The influence of leaf area and various variants of manual defoliation on the phenolic profile of the Prokupac variety grape berry were investigated in the agroecological conditions of southern Serbia. The following four trial variants of manual defoliation were assessed: Early defoliation—variant I (flowering stage, 50% open flowers); early defoliation—variant II (grape size 3–5 mm); late defoliation—variant III (onset of grape ripening, veraison); and control (no defoliation). The first six leaves of each primary shoot were removed from all defoliated vines. The greatest assimilation area of primary and lateral shoots during the study was observed in the control, i.e., the trial variant with no defoliation. Defoliation significantly decreased the grape yield of the all three defoliated variants in regard to the control. The phenolic profile of the three variants and control was established by analyzing the grape seeds and skin. Based on the collected results for the Prokupac variety, significant differences between the trial variants were established regarding the content of phenols and total polyphenols, as well as radical scavenging activity. Defoliation variants showed a significant effect on the total phenols content of grape skin. In all defoliation variants, as well as in the control, high amounts of ellagic acid were measured. Resveratrol was identified only in grape skin samples of the control variant. The removal of leaves increased the concentration of phenolic compounds in variants where early defoliation was applied. The highest total anthocyanins content was found in 2015 in variant I, where leaves were removed during the full flowering stage.

Biperiden, an antiparkinsonian anticholinergic drug, has been found to inhibit enzymatic hydrolysis of butyrylthiocholine iodide, which is catalyzed by serum cholinesterase. By measuring the difference in the basic and inhibitory hydrolysis reaction rates, in the presence of biperiden as an inhibitor, it is possible to develop a kinetic method for its determination. Both systems, enzyme-substrate-chromogen and enzyme-substrate-chromogen-inhibitor, were characterized by biochemical parameters (KM = 0.326 ? 0.330 mmol dm-3; Vmax = 40 - - 99 ?mol dm-3 min-1), while inhibition was defined as non-competitive with the constant of inhibition Ki = 6.142 ?mol dm-3. The reaction conditions have been optimized followed by determination of the calibration curve, the corresponding equation and the limits of detection and quantification yielding 3.84 and 12.80 nmol dm-3, respectively. Using the calibration chart, it is possible to determine biperiden in different samples in the concentration range of 0.035?35.940 ?mol dm-3. Influence of a number of substances, found in the sample, on the reaction rate was also examined. The optimized method was applied for determination of biperiden in pharmaceutical preparations. Accuracy of the method was tested using the standard addition method. The proposed method has good sensitivity, selectivity, it is simple and fast, and above all easily accessible, and thus applicable in biochemical and pharmaceutical laboratories.

L-ascorbic acid is one of the essential nutrients and most common food supplements, fortificants, and preservatives. It is commercially available as solutions, drops, tablets, capsules, crystal powder, beverage mixtures, multivitamin formulations, and multi antioxidant formulations. The usual daily dose is from 25 mg to 1.5 g. Ascorbic acid is a distinctly reducing agent with low redox potential (0.18 and 0.08 V at pH 4.5 and 6.4, respectively). Based on ascorbate property, numerous methods for its quantitative determination are developed, from titrimetric, electrochemical, and chromatographic methods, to fluorometric and kinetic ones. Enzyme peroxidase is interfered with by ascorbic acid, which decreases the oxidation speed of its co-substrates during hydrogen peroxide decomposition by peroxidase. Absorbance changes at the wavelength of corresponding reagents are in correlation with ascorbate concentration. During this study, benzidine and o-tolidine have been used as chromogenic reagents. Reaction conditions were optimized for various buffer systems, calibration curves were constructed, and limits of detection (0.04 mmol/L) and quantification (0.12 mmol/L) were calculated. Using calibration charts, it was possible to detect ascorbic acid within limits from 0.4 to 10 mmol/L. The optimized method was applied for the determination of ascorbic acid in pharmaceutical products. The method was characterized by exceptional sensitivity and accuracy, but only for preparations not containing substances that affect enzyme peroxidase.

This paper deals with the resistance of twenty commercial winter wheat cultivars to common bunt causal agent (Tilletia tritici). Significant differences among the cultivars concerning the infection percent were observed, as well as the differences in the level of commercial cultivars' resistance to T. tritici. Most of the studied cultivars belonged to susceptible categories, and just few of them to the resistant ones. Cultivar Lasta was classified as highly resistant during the both investigation years in Kragujevac, while in Leposavic Lasta and Tiha were classified as resistant. The other studied cultivars were more or less susceptible.

Primary role of peroxidase enzyme is to decompose endogenous hydrogen peroxide, when oxygen radical is being replaced by a less potent radical, which is its cosubstrates oxidized form. During this study, catalytic activity of horseradish peroxidase has been observed in the presence of antioxidants from vitamin group, such as C, E and A, i.e. their water-soluble forms. It was found that vitamin E showed no effect on the enzyme activity and fate of cosubstrate radicals from the group of benzidine derivatives. Vitamin C proceeds enzymatic reaction showing its antioxidative character, and absorbs electrons from radicals, bringing cosubstrate back to its relaxed state. On the other hand, vitamin A plays a role of uncompetitive peroxidase inhibitor, which is visible through decreasing initial rate of catalytic reaction, and is reflected as virtual decrease of enzyme concentration. Furthermore, it prolongs life of endogenous hydrogen peroxide, which could potentially lead to oxidative stress of cells. This inhibitory effect can be used in analytical purpose, for determination of retinol acetate content in a sample.