Design and Development of Advanced Glucose Biosensors via Tuned Interactions between Marine Polysaccharides and Diagnostic Elements – A Survey

Extensive scientific analysis on the susceptibility of different populations to COVID-19 highlights that compared to populations with no co-morbidities, people with co-morbidities such as diabetes mellitus (DM) are at a significantly higher risk of serious infection, hospitalization, or even death. This underscores the importance of controlling DM through self-monitoring of blood glucose (SMBG) or continuous glucose monitoring (CGM) using biosensors. These biosensors, which can be either enzymatic or non-enzymatic, have undergone several rounds of development in terms of the materials used for device construction. In terms of the immobilization agent needed to anchor enzymatic or non-enzymatic detection elements to the electrode surface, marine polysaccharides, such as chitosan and alginate, hold a distinct advantage. This, in turn, can be ascribed to their biocompatibility, chemical stability, film-forming ability, capacity to bind with enzymes/diagnostic elements, and easy availability. In this review, we focus extensively on the use of cationic chitosan and anionic alginate in the past decade for designing advanced glucose biosensors. Their role in enhancing sensor response via physical/chemical interactions with the conducting and diagnostic elements is analyzed in detail from a structural point of view. In addition, the possibility of using these polysaccharides in non-invasive CGM sensors is discussed and several potential future research avenues are presented.

Effects of Juglone on Neutrophil Degranulation and Myeloperoxidase Activity Related to Equine Laminitis

Experimental laminitis, characterized by a failure of the dermal–epidermal interface of the foot, can be induced in horses by the oral administration of a black walnut extract (BWE). In the early phase of this severe and painful disease, an activation of neutrophil occurs, with the release of myeloperoxidase (MPO), a pro-oxidant enzyme of neutrophils, in plasma, skin, and laminar tissue. Juglone, a naphthoquinone derivative endowed with redox properties, is found in walnuts and has been incriminated in this neutrophil activation. We report for the first time the inhibitory activity of juglone on the degranulation of neutrophils induced by cytochalasin B and formyl-methionyl-leucyl-phenylalanine as monitored by the MPO release (>90% inhibition for 25 and 50 μM). Moreover, it also acts on the peroxidase activity of MPO by interacting with the intermediate “π cation radical,” as evidenced by the classical and specific immunological extraction followed by enzymatic detection (SIEFED) assays. These results are confirmed by a docking study showing the perfect positioning of juglone in the MPO enzyme active site and its interaction with one of the amino acids (Arg-239) of MPO apoprotein. By chemiluminescence and electron paramagnetic resonance techniques, we demonstrated that juglone inhibited reactive oxygen species (ROS) and superoxide anion free radical produced from phorbol myristate acetate (PMA)-activated polymorphonuclear neutrophils (PMNs). These results indicate that juglone is not the trigger for equine laminitis, at least if we focus on the modulation of neutrophil activation.

An Enzyme-Based Biosensor for the Detection of Organophosphate Compounds Using Mutant Phosphotriesterase Immobilized onto Reduced Graphene Oxide

Enzymatic detection of organophosphate (OP) compounds can be tailored using highly sensitive and selective enzymes in the development of biosensors. Previously, mutant (YT) phosphotriesterase (PTE) was reported to efficiently hydrolyze Sp and Rp enantiomers of phosphotriester. This study reports the use of phosphotriesterase mutant YT (YT-PTE) immobilized onto reduced graphene oxide (rGO) and fabricated onto a screen-printed carbon electrode (SPCE) for electrochemical detection of OP compounds. Immobilization of YT-PTE onto rGO was secured using N-hydroxysuccinimide (NHS) and N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide (EDC) cross-linker, and the resulting immobilized enzyme was able to retain up to 90% of its activity. Electrochemical analysis of the SPCE/rGO/YT-PTE showed detection of paraoxon in a linear range of 1 mM–0.005 μM with its limit of detection as low as 0.11 μM. SPCE/rGO/YT-PTE exhibited high selectivity towards paraoxon and parathion and have good reproducibility. Furthermore, detection of paraoxon was also possible in a real water sample with only minor interferences.

Simultaneous Detection of Glucose and Fructose in Synthetic Musts by Multivariate Analysis of Silica-Based Amperometric Sensor Signals

Silica-based electrodes which permanently include a graphite/Au nanoparticles composite were tested for non-enzymatic detection of glucose and fructose. The composite material showed an effective electrocatalytic activity, to achieve the oxidation of the two analytes at quite low potential values and with good linearity. Reduced surface passivation was observed even in presence of organic species normally constituting real samples. Electrochemical responses were systematically recorded in cyclic voltammetry and differential pulse voltammetry by analysing 99 solutions containing glucose and fructose at different concentration values. The analysed samples consisted both in glucose and fructose aqueous solutions at pH 12 and in solutions of synthetic musts of red grapes, to test the feasibility of the approach in a real frame. Multivariate exploratory analyses of the electrochemical signals were performed using the Principal Component Analysis (PCA). This gave evidence of the effectiveness of the chemometric approach to study the electrochemical sensor responses. Thanks to PCA, it was possible to highlight the different contributions of glucose and fructose to the voltammetric signal, allowing their selective determination.