Search Results (14)

Alzheimer’s disease (AD) early screening requires non-invasive, high-sensitivity detection of low-abundance biomarkers in complex biofluids like saliva. In this study, we present a miniaturized, silicon-based electrochemical sensor for sequential detection of two AD salivary biomarkers, lactoferrin (Lf) and amyloid β-protein 1-42 (Aβ_1-42), on a single reusable electrode. The sensor features a three-electrode system fabricated by sputter-coating a quartz substrate with gold (Au) sensing electrodes, which are further modified with gold nanoparticles (AuNPs) to form 3D dendritic structures that enhance surface area and electron transfer. To improve specificity, immunomagnetic beads (MBs) are employed to selectively capture and isolate target biomarkers from saliva samples. These MB–biomarker complexes are introduced into a polydimethylsiloxane chamber aligned with Au sensing electrodes, where a detachable magnet localizes the complexes onto the electrode surface to amplify redox signals. The AuNPs/MBs sensor achieves detection limits of 2 μg/mL for Lf and 0.1 pg/mL for Aβ_1-42, outperforming commercial ELISA kits (37.5 pg/mL for Aβ_1-42) and covering physiological salivary concentrations. After the MBs capture the biomarkers, the sensor can output the result within one minute. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) measurements confirm enhanced electron transfer kinetics on AuNP-decorated surfaces, while linear correlations (R² > 0.95) validate quantitative accuracy across biomarker ranges. The compact and integrated design eliminates reliance on bulky instrumentation and enables user-friendly operation, establishing a promising platform for portable, cost-effective AD screening and monitoring. Full article

The rapid and timely evaluation of the mental health of emergency rescuers can effectively improve the quality of emergency rescues. However, biosensors for mental health evaluation are now facing challenges, such as the rapid and portable detection of multiple mental biomarkers. In this study, a non-invasive, flexible, wearable electrochemical biosensor was constructed based on the self-assembly of nanomagnetic beads for the rapid detection of cortisol in interstitial fluid (ISF) to assess the mental stress of emergency rescuers. Based on a one-step reduction, gold nanoparticles (AuNPs) were functionally modified on a screen-printed electrode to improve the detection of electrochemical properties. Afterwards, nanocomposites of MXene and multi-wall carbon nanotubes were coated onto the AuNPs layer through a physical deposition to enhance the electron transfer rate. The carboxylated nanomagnetic beads immobilized with a cortisol antibody were treated as sensing elements for the specific recognition of the mental stress marker, cortisol. With the rapid attraction of magnets to nanomagnetic beads, the sensing element can be rapidly replaced on the electrode uniformly, which can lead to extreme improvements in detection efficiency. The detected linear response to cortisol was 0–32 ng/mL. With the integrated reverse iontophoresis technique on a flexible printed circuit board, the ISF can be extracted non-invasively for wearable cortisol detection. The stimulating current was set to be under 1 mA for the extraction, which was within the safe and acceptable range for human bodies. Therefore, based on the positive correlation between cortisol concentration and mental stress, the mental stress of emergency rescuers can be evaluated, which will provide feedback on the psychological statuses of rescuers and effectively improve rescuer safety and rescue efficiency. Full article

Background/Objectives: Focused ultrasound has advantages as an external stimulus for drug delivery as it is non-invasive, has high precision and can penetrate deep into tissues. Here, we report a gold-plated alginate (ALG) hydrogel system that retains highly water-soluble small-molecule fluorescein for sharp off/on release after ultrasound exposure. Methods: The ALG is crosslinked into beads with calcium chloride and layered with a polycation to adjust the surface charge for the adsorption of catalytic platinum nanoparticles (Pt NPs). The coated bead is subject to electroless plating, forming a gold shell. Ultrasound is applied to the gold-plated ALG beads and the release of fluorescein with or without ultrasound stimulation is quantified. Results: Polyethylenimine (PEI), not poly-L-lysine (PLL), is able to facilitate Pt NP adsorption. Gold shell thickness is proportional to the duration of electroless plating and can be controlled. Gold-plated ALG beads are impermeable to the fluorescein cargo and have nearly zero leakage. Exposure to focused ultrasound initiated the release of fluorescein with full release achieved after 72 h. Conclusions: The gold-plated ALG hydrogel is a new material platform that can retain highly water-soluble molecules with a sharp off/on release initiated by focused ultrasound. Full article

The association between microRNAs and various diseases, especially cancer, has been established in recent years, indicating that miRNAs can potentially serve as biomarkers for these diseases. Determining miRNA concentrations in biological samples is crucial for disease diagnosis. Nevertheless, the stem-loop reverse transcription quantitative PCR method, the gold standard for detecting miRNA, has great challenges in terms of high costs and enzyme limitations when applied to clinical biological samples. In this study, an isothermal signal amplification method based on a duplex-specific nuclease (DSN) enzyme-driven DNA walker and an improved catalytic hairpin assembly (CHA) was designed for miRNA detection. First, biotin–triethylene glycol-modified trigger-releasable DNA probes were conjugated to the streptavidin-coated magnetic beads for recognizing the target miRNA. The DSN enzyme specifically hydrolyzes DNA strands when the DNA probe hybridizes with the targeted miRNA. This recycling process converts the input miRNA into short trigger fragments (catalysts). Finally, three hairpins of improved CHA are driven by this catalyst, resulting in the three-armed CHA products and a fluorescence signal as the output. This dual-cycle biosensor shows a good linear relationship in the detection of miR-21 and miR-141 over the final concentration range of 250 fM to 50 nM, presenting an excellent limit of detection (2.95 amol). This system was used to detect miR-21 and miR-141 in MCF-7 and 22RV1 cells, as well as in 1% human serum. This system can be used to evaluate the expression levels of miRNAs in different biological matrices for the clinical diagnosis and prognosis of different cancers. Full article

The early detection of procalcitonin (PCT) is crucial for diagnosing bacterial infections due to its high sensitivity and specificity. While colloidal gold colorimetric and immune-chemiluminescence methods are commonly employed in clinical detection, the former lacks sensitivity, and the latter faces challenges with a brief luminescence process and an elevated background. Here, we introduce a novel approach for the quantitative analysis of PCT using surface-enhanced Raman spectroscopy (SERS), leveraging the enhanced properties of metal nanoparticles. Simultaneously, we employed a magnetic nanoparticle coating and surface biofunctionalization modification to immobilize PCT-trapping antibodies, creating the required immune substrates. The resulting magnetic nanoparticles and antibody complexes, acting as carriers and recognition units, exhibited superparamagnetism and the specific recognition of biomarkers. Then, this complex efficiently underwent magnetic separation with an applied magnetic field, streamlining the cumbersome steps of traditional ELISA and significantly reducing the detection time. In conclusion, the exploration of immunomagnetic bead detection technology based on surface-enhanced Raman spectroscopy holds crucial practical significance for the sensitive detection of PCT. Full article

This work reports the development of a functional photocatalytic coating based on a combination of polymeric electrospun fibres and nanoparticles that is intended to be activated in the visible light range. In this sense, the resulting fibres can act as an effective matrix for the incorporation of titanium dioxide (TiO₂) particles, which are covered by gold nanoparticles (AuNPs), in the outer surface of the metal oxide precursor. In the first step of the process, the optical properties of the nanoparticles were determined by UV-Vis spectroscopy. The extension of the visible absorption can be associated to the localized surface plasmon resonance (LSPR) of the metallic AuNPs. In addition, the resultant particle size distribution and average particle diameter was evaluated by dynamic light scattering (DLS) measurements. Furthermore, the phase composition and porosity of the functional particle powder were analysed by an XRD and N₂ adsorption test. In the second step, these synthesized particles have been successfully immobilized into a PAA + β-CD electrospun fibre matrix by using the two different deposition methods of dip-coating and solution-casting, respectively. The morphological characterization of the samples was implemented by means of scanning electron microscopy (SEM), showing uniform and homogeneous, free-beaded fibres with a random distribution of the synthesized particles deposited onto the electrospun fibres. Then, the functional coatings were removed from the substrate, and a thermogravimetric (TGA) analysis was carried out for each sample in order to obtain the precursor mass immobilized in the coating. Once the overall mass of precursor was obtained, the percentage of TiO₂ particles and AuNPs in the precursor was calculated by using inductively coupled plasma atomic emission spectrometry (ICP-AES). Finally, the photocatalytic activity of both functional solution and electrospun coatings were evaluated in terms of a gradual degradation of rhodamine B (RhB) dye after continuous exposition to a visible-light lamp. Full article

The gold standard for diagnostics of SARS-CoV-2 (COVID-19) virus is based on real-time polymerase chain reaction (RT-PCR) using centralized PCR facilities and commercial viral RNA extraction kits. One of the key components of these kits are magnetic beads composed of silica coated magnetic iron oxide (Fe₂O₃ or Fe₃O₄) nanoparticles, needed for the selective extraction of RNA. At the beginning of the pandemic in 2019, due to a high demand across the world there were severe shortages of many reagents and consumables, including these magnetic beads required for testing for SARS-CoV-2. Laboratories needed to source these products elsewhere, preferably at a comparable or lower cost. Here, we describe the development of a simple, low-cost and scalable preparation of magnetic nanoparticles (MNPs) from biowaste and demonstrate their successful application in viral RNA extraction and the detection of COVID-19. These MNPs have a unique nanoplatelet shape with a high surface area, which are beneficial features, expected to provide improved RNA adsorption, better dispersion and processing ability compared with commercial spherical magnetic beads. Their performance in COVID-19 RNA extraction was evaluated in comparison with commercial magnetic beads and the results presented here showed comparable results for high throughput PCR analysis. The presented magnetic nanoplatelets generated from biomass waste are safe, low-cost, simple to produce in large scale and could provide a significantly reduced cost of nucleic acid extraction for SARS-CoV-2 and other DNA and RNA viruses. Full article

A simple and sensitive AuNP-coated magnetic beads (AMB)-based electrochemical biosensor platform was fabricated for bioassay. In this study, AuNP-conjugated magnetic particles were successfully prepared using biotin–streptavidin conjugation. The morphology and structure of the nanocomplex were characterized by scanning electron microscopy (SEM) with energy-dispersive X-ray analysis (EDX) and UV–visible spectroscopy. Moreover, cyclic voltammetry (CV) was used to investigate the effect of AuNP-MB on alkaline phosphatase (ALP) for electrochemical signal enhancement. An ALP-based electrochemical (EC) immunoassay was performed on the developed AuNP-MB complex with indium tin oxide (ITO) electrodes. Subsequently, the concentration of capture antibodies was well-optimized on the AMB complex via biotin–avidin conjugation. Lastly, the developed AuNP-MB immunoassay platform was verified with extracellular vesicle (EV) detection via immune response by showing the existence of EGFR proteins on glioblastoma multiforme (GBM)-derived EVs (10⁸ particle/mL) spiked in human plasma. Therefore, the signal-enhanced ALP-based EC biosensor on AuNP-MB was favorably utilized as an immunoassay platform, revealing the potential application of biosensors in immunoassays in biological environments. Full article

In 1914, a magnificent reliquary cross dating from the early XIIIth century was discovered in a safe from the Liège Cathedral. This double-arm cross shows a wooden structure, covered by gold-coated copper on the front, and by carved silver plates on the back. Its total length is 34 cm, and it is covered by filigrees, gems, glass beads, and pearls on its front. The reliquary cross was analysed by Raman spectrometry and X-ray fluorescence spectrometry (pXRF) to determine the mineralogical and chemical compositions of gems, glass beads, and metals that have been used to decorate it. The results confirm the identification of twenty-five turquoises from Egypt, one garnet from Sri Lanka, as well as six quartz and one opal whose origin is difficult to certify. Twelve glass beads, showing green, blue, or amber tints, were also identified. Their compositions either correspond to soda lime glasses with natron or to potash–lead glasses similar to those of Central Europe. Moreover, a small polished red cross and a green stone appear to be constituted by nice doublets, composed of coloured glass covered by quartz. The filigrees contain Au and Cu, while carved plates covering the edges and the back of the cross are made of silver. Full article

This review describes and appraises a novel protein-based antigen detection test for visceral leishmaniasis (VL). The test detects in patient’s urine six proteins from Leishmania infantum (chagasi) and Leishmania donovani, the etiological agents of VL. The gold standard test for VL is microscopic observation of the parasites in aspirates from spleen, liver, or bone marrow (and lymph node for dogs). Culture of the parasites or detection of their DNA in these aspirates are also commonly used. Serological tests are available but they cannot distinguish patients with active VL from either healthy subjects exposed to the parasites or from subjects who had a successful VL treatment. An antigen detection test based on the agglutination of anti-leishmania carbohydrates antibody coated latex beads has been described. However, the results obtained with this carbohydrate-based test have been conflicting. Using mass spectrometry, we discovered six L. infantum/L. donovani proteins excreted in the urine of VL patients and used them as markers for the development of a robust mAb-based antigen (protein) detection test. The test is assembled in a multiplexed format to simultaneously detect all six markers. Its initial clinical validation showed a sensitivity of 93% and specificity of 100% for VL diagnosis. Full article

A new generation of magnetic lateral flow immunoassays is emerging as powerful tool for diagnostics. They rely on the use of magnetic nanoparticles (MNP) as detecting label, replacing conventional gold or latex beads. MNPs can be sensed and quantified by means of external devices, allowing the development of immunochromatographic tests with a quantitative capability. Moreover, they have an added advantage because they can be used for immunomagnetic separation (IMS), with improvements in selectivity and sensitivity. In this paper, we have reviewed the current knowledge on magnetic-lateral flow immunoassay (LFIA), coupled with both research and commercially available instruments. The work in the literature has been classified in two categories: optical and magnetic sensing. We have analysed the type of magnetic nanoparticles used in each case, their size, coating, crystal structure and the functional groups for their conjugation with biomolecules. We have also taken into account the analytical characteristics and the type of transduction. Magnetic LFIA have been used for the determination of biomarkers, pathogens, toxins, allergens and drugs. Nanocomposites have been developed as alternative to MNP with the purpose of sensitivity enhancement. Moreover, IMS in combination with other detection principles could also improve sensitivity and limit of detection. The critical analysis in this review could have an impact for the future development of magnetic LFIA in fields requiring both rapid separation and quantification. Full article

In this study, a passive sampler for gaseous elemental mercury (GEM) was developed and applied to field monitoring. Three Radiello^® diffusive bodies with gold-coated beads as Hg adsorbent were installed in an acrylic external shield. Hg uptake mass linearly increased as the deployment time increased until 8 weeks with an average gaseous Hg concentration of 2 ng m⁻³. The average of the experimental sampling rate (SR) was 0.083 m³ day⁻¹ and showed a good correlation with theoretical SRs, indicating that a major adsorption mechanism was molecular diffusion. Nonetheless, the experimental SR was approximately 33% lower than the modeled SR, which could be associated with inefficient uptake of GEM in the sampler or uncertainty in constraining model parameters. It was shown that the experimental SR was statistically affected by temperature and wind speed but the calibration equation for the SR by meteorological variables should be obtained with a wider range of variables in further investigation. When the uptake rates were compared to the active Hg measurements, the correlation was not significant because the passive sampler was not sufficiently adept at detecting a small difference in the GEM concentration of from 1.8 to 2.0 ng m⁻³. However, the results for spatial Hg concentrations measured near cement plants in Korea suggest a possible application in field monitoring. Future research is needed to fully employ the developed passive sampler in quantitative assessment of Hg concentrations. Full article

A large number of low-resolution models have been proposed in the last decades to reduce the computational cost of molecular dynamics simulations for bio-nano systems, such as those involving the interactions of proteins with functionalized nanoparticles (NPs). For the proteins, “minimalist” models at the one-bead-per residue (Cα-based) level and with implicit solvent are well established. For the gold NPs, widely explored for biotechnological applications, mesoscale (MS) models treating the NP core with a single spheroidal object are commonly proposed. In this representation, the surface details (coating, roughness, etc.) are lost. These, however, and the specificity of the functionalization, have been shown to have fundamental roles for the interaction with proteins. We presented a mixed-resolution coarse-grained (CG) model for gold NPs in which the surface chemistry is reintroduced as superficial smaller beads. We compared molecular dynamics simulations of the amyloid β2-microglobulin represented at the minimalist level interacting with NPs represented with this model or at the MS level. Our finding highlights the importance of describing the surface of the NP at a finer level as the chemical-physical properties of the surface of the NP are crucial to correctly understand the protein-nanoparticle association. Full article

Viscosity is an important property of liquids. A viscosity change of aqueous substances that deviates from their normal levels usually implies a compromise in quality due to degradation or microorganism proliferation. Monitoring of macro-scale viscosity can be simply realized by various conventional tools, such as rotational viscometers, capillary tubes, falling bodies, and so forth. Nevertheless, today, micro-volume viscosity measurement remains a challenging endeavor, resulting in rare, expensive, or difficult-to-obtain samples not very well studied. For this reason, a novel technique for micro-viscosity based on rotational Brownian motion is presented in this paper. Janus microbeads were made by coating fluorescent polystyrene beads with gold film. Taking advantage of the bead configuration of half gold/half fluorescence, the rotational Brownian signal was expressed in terms of blinking fluorescent intensity. The characteristic correlation time was derived from the blinking intensity of trace amounts of a selected medium over a certain time period, and results were correlated with viscosity. Given a volume of only 2 μL for each measurement, calibration of a series of glycerol–water mixtures (100%–1% (v/v) water content) yielded good agreement with the expected viscosity predictions over the range of 0.8–574.8 cP. Five common oil products, including lubricant oil, baby oil, food oil, olive oil, and motor oil, were further investigated to demonstrate the feasibility and practicability of the proposed technique. Data measured by the rotational Brownian motion-based diffusometer were comparable with those measured by a commercial rotational viscometer. The method also explicitly showed viscosity degradation after the oils were heated at a high temperature of over 100 °C for 10 min. Evaluation proved the proposed Janus microbead-enabled rotational diffusometric technique to be a promising approach for rapid and micro-scale viscosity measurement. Full article