Penicillium expansum Impact and Patulin Accumulation on Conventional and Traditional Apple Cultivars

Penicillium expansum is a necrotrophic plant pathogen among the most ubiquitous fungi disseminated worldwide. It causes blue mould rot in apples during storage, transport and sale, threatening human health by secreting patulin, a toxic secondary metabolite that contaminates apples and apple-derived products. Nevertheless, there is still a lack of sufficient data regarding the resistance of different apple cultivars to P. expansum, especially ancient ones, which showed to possess certain resistance to plant diseases. In this work, we investigated the polyphenol profile of 12 traditional and 8 conventional apple cultivar and their resistance to P. expansum CBS 325.48. Eight polyphenolic compounds were detected; the most prominent were catechin, epicatechin and gallic acid. The highest content of catechin was detected in ‘Apistar’—91.26 mg/100 g of fresh weight (FW), epicatechin in ‘Bobovac’—67.00 mg/100 g of FW, and gallic acid in ‘Bobovac’ and ‘Kraljevčica’—8.35 and 7.40 mg/100 g of FW, respectively. The highest content of patulin was detected in ‘Kraljevčica’ followed by ‘Apistar’—1687 and 1435 µg/kg, respectively. In apple cultivars ‘Brčko’, ‘Adamčica’ and ‘Idared’, patulin was not detected. Furthermore, the patulin content was positively correlated with gallic acid (r = 0.4226; p = 0.002), catechin (r = 0.3717; p = 0.008) and epicatechin (r = 0.3305; p = 0.019). This fact indicates that higher contents of gallic acid, catechin and epicatechin negatively affected and boost patulin concentration in examined apple cultivars. This can be related to the prooxidant activity of polyphenolic compounds and sensitivity of P. expansum to the disturbance of oxidative status.

Ethanolic Extracts from Agro-Industrial Co-Products Enhance Oxidative Stability of Candelilla Wax or Celluloses Derivatives Oleogels

Ethanol oleoresins obtained from orange peel, pea pod, or pomegranate peel were employed to enrich soybean oil before elaborating two types of oleogels, one made with candelilla wax, another made with a mixture of celluloses derivatives, in order to determine their effect on oleogel oxidative stability. Orange peel oleoresin obtained a higher amount of polyphenols as catechol acid equivalent (9.09 meq/g), as compared to pea pod oleoresin or pomegranate peel (8.80 and 8.55, respectively), although pomegranate peel presented the higher TEAC (1.67, twice than the other samples). Oleogels elaborated with celluloses were presented better oxidative stability (oxidative rancidity and peroxide index) as compared to samples elaborated with candelilla wax, since waxes minor constituents employed as oleogelators could promote prooxidant activity. Samples with pomegranate oleoresin presented enhanced oxidative stability. Candelilla wax has a more marked effect on the oleogels thermal properties of due to their influence on fatty acids crystallization, because in celluloses oleogels the mechanism of gelation does not imply the formation of a highly ordered secondary structure. This finding opens the possibility to, on one hand, use oleoresins to enhance the oxidative stability of oleogels; and on the other hand, to select the oleogelator, waxes, or celluloses, based on thermal properties and other aspects, depending on the further application of oleogel thinking in oleogel thermoreversible capacity.

Concentration Dependence of the Antioxidant and Prooxidant Activity of Trolox in HeLa Cells: Involvement in the Induction of Apoptotic Volume Decrease

Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), a hydrophilic analog of vitamin E, is known for its strong antioxidant activity, being a high radical scavenger of peroxyl and alkoxyl radicals. Under particular conditions, Trolox may also exhibit prooxidant properties. The present work aimed at studying the dual antioxidant/prooxidant behavior of Trolox over a wide range of concentrations (from 2.5 to 160 µM) in HeLa cells. In particular, the study addressed the dose-dependent effects of Trolox on the oxidative cell status and vitality of HeLa cells, focusing on the potential role of the vitamin E analog in the induction of one of the first steps of the apoptotic process, Apoptotic Volume Decrease (AVD). In HeLa cells, Trolox showed significant antioxidant activity, expressed as the ability to reduce the endogenous ROS production detected by the ROS-sensitive probe 5-(and-6)-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate (CM-H2DCFDA), at low concentrations (range: 2.5–15 µM), but exerted a dose-dependent prooxidant effect at higher concentrations after 24 h exposure. The prooxidant effect was paralleled by the reduction in cell viability due to the induction of the apoptotic process. The dual behavior, antioxidant at lower concentrations and prooxidant at higher concentrations, was evident also earlier after 2 h incubation, and it was paralleled by the isotonic shrinkage of the cells, ascribed to AVD. The use of SITS, known Cl− channel blocker, was able to completely inhibit the Trolox-induced isotonic cell shrinkage, demonstrating the involvement of the vitamin E analog in the alteration of cell volume homeostasis and, in turn, in the AVD induction. In conclusion, the study shed light on the concentration dependence of the Trolox antioxidant/prooxidant activity in HeLa cells and revealed its role in the induction of one of the first events of apoptosis, AVD, at high concentrations.

ANTIOXIDANT AND CYTOTOXIC ACTIVITY OF A SERIES OF O- AND S-CONTAINING MACROCYCLES

The significant interest in the search and study of antioxidants, both natural and synthetic, is explained by the possibility of preventing these harmful effects of free radicals in the human body. The similarity of macroheterocyclic derivatives to endogenous biomolecules makes them promising for research as antioxidant agents. By the interaction of 1,3-oxoheterocycloalkanes with ethyl 2-diazo-3-oxobutanoate, previously unexplored polyfunctional oxygen and sulfur-containing macroheterocycles are obtained. The relative antioxidant activity of new macroheterocycles in two model systems: generating active forms of oxygen and modulating lipid peroxidation reaction, was revealed active forms of oxygen and described by the method of recording luminol-dependent chemiluminescence lipid peroxidation reaction. The in vitro cytotoxic activity of the studied compounds was studied on Jurkat (human T-lymphoblastic leukemia cells), HepG2 (human liver carcinoma cells), HEK293 (human embryonic kidney cells) cell lines. The data obtained indicate practical significance. So, reagents 1-7 in the active forms of oxygen generating system have anti- and prooxidative properties, and in the system modeling lipid peroxidation reaction, they show only prooxidant activity. The influence of heterocycles 1–7 on the processes of free radical oxidation was determined by the intensity of the maximum flash (Imax, у.е.) and the luminosity of light (S, у.е.). Compounds with maximum anti- (4) and prooxidative (6) properties in the active forms of oxygen generating system and reagent 3 with the highest prooxidant activity in the lipid peroxidation reaction modeling system do not possess cytotoxic activity in vitro on cell lines HEK293 and HepG2 cultured in DMEM medium (Biolot, Russia), Jurkat in RPMI medium (Biolot, Russia) in the presence of 10% fetal calf serum (Invitrogen, USA), 2 mM L-glutamine and 50 μg / ml gentamicin sulfate.