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1.
Nano Lett ; 2024 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-39360805

RESUMO

Microrobots possessing multifunctional integration are desired for therapeutics and biomedicine applications. However, existing microrobots with desired functionalities need to be fabricated through complex procedures due to their constrained volume, limited manufacturing processes, and lack of effective in vivo observation methods. Inspired by bubbles exhibiting various abilities, we report magnetic air bubble microrobots with simpler structures to simultaneously integrate multiple functions, including microcargo delivery, multimode locomotion, imaging, and biosensing. Contributed by buoyancy and magnetic actuation to overcome obstacles, flexible three-dimensional locomotion is implemented, guaranteeing the integrity of micro-objects adsorbed on the surface of the air bubble microrobot. Introducing air microbubbles enhances the ultrasound imaging capability of microrobots in the vascular system of mice in vivo, facilitating ample medical applications. Moreover, air-liquid reactions endow microrobots with rapid pH biosensing. This work provides a unique strategy to utilize relatively simple air bubbles to achieve the complex functions of microrobots for biomedical applications.

2.
Bull Environ Contam Toxicol ; 113(4): 42, 2024 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-39306638

RESUMO

In the present paper, the assessment of vortex-assisted dispersive magnetic solid-phase extraction using amino-functionalized mesoporous combined with direct magnetic sorbent sampling (DMSS) in flame or furnace atomic absorption spectrometry (FAAS or FF-AAS) was demonstrated for highly sensitive silver determination in water samples. The developed method showed significant enrichment factors compared to conventional pneumatic nebulization by FAAS, 607 for DMSS-FF-AAS and 114 for DMSS-FAAS. The analytical curve showed linearity in the range from 5.0 to 70.0 µg L- 1 and 1.0 to 15.0 µg L- 1 and limits of detection of 0.59 and 0.09 µg L- 1 for DMSS-FAAS and DMSS-FF-AAS, respectively. The intra and inter-day precision evaluated as a percentage of the relative standard deviation (RSD,%) ranged from 1.89 to 4.71% for levels of 25.0 and 65.0 µg L- 1. The method was applied in different kinds of water samples without matrix effects, yielding recovery values from 90 to 110%.


Assuntos
Prata , Extração em Fase Sólida , Espectrofotometria Atômica , Poluentes Químicos da Água , Extração em Fase Sólida/métodos , Prata/química , Poluentes Químicos da Água/análise , Monitoramento Ambiental/métodos , Limite de Detecção
3.
Biosensors (Basel) ; 14(9)2024 Aug 23.
Artigo em Inglês | MEDLINE | ID: mdl-39329783

RESUMO

Due to the extremely low Reynolds number, the mixing of substances in laminar flow within microfluidic channels primarily relies on slow intermolecular diffusion, whereas various rapid reaction and detection requirements in lab-on-a-chip applications often necessitate the efficient mixing of fluids within short distances. This paper presents a magnetic pillar-shaped particle fabrication device capable of producing particles with planar shapes, which are then utilized to achieve the rapid mixing of multiple fluids within microchannels. During the particle fabrication process, a degassed PDMS chip provides self-priming capabilities, drawing in a UV-curable adhesive-containing magnetic powder and distributing it into distinct microwell structures. Subsequently, an external magnetic field is applied, and the chip is exposed to UV light, enabling the mass production of particles with specific magnetic properties through photo-curing. Without the need for external pumping, this chip-based device can fabricate hundreds of magnetic particles in less than 10 min. In contrast to most particle fabrication methods, the degassed PDMS approach enables self-priming and precise dispensing, allowing for precise control over particle shape and size. The fabricated dual-layer magnetic particles, featuring fan-shaped blades and disk-like structures, are placed within micromixing channels. By manipulating the magnetic field, the particles are driven into motion, altering the flow patterns to achieve fluid mixing. Under conditions where the Reynolds number in the chip ranges from 0.1 to 0.9, the mixing index for substances in aqueous solutions exceeds 0.9. In addition, experimental analyses of mixing efficiency for fluids with different viscosities, including 25 wt% and 50 wt% glycerol, reveal mixing indices exceeding 0.85, demonstrating the broad applicability of micromixers based on the rapid rotation of magnetic particles.


Assuntos
Dispositivos Lab-On-A-Chip , Técnicas Analíticas Microfluídicas , Microfluídica , Dimetilpolisiloxanos/química , Magnetismo
4.
Int J Pharm ; 662: 124503, 2024 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-39043284

RESUMO

In the study, we aimed to investigate the activity of nanoformulations containing 5-fluorouracil and polymer-magnetic hybrids bearing membrane-penetrating and ligand-receptor-recognizing agents against colorectal cancer cells. The formation and characterization of iron oxide particles covered with polymeric shells comprising lithocholic acid and folic acid moieties are presented. The efficiency of nanoformulations combined by the simple mixing of low doses of 5-fluorouracil with the obtained hybrids was demonstrated against DLD-1 and HT-29 colon cancer cells. The most pronounced cytotoxic potential against HT-29 cells was observed in the cases of particles based on block and randomly arranged copolymers functionalized by FA motifs with depletion of viable cells by approximately 50 % compared to control cells and cells treated by 5-FU applied in free form. In the case of the DLD-1 cell line, the percentage of viable DLD-1 cells decreased by about 30 to 40% after treatment with the block and randomly arranged copolymer decorated by FA-moiety, when compared to 5-FU at the free form and the untreated control. The induction of apoptosis associated with PS-translocation was determined to be the main mechanism of their cytotoxic effects. Moreover, the safety profiles of the nanoformulations were established through hemolysis assay and the analysis of the viability of human colorectal fibroblasts. It was indicated that all tested nanoparticles met the compatibility requirements at the in vitro level. It should be emphasized that in many cases, there was a significant improvement in the compatibility of hybrids with the FA motif compared to non-functionalized hybrids with the addition of 5-FU. These findings suggest that the presence of FA might modulate the toxicity of chemotherapeutic agents.


Assuntos
Apoptose , Sobrevivência Celular , Neoplasias do Colo , Fluoruracila , Ácido Fólico , Ácido Litocólico , Polímeros , Humanos , Ácido Fólico/química , Ácido Fólico/administração & dosagem , Ácido Litocólico/química , Fluoruracila/administração & dosagem , Fluoruracila/química , Neoplasias do Colo/tratamento farmacológico , Sobrevivência Celular/efeitos dos fármacos , Células HT29 , Polímeros/química , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Hemólise/efeitos dos fármacos , Portadores de Fármacos/química , Nanopartículas de Magnetita/química
5.
Artigo em Inglês | MEDLINE | ID: mdl-38936266

RESUMO

Hyaluronic acid (HA), a glycosaminoglycan, is comprised of alternating units of D-glucuronic acid and N-acetylglucosamine. This compound harbors numerous biomedical applications, including its use in pharmaceuticals, wound healing, osteoarthritis treatment, and drug delivery. Its unique composition and exceptional features, such as its high water-absorbing and retaining capacity, have also led to its use in the cosmetics industry. The employment of this biopolymer has given rise to an escalation in the request for its manufacture. The present investigation has explored the correlation between hyaluronic acid and chitosan and silica for the purpose of separation. Consequently, Iron oxide magnetic nano particles and micro particles were produced via co-precipitation method and were layered with chitosan and silica to purify the hyaluronic acid from the fermentation broth that was generated by Streptococcus Zooepidemicus. The size distribution and zeta potentials of the two kinds of particles were gauged with the aid of a dynamic laser light scattering apparatus and zeta potential meter (Malvern, Zeta master) respectively. The confirmation of the chemical structure of the Fe3O4 nanoparticles and Fe3O4 particles conjugated with chitosan and silica was accomplished through the utilization of Fourier Transform Infrared Spectroscopy (FT-IR). Protein contamination was thoroughly characterized by means of sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Nanodrop 2000/2000c spectrophotometers protein estimation method. The maximum HA adsorption capacity, under optimal pH conditions of 4, was determined to be 87 mg/g, 112 mg/g, 51 mg/g, and 44 mg/g for Fe3O4 -chitosan nanoparticle, Fe3O4 -chitosan micro particle, Fe3O4 -silica microparticle, and Fe3O4 -silica nanoparticle, respectively.


Assuntos
Quitosana , Ácido Hialurônico , Nanopartículas de Magnetita , Dióxido de Silício , Quitosana/química , Ácido Hialurônico/química , Ácido Hialurônico/isolamento & purificação , Dióxido de Silício/química , Nanopartículas de Magnetita/química , Tamanho da Partícula
6.
Bioorg Chem ; 149: 107529, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38850780

RESUMO

Trace natural products (TNPs) are still the vital source of drug development. However, the mining of novel TNPs is becoming increasingly challenging due to their low abundance and complex interference. A comprehensive strategy was proposed in which the functionalized magnetic particles integrated with LC-MS for TNPs discovery. Under the guidance of the approach, fifteen trace Nuphar alkaloids including seven new ones, cyanopumiline A sulfoxide (1), cyanopumiline C sulfoxide (8) and cyanopumilines A-E (4-5, 10, 12-13) featuring an undescribed nitrile-containing 6/6/5/6/6 pentacyclic ring system were isolated from the rhizomes of Nuphar pumila. Their structures and absolute configurations were determined on the basis of detailed spectroscopic data analysis and single-crystal X-ray diffraction analysis. Notably, a concise method based on 13C NMR spectroscopy was established to determine the relative configurations of spiroatoms. Biologically, compounds 1-12 exhibited potent immunosuppressive activities with IC50 values ranging from 0.1-12.1 µM against anti-CD3/CD28 induced human peripheral T cell proliferation. Mechanistic studies revealed that 4 could dose-dependently decrease pro-inflammatory cytokines and the expression levels of CD25 and CD71.


Assuntos
Alcaloides , Proliferação de Células , Relação Dose-Resposta a Droga , Imunossupressores , Humanos , Proliferação de Células/efeitos dos fármacos , Imunossupressores/farmacologia , Imunossupressores/química , Imunossupressores/isolamento & purificação , Estrutura Molecular , Alcaloides/química , Alcaloides/farmacologia , Alcaloides/isolamento & purificação , Relação Estrutura-Atividade , Cromatografia Líquida , Descoberta de Drogas , Linfócitos T/efeitos dos fármacos , Espectrometria de Massas , Espectrometria de Massa com Cromatografia Líquida
7.
Soft Robot ; 2024 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-38836749

RESUMO

Suction grippers offer a distinct advantage in their ability to handle a wide range of items. However, attaching these grippers to irregular and rough surfaces presents an ongoing challenge. To address this obstacle, this study explores the integration of magnetic intelligence into a soft suction gripper design, enabling fast external magnetic actuation of the attachment process. Additionally, miniaturization options are enhanced by implementing a compliant deploying mechanism. The resulting design is the first-of-its-kind magnetically-actuated deployable suction gripper featuring a thin magnetic membrane (Ø 50 mm) composed of carbonyl iron particles embedded in a silicone matrix. This membrane is supported by a frame made of superelastic nitinol wires that facilitate deployment. During experiments, the proof-of-principle prototype demonstrates successful attachment on a diverse range of curved surfaces in both dry and wet environments. The gripper achieves attachment on curved surfaces with radii of 50-75 mm, exerting a maximum attachment force of 2.89 ± 0.54 N. The current gripper design achieves a folding percentage of 75%, enabling it to fit into a Ø 12.5 mm tube and access hard-to-reach areas while maintaining sufficient surface area for attachment forces. The proposed prototype serves as a foundational steppingstone for further research in the development of reliable and effective magnetically-actuated suction grippers across various configurations. By addressing the limitations of attachment to irregular surfaces and exploring possibilities for miniaturization and precise control, this study opens new avenues for the practical application of suction grippers in diverse industries and scenarios.

8.
Nanomaterials (Basel) ; 14(10)2024 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-38786825

RESUMO

Nanocrystalline carbonyl iron (CI) particles are promising microwave absorbents at elevated temperature, whereas their excessive grain boundary energy leads to the growth of nanograins and a deterioration in permeability. In this work, we report a strategy to enhance the thermal stability of the grains and microwave absorption of CI particles by doping a SiBaFe alloy. Grain growth was effectively inhibited by the pinning effect of SiBaFe alloy nanoparticles at the grain boundaries. After heat treatment at 600 °C, the grain size of CI particles increased from ~10 nm to 85.1 nm, while that of CI/SiBaFe particles was only 32.0 nm; with the temperature rising to 700 °C, the grain size of CI particles sharply increased to 158.1 nm, while that of CI/SiBaFe particles was only 40.8 nm. Excellent stability in saturation magnetization and microwave absorption was also achieved in CI/SiBaFe particles. After heat treatment at 600 °C, the flaky CI/SiBaFe particles exhibited reflection loss below -10 dB over 7.01~10.11 GHz and a minimum of -14.92 dB when the thickness of their paraffin-based composite was 1.5 mm. We provided a low-cost and efficient kinetic strategy to stabilize the grain size in nanoscale and microwave absorption for nanocrystalline magnetic absorbents working at elevated temperature.

9.
Bioengineering (Basel) ; 11(4)2024 Mar 26.
Artigo em Inglês | MEDLINE | ID: mdl-38671738

RESUMO

The flow and heat transfer of a steady, viscous biomagnetic fluid containing magnetic particles caused by the swirling and stretching motion of a three-dimensional cylinder has been investigated numerically in this study. Because fluid and particle rotation are different, a magnetic field is applied in both radial and tangential directions to counteract the effects of rotational viscosity in the flow domain. Partial differential equations are used to represent the governing three-dimensional modeled equations. With the aid of customary similarity transformations, this system of partial differential equations is transformed into a set of ordinary differential equations. They are then numerically resolved utilizing a common finite differences technique that includes iterative processing and the manipulation of tridiagonal matrices. Graphs are used to depict the physical effects of imperative parameters on the swirling velocity, temperature distributions, skin friction coefficient, and the rate of heat transfer. For higher values of the ferromagnetic interaction parameter, it is discovered that the axial velocity increases, whereas temperature and tangential velocity drop. With rising levels of the ferromagnetic interaction parameter, the size of the axial skin friction coefficient and the rate of heat transfer are both accelerated. In some limited circumstances, a comparison with previously published work is also handled and found to be acceptably accurate.

10.
Cell Rep Phys Sci ; 5(4): 101923, 2024 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-38680545

RESUMO

Nature has already suggested bioinspired functions. Beyond them, adaptive and trainable functions could be the inspiration for novel responsive soft matter beyond the state-of-the-art classic static bioinspired, stimulus-responsive, and shape-memory materials. Here, we describe magnetic assembly/disassembly of electrically conducting soft ferromagnetic nickel colloidal particles into surface topographical pillars for bistable electrical trainable memories. They allow magnetic sensing with adaptable and rescalable sensitivity ranges, enabled by bistable memories and kinetic concepts inspired by biological sensory adaptations. Based on the soft ferromagnetism of the nanogranular composition and the resulting rough particle surfaces prepared via a solvothermal synthesis, triggerable structural memory is achieved by the magnetic field-driven particle assembly and disassembly, promoted by interparticle jamming. Electrical conversion from current to frequency for electrical spikes facilitates rescalable and trainable frequency-based sensitivity on magnetic fields. This work suggests an avenue for designing trainable and adaptable life-inspired materials, for example, for soft robotics and interactive autonomous devices.

11.
Sensors (Basel) ; 24(8)2024 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-38676098

RESUMO

This paper designed and developed an online digital imaging excitation sensor for wind power gearbox wear condition monitoring based on an adaptive deep learning method. A digital imaging excitation sensing image information collection architecture for magnetic particles in lubricating oil was established to characterize the wear condition of mechanical equipment, achieving the real-time online collection of wear particles in lubricating oil. On this basis, a mechanical equipment wear condition diagnosis method based on online wear particle images is proposed, obtaining data from an engineering test platform based on a wind power gearbox. Firstly, a foreground segmentation preprocessing method based on the U-Net network can effectively eliminate the interference of bubbles and dark fields in online wear particle images, providing high-quality segmentation results for subsequent image processing, A total of 1960 wear particle images were collected in the experiment, the average intersection union ratio of the validation set is 0.9299, and the accuracy of the validation set is 0.9799. Secondly, based on the foreground segmentation preprocessing of wear particle images, by using the watered algorithm to obtain the number of particles in each size segment, we obtained the number of magnetic particle grades in three different ranges: 4-38 µm, 39-70 µm, and >70 µm. Thirdly, we proposed a method named multidimensional transformer (MTF) network. Mean Square Error (MSE), Root Mean Square Error (RMSE), and Mean Absolute Error (MAE) are used to obtain the error, and the maintenance strategy is formulated according to the predicted trend. The experimental results show that the predictive performance of our proposed model is better than that of LSTM and TCN. Finally, the online real-time monitoring system triggered three alarms, and at the same time, our offline sampling data analysis was conducted, the accuracy of online real-time monitoring alarms was verified, and the gearbox of the wind turbine was shut down for maintenance and repair.

12.
Bioprocess Biosyst Eng ; 47(6): 851-862, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38676738

RESUMO

In this paper, a magnetic sequencing batch reactor (SBR) was constructed, and the influence rule of magnetic particle dosing performance of denitrification was investigated. The diversity, structure, and potential functions of the microbial community were comprehensively explored. The results showed that the particle size and the dosage of Fe3O4 magnetic particles were the main parameters affecting the sedimentation performance of activated sludge. The start-up phase of the SBR reactor with Fe3O4 magnetic particles was 5 days less than the control. Moreover, total nitrogen removal efficiency during the start-up phase was improved, with the maximum value reaching 91.93%, surpassing the control by 9.7% with the Fe3O4 dosage of 1.2 g L-1. In addition, the activated sludge concentration and dehydrogenase activity were improved, compared to the control. High-throughput sequencing showed that the denitrifying bacterium Saccharimonadales dominated the reactor and was enriched by magnetic particles. According to predicted functions, the abundance of genes for denitrification increased with the addition of magnetic particles, suggesting the capacity of nitrogen removal was enhanced in the microbial community. Overall, the Fe3O4 magnetic particles provide great potential for enhanced wastewater nitrogen removal.


Assuntos
Reatores Biológicos , Desnitrificação , Nitrogênio , Nitrogênio/química , Nitrogênio/metabolismo , Esgotos/microbiologia , Bactérias/metabolismo , Bactérias/genética , Águas Residuárias/microbiologia , Águas Residuárias/química , Nanopartículas de Magnetita/química
13.
Mikrochim Acta ; 191(5): 235, 2024 04 03.
Artigo em Inglês | MEDLINE | ID: mdl-38570380

RESUMO

A fast and accurate identification of Listeria monocytogenes. A new quartz crystal microbalance (QCM) aptasensor was designed for the specific and rapid detection of L. monocytogenes. Before detection of the target bacterium from samples in the QCM aptasensor, a magnetic pre-enrichment system was used to eliminate any contaminant in the samples. The prepared magnetic system was characterized using ATR-FTIR, SEM, VSM, BET, and analytical methods. The saturation magnetization values of the Fe3O4, Fe3O4@PDA, and Fe3O4@PDA@DAPEG particles were 57.2, 40.8, and 36.4 emu/g, respectively. The same aptamer was also immobilized on the QCM crystal integrated into QCM flow cell and utilized to quantitatively detect L. monocytogenes cells from the samples. It was found that a specific aptamer-magnetic pre-concentration system efficiently captured L. monocytogenes cells in a short time (approximately 10 min). The Fe3O4@PDA@DA-PEG-Apt particles provided selective isolation of L. monocytogenes from the bacteria-spiked media up to 91.8%. The immobilized aptamer content of the magnetic particles was 5834 µg/g using 500 ng Apt/mL. The QCM aptasensor showed a very high range of analytical performance to the target bacterium from 1.0 × 102 and 1.0 × 107 CFU/mL. The limit of detection (LOD) and limit of quantitation (LOQ) were 148 and 448 CFU/mL, respectively, from the feeding of the QCM aptasensor flow cell with the eluent of the magnetic pre-concentration system. The reproducibility of the aptasensor was more than 95%. The aptasensor was very specific to L. monocytogenes compared to the other Listeria species (i.e., L. ivanovii, L. innocua, and L. seeligeri) or other tested bacteria such as Staphylococcus aureus, Escherichia coli, and Bacillus subtilis. The QCM aptasensor was regenerated with NaOH solution, and the system was reused many times.


Assuntos
Aptâmeros de Nucleotídeos , Listeria monocytogenes , Técnicas de Microbalança de Cristal de Quartzo , Reprodutibilidade dos Testes , Aptâmeros de Nucleotídeos/química , Escherichia coli , Fenômenos Magnéticos
14.
Talanta ; 275: 126115, 2024 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-38663068

RESUMO

High-quality DNA is an important guarantee to start downstream experiments in many biological and medical research areas. Magnetic particle-based DNA extraction methods from blood mainly depend on electrostatic adsorption in a low-pH environment. However, the strong acidic environment can influence the DNA stability. Herein, a polydopamine-functionalized magnetic particle (PDA@Fe3O4)-based protocol was developed for DNA extraction from whole blood samples. In the protocol, Mg2+ and Ca2+ were utilized to bridge the adsorption of DNA by PDA@Fe3O4 via the metal-mediated coordination. Isopropanol was found to efficiently promote DNA adsorption by triggering the change of the conformation of DNA from B-form to more compact A-form. In 50 % isopropanol solution, the DNA adsorption efficiency was nearly 100 % in the presence of 0.5 mM Ca2+ or 1.5 mM Mg2+. The role of metal ions and isopropanol in DNA adsorption was explored. The protocol averts the strong acidic environment and PCR inhibitors, such as high concentrations of salt or polyethylene glycol. It demonstrates superiority in DNA yield (59.13 ± 3.63 ng µL-1) over the commercial kit (27.33 ± 4.98 ng µL-1) and phenol-chloroform methods (37.90 ± 0.47 ng µL-1). In addition, to simplify the operastion, an automated nucleic acid extraction device was designed and fabricated to extract whole genomic DNA from blood. The feasibility of the device was verified by extracting DNA from cattle and pig blood samples. The extracted DNA was successfully applied to discriminate the beef authenticity by a duplex PCR system. The results demonstrate that the DNA extraction protocol and the automated device have great potential in blood samples.


Assuntos
2-Propanol , DNA , Indóis , Polímeros , Polímeros/química , 2-Propanol/química , DNA/química , DNA/isolamento & purificação , DNA/sangue , Indóis/química , Adsorção , Magnésio/química , Animais , Cálcio/química , Cálcio/sangue , Bovinos , Nanopartículas de Magnetita/química
15.
ACS Nano ; 18(8): 6359-6372, 2024 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-38363638

RESUMO

On-demand droplet transportation is of great significance for numerous applications. Although various strategies have been developed for droplet transportation, out-of-surface three-dimensional (3D) transportation of droplets remains challenging. Here, a versatile droplet transportation strategy based on magnetic-actuated jumping (MAJ) of droplets on superhydrophobic grooved surfaces (SHGSs) is presented, which enables 3D, remote, and precise manipulation of droplets even in enclosed narrow spaces. To trigger MAJ, an electromagnetic field is utilized to deform the droplet on the SHGS with the aid of an attached magnetic particle, thereby the droplet acquires excess surface energy. When the electromagnetic field is quickly removed, the excess surface energy is partly converted into kinetic energy, allowing the droplet to jump atop the surface. Through high-speed imaging and numerical simulation, the working mechanism and size matching effect of MAJ are unveiled. It is found that the MAJ behavior can only be observed if the sizes of the droplets and the superhydrophobic grooves are matched, otherwise unwanted entrapment or pinch-off effects would lead to failure of MAJ. A regime diagram which serves as a guideline to design SHGSs for MAJ is proposed. The droplet transportation capacities of MAJ, including in-surface and out-of-surface directional transportation, climbing stairs, and crossing obstacles, are also demonstrated. With the ability to remotely manipulate droplets in enclosed narrow spaces without using any mechanical moving parts, MAJ can be used to design miniaturized fluidic platforms, which exhibit great potential for applications in bioassays, microfluidics, droplet-based switches, and microreactions.

16.
Talanta ; 271: 125718, 2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38301374

RESUMO

Most electroanalytical detection schemes for DNA markers require considerable time and effort from expert personnel to thoroughly follow the analysis and obtain reliable outcomes. This work aims to present an electrochemical assay performed inside a small card-based platform powered by microfluidic manipulation, requiring minimal human intervention and consumables. The assay couples a sample/signal dual amplification and DNA-modified magnetic particles for the detection of DNA amplification products. Particularly, the sul1 and sul4 genes involved in the resistance against sulfonamide antibiotics were analyzed. As recognized by the World Health Organization, antimicrobial resistance threatens global public health by hampering medication efficacy against infections. Consequently, analytical methods for the determination of such genes in environmental and clinical matrices are imperative. Herein, the resistance genes were extracted from E. coli cells and amplified using an enzyme-assisted isothermal amplification at 37 °C. The amplification products were analyzed in an easily-produced, low-cost, card-based set-up implementing a microfluidic system, demanding limited manual work and small sample volumes. The target amplicon was thus captured and isolated using versatile DNA-modified magnetic beads injected into the microchannel and exposed to the various reagents in a continuously controlled microfluidic flow. After the optimization of the efficiency of each phase of the assay, the platform achieved limits of detections of 44.2 pmol L-1 for sul1 and 48.5 pmol L-1 for sul4, and was able to detect down to ≥500-fold diluted amplification products of sul1 extracted from E. coli living cells in around 1 h, thus enabling numerous end-point analyses with a single amplification reaction.


Assuntos
Escherichia coli , Microfluídica , Humanos , Microfluídica/métodos , Escherichia coli/genética , DNA , Técnicas de Amplificação de Ácido Nucleico/métodos , Sulfonamidas/farmacologia
17.
J Biotechnol ; 385: 13-22, 2024 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-38423470

RESUMO

BACKGROUND: Owing to improvement of the molecular diagnostic methods using purified preparations of nucleic acids (NAs), the development of effective methods providing the isolation of DNA is still relevant. The sorption properties of magnetic multi-walled carbon nanotubes (MWCNTs), oxidized MWCNTs and MWCNTs (pristine and oxidized) modified with polydiallyldimethylammonium chloride (pDADMAC) with respect to double strained DNA have been studied. RESULTS: It was shown that in the presence of MWCNTs/pDADMAC particles the DNA molecules were reversibly retained by the particle's surface. The optimal conditions for each step of DNA extraction from model solutions using the obtained material were selected. A comparative evaluation of the effectiveness of the proposed method for DNA isolation based on the results of spectrophotometry and real-time PCR was carried out. It was shown that the desorbed DNA was efficiently amplified in PCR, inhibition of polymerase did not occurred. Probable mechanisms of DNA retention due to the influence of residual impurities of catalysts in the MWCNT composition, as well as the surface charge of nanotubes are proposed. CONCLUSION: Sequentially oxidized and coated with pDADMAC magnetically susceptible CNTs are seemed to be a promising material for development of low-cost systems proving an easy isolation, storage, and subsequent use of dsDNA in molecular diagnostics. The sorption properties of such systems are determined with highly developed specific surface area and their chemical composition.


Assuntos
Nanotubos de Carbono , Polietilenos , Compostos de Amônio Quaternário , Nanotubos de Carbono/química , Adsorção , DNA , Fenômenos Magnéticos
18.
Mikrochim Acta ; 191(2): 82, 2024 01 09.
Artigo em Inglês | MEDLINE | ID: mdl-38191940

RESUMO

A novel approach is presented that combines filtration and the direct immunomagnetic separation of the retained bacteria Legionella in filters, for further electrochemical immunosensing. This strategy allows for the separation and preconcentration of the water-borne pathogen from high-volume samples, up to 1000 mL. The limit of detection of the electrochemical immunosensor resulted in 100 CFU mL-1 and improved up to 0.1 CFU mL-1 when the preconcentration strategy was applied in 1 L of sample (103-fold improvement). Remarkably, the immunosensor achieves the limit of detection in less than 2.5 h and simplified the analytical procedure. This represents the lowest concentration reported to date for electrochemical immunosensing of Legionella cells without the need for pre-enrichment or DNA amplification. Furthermore, the study successfully demonstrates the extraction of bacteria retained on different filtering materials using immunomagnetic separation, highlighting the high efficiency of the magnetic particles to pull out the bacteria directly from solid materials. This promising feature expands the applicability of the method beyond water systems for detecting bacteria retained in air filters of air conditioning units by directly performing the immunomagnetic separation in the filters.


Assuntos
Técnicas Biossensoriais , Legionella , Separação Imunomagnética , Imunoensaio , Bactérias , Água
19.
Curr Protein Pept Sci ; 25(4): 286-306, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38178676

RESUMO

The separation of proteins in biological samples plays an essential role in the development of disease detection, drug discovery, and biological analysis. Protein imprinted polymers (PIPs) serve as a tool to capture target proteins specifically and selectively from complex media for separation purposes. Whereas conventional molecularly imprinted polymer is time-consuming in terms of incubation studies and solvent removal, magnetic particles are introduced using their magnetic properties for sedimentation and separation, resulting in saving extraction and centrifugation steps. Magnetic protein imprinted polymers (MPIPs), which combine molecularly imprinting materials with magnetic properties, have emerged as a new area of research hotspot. This review provides an overview of MPIPs for proteins, including synthesis, preparation strategies, and applications. Moreover, it also looks forward to the future directions for research in this emerging field.


Assuntos
Microesferas , Impressão Molecular , Polímeros Molecularmente Impressos , Proteínas , Impressão Molecular/métodos , Polímeros Molecularmente Impressos/química , Proteínas/química , Humanos , Polímeros/química
20.
Int J Biol Macromol ; 256(Pt 2): 128418, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38029902

RESUMO

The objective of this study was to immobilize a recombinant ß-galactosidase (Gal) tagged with a cellulose-binding domain (CBD) onto a magnetic core-shell (CS) cellulose system. After 30 min of reaction, 4 U/capsule were immobilized (CS@Gal), resulting in levels of yield and efficiency exceeding 80 %. The optimal temperature for ß-galactosidase-CBD activity increased from 40 to 50 °C following oriented immobilization. The inhibitory effect of galactose decreased in the enzyme reactions catalyzed by CS@Gal, and Mg2+ increased the immobilized enzyme activity by 40 % in the magnetic CS cellulose system. The relative enzyme activity of the CS@Gal was 20 % higher than that of the soluble enzyme activity after 20 min at 50 °C. The CS support and CS@Gal capsules exhibited an average size of 8 ± 1 mm, with the structure of the shell (alginate-pectin-cellulose) enveloping and isolating the magnetic core. The immobilized ß-galactosidase-CBD within the magnetic CS cellulose system retained ∼80 % of its capacity to hydrolyze lactose from skim milk after 10 reuse cycles. This study unveils a novel and promising support for the oriented immobilization of recombinant ß-galactosidase using a magnetic CS system and a CBD tag. This support facilitates ß-galactosidase reuse and efficient separation, consequently enhancing the catalytic properties of the enzyme.


Assuntos
Celulose , Enzimas Imobilizadas , Celulose/química , Enzimas Imobilizadas/química , Catálise , beta-Galactosidase/química , Fenômenos Magnéticos
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