Magnetic Microparticles Research Articles

Retraction Note: Biocompatibility and Antimicrobial Investigation of Agar-Tannic Acid Hydrogel Reinforced with Silk Fibroin and Zinc Manganese Oxide Magnetic Microparticles

Retraction Note: Biocompatibility and Antimicrobial Investigation of Agar-Tannic Acid Hydrogel Reinforced with Silk Fibroin and Zinc Manganese Oxide Magnetic Microparticles

Actuating Extracellular Matrices Decouple the Mechanical and Biochemical Effects of Muscle Contraction on Motor Neurons.

Emerging in vivo evidence suggests that repeated muscle contraction, or exercise, impacts peripheral nerves. However, the difficulty of isolating the muscle-specific impact on motor neurons in vivo, as well as the inability to decouple the biochemical and mechanical impacts of muscle contraction in this setting, motivates investigating this phenomenon in vitro. This study demonstrates that tuning the mechanical properties of fibrin enables longitudinal culture of highly contractile skeletal muscle monolayers, enabling functional characterization of and long-term secretome harvesting from exercised tissues. Motor neurons stimulated with exercised muscle-secreted factors significantly upregulate neurite outgrowth and migration, with an effect size dependent on muscle contraction intensity. Actuating magnetic microparticles embedded within fibrin hydrogels enabledynamically stretching motor neurons and non-invasively mimicking the mechanical effects of muscle contraction. Interestingly, axonogenesis is similarly upregulated in both mechanically and biochemically stimulated motor neurons, but RNA sequencing reveals different transcriptomic signatures between groups, with biochemical stimulation having a greater impact on cell signaling related to axonogenesis and synapse maturation. This study leverages actuating extracellular matrices to robustly validate a previously hypothesized role for muscle contraction in regulating motor neuron growth and maturation from the bottom-up through both mechanical and biochemical signaling.

Driving-sensing integrated magnetic soft robots via laser thermal printing

The integration of driving and sensing can effectively promote the intelligence and miniaturization of magnetic soft robots. A highly adaptable, efficient, and flexible method for constructing functional structures of composites is crucial for magnetic soft robots with sensing capabilities. In this work, a novel laser thermal printing (LTP) process is proposed to flexibly and efficiently prepare thermosetting composite based magnetic soft robots with integrated sensing capabilities. By cleverly utilizing the laser photothermal effect, CNTs (Carbon Nanotubes) photothermal absorbers and PDMS (Polydimethylsiloxane) thermosetting properties, LTP process effectively achieves rapid thermoforming of thermosetting resin PDMS. On this basis, we present the first-ever application of the process for the synthesis of butterfly-shaped, inchworm-shaped and claw-shaped magnetic soft robots. Due to NdFeB magnetic microparticles within PDMS matrix and a CNTs conductive layer on PDMS surface, the obtained biomimetic robots achieve seamless integration of driving and sensing capabilities. In applications such as circuit repair and object grasping, real-time and in-situ changes in sensing signals provide feedback on the posture changes and motion processes of these robots, highlighting their intelligent features. Therefore, LTP process provides a novel approach for the efficient fabrication of miniaturized, intelligent soft robots.

Magneto-Thermal Hydrogel Swarms for Targeted Lesion Sealing.

Magnetic microswarms capable of performing navigation to targeted lesions show great potential for in vivo medical applications. However, using the swarms for lesion cavity filling encounters challenges from precise delivery and sealing. Herein, this work develops a magneto-thermal hydrogel swarm consisting of magnetic hydrogel particles, which can perform phase transition induced by temperature change. The particles are prepared using a temperature-responsive hydrogel matrix, tissue adhesive monomers, and magnetic microparticles. The swarms can be remolded to various shapes, and it can be used to seal perforation in phantom and gastric tissue. The swarms can also serve as drug carriers, and their drug release profiles induced by temperature changes are characterized. Finally, the targeted delivery, adaptive filling, and sealing of a gastric ulcer using the swarms are achieved in ex vivo and in vivo environments.

Synergistic Dual Antibacterial Activity of Magnetite Hydrogels Doped with Silver.

In this work, we utilized poly-N-isopropylacrylamide (NIPAM), magnetic nanoparticles (MNPs), and silver nitrate to prepare magnetic hydrogel microparticles doped with silver, which exhibited a dual antimicrobial effect. The antibacterial effect of these composites was mediated by the antimicrobial activity of silver and the magnetic hyperthermic induction, which we believe increased biofilm disruption and silver release into the surrounding bacterial biofilms. The prepared particles were characterized by using several analytical techniques. The particles exhibited a porous morphology impregnated evenly with silver nanoparticles, as observed by scanning electron microscopy (SEM). Furthermore, we examined the antibacterial activity of our microparticles against Escherichia coli by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Our findings revealed that the composites demonstrated significant antibacterial activity of up to 81% under magnetic hyperthermia as compared to 45% when samples were heated to the same temperature in a water bath at constant silver concentration. This demonstrates the distinctive inhibitory features of MNPs in enhancing bacterial killing when a magnetic field is applied. The findings of this study lay the groundwork for further exploration of microparticle-based antimicrobial therapies, which can contribute to the development of more advanced wound healing devices and better sterilization methods for medical devices.

Development of a chemiluminescent immunoassay based on magnetic solid phase for quantification of homocysteine in human serum

BackgroundHomocysteine (HCY) is a sulfur-containing amino acid that is an independent or important risk factor for the occurrence of many chronic diseases and is one of the most important indicators for determining health risks. However, existing HCY detection methods do not meet the requirements of clinical diagnosis. Therefore, there is an urgent need to establish new detection methods to meet the needs of clinical detection.ResultsIn this study, we used the principle of competitive method to establish a new method for the determination of HCY in human serum using a chemiluminescent enzyme immunoassay in conjunction with a chemiluminescent assay instrument that uses magnetic microparticles as the solid phase of the immunoreaction. The established method achieved satisfactory results in terms of minimum detection limit, specificity, accuracy, and clinical application. The limit of detection was 0.03 ng/mL. The intra-assay coefficient of variation (CV) was 1.94–5.05%, the inter-assay CV was 2.29–6.88%, and the recovery rate was 88.60–93.27%. Cross-reactivity with L-cysteine ranged from 0.0100 to 0.0200 μmol/L, and cross-reactivity with glutathione ranged from 0.0100 to 0.200 μmol/L, all of which were less than the limit of detection (LoD) of this method. The linear factor R of this method was greater than 0.99.ConclusionsIn summary, the developed method showed a good correlation with the product from Abbott. A total of 996 clinical patients with cardiovascular diseases were evaluated using the method developed in this study.

Magnetically Driven Hydrogel Surfaces for Modulating Macrophage Behavior.

During the host response toward implanted biomaterials, macrophages can shift phenotypes rapidly upon changes in their microenvironment within the host tissue. Exploration of this phenomenon can benefit significantly from the development of adequate tools. Creating cell microenvironment alterations on classical hydrogel substrates presents challenges, particularly when integrating them with cell cultivation and monitoring processes. However, having the capability to dynamically manipulate the cell microenvironment on biomaterial surfaces holds significant potential. We introduce magnetically actuated hydrogels (MadSurface) tailored to induce reversible stiffness changes on polyacrylamide hydrogel substrates with embedded magnetic microparticles in a time-controllable manner. Our investigation focused on exploring the potential of magnetic fields and MadSurfaces in dynamically modulating macrophage behavior in a programmable manner. We achieved a consistent modulation by subjecting the MadSurface to a pulsed magnetic field with a frequency of 0.1 Hz and a magnetic field flux density of 50 mT and analyzed exposed cells using flow cytometry and ELISA. At the single-cell level, we identified a subpopulation for which the dynamic stiffness conditions in conjunction with the pulsed magnetic field increased the expression of CD206 in M1-activated THP-1 cells, indicating a consistent shift toward the M2 anti-inflammatory phenotype on MadSurface. At the population level, this effect was mostly hindered in the culture period utilized in this work. The MadSurface approach advances our understanding of the interplay between magnetic field, cell microenvironment alterations, and macrophage behavior.

Navigation of superparamagnetic particles with a pre-magnetized high-temperature superconducting bulk in a weak background field

Abstract Recent research has shown significant progress towards a full body magnetic drug delivery (MDD) system for use in targeted cancer treatments. Although many different MDD systems have been proposed to generate strong remote forces capable of rapidly changing directions at distances greater than 10 cm, current state-of-the-art technologies lack in force strength and/or degrees of freedom. Knowing that high temperature superconducting (HTS) bulks can achieve trapped fields an order of magnitude larger than ferromagnets, this work aims at numerically and experimentally evaluating the forces that can be produced by HTS bulks in a uniform magnetic field. We first use Hall probe measurements and finite element simulations to determine the magnetic field generated by an HTS pellet and show that both results are in good agreement. Using a combination of simulations and experiments, we then show that for a 14 × 6 mm YBa2Cu3O 7 − x pellet magnetized at 2 T, remote forces on superparamagnetic microparticles are maximized at background fields of ∼50 mT. In addition, the direction of the magnetic forces can be flipped by changing the direction of the applied field relative to the HTS’s magnetization. The HTS bulk was successfully used to navigate magnetic microparticles in a glass bifurcation mimicking the hepatic artery of the human liver. Finally, we show by simulation that a large HTS pellet magnetized at 5 T in a field of ∼250 mT can generate stronger forces with more degrees of freedom than the strongest forces achievable in current MDD technologies.

DNA polymerase mediated CRISPR/Cas12a trans-cleavage for dual-mode quantification of uracil DNA glycosylase activity

Since an unusual expression of uracil-DNA glycosylase (UDG) is often associated with the pathogenesis of numerous disorders, the detection of UDG activity is regarded as a promising application in disease diagnosis. Here, we develop a DNA polymerase-mediated CRISPR/Cas12a trans cleavage strategy, which can achieve dual-mode determination of UDG activity. By introducing a hairpin DNA probe containing a single uracil base, the probe undergoes specific cleavage and elongation under the existence of UDG only, thus activating the trans cleavage of ssDNA regardless of its length and sequence. To accommodate different detection modes, the ssDNA was further modified by fluorophore-quencher pairs or designed for connecting magnetic microparticles (MMPs) and polystyrene microparticles (PMPs). Finally, the UDG activity is quantified by fluorescence signal and microparticle accumulation length on a microfluidic chip visible to the naked eye. This strategy provides a detectable minimum UDG concentration of 0.00047 U/mL for fluorescent mode and 0.0048 U/mL for microfluidic mode. Furthermore, we performed the UDG inhibition test and detected UDG activity in cell lysates, suggesting its potential for inhibitor screening and detection of UDG in biological samples.

Soft Magnetoelastic Tactile Multi-Sensors with Energy-Absorbing Properties for Self-Powered Human-Machine Interfaces.

Tactile sensors play a key role in human-machine interfaces (HMIs) for augmented and virtual reality, point-of-care devices, and human-robot collaboration, which show the promise of revolutionizing our ways of life. Here, we present a sensor (EMTS) that utilizes the magnetoelastic effect in a soft metamaterial to convert mechanical pressure into electrical signals. With this unique mechanism, the proposed EMTS simultaneously possesses self-powering, waterproof, and compliant features. The soft metamaterial is essentially a porous magnetoelastomer structure designed based on the Fourier series expansion, which allows for programmable mechanical response and sensing performance of the EMTS. Fabricated by simple 3D-printed molds, the EMTS also holds potential for low-cost production. Particularly, the porous magnetoelastomer structure comes with selectable buckling instabilities that can significantly enhance biomechanical-to-electrical energy conversion. Also, with the embedded magnetic microparticles, the energy-absorbing performance of the sensor is greatly improved, which is highly beneficial to HMIs. To pursue practical applications, the EMTSs are further integrated with two systems as control and perception modules. It is demonstrated that the EMTS is able to identify different hand gestures to control a lighting system even in a high-humidity environment. Also, the EMTS stands out for its superior capability of simultaneous impact perception and energy absorption in drop tests. Overall, with its compelling array of features, the presented EMTS gives impetus to multi-sensing technology and practically enables a variety of HMI applications.

Portable cell imprinted polymer-based microfluidic sensor for bacteria detection in real water

Cell-imprinted polymer (CIP) based optical biosensors have transformed point-of-care detection. However, challenges remain in their portability and detection sensitivity, time, and cost. Herein, we present an imprinted polymer-based low-cost microfluidic device integrated into a portable enclosure that enables rapid and sensitive bacteria detection in real water. A portable 3D-printed platform was custom-designed, housing all essential detection components, i.e., pumping and fluorescent imaging units and the microfluidic sensor. CIP coated magnetic microparticles (MPs) with affinity to bacteria were manipulated inside the magnetophoretic microfluidic device at an optimized flow rate of 0.01 mL/min for bacteria capturing. Fluorescent imaging pre- and post-bacteria capture facilitated quantification of fluorescence intensity changes as bacteria were trapped by the CIP-MPs. The sensor’s dose–response curve established limits of detection (LOD) and quantification (LOQ) at 8 × 103 and 6 × 105 CFU/mL, respectively, within a dynamic range of 103 to 109 CFU/mL. It specifically detected E. coli, distinguishing it from non-specific bacteria like Salmonella and Sarcina. In real pond water tests, our sensor detected 2 × 10⁶ CFU/mL, matching a central lab’s result of 2.33 × 10⁶ CFU/mL, demonstrating its effectiveness for real-water monitoring. While further enhancements are needed for improving the specificity in complex environmental matrices and broader bacterial strain detection, the sensor’s simplicity and portability highlight its potential for practical potential.

Association of MTHFR C677T, MTHFRA1298C, and MTRRA66G Gene Polymorphisms with Hyperhomocysteinemia and Its Modulation by the Combined Effect of Vitamin B12 and Folate in Chinese Population with Hypertension

BackgroundIn China, the MTHFR 677T allele, unlike in most Western populations, is a rare genetic variant linked to various disorders. The contributing nutritional and genetic factors to this genetic risk remain unclear. ObjectiveThis study aimed to elucidate the interactions between genetic variations in total homocysteine (tHcy) pathway genes, serum tHcy concentrations, and nutritional factors in a Chinese population with hypertension. MethodsThis study analyzed 1304 Chinese adults with hypertension aged ≥18 y enrolled in the China Precision Nutrition and Health KAP Real World Study (CPNAS). Serum concentrations of vitamin B12 and folate were measured using the magnetic microparticle chemiluminescence method, and tHcy concentrations were measured using Hcy Assay kits. Identification of the MTHFR C677T, MTHFR A1298C, and MTRR A66G polymorphisms was performed via time-of-flight nucleic spectrometry. ResultsOur findings revealed significant sex differences in tHcy concentrations, with males exhibiting higher tHcy concentrations than females (13.95 μmol/L vs. 11.15 μmol/L, P < 0.001). Individuals deficient in both vitamin B12 and folate had an increased risk of hyperhomocysteinemia (H-Hcy) (57.4%). In contrast, the prevalence of H-Hcy was lower among those deficient in either vitamin B12 (31.1%) or folate (23.2%) alone. Significant associations were identified between the MTHFR C677T and A1298C polymorphisms and elevated serum tHcy concentrations, particularly in individuals homozygous for the T allele. Conversely, the MTRR A66G genotype did not show a significant correlation with tHcy concentrations. Optimal vitamin B12 concentrations significantly modulated the genotypic effect on tHcy concentrations, with individuals having adequate vitamin B12 and folate exhibiting low tHcy concentrations, even among high-risk genotypes (TT). ConclusionsAdequate concentrations of folate and vitamin B12 significantly reduce serum tHcy concentrations and mitigate the genotypic impact on tHcy concentrations, highlighting the potential for targeted nutritional interventions to manage cardiovascular risks associated with H-Hcy.This trial was registered at clinicaltrials.gov as ChiCTR2100051983.

Comparative study on the effects of anionic, cationic, and nonionic polyacrylamide surface modified magnetic micro-particles (MMP) for anaerobic digestion treatment of vegetable waste water (VWW)

Anaerobic digestion provides a solution for the treatment of vegetable waste water (VWW), but there are currently limited targeted treatment methods available. Building upon previous studies, this research investigated the effects of polyacrylamide-modified magnetic micro-particles (MMP) on anaerobic digestion (AD) of VWW. Three variations of these particles were created by grafting anionic, cationic, and non-ionic polyacrylamide (PAM) onto the MMPs’ surfaces, resulting in aPAM-MMP, cPAM-MMP, and nPAM-MMP, respectively. In AD experiments, the addition of aPAM-MMP notably enhanced the degradation of chemical oxygen demand (COD) in VWW. COD decreased to 1290 mg/L in the reactor with aPAM-MMP by day 12 and remained low, while the other reactors had COD concentrations of 4137.5, 5510, and 3010 mg/L on the same day, decreasing thereafter. This modification also improved the production and utilization of hydrogen gas and volatile fatty acids (VFAs), along with the conversion of methane. When tested for bioaffinity using fluorescent GFP-E.coli bacteria, the aPAM-MMP, cPAM-MMP, and nPAM-MMP demonstrated increases in fluorescence intensity by 51.66%, 36.13%, and 37.02%, respectively, compared to unmodified MMP when attached with GFP-E.coli. Further analyses of microbial community revealed that the reactor with aPAM-MMP had the highest microbial richness and enriched bacteria capable of organic matter degradation, such as Bacteroidota, Synergistota, Chloroflexi, Halobacterota phyla, and Parabacteroides, Muribaculaceae, and Azotobacter genera. In conclusion, our experiment verifies that APAM-MMP promotes anaerobic treatment of VWW and provides a novel reference point for enhancing VWW degradation.

Amplification-Free Strategy for miRNA Quantification in Human Serum Using Single Particle ICP-MS and Gold Nanoparticles as Labels.

MicroRNAs (miRNAs), which are short single-stranded RNA sequences between 18 and 24 nucleotides, are known to play a crucial role in gene expression. Changes in their expression are not only involved in many diseases but also as a response to physiological changes, such as physical exercise. In this work, a new analytical strategy for the sensitive and specific analysis of miRNA sequences in human plasma is presented. The developed strategy does not depend on any nucleic acid amplification process and can be obtained in direct correlation to the number of events obtained by using single-particle ICP-MS measurements. The high selectivity of the assay (up to single nucleotide polymorphisms) can be achieved by a double hybridization process of the target miRNA with a complementary capture oligonucleotide that is conjugated to a magnetic microparticle and simultaneously with a complementary reporter oligonucleotide conjugated to a gold nanoparticle. Thanks to the novel approach followed in this method, the stoichiometry of the oligonucleotide-nanoparticle conjugates does not need to be addressed for the quantification of the target miRNA, which also represents a big advantage over other similar methods. The optimized method is applied to the determination of a miRNA as a biomarker of physical exercise in non-spiked human serum samples, and the results are validated against rt-qPCR. The achieved sensitivity permits the direct differentiation among sedentary and sportive subjects. This general platform can be easily applied to any other sequence by only modifying the capture and reporter oligonucleotides, paving the way for multiple clinically interesting applications.

Microfluidic production, stability and loading of synthetic giant unilamellar vesicles

In advanced drug delivery, versatile liposomal formulations are commonly employed for safer and more accurate therapies. Here we report a method that allows a straightforward production of synthetic monodisperse (~ 100 μm) giant unilamellar vesicles (GUVs) using a microfluidic system. The stability analysis based on the microscopy imaging showed that at ambient conditions the produced GUVs had a half-life of 61 ± 2 h. However, it was observed that ~ 90% of the calcein dye that was loaded into GUVs was transported into a surrounding medium in 24 h, thus indicating that the GUVs may release these small dye molecules without distinguishable membrane disruption. We further demonstrated the feasibility of our method by loading GUVs with larger and very different cargo objects; small soluble fluorescent proteins and larger magnetic microparticles in a suspension. Compared to previously reported microfluidics-based production techniques, the obtained results indicate that our simplified method could be equally harnessed in creating GUVs with less cost, effort and time, which could further benefit studying closed membrane systems.

Spiky Magnetic Microparticles Synthesized from Microrod-Stabilized Pickering Emulsion.

Tailoring the microstructure of magnetic microparticles is of vital importance for their applications. Spiky magnetic particles, such as those made from sunflower pollens, have shown promise in single cell treatment and biofilm removal. Synthetic methods that can replicate or extend the functionality of such spiky particles would be advantageous for their widespread utilization. In this work, a wet-chemical method is introduced for spiky magnetic particles that are templated from microrod-stabilized Pickering emulsions. The spiky morphology is generated by the upright attachment of silica microrods at the oil-water interface of oil droplets. Spiky magnetic microparticles with control over the length of the spikes are obtained by dispersing hydrophobic magnetic nanoparticles in the oil phase and photopolymerizing the monomer. The spiky morphology dramatically enhances colloidal stability of these particles in high ionic strength solutions and physiologic media such as human saliva and saline-based biofilm suspension. To demonstrate their utility, the spiky magnetic particles are applied for magnetically controlled removal of oral biofilms and retrieval of bacteria for diagnostic sampling. This method expands the toolbox for engineering microparticle morphology and could promote the fabrication of functional magnetic microrobots.

Multispectral 3D DNA Machine Combined with Multimodal Machine Learning for Noninvasive Precise Diagnosis of Bladder Cancer.

Extracellular vesicle (EV) molecular phenotyping offers enormous opportunities for cancer diagnostics. However, the majority of the associated studies adopted biomarker-based unimodal analysis to achieve cancer diagnosis, which has high false positives and low precision. Herein, we report a multimodal platform for the high-precision diagnosis of bladder cancer (BCa) through a multispectral 3D DNA machine in combination with a multimodal machine learning (ML) algorithm. The DNA machine was constructed using magnetic microparticles (MNPs) functionalized with aptamers that specifically identify the target of interest, i.e., five protein markers on bladder-cancer-derived urinary EVs (uEVs). The aptamers were hybridized with DNA-stabilized silver nanoclusters (DNA/AgNCs) and a G-quadruplex/hemin complex to form a sensing module. Such a DNA machine ensured multispectral detection of protein markers by fluorescence (FL), inductively coupled plasma mass spectrometry (ICP-MS), and UV-vis absorption (Abs). The obtained data sets then underwent uni- or multimodal ML for BCa diagnosis to compare the analytical performance. In this study, urine samples were obtained from our prospective cohort (n = 45). Our analytical results showed that the 3D DNA machine provided a detection limit of 9.2 × 103 particles mL-1 with a linear range of 4 × 104 to 5 × 107 particles mL-1 for uEVs. Moreover, the multimodal data fusion model exhibited an accuracy of 95.0%, a precision of 93.1%, and a recall rate of 93.2% on average, while those of the three types of unimodal models were no more than 91%. The elevated diagnosis precision by using the present fusion platform offers a perspective approach to diminishing the rate of misdiagnosis and overtreatment of BCa.

Non-targeted analysis of the plasma proteome between early and late rugby playing career

Plasma proteomics can offer objective biological insight into the physiological responses of athletes. Rugby players may be repeatedly exposed to high-impact collisions that could predispose them to a higher risk of neurodegenerative conditions. The potential mechanisms underlying the heightened risk are unclear and discovery proteomics could provide unbiased insight into the pathways involved. Plasma extracellular vesicles (EV) are a novel system of intercellular communication which have been implicated in the propagation of misfolded proteins in the brain and could carry putative diagnostic biomarkers. This project investigates whether differences in plasma EV proteome exist between early and late career rugby players. Twenty-four male participants were recruited, including 8 academy players (18 ± 1 years), 8 rugby players (33 ± 5 years) with 10-year professional rugby career and 8 CrossFit athletes (32 ± 5 years) with no history of collision-related sports injuries. Participants gave their informed consent to the ethically approved procedures. Venous blood samples were collected in EDTA blood sample tubes, according to standard operating procedures. Hyper-porous strong-anion exchange magnetic microparticles (MagReSyn® SAX) were used to capture membrane-bound particles (i.e. EV) from plasma. EV-enriched samples were digested with trypsin and analysed using nano-flow liquid chromatography and high-resolution tandem mass spectrometry (Thermo, U3000 and Q-Exactive mass spectrometer). Label free quantitation was performed using Progenesis QI-P (Waters Corp) and proteins were identified by Mascot (Matrix Sci) searches of the UniProt Knowledge base. Statistical differences were investigated by one-way analysis of variance (with correction for multiple testing) in R (v4.3.2). Functional enrichment analysis was performed using ShinyGO (0.80). In total, 449 proteins were confidently identified (false discovery rate; FDR &lt;1%) and gene ontology profiling confirmed 414 of these proteins were of EV origin (Jensen COMPARTMENTS). One-way ANOVA highlighted 192 significantly (P &lt; 0.05, q &lt; 0.02) different proteins across the 3 participant groups, of which 24 proteins were specific to professional rugby players. Functional enrichment analysis revealed key pathways related to Vitamin digestion and absorption, complement and coagulation cascade, inflammatory response, neuron death and negative regulation of cell death (FDR=5%).In conclusion, proteomic profiling is able to detect robust differences between the plasma EV proteome of long-serving professional rugby players compared to younger rugby players or age-matched athletes that did not have a history of collision-related sports injuries. Our findings shed new light on processes affected by a professional rugby playing career, further exploitation of this type of analysis could be used to discover sport-specific biomarkers.

A sandwich chemiluminescent magnetic microparticle immunoassay for cryptococcal antigen detection

ABSTRACT Background Cryptococcosis is a global invasive mycosis associated with significant morbidity and mortality. Cryptococcal antigen (CrAg) testing from serum and cerebrospinal fluid (CSF) has been regarded as a gold standard for early diagnosis. This study aimed to develop and validate a rapid and sensitive sandwich chemiluminescent magnetic microparticle immunoassay (CMIA) for quantitative detection of CrAg in sera. Research design and methods CMIA is based on magnetic beads modified with capture antibodies and biotinylated antibodies and Streptavidin-polyHRP, where biotinylated antibodies functioned as the recognition element and Streptavidin-polyHRP as the signal component. Assay parameters were first optimized, and then assay performances were evaluated. Results Under optimized conditions, the total runtime of the CMIA was 22 min. The assay had a wide linear range (2 –10,000 ng/mL) and high analytical sensitivity (0.24 ng/mL), together with acceptable reproducibility, accuracy, and stability. Besides, it exhibited no cross-reactivity with other pathogens. Importantly, the assay showed 92.91% (95% CI, 80.97–93.02%) overall qualitative agreement with a commercial ELISA kit in a retrospective cohort of 55 cases with confirmed cryptococcal infection, and 72 controls without evidence of invasive fungal disease (IFD). Conclusion These results demonstrated that the present study paved a novel strategy for reliable quantitative detection of CrAg in sera.

Towards cost-effective drug discovery: Reusable immobilized enzymes for neurological disease research

Enzyme handling and utilization bears many challenges such as their limited stability, intolerance of organic solvents, high cost, or inability to reuse. Most of these limitations can be overcome by enzyme immobilization on the surface of solid support. In this work, the recombinant form of human cholinesterases and monoamine oxidases as important drug targets for neurological diseases were immobilized on the surface of magnetic non-porous microparticles by a non-covalent bond utilizing the interaction between a His-tag terminus on the recombinant enzymes and cobalt (Co2+) ions immobilized on the magnetic microparticles. This type of binding led to targeted enzyme orientation, which completely preserved the catalytic activity and allowed high reproducibility of immobilization. In comparison with free enzymes, the immobilized enzymes showed exceptional stability in time and the possibility of repeated use. Relevant Km, Vmax, and IC50 values using known inhibitors were obtained using particular immobilized enzymes. Such immobilized enzymes on magnetic particles could serve as an excellent tool for a sustainable approach in the early stage of drug discovery.