RPA/CRISPR/Cas12a-Based On-Site and Rapid Nucleic Acid Detection of Toxoplasma gondii in the Environment.

Toxoplasma gondii is an opportunistic pathogen widely distributed within the world, poses a huge threat to human health, and causes significant economic losses to the livestock industry. Herein, we developed a portable one-pot detection of T. gondii by combining recombinase polymerase amplification (RPA) and a clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a system. A glass microfiber filter device used for the first step can efficiently extract T. gondii from low-concentration samples. The lyophilized RPA reagents and Cas12a/crRNA reagents are prestored in one Eppendorf tube, and both reactions can be performed on a low-cost thermal controller (∼37 °C), avoiding the drawbacks of the step-by-step addition of components. The developed RPA/CRISPR/Cas12a system exhibits a high selectivity toward the B1 gene amplicon of T. gondii over other parasites with a limit of detection of 3.3 copies/µL. The visual signal readout can be easily realized by a fluorometer or lateral-flow strip. A portable suitcase containing the minimum equipment and lyophilized reagents was adopted for the rapid determination of T. gondii in heavily polluted landfill leachate. This system presents rapidness, robustness and on-site features for the detection of nucleic acids of the parasite, making it a promising tool for field applications in remote areas.

Rearrangement of protein structures on a gold nanoparticle surface is regulated by ligand adsorption modes.

With development of the nanomedicine field and increasing hazards of exposure to nanobiological materials, research on the protein corona is urgently required. In particular, the understanding of the mechanism of structural changes of protein on a nanosurface should be improved. Herein, we focus on exploring the role of ligand adsorption modes (physiosorbed citrates or chemisorbed GSH) in the regulation of conformational rearrangement of three blood proteins (serum albumin, globulin, and fibrinogen) on the surface of gold nanoparticles. Through experimental measurements, protein adsorption features (thermodynamics, kinetics, adsorption orientation, and structural changes) were estimated. Molecular dynamics simulations further indicated that physiosorbed citrates could be gradually peeled off by approaching proteins and that the bare Au surface provided a stronger interface interaction than the chemisorbed GSH layer. Protein structure rearrangements were due to the reduction in protein internal energy, with an increase in H-bond formation involving a decrease in the α-helical content and an increase in the ß-sheet content, to offset the high interfacial energy. Rearrangement of protein structures could occur either intramolecularly or intermolecularly. These findings enhanced our understanding of nano-protein interaction in the biological milieu and facilitate biomedical exploration of engineered nanomaterials.

Platinum Nanoparticle Encapsulated Metal-Organic Frameworks for Colorimetric Measurement and Facile Removal of Mercury(II).

Pt nanoparticle (Pt NP)@UiO-66-NH2 composites were synthesized and encompassed the benefits of permanent porosity, high thermal and chemical stability of metal-organic frameworks (MOFs), together with the functional behavior of isolated Pt NPs. The PVP-stabilized Pt NPs with the average diameter of 2.48 nm were well dispersed and confined within the framework of UiO-66-NH2. Pt NPs possess highly peroxidase-like activities and make the composites oxidize 3,3',5,5'-tetramethylbenzidine in the presence of H2O2. Moreover, the specific interaction between Hg2+ and Pt NPs leads to the effective suppression of the peroxidase-like activity of Pt NP@UiO-66-NH2, which endows excellent selectivity for Hg2+ measurement over the interfering metal ions. Based on the colorimetric sensing system, Hg2+ is linearly measured over the range from 0 to 10 nM with a detection limit of 0.35 nM. Moreover, the as-obtained Pt NP@UiO-66-NH2 nanocomposites exhibit high capacity and good selectivity for Hg2+ adsorption, which is successfully applied to treat Hg2+ in water with removal efficiency over 99%. With these findings, Pt NP@UiO-66-NH2 composites can be used to develop a simple and rapid colorimetric sensing system and are utilized as nanoadsorbents for facile removal of Hg2+. This work not only expands the scientific researches on MOFs but also provides practical application in environmental, biological, and relative fields.