Aneta Pop

The role of antioxidants in the chemistry of oxidative stress: A review

This Review Article is focused on the action of the reactive oxygenated species in inducing oxidative injury of the lipid membrane components, as well as on the ability of antioxidants (of different structures and sources, and following different mechanisms of action) in fighting against oxidative stress. Oxidative stress is defined as an excessive production of reactive oxygenated species that cannot be counteracted by the action of antioxidants, but also as a perturbation of cell redox balance. Reactive oxygenated/nitrogenated species are represented by superoxide anion radical, hydroxyl, alkoxyl and lipid peroxyl radicals, nitric oxide and peroxynitrite. Oxidative stress determines structure modifications and function modulation in nucleic acids, lipids and proteins. Oxidative degradation of lipids yields malondialdehyde and 4-hydroxynonenal, but also isoprostanes, from unsaturated fatty acids. Protein damage may occur with thiol oxidation, carbonylation, side-chain oxidation, fragmentation, unfolding and misfolding, resulting activity loss. 8-hydroxydeoxyguanosine is an index of DNA damage. The involvement of the reactive oxygenated/nitrogenated species in disease occurrence is described. The unbalance between the oxidant species and the antioxidant defense system may trigger specific factors responsible for oxidative damage in the cell: over-expression of oncogene genes, generation of mutagen compounds, promotion of atherogenic activity, senile plaque occurrence or inflammation. This leads to cancer, neurodegeneration, cardiovascular diseases, diabetes, kidney diseases. The concept of antioxidant is defined, along with a discussion of the existent classification criteria: enzymatic and non-enzymatic, preventative or repair-systems, endogenous and exogenous, primary and secondary, hydrosoluble and liposoluble, natural or synthetic. Primary antioxidants are mainly chain breakers, able to scavenge radical species by hydrogen donation. Secondary antioxidants are singlet oxygen quenchers, peroxide decomposers, metal chelators, oxidative enzyme inhibitors or UV radiation absorbers. The specific mechanism of action of the most important representatives of each antioxidant class (endogenous and exogenous) in preventing or inhibiting particular factors leading to oxidative injury in the cell, is then reviewed. Mutual influences, including synergistic effects are presented and discussed. Prooxidative influences likely to occur, as for instance in the presence of transition metal ions, are also reminded.

Determination of Ascorbic Acid Content of Some Fruit Juices and Wine by Voltammetry Performed at Pt and Carbon Paste Electrodes

A method was developed for assessing ascorbic acid concentration in fruit juices and wine by differential pulse voltammetry. The oxidation peak for ascorbic acid occurs at about 530 mV (versus SCE) on a Pt strip working electrode and at about 470 mV on a carbon paste working electrode. The influence of the operational parameters like the pulse amplitude and the pulse period on the analytical signal was investigated. The obtained calibration graph shows a linear dependence between the peak height and ascorbic acid concentration within the range 0.31-20 mM with a Pt working electrode, and within the range 0.07-20 mM with a carbon paste working electrode. The equation of the calibration graph was y = 21.839x + 35.726, r2 = 0.9940, when a Pt strip electrode was used (where y represents the value of the current intensity measured for the peak height, expressed as µA and x the analyte concentration, as mM). R.S.D. = 2.09%, n = 10, Cascorbic acid = 2.5 mM. The equation of the calibration graph was y = 3.4429x + 5.7334, r2 = 0.9971, when a carbon paste electrode was used (where y represents the value of intensity measured for the peak height, expressed as µA and x the analyte concentration, as mM). R.S.D. = 2.35%, n = 10, Cascorbic acid = 2.5 mM. The developed method was applied to ascorbic acid assessment in fruit juices and wine. The ascorbic acid content determined ranged between 6.83 mg/100 mL juice for soft drinks (Fanta Madness) and 54.74 mg/100 mL for citrus (lemon) juices obtained by squeezing fruit. Different ascorbic acid concentrations (from standard solutions) were added to the analysed samples, the degree of recovery being comprised between 94.74 and 104.97%. The results of ascorbic acid assessment by differential pulse voltammetry were compared with those obtained by cyclic voltammetry. The results obtained by the two methods were in good agreement.

Antimicrobial and Antiparasitic Activity of Lectins

Antibiotic resistance is a major problem in current contemporary medicine and it has become a major concern of the 21st century. New resistance mechanisms developed by microorganisms spread greatly, threatening the ability to treat numerous infectious diseases, and increasing the number of nosocomial infections. Besides the role in immunology and glycobiology where they are used as hemaglutinine and identification of complex carbohydrates and glycoconjugates, lectins proved to mediate diversified biological functions like cytotoxicity, complement activation, cell-to-cell and host-pathogen communications, innate immune response, and cell-to-cell signalling. Recently, great interest has been developed for the research and applications of lectins in agriculture and medicine due to their antiparasitic and antimicrobial potentials. This review focuses on the recent data regarding the antimicrobial and antiparasitic activities of lectins, by presenting the role of lectins in host-pathogen interaction and also the cytotoxic effects on microorganisms and parasites. Identification and characterisation of new lectins with antimicrobial activity could serve as a natural alternative for the treatment of infections caused by antibiotic-resistant microorganisms and parasites.

Comparative Assessment of the Analytical Parameters in Ascorbic Acid and Sulphite Assay at a Spectrographic Carbon Working Electrode

Abstract The aim of this study is the comparative investigation of spectrographic carbon electrode’s viability as working electrode, in ascorbic acid and sulphite asssessment. Cyclic voltammetry involves a linear sweeping of the potential, the analytical signal being represented by the anodic oxidation /cathodic reduction peak of the analyte. For both analytes, the electro-oxidation resulted in an anodic peak, correlable with ascorbic acid / sulphite concentration. The analytical range of linear response corresponded to 0.07 - 10 mM for ascorbic acid and to 15.5 mg/L - 4 g/L for sulphite. The relative standard deviation RSD (%) was 2.71 % for ascorbic acid and 2.88 % for sulphite. The sensitivities, given by the slopes of the calibration graphs were 88.88 μA/mmole/L for ascorbic acid and 477.37 μA/g/L for sulphite.

Identification of Genetically Modified Foods

Modern biotechnology is trying to create new genetically modified (GM) foods that express useful traits, such as insect resistance, herbicide tolerance, infectious-agent resistance, and resistance to environmental changes. The traceability of GM organisms (GMOs) is ensured through the use of strategies and regulations based on molecular detection methods. These methods can be categorized as indirect (e.g., immunologic) or direct (e.g., genetic). Among these analytical methods, real-time PCR is considered as the “gold standard” technique. This chapter aims to discuss the advantages and disadvantages of these techniques and provide information on new technologies that offer high-throughput screening of GM foods. Furthermore, the sample collection and validation criteria presented highlight their crucial role in obtaining reliable results.

Nanoencapsulation techniques for compounds and products with antioxidant and antimicrobial activity - A critical view

Oxidative decay and microbial spoilage are issues of concern, as they constitute threats to human health. Natural antioxidants and antimicrobials hamper the negative impact of synthetic compounds and they need appropriate delivery systems. Different nanostructures can be developed: association colloids-based nanostructures, nanoemulsions, nanoliposomes, nanolaminates, nanofibers, carbon nanotubes, nanocomposites. The main nanoencapsulation techniques applied to antioxidants and antimicrobials are described: association colloid-based nanoincorporation, lipid-based nanoencapsulation techniques, encapsulation techniques based on biologically-derived polymeric nanocarriers, encapsulation techniques based on non-biological polymeric nanocarriers, cyclodextrin incorporation, electrospraying and electrospinning, carbon nanotubes and nanocomposite encapsulation. Several nanoencapsulation methods can be followed by freeze-drying or spray-drying. Protection of bioactive compounds and controlled release are achieved, but the impact of the nanomaterials on human health and on the environment should be considered. The influence of the nanoencapsulation techniques on the antioxidant/antimicrobial activity is discussed. The choice of the appropriate encapsulation method is vital. Bioactivity increase, preservation or decrease, depend on the interactions established between the functional groups of encapsulated compound and the encapsulating nanomaterial.

Impact of Pseudomonas aeruginosa quorum sensing signaling molecules on adhesion and inflammatory markers in endothelial cells

Pseudomonas aeruginosa relies on the quorum sensing (QS) signaling system as a central regulator mechanism of virulence expression that contributes to the formation and maintenance of biofilms and tolerance to conventional antimicrobials. QS Signaling molecules (QSSMs) may be recognized and may function also within the host cells, being potentially involved in the progression of the infectious process. In this study we evaluate the expression of adhesion and inflammatory molecules in endothelial cells treated with P. aeruginosa QSSMs, in order to bring new insights on the mechanisms involved in the interaction of P. aeruginosa with host cells during the infectious process. Endothelial cells were stimulated with 20 µM of main P. aeruginosa QSSMs (OdDHL = N-(3-oxododecanoyl)-L-homoserine lactone, C4HSL = N-butyryl-L-homoserine lactone, PQS = 2-heptyl-3-hydroxy-4(1H)-quinolone and HHQ = 2-heptyl-4-quinolone). Adherence to endothelial cells, inert substratum and biofilm formation was evaluated. The expression of adhesion molecules (VE-cadherin, PECAM-1, ICAM-1, and P-selectin) and inflammatory response molecules (IL-1β, IL-6, TNFα, TGFβ, and eNOS) was assessed by qRT-PCR and flow cytometry. Our results showed that bacterial adherence to inert substratum and biofilm were decreased in the presence of all tested QSSMs. The adherence index of PAO1 laboratory strain to host cells was decreased between 10–40% in the presence of QSSMs, as compared to untreated control. Expression of eukaryotic cells adhesion molecules ICAM-1 and P-selectin was stimulated by QSSMs, whereas VE-cadherin and PECAM-1 levels were increased only by C4HSL. The inflammatory response of endothelial cells was also modulated, as observed by the modified expression of IL-1β (for C4HSL, PQS and HHQ), IL-6 (for C4HSL and HHQ), TNFα (for C4HSL and HHQ), TGFβ, and eNOS factors. Our results demonstrate that the main pseudomonadal QSSMs differentially modulate endothelial cells adhesion and proinflammatory cytokine expression. These observations provide new insights in the mechanisms by which different QSSMs activate endothelial cells and modulate the infectious process, and support the importance of recent studies aiming to develop anti-QS therapeutic strategies to fight against P. aeruginosa infections.

E-learning tool for modern veterinary teaching.

The aims of the project was to develop an e-learning platform to offer a teaching package forthe students instruction before attending practical training stages.