fruit tomato cultivars. Results: Genetic analysis revealed that the white fruit color trait is controlled by a single gene that dominates both red and yellow fruits. Bulk segregant RNA sequencing provided a preliminary map of a 3.17 Mb region on chromosome 3 associated with the white color trait. Based on kompetitive allele-specific PCR (KASP) markers, we narrowed the candidate gene region to 819 kb. Within this region, we identified a 4906-bp sequence absence variation near Phytoene Synthase 1 (SlPSY1) specific to white-colored tomatoes. Genotyping of the progeny and natural populations using a single nucleotide polymorphism adjacent to this absence of variation confirmed its key role in white fruit formation. Discussion: Collectively, our findings provide insights into white fruit trait formation in tomatoes, enabling tomato breeders to precisely introduce white fruit traits for commercial exploitation.

3.
Biosens Bioelectron ; 266: 116738, 2024 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-39241336

RESUMEN

An in-situ nanozyme signal tag combined with a DNA-mediated universal antibody-oriented strategy was proposed to establish a high-performance immunosensing platform for Alzheimer's disease (AD)-related biomarker detection. Briefly, a Zr-based metal-organic framework (MOF) with peroxidase (POD)-like activity was synthesized to encapsulating the electroactive molecule methylene blue (MB), and subsequently modified with a layer of gold nanoparticles on its surface. This led to the creation of double POD-like activity nanozymes surrounding the MB molecule to form a nanozyme signal tag. A large number of hydroxyl radicals were generated by the nanozyme signal tag with the help of H2O2, which catalyzed MB molecules in situ to achieve efficient signal amplification. Subsequently, a DNA-aptamer-mediated universal antibody-oriented strategy was proposed to enhance the binding efficiency for the antigen (target). Meanwhile, a poly adenine was incorporated at the end of the aptamer, facilitating binding to the gold electrode and providing anti-fouling properties due to the hydrophilicity of the phosphate group. Under optimal conditions, this platform was successfully employed for highly sensitive detection of AD-associated tau protein and BACE1, achieving limits of detection with concentrations of 3.34 fg/mL and 1.67 fg/mL, respectively. It is worth mentioning that in the tau immunosensing mode, 20 clinical samples from volunteers of varying ages were analyzed, revealing significantly higher tau expression levels in the blood samples of elderly volunteers compared to young volunteers. This suggests that the developed strategy holds great promise for early AD diagnosis.


Asunto(s)
Enfermedad de Alzheimer , Aptámeros de Nucleótidos , Biomarcadores , Técnicas Biosensibles , Técnicas Electroquímicas , Oro , Nanopartículas del Metal , Proteínas tau , Técnicas Biosensibles/métodos , Humanos , Enfermedad de Alzheimer/diagnóstico , Enfermedad de Alzheimer/sangre , Técnicas Electroquímicas/métodos , Oro/química , Aptámeros de Nucleótidos/química , Biomarcadores/sangre , Nanopartículas del Metal/química , Proteínas tau/sangre , Estructuras Metalorgánicas/química , Inmunoensayo/métodos , Límite de Detección , Secretasas de la Proteína Precursora del Amiloide , Azul de Metileno/química , Ácido Aspártico Endopeptidasas/sangre , Peróxido de Hidrógeno/química , Catálisis
4.
Nano Lett ; 24(28): 8650-8657, 2024 Jul 17.
Artículo en Inglés | MEDLINE | ID: mdl-38949785

RESUMEN

The ion permeability and selectivity of membranes are crucial in nanofluidic behavior, impacting industries ranging from traditional to advanced manufacturing. Herein, we demonstrate the engineering of ion-conductive membranes featuring angstrom-scale ion-transport channels by introducing ionic polyamidoamine (PAMAM) dendrimers for ion separation. The exterior quaternary ammonium-rich structure contributes to significant electrostatic charge exclusion due to enhanced local charge density; the interior protoplasmic channels of PAMAM dendrimer are assembled to provide additional degrees of free volume. This facilitates the monovalent ion transfer while maintaining continuity and efficient ion screening. The dendrimer-assembled hybrid membrane achieves high monovalent ion permeance of 2.81 mol m-2 h-1 (K+), reaching excellent mono/multivalent selectivity up to 20.1 (K+/Mg2+) and surpassing the permselectivities of state-of-the-art membranes. Both experimental results and simulating calculations suggest that the impressive ion selectivity arises from the significant disparity in transport energy barrier between mono/multivalent ions, induced by the "exterior-interior" synergistic effects of bifunctional membrane channels.

5.
Anal Chem ; 96(21): 8782-8790, 2024 05 28.
Artículo en Inglés | MEDLINE | ID: mdl-38728110

RESUMEN

Sensitive and on-site discrimination of live and dead foodborne pathogenic strains remains a significant challenge due to the lack of appropriate assay and signal probes. In this work, a versatile platinum nanoparticle-decorated phage nanozyme (P2@PtNPs) that integrated recognition, bacteriolysis, and catalysis was designed to establish the bioluminescence/pressure dual-mode bioassay for on-site determination of the vitality of foodborne pathogenic strains. Benefiting from the bacterial strain-level specificity of phage, the target Salmonella typhimurium (S.T) was specially captured to form sandwich complexes with P2@PtNPs on another phage-modified glass microbead (GM@P1). As the other part of the P2@PtNPs nanozyme, the introduced PtNPs could not only catalyze the decomposition of hydrogen peroxide to generate a significant oxygen pressure signal but also produce hydroxyl radicals around the target bacteria to enhance the bacteriolysis of phage and adenosine triphosphate release. It significantly improved the bioluminescence signal. The two signals corresponded to the total and live target bacteria counts, so the dead target could be easily calculated from the difference between the total and live target bacteria counts. Meanwhile, the vitality of S.T was realized according to the ratio of live and total S.T. Under optimal conditions, the application range of this proposed bioassay for bacterial vitality was 102-107 CFU/mL, with a limit of detections for total and live S.T of 30 CFU/mL and 40 CFU/mL, respectively. This work provides an innovative and versatile nanozyme signal probe for the on-site determination of bacterial vitality for food safety.


Asunto(s)
Bacteriófagos , Mediciones Luminiscentes , Nanopartículas del Metal , Platino (Metal) , Salmonella typhimurium , Platino (Metal)/química , Nanopartículas del Metal/química , Salmonella typhimurium/aislamiento & purificación , Salmonella typhimurium/virología , Salmonella typhimurium/química , Catálisis , Bacteriófagos/química , Microbiología de Alimentos , Bioensayo/métodos , Técnicas Biosensibles/métodos , Presión , Peróxido de Hidrógeno/química
6.
Anal Chem ; 96(17): 6674-6682, 2024 04 30.
Artículo en Inglés | MEDLINE | ID: mdl-38642044

RESUMEN

Photodynamic therapy (PDT) is a significant noninvasive therapeutic modality, but it is often limited in its application due to the restricted tissue penetration depth caused by the wavelength limitations of the light source. Two-photon (TP) fluorescence techniques are capable of having an excitation wavelength in the NIR region by absorbing two NIR photons simultaneously, which offers the potential to achieve higher spatial resolution for deep tissue imaging. Thus, the adoption of TP fluorescence techniques affords several discernible benefits for photodynamic therapy. Organic TP dyes possess a high fluorescence quantum yield. However, the biocompatibility of organic TP dyes is poor, and the method of coating organic TP dyes with silica can effectively overcome the limitations. Herein, based on the TP silica nanoparticles, a functionalized intelligent biogenic missile TP-SiNPs-G4(TMPyP4)-dsDNA(DOX)-Aptamer (TGTDDA) was developed for effective TP bioimaging and synergistic targeted photodynamic therapy and chemotherapy in tumors. First, the Sgc8 aptamer was used to target the PTK7 receptor on the surface of tumor cells. Under two-photon light irradiation, the intelligent biogenic missile can be activated for TP fluorescence imaging to identify tumor cells and the photosensitizer assembled on the nanoparticle surface can be activated for photodynamic therapy. Additionally, this intelligent biogenic missile enables the controlled release of doxorubicin (DOX). The innovative strategy substantially enhances the targeted therapeutic effectiveness of cancer cells. The intelligent biogenic missile provides an effective method for the early detection and treatment of tumors, which has a good application prospect in the real-time high-sensitivity diagnosis and treatment of tumors.


Asunto(s)
Imagen Óptica , Fotoquimioterapia , Fotones , Fármacos Fotosensibilizantes , Humanos , Animales , Fármacos Fotosensibilizantes/química , Fármacos Fotosensibilizantes/farmacología , Fármacos Fotosensibilizantes/uso terapéutico , Ratones , Nanopartículas/química , Doxorrubicina/química , Doxorrubicina/farmacología , Doxorrubicina/uso terapéutico , Dióxido de Silicio/química , Aptámeros de Nucleótidos/química , Colorantes Fluorescentes/química , Neoplasias/tratamiento farmacológico , Neoplasias/diagnóstico por imagen , Antineoplásicos/química , Antineoplásicos/farmacología , Ratones Desnudos , Línea Celular Tumoral , Ratones Endogámicos BALB C
7.
Colloids Surf B Biointerfaces ; 238: 113877, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38615390

RESUMEN

An ideal bone metastasis animal model is critical and fundamental for mechanistic research and following development of new drug and treatment. Caudal artery (CA) injection allows bone metastasis in the hindlimb, while in-depth targeted and quantitative studies of bone metastasis require a new model to overcome its limitations. Here, we developed a targeted, quantitative, and highly consistent method for the modeling of bone metastasis with cell-based magnetic micro-living-motor (MLM) system created by effectively combining Fe3O4-PDA-Au with biosafety. The MLM system can achieve efficient migration, target site colonization and control tumorigenesis in bone precisely with the application of a magnetic field. In vivo, day 3 post cell injection, tumor bone metastasis signals were observed locally in the injected femur among 82.76% mice of the MLM group as compared to the 56.82% in the CA group, and the signal intensity was 45.1 and 95.9 times stronger than that in the left and right lower limbs of the CA group, respectively. Post-injection day 28, metastasis in vital organs was reduced by approximately 90% in the MLM group compared to the CA group. Our innovative use of the MLM system in the field of tumor modeling opens a new avenue for exploring the mechanisms of tumor bone metastasis, recurrence and drug resistance.


Asunto(s)
Neoplasias Óseas , Animales , Neoplasias Óseas/secundario , Neoplasias Óseas/patología , Ratones , Modelos Animales de Enfermedad , Línea Celular Tumoral , Humanos , Femenino , Campos Magnéticos , Ratones Endogámicos BALB C
8.
Talanta ; 275: 126067, 2024 Aug 01.
Artículo en Inglés | MEDLINE | ID: mdl-38640522

RESUMEN

The detection of pathogen viability is critically important to evaluate its infectivity. In the study, an integrated microfluidic chip based on dual-mode analytical strategy was developed to rapidly realize detection of bacteria activity (with Salmonella typhimurium, S.T, as a model analyte). Firstly, the composite probes, including deactivated phage modified magnetic beads and nano Pt-antimicrobial peptide (AMP) which can specifically recognize Gram-negative bacteria as nanozyme were prepared. When the composite probes are introduced into the chip together with target bacteria, after enrichment, oscillating and magnetic separation, they will conjugate with S.T and produce a magnetic sandwich complex. The complex can catalyze tetramethylbenzidine (TMB)-H2O2 to produce visible colorimetric signals which is correspondent to the total S.T content. Simultaneously, PtNPs in the complex can produce hydroxyl radical oxidation (∙OH) by decomposing H2O2. Under the synergistic action of ∙OH and AMP, the captured live S.T can be lysed to release ATP and emit bioluminescence signals which corresponds to the live S.T concentration. Therefore, the chip can simultaneously detect and image S.T at different viability in one test. The dual-mode assay demonstrated high sensitivity (≤33 CFU/mL), high specificity (identifying strain), signal amplification (5 folds) and short time (≤40min). The chip array can detect four samples in one test and exhibited advantages of high-integration, -sensitivity, -specificity and miniaturization, which are suitable to rapidly detect and image pathogen's viability in trace level. The replacement of phage probes can detect other bacteria. It has a wide prospect in pathogens screening.


Asunto(s)
Dispositivos Laboratorio en un Chip , Platino (Metal) , Salmonella typhimurium , Platino (Metal)/química , Salmonella typhimurium/aislamiento & purificación , Peróxido de Hidrógeno/química , Peróxido de Hidrógeno/análisis , Viabilidad Microbiana , Bencidinas/química , Bacteriófagos/química , Colorimetría/métodos , Nanopartículas del Metal/química
9.
Pak J Med Sci ; 40(4): 747-752, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38544986

RESUMEN

Objective: To explore the effects of clinical nursing pathway (CNP) on the postoperative satisfaction and quality of life (QOL) of patients with subarachnoid hemorrhage (SAH). Methods: This is a retrospective study. Eighty patients with SAH admitted to Baoding No.1 Central Hospital from June 2021 to January 2023 were prospectively divided into a observation group and a control group by random numbers. The control group was given routine nursing, and the observation group was additional given CNP. The prognosis, cognitive function, QOL, self-care ability, nursing satisfaction and the incidence of complications were compared between the two groups. Results: After CNP nursing, the GCS and MMSE scores in the observation group were higher than those in the control group 14 days, one month and six months after the operation; and the difference was statistically significant (p< 0.05). Six months after the operation, the SS-QOL and Ability of daily living (ADL) scores in both groups were significantly improved compared with those before the intervention; and the improvement in the observation group was significantly better than that in the control group; and the difference was statistically significant(p<0.05). The nursing satisfaction score in the observation group was significantly higher than in the control group. The total incidence of complications in the observation group was lower than that in the control group. Conclusions: The CNP intervention in perioperative period of SAH patients has remarkable clinical effect, can improve the pertinence and efficiency of nursing, promote patients to recover as soon as possible, significantly improve the QOL of patients,and is worthy of clinical popularization.

10.
Analyst ; 149(3): 876-884, 2024 Jan 29.
Artículo en Inglés | MEDLINE | ID: mdl-38175666

RESUMEN

Specific and rapid detection of live Staphylococcus aureus (S.A) in environmental and food samples is critically important for protecting human health. In order to fulfill this purpose, two kinds of novel egg yolk antibody (IgY) immobilized immunomagnetic beads (IMBs; mSiO2-IgY and mMOF-IgY), with core-shell mSiO2 and mMOF as substrate, were prepared for selectively enriching S.A from samples. Furthermore, the IMBs with captured S.A were collected and re-dissolved in 0.5 mL PBS. After that, a cotton swab coated with sodium dodecylsulfate (SDS) was put in the solution to lyse S.A cells and emit ATP bioluminescence of the luciferin/luciferase system. Finally, a portable bioluminescence detector was used for quantification of ATP corresponding to S.A concentration. The results demonstrated that mMOF-IgY can enrich more S.A than mSiO2-IgY and emit a stronger signal. The reasons may be due to the higher immobilization amount of IgY on the IMBs. Under optimal conditions, the calibration line of S.A concentration was 10-105 CFU mL-1 by mMOF-IgY within 30 min. The low detection limit of S.A was 3 CFU mL-1. The results demonstrated that the assay takes much shorter time than plate counting. Its portability and excellent detection capability are suitable for rapid monitoring of specific pathogens in foods.


Asunto(s)
Estructuras Metalorgánicas , Staphylococcus aureus , Humanos , Animales , Yema de Huevo , Anticuerpos , Inmunoglobulinas , Fenómenos Magnéticos , Adenosina Trifosfato , Pollos
11.
Anal Chim Acta ; 1287: 342053, 2024 Jan 25.
Artículo en Inglés | MEDLINE | ID: mdl-38182365

RESUMEN

Rapid and specific detection of virulent bacterial strains is a great challenge for food safety regarding large amounts of contaminated samples. Herein, a dual-mode hydrogel array biosensor was constructed to simultaneously rapidly screen and precisely quantitatively detect virulent Escherichia coli O157:H7 (E. coli O157:H7) based on a novel DNA-modified phage probe. First, E. coli O157:H7 was incubated with alginate to form the E. coli O157:H7/hydrogel premix complex. Subsequently, hydrogel formation by cross-linking upon the addition of calcium ions and phages for E. coli O157:H7 modified with a DNA primer (phage-DNA) was added to the alginate hydrogel. The DNA on the complex could trigger rolling circle amplification (RCA) to form a phage probe containing a long-chain DNA skeleton (phage@RCA-DNA). The RCA-DNA was then hybridized with the complementary DNA (cDNA) to form double-stranded DNA fragments (phage@RCA-dsDNA), which could be stained by the SYBR Green dye to emit visual green fluorescence (FL) and determined by a smartphone for rapid screening. Meanwhile, the unreacted cDNA in the supernatant could be quantitatively detected by microfluidic chip electrophoresis (MCE). The signal decrement was also proportional to the bacterial concentration. The detection limit values of E. coli O157:H7 were 50 CFU mL-1 by the FL signal and 6 CFU mL-1 by the MCE signal. The two results could be mutually corrected to decrease the false-positive results. This assay was also employed to detect virulent Salmonella Typhimurium (S. Typhimurium) using the corresponding S. Typhimurium phage@RCA-DNA probe. All these results demonstrated that the universal bioassay was suitable for simultaneous rapid screening and precisely quantitative detection of virulent bacterial strains.


Asunto(s)
Bacteriófagos , Escherichia coli O157 , ADN Complementario , Hidrogeles , Microfluídica , Sondas de ADN , Alginatos , Colorantes , Electroforesis
12.
Heliyon ; 9(12): e23048, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-38144276

RESUMEN

Porphyromonas gingivalis (P. gingivalis) is one of the keystone pathogenic bacteria of periodontitis and peri-implantitis. This study aimed to investigate the antibacterial effects and molecular mechanisms of trans-cinnamaldehyde (TC), a safe extract from natural plants, on P. gingivalis. Minimum inhibitory and minimum bactericidal concentrations (MIC and MBC) of TC were determined, and scanning and transmission electron microscopies were used to assess the morphological changes. The overall biomass was estimated, and the metabolic activity of biofilms was determined at different TC concentrations. A microarray-based bioinformatics analysis was performed to elucidate the underlying molecular mechanisms of TC-inhibited P. gingivalis, and significant differences among groups were determined. TC showed an inhibitory effect on the proliferation and survival of planktonic P. gingivalis, of which the MIC and MBC were 39.07 µg/mL and 78.13 µg/mL, respectively. TC also significantly suppressed the formation and metabolic activity of P. gingivalis biofilm. The results of the significant pathways and gene ontology (GO) analyses revealed that TC treatment inhibited two metabolic pathways, accompanied by the downregulation of relative genes of nitrogen metabolism (NrfA, NrfH, and PG_2213) and starch and sucrose metabolism (PG_1681, PG_1682, and PG_1683). Thus, this study confirmed TC to be a natural antimicrobial agent against P. gingivalis and further demonstrated that TC suppressed the microbial activity on P. gingivalis through the disruption of physiological metabolism, which might inhibit the growth and the biofilm formation of P. gingivalis.

13.
Micromachines (Basel) ; 14(12)2023 Nov 27.
Artículo en Inglés | MEDLINE | ID: mdl-38138331

RESUMEN

In this work, high-frequency forced vibrations of lateral field excitation (LFE) devices with stepped electrodes based on monoclinic crystals GdCOB are modeled, and the influence laws of the device parameters (the step number, size, and thickness of the stepped electrodes) on the energy-trapping effects of the device are revealed. The results show that the step number has a significant effect on the energy-trapping effect of the device: with the increase in the step number, the stronger energy-trapping effect of the device can be obtained; with the increase in the thickness difference of two layers of electrodes, the energy-trapping effect of the device becomes stronger; with the increase in the difference of the electrode radius, the energy-trapping effect of the device is enhanced gradually. The results of this work can provide an important theoretical basis for the design of stepped-electrode LFE resonators and sensors with high-quality factors based on monoclinic crystals.

14.
Pak J Med Sci ; 39(6): 1737-1741, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37936735

RESUMEN

Objective: To investigate the effect of butylphthalide combined with fasudil in the treatment of subarachnoid hemorrhage (SAH) with cerebral vasospasm (CVS) on inflammatory factors, cognitive function and vascular endothelial function. Methods: It is a retrospective study in which a total of 104 patients with SAH with CVS admitted to Baoding First Central Hospital from July 2020 to February 2022 were selected and randomly divided into two groups by drawing lots. Patients in the control group were treated with basic symptomatic treatment, while those in the observation group were treated with butylphthalide soft capsule combined with fasudil hydrochloride injection on the basis of the control group. Before and after treatment, serum neuron specific enolase (NSE), tumor necrosis factor-α(TNF-α), interleukin-8(IL-8), hypersensitive C-reactive protein (CRP), Birmingham Cognitive Screen test (BCoS) score, serum soluble intercellular adhesion molecule-1(ICAM-1), serum endothelin-1(ET-1), vascular endothelial growth factor (VEGF) levels, and endothelium-dependent vasodilation function (FMD) in the two groups were compared. Results: After treatment, the expression levels of NSE, TNF-α, IL-8 and CRP in the two groups were significantly decreased, and the expression levels of all indicators in the observation group were lower than that in the control group (p<0.05). After treatment, the scores of orientations, attention, memory, language, practice and action in the two groups were significantly increased, and the scores of all dimensions in the observation group were higher than those in the control group (p<0.05). After treatment, S-ICAM-1, ET-1, VEGF, FMD decreased in both groups, and all indicators of the observation group were lower than those of the control group, with statistically significant differences (p<0.05). Conclusion: Butylphthalide combined with fasudil therapy was found as effective in reducing inflammatory factors, ameliorating cognitive function and vascular endothelial function in patients with subarachnoid hemorrhage complicated with cerebral vasospasm.

15.
Anal Chim Acta ; 1280: 341868, 2023 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-37858568

RESUMEN

Sensitive and precise determination of virulent foodborne pathogens is significant for food safety. Herein, an ultrasensitive photoelectrochemical (PEC) bioanalysis was developed using the endogenous adenosine triphosphate (ATP)-responded Au@Cu2O core-shell nanocubes (Au@Cu2O NCs) to measure Escherichia coli O157: H7 (E. coli O157:H7) in food. Briefly, the phage-functionalized gold wire was used to specifically recognize the target pathogen. With the bacteriolysis of lysozyme, the endogenous ATP molecules were emitted from the captured target bacteria and enriched by another ATP aptamer-modified gold wire. Following the exchange with complementary DNA (cDNA) chains, the bonded ATP would be released. It could simultaneously etch the Au@Cu2O NCs and compete with external circuit electrons to combine photogenerated holes on the Au@Cu2O NCs-modified screen-printed electrode. With the synergy of the two signal amplification mechanisms, a significant attenuation of photocurrent signal appeared even with femtomolar ATP. Therefore, the purpose of ultrasensitive determination of E. coli O157:H7 was realized, which depended on the endogenous ATP rather than exogenous signal probes. The proposed biosensor presented a good analysis performance within 10-106 CFU/mL with a detection limit of 5 CFU/mL. Besides, its specificity, repeatability, and stability were also investigated and acceptable. The detection results for food samples matched well with the results detected by the plate counting method. This work gives an innovative and sensitive signal amplification strategy for PEC bioassays in foodborne pathogens detection.


Asunto(s)
Técnicas Biosensibles , Escherichia coli O157 , Escherichia coli O157/genética , Adenosina Trifosfato , Oligonucleótidos , Oro/química , Técnicas Biosensibles/métodos , Microbiología de Alimentos
16.
Anal Chem ; 95(40): 14925-14933, 2023 Oct 10.
Artículo en Inglés | MEDLINE | ID: mdl-37769239

RESUMEN

Bioimaging is widely used in various fields of modern medicine. Fluorescence imaging has the advantages of high sensitivity, high selectivity, noninvasiveness, in situ imaging, and so on. However, one-photon (OP) fluorescence imaging has problems, such as low tissue penetration depth and low spatiotemporal resolution. These disadvantages can be solved by two-photon (TP) fluorescence imaging. However, TP imaging still uses fluorescence intensity as a signal. The complexity of organisms will inevitably affect the change of fluorescence intensity, cause false-positive signals, and affect the accuracy of the results obtained. Fluorescence lifetime imaging (FLIM) is different from other kinds of fluorescence imaging, which is an intrinsic property of the material and independent of the material concentration and fluorescence intensity. FLIM can effectively avoid the fluctuation of TP imaging based on fluorescence intensity and the interference of autofluorescence. Therefore, based on silica-coated gold nanoclusters (AuNCs@SiO2) combined with nucleic acid probes, the dual-mode nanoprobe platform was constructed for TP and FLIM imaging of intracellular endogenous miRNA-21 for the first time. First, the dual-mode nanoprobe used a dual fluorescence quencher of BHQ2 and graphene oxide (GO), which has a high signal-to-noise ratio and anti-interference. Second, the dual-mode nanoprobe can detect miR-21 with high sensitivity and selectivity in vitro, with a detection limit of 0.91 nM. Finally, the dual-mode nanoprobes performed satisfactory TP fluorescence imaging (330.0 µm penetration depth) and FLIM (τave = 50.0 ns) of endogenous miR-21 in living cells and tissues. The dual-mode platforms have promising applications in miRNA-based early detection and therapy and hold much promise for improving clinical efficacy.

17.
Mol Biotechnol ; 2023 Aug 09.
Artículo en Inglés | MEDLINE | ID: mdl-37556107

RESUMEN

The blood-brain barrier (BBB) is a highly selective semi-permeable barrier that separates circulating blood from the extracellular fluid of the brain and central nervous system, which is crucial for maintaining brain homeostasis. This study aimed to explore the role of propofol in BBB damage and further evaluate the underlying molecular mechanism. Lipopolysaccharide (LPS) was administered to mice to create an in vivo BBB damage mice model. Additionally, hCMEC/D3 cells as brain microvascular endothelial cells (BMECs) were treated with LPS to establish the in vitro BBB damage cell model. Subsequently, propofol was used for the BBB damage model. Evans blue staining and fluorescein sodium were utilized in the in vivo experiments to demonstrate BBB leakage and BBB permeability. Cell counting kit-8 (CCK-8) assay was used to assess cell viability and the trans-endothelial electrical resistance (TEER) value was measured using an epithelial voltmeter. Furthermore, enzyme-linked immunosorbent assay was performed to measure the levels of the inflammatory cytokines such as interleukin-1ß (IL-1ß) and tumor necrosis factor-alpha (TNF-α). The levels of miR-130a-5p and zonula occludens-1 (ZO-1) in brain tissues and cells were detected using reverse transcription-quantitative polymerase chain reaction, western blot, or immunofluorescence staining. Furthermore, a dual-luciferase reporter assay was used to demonstrate the association between miR-130a-5p and ZO-1. Propofol treatment suppressed BBB leakage, the amount of fluorescein sodium, and the levels of IL-1ß and TNF-α in the LPS-induced BBB damage mice model. Meanwhile, propofol treatment increased the TEER value in the LPS-induced hCMEC/D3 cells. Additionally, propofol treatment significantly down-regulated miR-130a-5p and up-regulated ZO-1. More importantly, miR-130a-5p directly targeted ZO-1 and negatively regulated ZO-1 expression in hCMEC/D3 cells. Furthermore, miR-130a-5p mimic partially reversed the effect of propofol on the TEER value and the levels of inflammatory cytokines such as IL-1ß and TNF-α in the LPS-induced hCMEC/D3 cells. Propofol suppressed LPS-induced BBB damage by regulating miR-130a-5p/ZO-1 axis. These findings suggested a potentially effective treatment approach for BBB damage.

18.
Anal Chem ; 95(30): 11211-11218, 2023 Aug 01.
Artículo en Inglés | MEDLINE | ID: mdl-37471313

RESUMEN

Rapid, specific, and on-site detection of virulent foodborne pathogenic strains plays a key role in controlling food safety. In this work, an ultrasensitive and specific Phage@DNAzyme signal probe was designed to detect foodborne pathogens. The proposed sensing probe was composed of the selected phage and functionalized DNAzyme, which realized the specific recognition of target foodborne pathogens at the strain level and the efficient catalysis of copper(II) based azide-alkyne cycloaddition (CuAAC) click reaction with fluorescent signal, respectively. As a proof of concept, the virulent Escherichia coli O157:H7 (E. coli O157:H7) as the representative analyte was first enriched and purified from the complex food samples by a 4-mercaptophenylboronic acid-modified gold slide. Following, the Phage@DNAzyme probes were specifically combined with the captured E. coli O157: H7 and catalyzed the click reaction between 3-azido-7-hydroxycoumarin and 3-butyn-1-ol with the assistance of Cu(II) to generate a visual fluorescent signal. Finally, the corresponding fluorescent signals were measured by a smartphone to quantify the target concentrations. Under optimized conditions, the bioassay exhibited a wide linear range from 102 to 108 CFU/mL and the detection limit was 50 CFU/mL (S/N = 3). It was further extended to the detection of another foodborne pathogen Salmonella typhimurium with satisfying sensing performances. This work gives a new path for developing rapid, specific, and on-site detection methods for trace levels of pathogenic strains in foods.

19.
Anal Chim Acta ; 1275: 341591, 2023 Sep 22.
Artículo en Inglés | MEDLINE | ID: mdl-37524477

RESUMEN

Sensitive and accurate detection of multiplex foodborne pathogens is crucial for food safety. In this work, a dual-mode and dual-target biosensor regulated by a Tesla valve was established for simultaneously determining Escherichia coli O157:H7 (E. coli) and Salmonella typhimurium (S. T). Two zeolitic imidazolate framework (ZIF-8) signal probes decorated with electroactive materials (ferrocene or methylene blue), DNAzyme, and different phages were synthesized to specifically recognize the targets and generate fluorescent/electrochemical dual-mode signals. In the presence of bacteria, they were captured and enriched on two individual working electrodes through the modified 4-mercaptophenylboric acid. The encoded signal probes added on different working electrodes could be conjugated with the corresponding target bacteria depending on the specificity of phages. Under the acidic condition, the DNAzyme could catalyze click chemistry for fluorescent signals. Simultaneously, the released ferrocene and methylene blue from ZIF-8 could generate electrochemical signals at different potentials. Benefiting from the flow regulation feature of the Tesla valve, the triggered fluorescent and electrochemical signals in the two individual electrodes would not influence each other, achieving simultaneous dual-mode and dual-target determination of foodborne pathogens. It depicted good linearity ranged 10-107 CFU mL-1. And the corresponding detection of limits were 5 CFU mL-1 and 8 CFU mL-1 for two bacteria, respectively. A low false positive was realized through the dual-mode strategy. The proposed biosensor can not only on-site, specifically, and sensitively determine E. coli and S. T, but also provide the wide prospect in rapid screening of other foodborne pathogens.


Asunto(s)
Bacteriófagos , Técnicas Biosensibles , ADN Catalítico , Zeolitas , Contaminación de Alimentos/análisis , Microbiología de Alimentos , Metalocenos , Escherichia coli , Azul de Metileno
20.
Micromachines (Basel) ; 14(6)2023 May 28.
Artículo en Inglés | MEDLINE | ID: mdl-37374721

RESUMEN

The relaxor ferroelectric single crystal (1-x)Pb(Mg1/3Nb2/3)O3-xPbTiO3 (PMN-PT) has high piezoelectric constants, and thus has a good application prospect in the field of highly sensitive piezoelectric sensors. In this paper, for relaxor ferroelectric single crystal PMN-PT, the bulk acoustic wave characteristics on pure- and pseudo-lateral-field-excitation (pure- and pseudo-LFE) modes are investigated. LFE piezoelectric coupling coefficients and acoustic wave phase velocities for PMN-PT crystals in different cuts and electric field directions are calculated. On this basis, the optimal cuts of pure-LFE and pseudo-LFE modes of relaxor ferroelectric single crystal PMN-PT are obtained, namely, (zxt)45° and (zxtl)90°/90°, respectively. Finally, finite element simulations are carried out to verify the cuts of pure-LFE and pseudo-LFE modes. The simulation results show that the PMN-PT acoustic wave devices in pure-LFE mode have good energy-trapping effects. For PMN-PT acoustic wave devices in pseudo-LFE mode, when the device is in air, no obvious energy-trapping emerges; when the water (as a virtual electrode) is added to the surface of the crystal plate, an obvious resonance peak and the energy-trapping effect appears. Therefore, the PMN-PT pure-LFE device is suitable for gas-phase detections. While the PMN-PT pseudo-LFE device is suitable for liquid-phase detections. The above results verify the correctness of the cuts of the two modes. The research results provide an important basis for the development of highly sensitive LFE piezoelectric sensors based on relaxor ferroelectric single crystal PMN-PT.

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