RESUMO
Scientists have established a connection between environmental exposure to toxins like ß-N-methylamino-l-alanine (BMAA) and a heightened risk of neurodegenerative disorders. BMAA is a byproduct from certain strains of cyanobacteria that are present in ecosystems worldwide and is renowned for its bioaccumulation and biomagnification in seafood. The sensitivity, selectivity, and reproducibility of the current analytical techniques are insufficient to support efforts regarding food safety and environment monitoring adequately. This work outlines the in vitro selection of BMAA-specific DNA aptamers via the systematic evolution of ligands through exponential enrichment (SELEX). Screening and characterization of the full-length aptamers was achieved using the SYBR Green (SG) fluorescence displacement assay. Aptamers BMAA_159 and BMAA_165 showed the highest binding affinities, with dissociation constants (Kd) of 2.2 ± 0.1 µM and 0.32 ± 0.02 µM, respectively. After truncation, the binding affinity was confirmed using a BMAA-conjugated fluorescence assay. The Kd values for BMAA_159_min and BMAA_165_min were 6 ± 1 µM and 0.63 ± 0.02 µM, respectively. Alterations in the amino proton region studied using solution nuclear magnetic resonance (NMR) provided further evidence of aptamer-target binding. Additionally, circular dichroism (CD) spectroscopy revealed that BMAA_165_min forms hybrid G-quadruplex (G4) structures. Finally, BMAA_165_min was used in the development of an electrochemical aptamer-based (EAB) sensor that accomplished sensitive and selective detection of BMAA with a limit of detection (LOD) of 1.13 ± 0.02 pM.
RESUMO
Polyphenols are natural compounds with strong antioxidant properties synthesized by plants and widely distributed in plant tissues. They compose a broad class of compounds that are commonly employed for multiple applications such as food, pharmaceutical, adhesives, biomedical, agricultural, and industrial purposes. Runoffs from these sources result in the introduction of polyphenols into aquatic environments where they further transform into highly toxic pollutants that can negatively affect aquatic ecosystems and humans. Therefore, the development of extraction and remediation methods for such compounds must be addressed. This study describes the identification and operation of a method to recover polyphenolic compounds from water environments by utilizing membrane-based separation. Composite membranes derived from electrospun cellulose acetate (CA) fibers and diblock copolymer (DiBCP) PEO-b-P4VP were prepared to evaluate the adsorption of polyphenolic compounds from aqueous environments. The highly porous CA fibers were developed using the electrospinning technique, and the fabricated DiBCP/CA membranes were characterized using scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), X-ray photoelectron spectroscopy (XPS), Fourier-transform infrared (FT-IR) spectroscopy, and tensile testing. Finally, the ability of the composite membranes to adsorb the soluble polyphenolic compounds catechol (CAT) and gallic acid (GA), from a wetland environment, was studied via batch adsorption experiments and by solid-phase extraction (SPE). Results revealed a successful recovery of both polyphenols, at concentrations within the parts per million (ppm) range, from the aqueous media. This suggests a novel approach to recover these compounds to prevent their transformation into toxic pollutants upon entrance to water environments.