Antioxidant activity of kafirin hydrolysates on UVB irradiated human keratinocyte cells and in silico identification.

BACKGROUND: Ultraviolet B (UVB) causes photoaging of the skin, the appearance of wrinkles, spots, and alteration of the skin barrier. The main cells in the most superficial layer of the skin are the keratinocytes; these cells play an important role in protecting this organ. OBJECTIVE: The present study aimed to investigate the antioxidant activity of the hydrolysates from kafirin to inhibit UVB-induced responses in human keratinocytes cells (HaCaT). METHODS: Kafirin hydrolysates were produced by enzymatic hydrolysis with alcalase. The activity of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), in the HaCaT cell line in the presence of UVB and the effects of the hydrolysates against the UVB-induced response were evaluated. Furthermore, the peptides that were generated by hydrolysis were identified in silico using the BIOPEP database. RESULTS: Two protein sequences were identified (α-kafirin and the precursor protein of α-kafirin), in the kafirin extract. A degree of hydrolysis of 18.8% was obtained by hydrolyzing the kafirin extract with alcalase. The kafirin hydrolysates avoided the decrease in endogenous antioxidant enzymes such as SOD, CAT, and GPx reducing the oxidative stress generated by UVB. Using the BIOPEP-UWM database, we found 102 peptide sequences, and it has shown that the peptides have a large amount of hydrophobic amino acids such as proline, alanine, and glutamine, and amino acids with high antioxidant capacity. CONCLUSION: These results suggest that the kafirin hydrolysates can be used as antioxidant agents to ameliorate UVB-induced skin keratinocytes cells' response in vitro, providing an alternative against UVB-induced photoaging.

Effect of Introducing Nonideal Capacitance in the Estimation of the Solution Resistance for Accurate Electrolytic Conductivity Measurements.

The standardization of secondary electrolytic conductivity cells requires the use of a certified reference material. The accepted certification method involves electrochemical impedance spectroscopy (EIS) to estimate the material's solution resistance. This method normally assumes that the impedance's imaginary component can be neglected; and hence, the measured impedance approximates the real impedance. Thus, a linear extrapolation of the impedance versus the period (inverse frequency) yields solution resistance. However, experimental impedance data usually do not exhibit a linear behavior over the spectra of frequency, which strongly suggest that the ideal capacitive assumption may not strictly apply. To account for the observed nonlinear behavior, we have proposed to introduce the concept of a constant phase element (CPE) to the analysis of impedance. This approach leads to the development of a relationship that improves the fitting of experimental data and improves the accuracy of the estimation, by establishing a critical frequency where extrapolation should be done. Finally, we are presenting simulated results to demonstrate how sizeable capacitive effects can influence the determination of solution resistance, and a final analysis to estimate the impact on constant cell or electrolytic conductivity values.