•Electrochemical oxidation of methionine residues in peptides.•Catalytic support of the neighboring carbonyl and amide groups.•Stabilization of electron-deficient reaction center and formation of ...S.·.O and S.·.N bonds.
The oxidation of methionine side chain residues in proteins provides a great diversity of possible oxidation mechanisms dictated, among others, by the oxidizing species, solvent properties, and protein structure. The oxidation behavior of a series of short acetylated synthetic peptides, Ac-GMG, Ac-GGMGG and Ac-GGGMGGG, was investigated by differential pulse and square wave voltammetry, in a wide pH range, at glassy carbon electrode. It was found that always the first oxidation step represents the one-electron oxidation of thioether moiety with the formation of a radical cation. Following this, depending on the experimental conditions and side chain position of methionine, the radical is stabilized by the nucleophilic attack of water or by catalytic support of the neighboring carbonyl and amide groups in an intermediate structure, finally converted in methionine sulfoxide which can be further oxidized, at more positive potential, into methionine sulfone. For Ac-GGGMGGG, at pH 8.0, the amino and amide groups are active involved in the oxidation process and the electrode reaction takes place with proton transfer
Bortezomib is an inhibitor of proteasomes and an anti-cancer drug. Although bortezomib is considered a safe drug, as confirmed by cytotoxicity assays, recent reports highlighted the possibility of ...interaction between bortezomib and cellular components, with detrimental long-term effects. The evaluation of the interaction between bortezomib and dsDNA was investigated in bulk solution and using a dsDNA electrochemical biosensor. The binding of bortezomib to dsDNA involved its electroactive centers and led to small morphological modifications in the dsDNA double helix, which were electrochemically identified through changes in the guanine and adenine residue oxidation peaks and confirmed by electrophoretic and spectrophotometric measurements. The redox product of bortezomib amino group oxidation was electrochemically generated in situ on the surface of the dsDNA electrochemical biosensor. The redox product of bortezomib was shown to interact primarily with guanine residues, preventing their oxidation and leading to the formation of bortezomib-guanine adducts, which was confirmed by control experiments with polyhomonucleotides electrochemical biosensors and mass spectrometry. An interaction mechanism between dsDNA and bortezomib is proposed, and the formation of the bortezomib redox product-guanine adduct explained.
Azathioprine (AZA) is a pharmacologic immunosuppressive agent administrated in various conditions such as autoimmune disease or to prevent the rejection of organ transplantation. The mechanism of ...action is based on its biologically active metabolite 6-mercaptopurine (6-MP), which is converted, among others, into thioguanine nucleotides capable of incorporating into replicating DNA, which may act as a strong UV chromophore and trigger DNA oxidation. The interaction between azathioprine and DNA, before and after exposure to solar simulator radiation, was investigated using UV-vis spectrometry and differential pulse voltammetry at a glassy carbon electrode. The results indicated that the interaction of AZA with UV radiation was pH-dependent and occurred with the formation of several metabolites, which induced oxidative damage in DNA, and the formation of DNA-metabolite adducts. Moreover, the viability assays obtained for the L929 cell culture showed that both azathioprine and degraded azathioprine induced a decrease in cell proliferation.
The electrochemical behavior and the interaction of the immunosuppressive drug azathioprine (AZA) with deoxyribonucleic acid (DNA) were investigated using voltammetric techniques, mass spectrometry ...(MS), and scanning electron microscopy (SEM). The redox mechanism of AZA on glassy carbon (GC) was investigated using cyclic and differential pulse (DP) voltammetry. It was proven that the electroactive center of AZA is the nitro group and its reduction mechanism is a diffusion-controlled process, which occurs in consecutive steps with formation of electroactive products and involves the transfer of electrons and protons. A redox mechanism was proposed and the interaction of AZA with DNA was also investigated. Morphological characterization of the DNA film on the electrode surface before and after interaction with AZA was performed using scanning electron microscopy. An electrochemical DNA biosensor was employed to study the interactions between AZA and DNA with different concentrations, incubation times, and applied potential values. It was shown that the reduction of AZA molecules bound to the DNA layer induces structural changes of the DNA double strands and oxidative damage, which were recognized through the occurrence of the 8-oxo-deoxyguanosine oxidation peak. Mass spectrometry investigation of the DNA film before and after interaction with AZA also demonstrated the formation of AZA adducts with purine bases.
Due to the great significance of amino acids, a substantial number of research studies has been directed toward the development of effective and reliable platforms for their evaluation, detection, ...and identification. In order to support these studies, a new electrochemical platform based on PANI/ZnO nanowires' modified carbon inks screen-printed electrodes was developed for qualitative analysis of electroactive amino acids, with emphasis on tyrosine (Tyr) and tryptophan (Trp). A comparative investigation of the carbon ink before and after modification with the PANI/ZnO was performed by scanning electron microscopy and by Raman spectroscopy, confirming the presence of PANI and ZnO nanowires. Electrochemical investigations by cyclic voltammetry and electrochemical impedance spectroscopy have shown a higher charge-transfer rate constant, which is reflected into lower charge-transfer resistance and higher capacitance values for the PANI/ZnO modified ink when compared to the simple carbon screen-printed electrode. In order to demonstrate the electrochemical performances of the PANI/ZnO nanowires' modified carbon inks screen-printed electrodes for amino acids analysis, differential pulse voltammograms were obtained in individual and mixed solutions of electroactive amino acids. It has been shown that the PANI/ZnO nanowires' modified carbon inks screen-printed electrodes allowed for tyrosine and tryptophan a peak separation of more than 100 mV, enabling their screening and identification in mixed solutions, which is essential for the electrochemical analysis of proteins within the proteomics research field.
Abstract
The present work reports a new configuration of soft artificial muscle based on a web of metal covered nylon 6/6 micrometric fibers attached to a thin polydimethylsiloxane (PDMS) film. The ...preparation process is simple and implies the attachment of metalized fiber networks to a PDMS sheet substrate while heating and applying compression. The resulting composite is versatile and can be cut in different shapes as a function of the application sought. When an electric current passes through the metallic web, heat is produced, leading to local dilatation and to subsequent controlled deformation. Because of this, the artificial muscle displays a fast and ample movement (maximum displacement of 0.8 cm) when applying a relatively low voltage (2.2 V), a consequence of the contrast between the thermal expanse coefficients of the PDMS substrate and of the web-like electrode. It was shown that the electrical current producing this effect can originate from both direct electric contacts, and untethered configurations i.e. radio frequency induced. Usually, for thermal activated actuators the heating is produced by using metallic films or conductive carbon-based materials, while here a fast heating/cooling process is obtained by using microfiber-based heaters. This new approach for untethered devices is an interesting path to follow, opening a wide range of applications were autonomous actuation and remote transfer of energy are needed.
This study presents the design and manufacture of metasurface lenses optimized for focusing light with 1.55 µm wavelength. The lenses are fabricated on silicon substrates using electron beam ...lithography, ultraviolet-nanoimprint lithography and cryogenic deep reactive-ion etching techniques. The designed metasurface makes use of the geometrical phase principle and consists of rectangular pillars with target dimensions of height h = 1200 nm, width w = 230 nm, length l = 354 nm and periodicity p = 835 nm. The simulated efficiency of the lens is 60%, while the master lenses obtained by using electron beam lithography are found to have an efficiency of 45%. The lenses subsequently fabricated via nanoimprint are characterized by an efficiency of 6%; the low efficiency is mainly attributed to the rounding of the rectangular nanostructures during the pattern transfer processes from the resist to silicon due to the presence of a thicker residual layer.
The quality of life of patients with chronic wounds can be extremely poor and, therefore, over the past decades, great efforts have been made to develop efficient strategies to improve the healing ...process and the social impact associated with these conditions. Cell based therapy, as a modern tissue engineering strategy, involves the design of 3D cell-scaffold bioconstructs obtained by preseeding drug loaded scaffolds with undifferentiated cells in order to achieve in situ functional de novo tissue. This paper reports on the development of bionanocomposites based on bacterial cellulose and magnetic nanoparticles (magnetite) for efficient chronic wounds healing. Composites were obtained directly in the cellulose bacterial culture medium by dispersing various amounts of magnetite nanoparticles during the biosynthesis process. After purification and drying, the membranes were characterized by Raman spectroscopy and X-ray diffraction to reveal the presence of magnetite within the bacterial cellulose matrix. Morphological investigation was employed through SEM and TEM analyses on bionanocomposites. The biocompatibility of these innovative materials was studied in relation to human adipose derived stem cells in terms of cellular morphology, viability, and proliferation as well as scaffolds cytotoxic potential.
A novel scaffold for in situ electrochemical detection of cell biomarkers was developed using electrospun nanofibers and commercial adhesive polymeric membranes. The electrochemical sensing of cell ...biomarkers requires the cultivation of the cells on/near the (bio)sensor surface in a manner to preserve an appropriate electroactive available surface and to avoid the surface passivation and sensor damage. This can be achieved by employing biocompatible nanofiber meshes that allow the cells to have a normal behavior and do not alter the electrochemical detection. For a better mechanical stability and ease of handling, nylon 6/6 nanofibers were collected on commercial polymeric membranes, at an optimal fiber density, obtaining a double-layered platform. To demonstrate the functionality of the fabricated scaffold, the screening of cellular stress has been achieved integrating melanoma B16-F10 cells and the (bio)sensor components on the transducer whereas the melanin exocytosis was successfully quantified using a commercial electrode. Either directly on the surface of the (bio)sensor or spatially detached from it, the integration of cell cultures in biosensing platforms based on electrospun nanofibers represents a powerful bioanalytical tool able to provide real-time information about the biomarker release, enzyme activity or inhibition, and monitoring of various cellular events.
Graphical Abstract
The paper is focused on designing a novel silk fibroin-magnetite biomaterial scaffold for wounds healing in terms of: synthesis, physico-chemical and morphological characterization and in vitro ...biological behaviour assessment. Magnetic scaffolds were prepared from silk fibroin solutions and magnetic nanoparticles with various concentrations by solvent casting technique. Specimens were investigated by FTIR-ATR spectroscopy and XRD measurements. Morphological investigation including internal structure was employed by AFM and TEM/HRTEM analyses. Biological assay was performed on human adipose derived stem cells isolated from subcutaneous adipose tissue. Results suggested that all the tested magnetic silk-magnetite scaffolds display a good biocompatibility in vitro on hASCs and could be promising candidates for further wound dressing testing.
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