Oxidation of anti-thyroid drugs and their selenium analogs by ABTS radical cation.

Lactoperoxidase was previously used as a model enzyme to test the inhibitory activity of selenium analogs of anti-thyroid drugs with 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) as a substrate. Peroxidases oxidize ABTS to a metastable radical ABTS•+, which is readily reduced by many antioxidants, including thiol-containing compounds, and it has been used for decades to measure antioxidant activity in biological samples. We showed that anti-thyroid drugs 6-n-propyl-2-thiouracil, methimazole, and selenium analogs of methimazole also reduced it rapidly. This reaction may explain the anti-thyroid action of many other compounds, particularly natural antioxidants, which may reduce the oxidized form of iodine and/or tyrosyl radicals generated by thyroid peroxidase thus decreasing the production of thyroid hormones. However, influence of selenium analogs of methimazole on the rate of hydrogen peroxide consumption during oxidation of ABTS by lactoperoxidase was moderate. Direct hydrogen peroxide reduction, proposed before as their mechanism of action, cannot therefore account for the observed inhibitory effects. 1-Methylimidazole-2-selone and its diselenide were oxidized by ABTS•+ to relatively stable seleninic acid, which decomposed slowly to selenite and 1-methylimidazole. In contrast, oxidation of 1,3-dimethylimidazole-2-selone gave selenite and 1,3-dimethylimidazolium cation. Accumulation of the corresponding seleninic acid was not observed.

In vitro oxidation of hispidin and gallic acid by horseradish peroxidase.

Gallic acid and hispidin have been previously described by us as inhibitors of horseradish peroxidase (Benarous, K., Benali, F. Z., Bekhaoua, I. C., and Yousfi, M. Journal of Biomolecular Structure & Dynamics, (2021) 39(18), 7168-7180). However, additional experiments have demonstrated that under the applied assay conditions both compounds are rapidly oxidized by this enzyme. After oxidation, the components of the reaction mixture undergo complex reactions giving products with much weaker absorption at the detection wavelength. This was interpreted by us as enzyme inhibition, which, however, is only apparent. In fact, the activity of horseradish peroxidase is not affected by these compounds, which was demonstrated by measurements of hydrogen peroxide consumption.Communicated by Ramaswamy H. Sarma.

Rifampicin is not an inhibitor of tyrosinase.

Rifampicin has been previously described as an inhibitor of tyrosinase (Chai et al., Int. J. Biol. Macromol. 102 (2017) 425-430). However, rifampicin contains a p-diphenol group and compounds with such a moiety have been shown before to reduce tyrosinase-generated o-quinones. Rifampicin also shows strong absorption in a region completely overlapping with the visible absorption band of dopachrome, the oxidation product of L-tyrosine and L-dopa, whose concentration is measured spectrophotometrically in the standard enzymatic assay to monitor the activity of tyrosinase. We have demonstrated that rifampicin is also rapidly oxidized by o-quinones generated from catechols by tyrosinase or by treatment with sodium periodate. Smaller changes of absorbance at 475 nm during oxidation of L-dopa by tyrosinase in the presence of rifampicin do not result from enzyme inhibition but from oxidation of rifampicin by dopaquinone, which leads to rapid decrease of rifampicin absorption in this range. The actual reaction rates are not affected, which we have demonstrated by measurements of oxygen consumption. Rifampicin behaves therefore as other compounds with reducing properties, such as ascorbic acid, hydroquinone, hydrazine derivatives, and flavonoids, some of which have also been incorrectly described before as inhibitors of tyrosinase.

Mechanisms of interference of p-diphenols with the Trinder reaction.

p-Diphenols, such as homogentisic acid, gentisic acid, etamsylate, and calcium dobesilate, interfere with diagnostic tests utilizing the Trinder reaction but the mechanisms of these effects are not fully understood. We observed substantial differences both in oxidation of p-diphenols by horseradish peroxidase and their influence on oxidation of 4-aminoantipyrine and various phenolic substrates. Homogentisic acid was rapidly oxidized by the enzyme and completely blocked chromophore formation. Enzymatic oxidation of the remaining p-diphenols was slow and they only moderately inhibited chromophore formation. However, in the presence of standard substrates all tested p-diphenols were rapidly converted to p-quinones. Hydrogen peroxide consumption was significantly accelerated by homogentisic acid but not much affected by the other p-diphenols. The magnitude and mechanisms of interference caused by p-diphenols therefore depend on their structure which determines their electrochemical properties - while for homogentisic acid with an electron-donating substituent and a lower reduction potential both enzymatic oxidation and reduction of the peroxidase-generated radicals occur, for p-diphenols with electron-withdrawing substituents and higher reduction potentials only the second mechanism is significant. Correlation of the effects on the Trinder reaction with reduction potentials of interfering compounds allows prediction of such properties for a wide range of other reducing compounds based on this parameter. It also explains why compounds with very different structures but strong reducing properties show such effects.

The BEM-FDM model of thermal processes proceeding in the domain of the human finger.

PURPOSE: The problem of the numerical modeling of thermal processes proceeding in the non-homogeneous domain of the human finger is discussed. The domain considered constitutes the assembling of soft and bone tissues and the system of supplying blood vessels (arteries and veins). The mathematical description of the process analyzed corresponds to the so-called vascular models. METHODS: At the stage of numerical modeling the algorithm being the composition of the boundary element method (BEM) and the finite difference method (FDM) is applied. RESULTS: The algorithm presented allows one to determine the steady state temperature field in the finger domain in natural convection conditions. To verify the effectiveness and exactness of the method of the problem solution, the thermal imaging measurements of the finger surface temperature have been done. CONCLUSIONS: The compatibility of numerical and experimental results (the natural convection conditions) has proved to be quite satisfactory. It is possible to use the algorithm proposed for the modeling of thermal processes proceeding in the conditions of low or high ambient temperatures and the big values of heat transfer coefficients. The impact of protective clothing on the temperature field in the domain of the finger can also be analyzed.