A modification of the ABTS• decolorization assay for plate readers is presented. In our modification, 200 µL of ABTS solution of absorbance 1.0 at 734 nm was added with an antioxidant and decreased absorbance resulted. For comparison of antioxidant activities in the kinetic assay of absorbance decrease, concentration dependence of absorbance decrease and of area under curve are recommended. “Fast” and “slow” antioxidants were distinguished: while the reactions of “fast” antioxidants ABTS• were completed within seconds, the reactions of “slow” antioxidants were not finished after 6 min. We recommend reaction time of 60 min for assays of such antioxidants, blood plasma and plant extracts. Sub-additive interactions between some antioxidants (ascorbate and Trolox, hispidulin and Trolox, and glutathione and ascorbate) were found in the ABTS• decolorization; possible reasons for such interactions are discussed.
Parkinson’s disease (PD) patients can benefit from antioxidant supplementation, and new efficient antioxidants are needed. The aim of this study was to evaluate the protective effect of selected nitroxide-containing redox nanoparticles (NRNPs) in a cellular model of PD. Antioxidant properties of NRNPs were studied in cell-free systems by protection of dihydrorhodamine 123 against oxidation by 3-morpholino-sydnonimine and protection of fluorescein against bleaching by 2,2-azobis(2-amidinopropane) hydrochloride and sodium hypochlorite. Model blood-brain barrier penetration was studied using hCMEC/D3 cells. Human neuroblastoma SH-SY5Y cells, exposed to 6-hydroxydopamine (6-OHDA), were used as an in vitro model of PD. Cells were preexposed to NRNPs or free nitroxides (TEMPO or 4-amino-TEMPO) for 2 h and treated with 6-OHDA for 1 h and 24 h. The reactive oxygen species (ROS) level was estimated with dihydroethidine 123 and Fluorimetric Mitochondrial Superoxide Activity Assay Kit. Glutathione level (GSH) was measured with ortho-phtalaldehyde, ATP by luminometry, changes in mitochondrial membrane potential with JC-1, and mitochondrial mass with 10-Nonyl-Acridine Orange. NRNP1, TEMPO, and 4-amino-TEMPO (25-150 μM) protected SH-SY5Y cells from 6-OHDA-induced viability loss; the protection was much higher for NRNP1 than for free nitroxides. NRNP1 were better antioxidants in vitro and permeated better the model BBB than free nitroxides. Exposure to 6-OHDA decreased the GSH level after 1 h and increased it considerably after 24 h (apparently a compensatory overresponse); NRNPs and free nitroxides prevented this increase. NRNP1 and free nitroxides prevented the decrease in ATP level after 1 h and increased it after 24 h. 6-OHDA increased the intracellular ROS level and mitochondrial superoxide level. Studied antioxidants mostly decreased ROS and superoxide levels. 6-OHDA decreased the mitochondrial potential and mitochondrial mass; both effects were prevented by NRNP1 and nitroxides. These results suggest that the mitochondria are the main site of 6-OHDA-induced cellular damage and demonstrate a protective effect of NRNP1 in a cellular model of PD.
Several Cu2O and TiO2 thin films and four additional TiO2/Cu2O structures were fabricated by direct current (DC) magnetron sputtering. The process parameters were selected on the basis of earlier studies and numerical simulations. We examined the morphology of a cross-section of the PV structures, roughness and topography, and the transmission spectra of the thin films. Additionally, the properties of the samples were determined by X-ray diffraction. Next, the morphology cross-sectional and layer compositions of the solar cells was evaluated by scanning electron microscopy. Only one of the TiO2/Cu2O structures appeared smooth and homogeneous with columnar-type growth. For the as-grown films, diffraction peaks were observed and identified as brookite, rutile, CuO, and Cu2O and the average roughness of the samples was 0.5, 1.2, 5.4, and 4.0 nm, respectively. Finally, the transmission spectra of the thin films were recorded. Transmission and reflection spectra of ultraviolet-visible spectroscopy were analyzed, and the optical band gap and absorption coefficient of the oxidized layers were calculated. In the region of 400 to 1000 nm, transmittance varied from 5% to 70% in the TiO2 samples, and from 15% to 40% in the Cu2O samples, and reflectance of the TiO2 and Cu2O samples ranged from 20% to 90%. In the region of 1.5 eV to 3.5 eV, the mean absorption coefficient varied from ∼105 1/cm to ∼3 · 105 1/cm for TiO2 thin film, and from ∼2 · 105 to ∼6 · 105 1/cm for Cu2O thin film. The optical band gap values of the samples shifted slightly toward bulk anatase-3.5 eV, bulk rutile-3.1 eV, and copper(I) oxide. Finally, silver contacts were used for the electrodes. One of the fabricated TiO2/Cu2O PV structures was found to be sensitive to electromagnetic radiance during the experiment.
Molybdenum disulphide (MoS2) is a layered material with interesting photocatalytic properties.
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