Adaptation of a Bacterial Bioluminescent Assay to Monitor Bioeffects of Gold Nanoparticles.

Our current study aimed to adapt a bioluminescent bacteria-based bioassay to monitor the bioeffects of gold nanoparticles (AuNPs). Luminous marine bacteria Photobacterium phosphoreum and AuNPs modified with polyvinylpyrrolidone were employed; low-concentration (≤10-3 g/L) bioeffects of AuNPs were studied. Bioluminescence intensity was used as an indicator of physiological activity in bacteria. Two additional methods were used: reactive oxygen species (ROS) content was estimated with a chemiluminescent luminol method, and bacterial size was monitored using electron microscopy. The bacterial bioluminescent response to AuNPs corresponded to the "hormesis" model and involved time-dependent bioluminescence activation, as well as a pronounced increase in the number of enlarged bacteria. We found negative correlations between the time courses of bioluminescence and the ROS content in bacterial suspensions, demonstrating the relationship between bioluminescence activation and bacterial ROS consumption. The combined effects of AuNPs and a beta-emitting radionuclide, tritium, revealed suppression of bacterial bioluminescent activity (as compared to their individual effects) and a reduced percentage of enlarged bacteria. Therefore, we demonstrated that our bacteria-based bioluminescence assay is an appropriate tool to study the bioeffects of AuNPs; the bioeffects can be further classified within a unified framework for rapid bioassessment.

Structure- and Interaction-Based Design of Anti-SARS-CoV-2 Aptamers.

Aptamer selection against novel infections is a complicated and time-consuming approach. Synergy can be achieved by using computational methods together with experimental procedures. This study aims to develop a reliable methodology for a rational aptamer in silico et vitro design. The new approach combines multiple steps: (1) Molecular design, based on screening in a DNA aptamer library and directed mutagenesis to fit the protein tertiary structure; (2) 3D molecular modeling of the target; (3) Molecular docking of an aptamer with the protein; (4) Molecular dynamics (MD) simulations of the complexes; (5) Quantum-mechanical (QM) evaluation of the interactions between aptamer and target with further analysis; (6) Experimental verification at each cycle for structure and binding affinity by using small-angle X-ray scattering, cytometry, and fluorescence polarization. By using a new iterative design procedure, structure- and interaction-based drug design (SIBDD), a highly specific aptamer to the receptor-binding domain of the SARS-CoV-2 spike protein, was developed and validated. The SIBDD approach enhances speed of the high-affinity aptamers development from scratch, using a target protein structure. The method could be used to improve existing aptamers for stronger binding. This approach brings to an advanced level the development of novel affinity probes, functional nucleic acids. It offers a blueprint for the straightforward design of targeting molecules for new pathogen agents and emerging variants.

Protein biosensor based on Schottky barrier nanowire field effect transistor.

A top-down nanofabrication approach involving molecular beam epitaxy and electron beam lithography was used to obtain silicon nanowire-based back gate field-effect transistors with Schottky contacts on silicon-on-insulator (SOI) wafers. The resulting device is applied in biomolecular detection based on the changes in the drain-source current (IDS). In this context, we have explained the physical mechanisms of charge carrier transport in the nanowire using energy band diagrams and numerical 2D simulations in TCAD. The results of the experiment and numerical modeling matched well and may be used to develop novel types of nanowire-based biosensors.

11C-radiolabeled aptamer for imaging of tumors and metastases using positron emission tomography- computed tomography.

Identification of primary tumors and metastasis sites is an essential step in cancer diagnostics and the following treatment. Positron emission tomography-computed tomography (PET/CT) is one of the most reliable methods for scanning the whole organism for malignancies. In this work, we synthesized an 11C-labeled oligonucleotide primer and hybridized it to an anti-cancer DNA aptamer. The 11C-aptamer was applied for in vivo imaging of Ehrlich ascites carcinoma and its metastases in mice using PET/CT. The imaging experiments with the 11C-aptamer determined very small primary and secondary tumors of 3 mm2 and less. We also compared 11C imaging with the standard radiotracer, 2-deoxy-2-[fluorine-18]fluoro-D-glucose (18F-FDG), and found better selectivity of the 11C-aptamer to metastatic lesions in the metabolically active organs than 18F-FDG. 11C radionuclide with an ultra-short (20.38 min) half-life is considered safest for PET/CT imaging and does not cause false-positive results in heart imaging. Its combination with aptamers gives us high-specificity and high-contrast imaging of cancer cells and can be applied for PET/CT-guided drug delivery in cancer therapies.