Antioxidants, Volume 8, Issue 1 (January 2019) – 28 articles

In the veterinary sector, many papers deal with the relationships between inflammation and oxidative stress. However, few studies investigate the mechanisms of action of oxidised molecules in the regulation of immune cells. Thus, authors often assume that these events, sometime leading to oxidative stress, are conserved among species. The aim of this review is to draw the state-of-the-art of the current knowledge about the role of oxidised molecules and dietary antioxidant compounds in the regulation of the immune cell functions and suggest some perspectives for future investigations in animals of veterinary interest. Full article

Loss-of-function mutations in the KRIT1 gene are associated with the pathogenesis of cerebral cavernous malformations (CCMs), a major cerebrovascular disease still awaiting therapies. Accumulating evidence demonstrates that KRIT1 plays an important role in major redox-sensitive mechanisms, including transcriptional pathways and autophagy, which play major roles in cellular homeostasis and defense against oxidative stress, raising the possibility that KRIT1 loss has pleiotropic effects on multiple redox-sensitive systems. Using previously established cellular models, we found that KRIT1 loss-of-function affects the glutathione (GSH) redox system, causing a significant decrease in total GSH levels and increase in oxidized glutathione disulfide (GSSG), with a consequent deficit in the GSH/GSSG redox ratio and GSH-mediated antioxidant capacity. Redox proteomic analyses showed that these effects are associated with increased S-glutathionylation of distinct proteins involved in adaptive responses to oxidative stress, including redox-sensitive chaperonins, metabolic enzymes, and cytoskeletal proteins, suggesting a novel molecular signature of KRIT1 loss-of-function. Besides providing further insights into the emerging pleiotropic functions of KRIT1, these findings point definitively to KRIT1 as a major player in redox biology, shedding new light on the mechanistic relationship between KRIT1 loss-of-function and enhanced cell sensitivity to oxidative stress, which may eventually lead to cellular dysfunctions and CCM disease pathogenesis. Full article

The aim of the current study is to characterize the bean varieties produced in the State of Oaxaca (Mexico) with the purpose of selecting varieties with the potential of being biofortified with micronutrients. Eleven bean varieties representative of the State of Oaxaca (Mexico) were harvested, the color was characterized and the physicochemical analysis, mineral content, antioxidant capacity, reducing power, and bioactive compounds were determined. Data obtained were subjected to a variance analysis for the difference between the means of the bean varieties studied through the 95% Tukey test. The results obtained show the significant difference was found among the 11 bean varieties harvested in Oaxaca (México). The analysis found five outstanding varieties with a larger quantity of favorable characteristics on the iron, zinc, protein content, antioxidant capacity and reducing power. The outstanding varieties relating Fe and Zn content were Michigan for Fe 67.4 ppm, Sangre de Toro 62.4 ppm, which showed the larger content of antioxidant capacity (82.12%) scavenging activity and Biche for Zn 34.7 ppm. The variety with a larger quantity of protein (26.66%) was Biche bean, followed by the Peruano bean with 24.91% of protein. The variety with larger content of reducing power 0.16% was Blanco Michigan bean. Lastly, it is also remarkable that such outstanding bean varieties are important to include in a biofortification program with micro-nutrients to improve the food safety and the nutrition in vulnerable communities of the urban and rural sector of Oaxaca (Mexico). Full article

The capacity of S-Allylmercapto-N-acetylcysteine (ASSNAC) to protect human retinal pigment epithelial (RPE) cells (line ARPE-19) and porcine lenses from oxidative stress was studied. Confluent ARPE-19 cultures were incubated with ASSNAC or N-acetyl-cysteine (NAC) followed by exposure to oxidants and glutathione level and cell survival were determined. Porcine lenses were incubated with ASSNAC and then exposed to H₂O₂ followed by lens opacity measurement and determination of glutathione (reduced (GSH) and oxidized (GSSG)) in isolated lens adhering epithelial cells (lens capsule) and fiber cells consisting the lens cortex and nucleus (lens core). In ARPE-19 cultures, ASSNAC (0.2 mM; 24 h) increased glutathione level by 2–2.5-fold with significantly higher increase in GSH compared to NAC treated cultures. Similarly, ex-vivo exposure of lenses to ASSNAC (1 mM) significantly reduced the GSSG level and prevented H₂O₂ (0.5 mM)-induced lens opacification. These results demonstrate that ASSNAC up-regulates glutathione level in RPE cells and protects them from oxidative stress-induced cell death as well as protects lenses from oxidative stress-induced opacity. Further validation of these results in animal models may suggest a potential use for ASSNAC as a protective therapy in retinal degenerative diseases as well as in attenuation of oxidative stress-induced lens opacity. Full article

This study investigated the effect of N-acetyl-cysteine (NAC) supplementation through drinking water on animal performance and the glutathione (GSH) redox system in weaned piglets, particularly in relation to the immediate post-weaning feed intake. To this end, 168 piglets were weaned and either fed ad libitum or fasted the first two days, and either or not administered 200 mg/L NAC via the drinking water until d14 post-weaning. Next to animal performance until day 42 (d42), the GSH redox system was measured in erythrocytes, small intestinal mucosa, liver, lung, and kidney tissue at d0, d2, and d14 post-weaning. Animal performance and GSH levels were not affected by NAC, nor by fasting. Irrespective of treatment, a significant drop in GSH at d2 post-weaning was found as compared to d0, in particular in liver (−69%), distal jejunal mucosa (−72%), and lung tissue (−80%). Post-weaning changes of the GSH redox status were strongly tissue-dependent. To conclude, this research indicates that GSH redox homeostasis was largely affected in multiple organs during the weaning transition. NAC supplementation did not increase GSH levels in any tissue, not even in fasted animals, questioning the fact if cysteine is the first or only limiting factor determining the rate of GSH synthesis in the early post-weaning phase. Full article

Membrane separation has brought about a significant change in the food processing industry because it could operate separation at low temperature without a reduction of nutrients and bioactive compounds. Citrus limon L. Burm. cv Femminello comune juice, an Italian IGP (Protected Geographical Indication) product, was subjected to the ultrafiltation (UF) process using a cellulose acetate membrane, with a cut-off of 100 kDa, subjected to different transmembrane pressures (TMP, 05–1.5 bar). Untreated and ultra-filtrated (UF) juices were investigated for physicochemical parameters including pH, titratable acidity (TA), total soluble solids (TSS) and ascorbic acid content. Total phenols (TPC) and flavonoids (TFC) contents were also determined. Rutin, hesperidin, eriocitrin, and neohesperidin were selected as markers and quantified by HPLC. Antioxidant potential was investigated by using DPPH, ABTS, and FRAP tests. RACI was used to identify the sample with highest antioxidant potential. The hypoglycemic activity was examined using carbohydrates hydrolyzing enzymes assay. The application of increasing pressures across the membrane led to a reduction in TSS without causing a loss of bioactive compounds in terms of TPC and TFC. UF juice obtained with TMP of 1.5 bar (J3) showed a significant amount of eriocitrin and hesperidin with concentrations of 15.8 and 10.5 mg/100 mL, respectively. This sample showed the highest antioxidant potential and exhibited a promising α-amylase and α-glucosidase inhibitory activity with IC₅₀ values of 31.1 and 35.3 mg/mL, respectively. Collectively our results support the use of cellulose acetate membrane to obtain an ultra-filtered juice with significant health potential. Full article

Popcorn, one of the most popular snack foods in the world, is known for being a high fiber, healthy food. Our research has found that commercial popcorn also contains significant amounts of the class of antioxidants known as phenolic acids. The total antioxidant capacity of raw and popped popcorn extract has been quantified using the Folin–Ciocalteu and FRAP assays. The polyphenols were found exclusively in the pericarp of the kernel completely bound to the oligosaccharide fiber matrix. An in vitro digestion study was also performed to predict the phenolic acids’ bioavailability. On average, nine commercial popcorn samples contain 5.93 ± 0.92 mg/g of total polyphenols after alkaline hydrolysis and 2.66 ± 0.15 mg/g after in vitro digestion as measured by the Folin–Ciocalteu assay. Furthermore, the popping process was found not to significantly decrease the antioxidant capacity. These results indicate that a considerable amount of the bound polyphenols are bioaccessible. Due to the high levels of bioaccessible polyphenols, popcorn may be a significant source of dietary polyphenol antioxidants. Full article

High neutrophil (PMN, Polymorphonuclear neutrophil) counts in the endometrium of cows affected by endometritis, suggests the involvement of oxidative stress (OS) among the causes of impaired fertility. Protein oxidation, in particular, advanced oxidation protein products (AOPP), are OS biomarkers linked to PMN activity. To test this hypothesis, the relationship between protein oxidation and uterus health was studied in thirty-eight dairy cows during the puerperium. The animals were found to be cycling, without any signs of disease and pharmacological treatments. PMN count was performed either through a cytobrush or a uterine horn lavage (UHL). Cows were classified into four groups, based on the uterine ultrasonographic characteristics and the PMN percentage in the uterine horns with a higher percentage of high neutrophil horn (HNH). They were classified as: Healthy (H); Subclinical Endometritis (SCE); Grade 1 Endometritis (EM1); and Grade 2 Endometritis (EM2). AOPP and carbonyls were measured in plasma and UHL. UHL samples underwent Western blot analysis to visualize the carbonyl and dityrosine formation. Plasma AOPP were higher (p < 0.05) in EM2. AOPP and carbonyl group concentrations were higher in the HNH samples (p < 0.05). Protein concentration in the UHL was higher in the EM2 (p < 0.05). Carbonyl and dityrosine formation was more intense in EM1 and EM2. Protein oxidation observed in the EM2 suggests the presence of an inflammatory status in the uterus which, if not adequately hindered, could result in low fertility. Full article

Dairy cows undergo various transition periods throughout their productive life, which are associated with periods of increased metabolic and infectious disease susceptibility. Redox balance plays a key role in ensuring a satisfactory transition. Nevertheless, oxidative stress (OS), a consequence of redox imbalance, has been associated with an increased risk of disease in these animals. In the productive cycle of dairy cows, the periparturient and neonatal periods are times of increased OS and disease susceptibility. This article reviews the relationship of redox status and OS with diseases of cows and calves, and how supplementation with antioxidants can be used to prevent OS in these animals. Full article

(1) Background: Paraoxonase 2 (PON2) is a ubiquitously expressed protein localized to endoplasmic reticulum and mitochondria. Previous studies have shown that PON2 exhibits anti-oxidant and anti-inflammatory functions, and PON2-deficient (PON2-def) mice are more susceptible to atherosclerosis. Furthermore, PON2 deficiency leads to impaired mitochondrial function. (2) Methods: In this study, we examined the susceptibility of PON2-def mice to diet-induced obesity. (3) Results: After feeding of an obesifying diet, the PON2-def mice exhibited significantly increased body weight due to increased fat mass weight as compared to the wild-type (WT) mice. The increased adiposity was due, in part, to increased adipocyte hypertrophy. PON2-def mice had increased fasting insulin levels and impaired glucose tolerance after diet-induced obesity. PON2-def mice had decreased oxygen consumption and energy expenditure. Furthermore, the oxygen consumption rate of subcutaneous fat pads from PON2-def mice was lower compared to WT mice. Gene expression analysis of the subcutaneous fat pads revealed decreased expression levels of markers for beige adipocytes in PON2-def mice. (4) Conclusions: We concluded that altered systemic energy balance, perhaps due to decreased beige adipocytes and mitochondrial dysfunction in white adipose tissue of PON2-def mice, leads to increased obesity in these mice. Full article

Myocardial infarction (MI) is the primary cause of cardiovascular mortality, and therapeutic strategies to prevent or mitigate the consequences of MI are a high priority. Cardiac progenitor cells (CPCs) have been used to treat cardiac injury post-MI, and despite poor engraftment, they have been shown to inhibit apoptosis and to promote angiogenesis through poorly understood paracrine effects. We previously reported that the direct injection of exosomes derived from CPCs (CPCexo) into mouse hearts provides protection against apoptosis in a model of acute ischemia/reperfusion injury. Moreover, we and others have reported that reactive oxygen species (ROS) derived from NADPH oxidase (NOX) can enhance angiogenesis in endothelial cells (ECs). Here we examined whether bioengineered CPCexo transfected with a pro-angiogenic miR-322 (CPCexo-322) can improve therapeutic efficacy in a mouse model of MI as compared to CPCexo. Systemic administration of CPCexo-322 in mice after ischemic injury provided greater protection post-MI than control CPCexo, in part, through enhanced angiogenesis in the border zones of infarcted hearts. Mechanistically, the treatment of cultured human ECs with CPCexo-322 resulted in a greater angiogenic response, as determined by increased EC migration and capillary tube formation via increased Nox2-derived ROS. Our study reveals that the engineering of CPCexo via microRNA (miR) programing can enhance angiogenesis, and this may be an effective therapeutic strategy for the treatment of ischemic cardiovascular diseases. Full article

NADP-dependent (Nicotinamide Adénine Dinucléotide Phosphate-dependent) isocitrate dehydrogenases (NADP-ICDH) are metabolic enzymes involved in 2-oxoglutarate biosynthesis, but they also supply cells with NADPH. Different NADP-ICDH genes are found in Arabidopsis among which a single gene encodes for a cytosolic ICDH (cICDH) isoform. Here, we show that cICDH is susceptible to oxidation and that several cysteine (Cys) residues are prone to S-nitrosylation upon nitrosoglutathione (GSNO) treatment. Moreover, we identified a single S-glutathionylated cysteine Cys363 by mass-spectrometry analyses. Modeling analyses suggest that Cys363 is not located in the close proximity of the cICDH active site. In addition, mutation of Cys363 consistently does not modify the activity of cICDH. However, it does affect the sensitivity of the enzyme to GSNO, indicating that S-glutathionylation of Cys363 is involved in the inhibition of cICDH activity upon GSNO treatments. We also show that glutaredoxin are able to rescue the GSNO-dependent inhibition of cICDH activity, suggesting that they act as a deglutathionylation system in vitro. The glutaredoxin system, conversely to the thioredoxin system, is able to remove S-nitrosothiol adducts from cICDH. Finally, NADP-ICDH activities were decreased both in a catalase2 mutant and in mutants affected in thiol reduction systems, suggesting a role of the thiol reduction systems to protect NADP-ICDH activities in planta. In line with our observations in Arabidopsis, we found that the human recombinant NADP-ICDH activity is also sensitive to oxidation in vitro, suggesting that this redox mechanism might be shared by other ICDH isoforms. Full article

Peroxiredoxin 6 (Prdx6) is a member of an evolutionary ancient family of peroxidase enzymes with diverse functions in the cell. Prdx6 is an important enzymatic antioxidant. It reduces a wide range of peroxide substrates in the cell, thus playing a leading role in the maintenance of the redox homeostasis in mammalian cells. Beside peroxidase activity, Prdx6 has been shown to possess an activity of phospholipase A2, an enzyme playing an important role in membrane phospholipid metabolism. Moreover, Prdx6 takes part in intercellular and intracellular signal transduction due to its peroxidase and phospholipase activity, thus facilitating the initiation of regenerative processes in the cell, suppression of apoptosis, and activation of cell proliferation. Being an effective and important antioxidant enzyme, Prdx6 plays an essential role in neutralizing oxidative stress caused by various factors, including action of ionizing radiation. Endogenous Prdx6 has been shown to possess a significant radioprotective potential in cellular and animal models. Moreover, intravenous infusion of recombinant Prdx6 to animals before irradiation at lethal or sublethal doses has shown its high radioprotective effect. Exogenous Prdx6 effectively alleviates the severeness of radiation lesions, providing normalization of the functional state of radiosensitive organs and tissues, and leads to a significant elevation of the survival rate of animals. Prdx6 can be considered as a potent and promising radioprotective agent for reducing the pathological effect of ionizing radiation on mammalian organisms. The radioprotective properties and mechanisms of radioprotective action of Prdx6 are discussed in the current review. Full article

The reactions of antioxidants with superoxide radical were studied by cyclic voltammetry (CV)—and hydrodynamic voltammetry at a rotating ring-disk electrode (RRDE). In both methods, the superoxide is generated in solution from dissolved oxygen and then measured after being allowed to react with the antioxidant being studied. Both methods detected and measured the radical scavenging but the RRDE was able to give detailed insight into the antioxidant behavior. Three flavonoids, chrysin, quercetin and eriodictyol, were studied, their scavenging activity of superoxide was assessed and the molecular structure of each flavonoid was related to its scavenging capability. From our improved and novel RRDE method, we determine the ability of these 3 antioxidants to react with superoxide radical in a more quantitative manner than the classical CV. Density Functional Theory (DFT) and single crystal X-ray diffraction data provide structural information that assists in clarifying the scavenging molecular mechanism. Hydroxyls associated with the A ring, as found in chrysin, scavenge superoxide in a different manner than those found in the B ring of flavonoids, as those in quercetin and eriodictyol. Full article

In plants, NADPH oxidase (NOX) is also known as a respiratory burst oxidase homolog (Rboh). This highly important enzyme, one of the main enzymatic sources of superoxide radicals (O₂•⁻), is involved in the metabolism of reactive oxygen and nitrogen species (ROS and RNS), which is active in the non-climacteric pepper (Capsicum annuum L.) fruit. We used sweet pepper fruits at two ripening stages (green and red) to biochemically analyze the O₂•⁻-generating Rboh activity and the number of isozymes during this physiological process. Malondialdehyde (MDA) content, an oxidative stress marker, was also assayed as an index of lipid peroxidation. In red fruits, MDA was observed to increase 2-fold accompanied by a 5.3-fold increase in total Rboh activity. Using in-gel assays of Rboh activity, we identified a total of seven CaRboh isozymes (I–VII) which were differentially modulated during ripening. CaRboh-III and CaRboh-I were the most prominent isozymes in green and red fruits, respectively. An in vitro assay showed that CaRboh activity is inhibited in the presence of nitric oxide (NO) donors, peroxynitrite (ONOO⁻) and glutathione (GSH), suggesting that CaRboh can undergo S-nitrosation, Tyr-nitration, and glutathionylation, respectively. In summary, this study provides a basic biochemical characterization of CaRboh activity in pepper fruits and indicates that this O₂•⁻-generating Rboh is involved in nitro-oxidative stress associated with sweet pepper fruit ripening. Full article

Members of the DJ-1 protein family are multifunctional enzymes whose loss increases the susceptibility of the cell to oxidative stress. However, little is known about the function of the plant DJ-1 homologs. Therefore, we analyzed the effect of oxidation on the structure and function of chloroplastic AtDJ-1B and studied the phenotype of T-DNA lines lacking the protein. In vitro oxidation of AtDJ-1B with H₂O₂ lowers its glyoxalase activity, but has no effect on its holdase chaperone function. Remarkably, upon oxidation, the thermostability of AtDJ-1B increases with no significant alteration of the overall secondary structure. Moreover, we found that AtDJ-1B transcript levels are invariable, and loss of AtDJ-1B does not affect plant viability, growth and stress response. All in all, two discrete functions of AtDJ-1B respond differently to H₂O₂, and AtDJ-1B is not essential for plant development under stress. Full article

Though the pathogenesis of Alzheimer’s Disease (AD) is not completely elucidated, it is generally accepted that the aggregation of toxic amyloid-β (Aβ) protein fibrils plays a major role in the disease’s onset and progression. Various phytoceutical compounds have been shown to attenuate Aβ toxicity and disrupt its aggregation, including various types of polyphenolic compounds. These polyphenolic compounds have also been found to demonstrate potent antioxidant activity, which may contribute to their anti-amyloidogenic properties. This study compares three plants, traditionally used for numerous medicinal purposes in Asian countries, including: Curcuma longa (Turmeric), Camellia sinensis (Green Tea), and Scoparia dulcis (Sweet Broomweed). Antioxidant effects of the crude, polyphenol rich phytoceutical extracts from these plants were analyzed using a 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The ability of these extracts to prevent Aβ fibril formation was then carried out in order to establish a relationship between antioxidant activity and Aβ aggregation. A positive correlation between antioxidant efficacy and prevention of Aβ aggregation was demonstrated, indicating that antioxidant activity may play some role in preventing Aβ aggregation. Full article

Studies have shown an increased risk for a variety of cancers, specifically brain cancer, in healthcare workers occupationally exposed to ionizing radiation. Although the mechanisms mediating these phenomena are not fully understood, ionizing radiation-mediated elevated levels of reactive oxygen species (ROS), oxidative DNA damage, and immune modulation are likely involved. A group of 20 radiation exposed workers and 40 sex- and age-matched non-exposed control subjects were recruited for the study. We measured superoxide (O₂•⁻) levels in whole blood of healthcare workers and all other measurements of cytokines, oxidative DNA damage, extracellular superoxide dismutase (EcSOD) activity and reduced/oxidized glutathione ratio (GSH/GSSG) in plasma. Levels of O₂•⁻ were significantly higher in radiation exposed workers compared to control. Similarly, a significant increase in the levels of interleukin (IL)-6, IL-1α and macrophage inflammatory protein (MIP)-1α in radiation exposed workers compared to control was observed, while there was no significance difference in the other 27 screened cytokines. A significant positive correlation was found between MIP-1α and O₂•⁻ levels with no correlation in either IL-6 or IL-1α. Further, a dose-dependent relationship with significant O₂•⁻ production and immune alterations in radiation exposed workers was demonstrated. There was no statistical difference between the groups in terms of oxidative DNA damage, GSH/GSSG levels, or EcSOD activity. Although the biologic significance of cytokines alterations in radiation exposed workers is unclear, further studies are needed for determining the underlying mechanism of their elevation. Full article

Peroxiredoxins have a long-established cellular function as regulators of redox metabolism by catalyzing the reduction of peroxides (e.g., H₂O₂, lipid peroxides) with high catalytic efficiency. This activity is also critical to the initiation and relay of both phosphorylation and redox signaling in a broad range of pathophysiological contexts. Under normal physiological conditions, peroxiredoxins protect normal cells from oxidative damage that could promote oncogenesis (e.g., environmental stressors). In cancer, higher expression level of peroxiredoxins has been associated with both tumor growth and resistance to radiation therapies. However, this relationship between the expression of peroxiredoxins and the response to radiation is not evident from an analysis of data in The Cancer Genome Atlas (TCGA) or NCI60 panel of cancer cell lines. The focus of this review is to summarize the current experimental knowledge implicating this class of proteins in cancer, and to provide a perspective on the value of targeting peroxiredoxins in the management of cancer. Potential biases in the analysis of the TCGA data with respect to radiation resistance are also highlighted. Full article

Thioredoxins (TRXs) are major protein disulfide reductases of the cell. Their redox activity relies on a conserved Trp-Cys-(Gly/Pro)-Pro-Cys active site bearing two cysteine (Cys) residues that can be found either as free thiols (reduced TRXs) or linked together by a disulfide bond (oxidized TRXs) during the catalytic cycle. Their reactivity is crucial for TRX activity, and depends on the active site microenvironment. Here, we solved and compared the 3D structure of reduced and oxidized TRX h1 from Chlamydomonas reinhardtii (CrTRXh1). The three-dimensional structure was also determined for mutants of each active site Cys. Structural alignments of CrTRXh1 with other structurally solved plant TRXs showed a common spatial fold, despite the low sequence identity. Structural analyses of CrTRXh1 revealed that the protein adopts an identical conformation independently from its redox state. Treatment with iodoacetamide (IAM), a Cys alkylating agent, resulted in a rapid and pH-dependent inactivation of CrTRXh1. Starting from fully reduced CrTRXh1, we determined the acid dissociation constant (pK_a) of each active site Cys by Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry analyses coupled to differential IAM-based alkylation. Based on the diversity of catalytic Cys deprotonation states, the mechanisms and structural features underlying disulfide redox activity are discussed. Full article

Oxidative damage is among the factors associated with the onset of chronic pathologies, such as neurodegenerative and metabolic diseases. Several classes of anti-oxidant compounds have been suggested as having a protective role against cellular stressors, but, in this perspective, peptides’ world represents a poorly explored source. In the present study, the free radical scavenging properties, the metal ion reducing power, and the metal chelating activity of a series of sulfurated amino acids and tripeptides were determined in vitro through canonical assays (DPPH, ABTS, CUPRAC, FRAP, PM, and EECC) and estimated in comparison with the corresponding activities of synthetic peptide semicarbazones, incorporating the peculiar non-proteinogenic amino acid, tert-leucine (tLeu). The compounds exhibited remarkable anti-oxidant properties. As expected, sulfurated compounds 1–5 were found to be the most efficient radical scavengers and strongest reductants. Nevertheless, tLeu-containing peptides 7 and 8 disclosed notable metal reducing and chelating activities. These unprecedented results indicate that tLeu-featuring di- and tripeptide backbones, bearing the semicarbazone chelating moiety, are compatible with the emergence of an anti-oxidant potential. Additionally, when tested against a panel of enzymes usually targeted for therapeutic purposes in neurodegenerative and metabolic disorders, all samples were found to be good inhibitors of tyrosinase. Full article

Reactive oxygen species (ROS) are produced as a natural byproduct of the normal metabolism of oxygen and play significant roles in cell signaling and homeostasis. Although ROS have been involved in pathological processes as diverse as cancer, cardiovascular disease, and aging, they may to exert an effect even in a physiological context. In the central nervous system, stem cells and hematopoietic stem cells are early progenitors that contain lower levels of ROS than their more mature progeny. These different concentrations have been reported to be crucial for maintaining stem cell function. Mammary gland remodeling has been proposed to be organized through the activation and regulation of cells with stemness, either considered real stem cells or primitive precursors. Given the state of oxidative stress in the mammary gland tissue induced by high milk production, in particular in highly productive dairy cows; several studies have focused on the relationship between adult mammary stem cells and the oxidative state of the gland. The oxidative state of the mammary gland appears to be involved in the initial development and metastasis of breast cancer through interference with mammary cancerous stem cells. This review summarizes some links between the mammary stem and oxidative state of the gland. Full article

The common bean is a good source of protein and bioactive substances giving it a large antioxidant capacity. The extensive variability of bean genotypes creates the need to characterize them with regard to their nutritional value as a tool in biofortification programs. The purpose of this study was to obtain the antioxidant capacity and phytonutrient content both in the seed coat and the cotyledon of 12 common bean varieties from different regions in Mexico. In the case of the whole seed, lightness (L*), a* (red-purple) and b* (yellow-purple) color coordinates were determined, as well as the chroma and hue angle. In the case of the seed coat and the cotyledon, the protein content, the phytonutrient content and the antioxidant capacity (2,2-diphenyl-1-picrylhydrazyl (DPPH)) were evaluated. A significant difference was observed (p ≤ 0.05) among bean varieties and between seed coat and cotyledon in all variables evaluated. Cotyledon showed a higher content of protein, H, Ni, Zn, Cu, N, P, K S and Mn, while the seed coat showed a higher content of Fe, Ca and Mg and a greater antioxidant capacity (59.99%). The Higuera Azufrado bean variety stood out as having a higher content of N, S and protein. We have concluded that the nutritional characterization performed on Mexican bean varieties represents a valuable tool for genetic enhancement programs and crop biofortification strategies. Full article

The expression of the phospholipase A₂ activity (aiPLA₂) of peroxiredoxin 6 (Prdx6) in the cell cytoplasm is physiologically relevant for the repair of peroxidized cell membranes, but aiPLA₂ assay in vitro indicates that, unlike assay at pH 4, activity at cytosolic pH is essentially absent with non-oxidized substrate. However, the addition of glutathione (GSH) to the assay medium significantly increased aiPLA₂ activity at cytosolic pH, while oxidized GSH (GSSG) and several other thiols had no effect. By mass spectroscopy (ESI MS), the addition of GSH to Prdx6 paradoxically led to oxidation of its conserved Cys47 residue to a sulfinic acid. The effect of GSH on PLA₂ activity was abolished by incubation under anaerobic conditions, confirming that auto-oxidation of the protein was the mechanism for the GSH effect. Analysis by circular dichroism (CD) and tryptophan fluorescence showed alterations of the protein structure in the presence of GSH. Independently of GSH, the oxidation of Prdx6 by exposure to H₂O₂ or the presence of oxidized phospholipid as substrate also significantly increased aiPLA₂ activity at pH 7. We conclude that the oxidation of the peroxidatically active Cys47 of Prdx6 results in an increase of aiPLA₂ activity at pH 7 without effect on the activity of the enzyme at pH 4. Full article

Light-induced oxidation of lipids and proteins provokes retinal injuries and results in progression of degenerative retinal diseases, such as, for instance, iatrogenic photic maculopathies. Having accumulated over years retinal injuries contribute to development of age-related macular degeneration (AMD). Antioxidant treatment is regarded as a promising approach to protecting the retina from light damage and AMD. Here, we examine oxidative processes induced in rabbit retina by excessive light illumination with or without premedication using mitochondria-targeted antioxidant SkQ1 (10-(6’-plastoquinonyl)decyltriphenyl-phosphonium). The retinal extracts obtained from animals euthanized within 1–7 days post exposure were analyzed for H₂O₂, malondialdehyde (MDA), total antioxidant activity (AOA), and activities of glutathione peroxidase (GPx) and superoxide dismutase (SOD) using colorimetric and luminescence assays. Oxidation of visual arrestin was monitored by immunoblotting. The light exposure induced lipid peroxidation and H₂O₂ accumulation in the retinal cells. Unexpectedly, it prominently upregulated AOA in retinal extracts although SOD and GPx activities were compromised. These alterations were accompanied by accumulation of disulfide dimers of arrestin revealing oxidative stress in the photoreceptors. Premedication of the eyes with SkQ1 accelerated normalization of H₂O₂ levels and redox-status of lipids and proteins, contemporarily enhancing AOA and, likely, sustaining normal activity of GPx. Thus, SkQ1 protects the retina from light-induced oxidative stress and could be employed to suppress oxidative damage of proteins and lipids contributing to AMD. Full article

The aim of this work is to investigate the in vitro acetylcholinesterase (AChE) and butyrycholinesterase (BChE) inhibitory activities of essential oils obtained by hydrodistillation from the leaves of Prunus armeniaca and P. domestica in relation to their composition, analysed by Gas Chromatography–Flame Ionization Detector (GC-FID) and Gas Chromatography-Mass Spectrometry (GC-MS) analyses, at different times. Moreover, considering the role of free radicals in the progression of neurodegenerative disorders, the antioxidant properties of essential oils were investigated by using, 2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and β-carotene bleaching tests. The relative antioxidant capacity index (RACI) was used to achieve more comprehensive comparison between analysed antioxidant effects of essential oils. P. armeniaca oils were more active than P. domestica oils against AChE. Against BChE, the most active was the essential oil from P. domestica leaves collected in August with an IC₅₀ value of 95.80 μg/mL. This oil exerted the highest inhibitory activity of lipid peroxidation with IC₅₀ values of 11.15 and 11.39 μg/mL after 30 and 60 min of incubation, respectively. All samples demonstrated a remarkable ABTS radicals scavenging activity, with IC₅₀ values in the range 0.45–0.57 μg/mL in comparison to the positive control, ascorbic acid. Full article

The present experiment sought to determine the effect of an eight-week, high antioxidant, whole-foods dietary supplement on Morris Water Maze performance in early and late middle-aged female rats. To improve ecological validity over past experimental studies, rats in the current study received antioxidants by consuming freeze-dried organic strawberries and spinach rather than by being given food extracts or antioxidant injections. Latency and path length measures both indicated that late middle-aged rats fed the high antioxidant diet performed on a par with the younger animals earlier in training than their standard diet counterparts (p < 0.05). Superior performance was not due to improved fitness in the antioxidant-supplemented rats. Thus, our model showed that a high antioxidant diet of relatively short duration mitigated the mild cognitive decline that was seen in control animals during the developmental period of late middle-age. The current results offer support for the promising role of dietary antioxidants in maintaining cognitive health in normal aging and extend past findings to females, who have been relatively neglected in experimental investigations. Moreover, the current model suggests that the period of transition from early to late middle age is a promising target for dietary intervention in healthy adults. Full article