A novel construction strategy is introduced for an ultrasensitive dynamic light scattering (DLS) immunosensor targeting alpha fetoprotein (AFP). This approach relies on a self-assembled heptamer fusion protein (A1-C4bpα), incorporating the dual functions of multivalent recognition and crosslinking aggregation amplification due to the presence of seven AFP-specific A1 nanobodies on the A1-C4bpα heptamer. Leveraging antibody-functionalized magnetic nanoparticles for target AFP capture and DLS signal output, the proposed heptamer-assisted DLS immunosensor offers high sensitivity, strong specificity, and ease of operation. Under the optimized conditions, the designed DLS immunosensor demonstrates excellent linear detection of AFP in the concentration range 0.06 ng mL-1 to 512 ng mL-1, with a detection limit of 15 pg mL-1. The selectivity, accuracy, precision, practicability, and reliability of this newly developed method were further validated through an assay of AFP levels in spiked and actual human serum samples. This work introduces a novel approach for constructing ultrasensitive DLS immunosensors, easily extendable to the sensitive determination of other targets via simply replacing the nanobody sequence, holding great promise in various applications, particularly in disease diagnosis.

Inorganic nanoparticles (NPs) have been widely recognized for their stability and biocompatibility, leading to their widespread use in biomedical applications. Our study introduces a novel approach that harnesses inorganic magnetic nanoparticles (MNPs) to stimulate apical-basal polarity and induce epithelial traits in cancer cells, targeting the hybrid epithelial/mesenchymal (E/M) state often linked to metastasis. We employed mesocrystalline iron oxide MNPs to apply an external magnetic field, disrupting normal cell polarity and simulating an artificial cellular environment. These led to noticeable changes in the cell shape and function, signaling a shift toward the hybrid E/M state. Our research suggests that apical-basal stimulation in cells through MNPs can effectively modulate key cellular markers associated with both epithelial and mesenchymal states without compromising the structural properties typical of mesenchymal cells. These insights advance our understanding of how cells respond to physical cues and pave the way for novel cancer treatment strategies. We anticipate that further research and validation will be instrumental in exploring the full potential of these findings in clinical applications, ensuring their safety and efficacy.

The present work reports the development and application of a new electrochemical sensor for the determination of low concentration levels of p-toluenediamine (PTD) in biological fluids and surface water samples. The proposed sensor was developed using a 3D-printed magnetic device as platform for carbon screen printed electrode (CSPE) modified by magnetic nanoparticles functionalized with carboxylic groups and l-cysteine (MNP-CA-CYS). The results obtained from the morphological and electrochemical characterizations of the sensing platform enabled us to confirm the success of the sensor functionalization with l-cysteine and to have a better understanding of the electrochemical behavior and preconcentration of PTD on the electrode surface. PTD oxidation occurred at 0.24V on MNP-CA-CYS and the mechanism recorded an increase of 51.0% in anodic peak current. Under optimized conditions, the square wave voltammograms obtained for the electrode modified by 40.0 μL MNP-CA-CYS suspension at 1.0 mg mL-1, with accumulation time of 3 min, presented an analytical curve with linear range of 8.00 × 10-7 to 8.00 × 10-5 mol L-1, represented by the equation Iap = (0.383 ± 0.011)[PTD] - (8.112 ± 0.07) × 10-8 (R2 = 0.9994), and detection and quantification limits of 8.53 × 10-8 and 2.56 × 10-7 mol L-1, respectively. Finally, the proposed method was validated through comparison with high performance liquid chromatography coupled to diode array detector (HPLC-DAD) technique and was successfully applied for PTD determination in samples of surface water, tap water, fetal bovine serum and artificial urine.

BACKGROUND: Lactate dehydrogenase (LDH) is one of the important enzyme systems for glycolysis and gluconeogenesis. It can catalyze the reduction and oxidation reaction between propionic acid and L-lactic acid, which is usually overexpressed in cancer cells. Therefore, inhibiting the activity of LDH is a promising way for the treatment of cancer. In this study, an effective method based on ligand fishing and ultra performance liquid chromatography-mass spectrum (UPLC-MS) was established to screen and identify active ingredients from Selaginella doederleinii with potential inhibitory activity for LDH.
METHODS: Firstly, LDH was immobilized on the magnetic nanoparticles (MNPs), three immobilization parameters including LDH concentration, immobilization time and pH were optimized by single factor and response surface methodology for maximum (max) immobilization yield. Then, a mixed model of galloflavin and chlorogenic acid (inhibitors and non-inhibitors of LDH) was used to verify the specificity of immobilized LDH ligand fishing, and the conditions of ligand fishing were further optimized. Finally, combined with UPLC-MS, immobilized LDH was used to simultaneously screen and identify potential LDH inhibitors from the ethyl acetate extract of Selaginella doederleinii.
RESULTS: The prepared fishing material was comprehensively characterized by scanning electron microscopy (SEM), transmission electron microscope (TEM), X-ray diffraction (XRD) and fourier transform infrared spectrometer (FT-IR). The optimal immobilization conditions were obtained as LDH concentration of 0.7 mg/mL, pH value of 4.5, and immobilization time of 3.5 h. Under these conditions, the max immobilization yield was (3.79 ± 0.08) × 103 U/g. The specificity analysis showed that immobilized LDH could recognize and capture ligands, and the optimal ligand fishing conditions included that the incubation time was 30 min, the elution time was 20 min, and the concentration of methanol as eluent was 80%. Finally, two LDH inhibitors, amentoflavone and robustaflavone, were screened by immobilized LDH from the ethyl acetate extract of Selaginella doederleinii.
CONCLUSIONS: The study provided a meaningful evidence for discovering the bioactive constituents in ethyl acetate extract of Selaginella doederleinii related to cancer treatment, and this ligand fishing method was feasible for screening enzyme inhibitors from similar complex mixtures.

Multifunctional nanocarriers, specifically for tumor targeting and traceable features, have been increasingly considered in cancer therapies. Herein, a novel targeting agent (TA), tryptophan (TRP), was proposed for the synthesis of functionalized (3-aminopropyl) triethoxysilane-iron oxide nanoparticles using two methods, creating a smart drug delivery system (DDS). In one method, two-step, glutaraldehyde (GA) as a linker, bonded TRP and amino-functionalized magnetite, and in the second method, one step, TRP binding was carried out by (3-dimethyl aminopropyl)-N\'-ethyl carbodiimide hydrochloride (EDC)/N-hydroxysuccinimide ester. The synthesis yield of the second method was 7% higher than the first method. After synthesizing DDS, 5-fluorouracil (5-FU) was loaded on nanocarriers and was observed TRP functionalized nanoparticles by GA have better loading efficiency, which was 50% greater than the product from the one-step method. A pH-sensitive release profile was also studied for 5-FU/DDS with the release of almost 75% and 50% at pH 5.5 and 7.4, respectively. To analyze the biological aspects of nanocarriers, human breast cancer, MCF-7, and embryonic kidney, HEK293, cell lines were used for cellular uptake and 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assays.In vitrostudies confirmed that TRP can act as a TA as its cellular uptake through cancerous cells was 40% greater than normal cells, and the MTT assay confirmed that using DDS can increase and decrease the cell viability of normal cells and cancerous cells, respectively, compared to free drug. Therefore, it was concluded that advanced nano-assembly is a great candidate for breast cancer cell-targeted delivery.

Insufficient acetylcholine (ACh) can cause cognitive and memory dysfunction, clinically known as, Alzheimer\'s disease (AD). Acetylcholinesterase (AChE) can hydrolyze ACh into acetic acid and inactivate choline. Therefore, inhibiting the activity of AChE would help to improve the effectiveness of AD treatment. Currently, the methods for rapid screening of AChE inhibitors are limited. This study reports the application of AChE-immobilized magnetic nanoparticles as a drug screening tool to screen AChE inhibitors for natural products. First, AChE was immobilized on a surface of amino-modified magnetic nanoparticles using covalent binding and the AChE concentration, and the pH as well as time was optimized to obtain the maximum enzyme immobilization yield (61.4 μg/mg), and the kinetic model indicated that AChE-immobilized magnetic nanoparticles and the substrate had the high affinity and specificity. Then, a ligand fishing experiment was carried out using a mixed model of tacrine (an inhibitor of AChE) and caffeic acid (a non-inhibitor of AChE) to verify the specificity of the immobilized AChE, and the conditions for ligand fishing were further optimized. Finally, the optimized immobilized AChE was combined with UPLC-MS to screen for AChE inhibitors in Selaginella doederleinii Hieron extracts. Four compounds were confirmed to be potent AChE inhibitors. Among the four compounds, amentoflavone had a stronger AChE inhibitory effect than tacrine (positive control) with an IC50 of 0.73 ± 0.009 μmol/L. The results showed that AChE-functionalized magnetic nanoparticles can be used in the discovery of target drugs from complex matrices.

The lipase from Bacillus licheniformis NCU CS-5 was immobilized onto β-cyclodextrin (CD) grafted and aminopropyl-functionalized chitosan-coated Fe3O4 magnetic nanocomposites (Fe3O4-CTS-APTES-GA-β-CD). Fourier transform infrared spectroscopy, thermogravimetry analysis, X-ray diffraction, scanning electron microscopy and transmission electron microscopy showed that not only the functionalized magnetic nanoparticles were synthesized but also the immobilized lipase was successfully produced. The immobilized lipase exhibited higher optimal pH value (10.5) and temperature (60℃) than the free lipase. The pH and thermal stabilities of the immobilized lipase were improved significantly compared to the free lipase. The immobilized lipase remained more than 80% of the relative activity at temperature of 60 ℃ and pH 12.0. The immobilized lipase also remained over 80% of its relative activity after 28 days of storage and 15 cycles of application. The application of the immobilized lipase in esterification of isoamyl acetate and pentyl valerate showed that maximum esterification efficiency was achieved in n-hexane having 68.0% and 89.2% respectively. Therefore, these results indicated that the Fe3O4-CTS-APTES-GA-β-CD nanoparticles are novel carriers for immobilizing enzyme, and the immobilized lipase can be used as an innovative green approach to the synthesis of fruity flavor esters in food industry.

Cascade reactions catalyzed by two or more enzymes have been widely used in industrial production and exhibited many advantages over the single-enzyme catalytic system. In this study, two components of hydroxylase monooxygenase (HpaBC) were first co-immobilized by Ni2+-nitrilotriacetic acid (Ni-NTA) functionalized magnetic silica nanoparticles (Ni-NTA/H2N-SiO2@Fe3O4) for enhancing the stability and activity of biocatalysts with multi-components. These two components, HpaB and HpaC, were modified with histidine-tag and employed to construct a bi-enzyme catalytic system. After co-immobilization, the activity of the bi-enzyme system was 2.6 times of free enzymes. Meanwhile, the co-immobilized system was more stable against high temperature and alkaline condition, and maintained 76.6% of the initial activity for storage 12 days. Moreover, the co-immobilized HpaBC remained more than 60% catalytic activity after 7 cycles. These results showed that co-immobilized multi-component enzymes based on functionalized magnetic nanoparticles without purification would play a great potential role in the field of industrial biocatalysis.

Here, an ultrasensitive non-enzymatic glucose sensor was fabricated using a facile and low price electro-deposition method. At first, thiol-functionalized magnetic nanoparticles cast onto the glassy carbon electrode (GCE) surface to provide a stable substrate with the high surface area. Then, CuO nanoparticles and Ag nanoparticles electrodeposit on the surface of Fe3O4-SH/GCE to obtain the final modified GCE. The characterization of electro-synthesized nanoparticles and the modified GCE was done by different related techniques such as field emission scanning electronic microscopy (FESEM), energy-dispersive X-ray spectroscopy (EDX), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The proposed electrode was applied to the electrochemical sensing of glucose. By employing the optimum conditions on the preparation of modified electrode such as time and potential for electrosynthesis of two different nanoparticles, high reproducibility of measurement and sensor preparation were achieved. The thus optimized simple glucose sensor could be provided stable responses in a wide linear range from 0.06 to 1000 μM with detection limit 15 nM, indicating its potential application for real biological samples.

Alarming growth of pharmaceutical residues in aquatic environment has elevated concerns about their potential impact on human health. Taking cognizance of this, the present study is focussed on the coating of cobalt ferrite nanoparticles with different functionalities and to use them as adsorbents for pharmaceutical waste. The thickness of the coating was analysed using Small angle X-ray scattering technique. Thorough study of the isotherms and kinetics were performed suggesting monolayer adsorption and pseudo kinetic order model, respectively. To get an insight of the interactions liable for adsorption of fluoroquinolones over the functionalized magnetic nanoparticles computational studies were undertaken. The results demonstrated substantial evidence proposing remarkable potential of these nanostructures as adsorbents for different pollutants with an additional advantage of stability and facile recoverability with a view to treat wastewater.