Protein cysteine thiol status is a major determinant of oxidative stress and oxidant signaling. The -SulfoBiotics- Protein Redox State Monitoring Kit provides a unique opportunity to investigate protein thiol states. This system adds a 15-kDa Protein-SHifter to reduced cysteine residues, and this molecular mass shift can be detected by gel electrophoresis. Even in biological samples, Protein-SHifter Plus allows the thiol states of specific proteins to be studied using Western blotting. Peroxiredoxin 6 (Prx6) is a unique one-cysteine peroxiredoxin that scavenges peroxides by utilizing conserved Cysteine-47. Human Prx6 also contains an additional non-conserved cysteine residue, while rat Prx6 only has the catalytic cysteine. In cultured cells, cysteine residues of Prx6 were found to be predominantly fully reduced. The treatment of human cells with hydrogen peroxide (H₂O₂) formed Prx6 with one cysteine reduced. Since catalytic cysteine becomes oxidized in rat cells by the same H₂O₂ treatment and treating denatured human Prx6 with H₂O₂ results in the oxidation of both cysteines, non-conserved cysteine may not be accessible to H₂O₂ in human cells. We also found that untreated cells contained Prx6 multimers bound through disulfide bonds. Surprisingly, treating cells with H₂O₂ eliminated these Prx6 multimers. In contrast, treating cell lysates with H₂O₂ promoted the formation of Prx6 multimers. Similarly, treating purified preparations of the recombinant cyclic nucleotide-binding domain of the human hyperpolarization-activated cyclic nucleotide-modulated channels with H₂O₂ promoted the formation of multimers. These studies revealed that the cellular environment defines the susceptibility of protein cysteines to H₂O₂ and determines whether H₂O₂ acts as a facilitator or a disrupter of disulfide bonds. Full article

The characterization of compounds with antioxidant activity is of great interest due to their ability to reduce reactive oxygen species production and, therefore, prevent some age-related diseases. Its antioxidant capacity can be analyzed by different methods both in vitro and in vivo. Caenorhabditis elegans is an in vivo model widely used in ageing research. Until now, available tests analyze functional effects in the worms, so the antioxidant activity of the compound is indirectly monitored. We have developed a simple and a reliable method to quantify internal antioxidant activity in vivo. To validate this method, we analyzed an aqueous green tea extract and two other compounds with a well-known antioxidant activity and without this activity. The results obtained (EC₅₀ green tea = 21.76 ± 1.28 µg/mL; EC₅₀ positive control = 8.50 ± 0.33 µg/mL; negative control EC₅₀ > 500 µg/mL) can help in the design of further in vivo experiments. Thus, our method can be used as a previous screening capable of reducing the gap between in vitro and in vivo assays. Full article

Feijoa fruit is becoming increasingly popular, yet limited studies have focused on the antioxidant capacity and phenolic profiling of its extracts. In this research, optimization of phenolic extraction from feijoa flesh, peel, and whole fruit from four New Zealand grown cultivars was conducted using orthogonal design. Antioxidant activities of the extracts were assessed, followed by phenolic profiling by a validated liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method. For feijoa flesh and whole fruit, the extraction was optimized using 70% ethanol, material to solvent ratio of 1:30, at extraction temperature of 50 °C for 30 min. For feijoa peel, extraction at 50 °C for 60 min using 50% ethanol with a material to solvent ratio of 1:30 were the optimized conditions. Results showed feijoa peel had higher total phenolic content (TPC) and antioxidant activities than the flesh and whole fruit. Overall, the Unique cultivar had a relatively higher TPC and antioxidant activity than the other cultivars tested. A total of 15 phenolic compounds were identified, and seven of them were reported for the first time in feijoa fruit. This is the first systematic investigation on the extraction method, phenolic content, antioxidant activity and phenolic profile of feijoa emphasis on comparison of sample types and cultivars. Full article

The zero-waste biorefinery concept inspired a green oleo-extraction of both natural volatile (e.g., borneol, camphor, o-cymene, eucalyptol, limonene, α-pinene, and terpinen-4-ol) and non-volatile (e.g., carnosol, carnosic, and rosmarinic acid) bioactive compounds from rosemary leaves with vegetable oils and their amphiphilic derivatives as simple food-grade solvents. It is noteworthy that soybean oil could obtain the highest total phenolic compounds (TPCs) among 12 refined oils including grapeseed, rapeseed, peanut, sunflower, olive, avocado, almond, apricot, corn, wheat germ, and hazelnut oils. Furthermore, the addition of oil derivatives to soybean oils, such as glyceryl monooleate (GMO), glyceryl monostearate (GMS), diglycerides, and soy lecithin in particular, could not only significantly enhance the oleo-extraction of non-volatile antioxidants by 66.7% approximately, but also help to remarkably improve the solvation of volatile aroma compounds (VACs) by 16% in refined soybean oils. These experimental results were in good consistency with their relative solubilities predicted by the more sophisticated COSMO-RS (COnductor like Screening MOdel for Real Solvents) simulation. This simple procedure of using vegetable oils and their derivatives as bio-based solvents for simultaneously improving the extraction yield of natural antioxidants and flavors from rosemary showed its great potential in up-scaling with the integration of green techniques (ultrasound, microwave, etc.) for zero-waste biorefinery from biomass waste to high value-added extracts in future functional food and cosmetic applications. Full article

Obesity is one of the major public health problems in the world because it is implicated in metabolic syndromes, such as type 2 diabetes, hypertension, and cardiovascular diseases. The objective of this study was to investigate whether Erigeron annuus (L.) Pers. (EAP) extract suppresses reactive oxygen species (ROS) production and fat accumulation in 3T3-L1 cells by activating an AMP-dependent kinase (AMPK) signaling pathway. Our results showed that EAP water extract significantly inhibits ROS production, adipogenesis, and lipogenesis during differentiation of 3T3-L1 preadipocytes. In addition, EAP decreased mRNA and protein levels of proliferator-activated receptor γ (PPARγ) and CCAAT/enhancer-binding protein alpha (C/EBPα). Moreover, EAP suppressed mRNA expressions of fatty acid synthase (FAS), lipoprotein lipase (LPL), adipocyte protein 2 (aP2) in a dose-dependent manner. Whereas, EAP upregulated adiponectin expression, phosphorylation levels of AMPK and carnitine palmitoyltransferase 1 (CPT-1) protein level during differentiation of 3T3-L1 preadipocytes. These results suggest that EAP water extract can exert ROS-linked anti-obesity effect through the mechanism that might involve inhibition of ROS production, adipogenesis and lipogenesis via an activating AMPK signaling pathway. Full article

The olive tree “Cellina di Nardò” (CdN) is one of the most widespread cultivars in Southern Italy, mainly grown in the Provinces of Lecce, Taranto, and Brindisi over a total of about 60,000 hectares. Although this cultivar is mainly used for oil production, the drupes are also suitable and potentially marketable as table olives. When used for this purpose, olives are harvested after complete maturation, which gives to them a naturally black color due to anthocyanin accumulation. This survey reports for the first time on the total phenolic content (TPC), anthocyanin characterization, and antioxidant activity of CdN olive fruits during ripening and after fermentation. The antioxidant activity (AA) was determined using three different methods. Data showed that TPC increased during maturation, reaching values two times higher in completely ripened olives. Anthocyanins were found only in mature olives and the concentrations reached up to 5.3 g/kg dry weight. AA was determined for the four ripening stages, and was particularly high in the totally black olive fruit, in accordance with TPC and anthocyanin amounts. Moreover, the CdN olives showed a higher TPC and a greater AA compared to other black table olives produced by cultivars commonly grown for this purpose. These data demonstrate the great potential of black table CdN olives, a product that combines exceptional organoleptic properties with a remarkable antioxidant capacity. Full article

Over the past few years, interest in health research has increased, making improved health a global goal for 2030. The purpose of such research is to ensure healthy lives and promote wellbeing across individuals of all ages. It has been shown that nutrition plays a key role in the prevention of some chronic diseases such as obesity, cardiovascular disease, diabetes, and cancer. One of the aspects that characterises a healthy diet is a high intake of vegetables and fruits, as both are flavonoid-rich foods. Flavonoids are one of the main subclasses of dietary polyphenols and possess strong antioxidant activity and anti-carcinogenic properties. Moreover, some population-based studies have described a relationship between cancer risk and dietary flavonoid intake. In this context, the goal of this review was to provide an updated evaluation of the association between the risk of different types of cancers and dietary flavonoid intake. We analysed all relevant epidemiological studies from January 2008 to March 2019 using the PUBMED and Web of Science databases. In summary, this review concludes that dietary flavonoid intake is associated with a reduced risk of different types of cancer, such as gastric, breast, prostate, and colorectal cancers. Full article

Majiayou pomelo (Citrus grandis L. Osbeck, MP) is a famous local red pulp pomelo from the Jiangxi province in China that is rich in natural active substances. In order to investigate the postharvest antioxidant capacities of MP pulp and determine the optimal harvesting time, fruits that were harvested at three different maturities (185, 200, and 215 days after full bloom) were observed for 180 days of preservation at ambient temperature. An abundance of ascorbic acid and lycopene in the MP pulp was found during storage, and in Harvest I, these substances were significantly higher than in Harvest II and Harvest III fruit (p < 0.05). The activity of ascorbate peroxidase (APX), peroxidase (POD), and catalases (CAT) in Harvest I and Harvest II were far higher after 90 days. The radical scavenging ability of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, superoxide anion radical (O₂⁻•), and hydroxyl radical (•OH) in Harvest I and Harvest II were higher. There was a significantly positive correlation (p < 0.01) between the antioxidant components (ascorbic acid, lycopene, carotenoids, total phenols, and total flavonoids), enzyme activity, and radical scavenging ability. The comprehensive scores determined by principal component analysis (PCA) in Harvest I and II were higher than those in Harvest III. Therefore, the optimal harvesting period of MP for each year is determined to be early November. The study provides a theoretical basis for the maintenance of the postharvest fruit value and the regulation of fruit functional components. Full article

Pulmonary arterial hypertension (PAH) is one of the diseases with a well-established gender dimorphism. The prevalence of PAH is increased in females with a ratio of 4:1, while poor survival prognosis is associated with the male gender. Nevertheless, the specific contribution of gender in disease development and progression is unclear due to the complex nature of the PAH. Oxidative and nitrosative stresses are important contributors in PAH pathogenesis; however, the role of gender in redox homeostasis has been understudied. This review is aimed to overview the possible sex-specific mechanisms responsible for the regulation of the balance between oxidants and antioxidants in relation to PAH pathobiology. Full article

Psoriasis is a chronic cutaneous condition characterized by the release of pro-inflammatory mediators and oxidative stress. The reduction of these factors is currently the most effective strategy to inhibit the symptoms of pathology. Antioxidants from natural sources are increasingly used to improve skin conditions. Dried red leaves from grapevine (Vitis vinifera L., cv Teinturiers) showed anti-inflammatory and anti-bacterial activities, but their possible effects on keratinocytes have not been previously investigated. In this study we tested the ability of a water extract from grapevine leaves (VVWE) to inhibit inflammatory conditions in human keratinocytes (HaCaT cells), challenged with proinflammatory (tumor necrosis factor-α (TNF-α) or lipopolysaccharide (LPS)) or prooxidant (ultraviolet B radiation (UVB) or H₂O₂) mediators. VVWE inhibited interleukin-8 (IL-8) secretion induced by proinflammatory stimuli, acting on the IL-8 promoter activity, but the effect was lower when prooxidant mediators were used. The effect was partly explained by the reduction of nuclear factor-κB (NF-κB)-driven transcription and nuclear translocation. Furthermore, vascular endothelial growth factor (VEGF) secretion, a regulator of angiogenesis, was inhibited by VVWE, but not matrix metalloproteinase-9 (MMP-9), a protease involved in matrix remodeling. VVWE, assayed on Franz diffusion cell system, showed a marked reduction of High Performance Liquid Chromatography (HPLC)-identified compounds. Pure molecules individually failed to reduce TNF-α-induced IL-8 release, suggesting synergistic effects or the presence of other bioactive compounds still unknown. Full article

Cis-dichlorodiammineplatinum II (CDDP) is a chemotherapeutic agent that induces nephrotoxicity by different mechanisms, including oxidative stress, mitochondrial dysfunction, autophagy, and endoplasmic reticulum stress. This study aimed to evaluate if the protective effects of the antioxidant alpha-mangostin (αM) in CDDP-induced damage in proximal tubule Lilly laboratory culture porcine kidney (LLC-PK1) cells, are related to mitochondrial function preservation. It was found that αM co-incubation prevented CDDP-induced cell death. Furthermore, αM prevented the CDDP-induced decrease in cell respiratory states, in the maximum capacity of the electron transfer system (E) and in the respiration associated to oxidative phosphorylation (OXPHOS). CDDP also decreased the protein levels of voltage dependence anion channel (VDAC) and mitochondrial complex subunits, which together with the reduction in E, the mitofusin 2 decrease and the mitochondrial network fragmentation observed by MitoTracker Green, suggest the mitochondrial morphology alteration and the decrease in mitochondrial mass induced by CDDP. CDDP also induced the reduction in mitochondrial biogenesis observed by transcription factor A, mitochondria (TFAM) decreased protein-level and the increase in mitophagy. All these changes were prevented by αM. Taken together, our results imply that αM’s protective effects in CDDP-induced toxicity in LLC-PK1 cells are associated to mitochondrial function preservation. Full article

Urinary dysfunction is a common pathological condition that can significantly decrease the quality of life. Bladder outlet obstruction (BOO) is a major cause of urinary dysfunction, and various lower urinary tract diseases including benign prostatic hyperplasia and urethral stricture disease cause BOO. According to the results of a variety of animal experiments on partial BOO (PBOO), there is a general agreement that ischemic conditions and repeated ischemia/reperfusion of the bladder are closely associated with BOO-induced bladder damage, and that increased oxidative stress by ischemia/reperfusion plays a crucial role in the pathological mechanisms underlying urinary dysfunction. Changes in biomarkers of oxidative stress in PBOO animal models support this association between oxidative stress and urinary dysfunction. Oxidative stress is defined as an imbalance between the production of pro-oxidants, such as free radicals and reactive species, and their elimination through protective mechanisms of antioxidants. Therefore, organizing the knowledge on the state of oxidative stress, changes in biomarkers, and biological roles of antioxidants in systemic and bladder tissues is essential to understand the detailed pathological characteristics of the urinary dysfunction caused by PBOO. Furthermore, information on drugs and supplements that have antioxidant effects is important for defining treatment strategies for urinary dysfunction with PBOO. In this review, we paid special attention to the following three issues; (1) changes in oxidative stress, including its biomarkers, (2) antioxidant status, and (3) previous reports on treatment strategies involving agents with antioxidative activity for urinary dysfunction caused by BOO. In particular, we provide systematic information on the detailed mechanisms underlying the antioxidative effects of agents used to treat PBOO. In addition, we show present research issues and research limitations, as well as suggest possible future antioxidant treatment strategies for patients with PBOO. Full article

Sorghum has a significant amount of proteins, especially kafirin; however, limited information is available on evaluating its potential for peptide antioxidants. The objectives of this study were to: (1) investigate the effects of two key variables, enzyme-to-substrate ratio and reaction time on kafirin hydrolysis using Alcalase; (2) evaluate the antioxidant performances of the hydrolysates and fractions from membrane ultrafiltration and gel filtration; and (3) identify peptide sequences in the antioxidant fraction using MALDI-TOF/TOF MS. Kafirin hydrolysates prepared at enzyme-to-substrate ratio of 0.4 Au/g and 4 h had a good balance of antioxidant activity, yield, and economic efficiency. Medium-sized fraction of hydrolysates (5–10 kDa) from membrane filtration possessed the highest antioxidant activities among various fractions. The fraction also unveiled a good inhibition effect against lipid oxidation in emulsion and ground meat systems. Smaller-sized fraction (F3) collected through gel-filtration chromatography had significantly stronger antioxidant activities than other fractions, and 26 representative peptide sequences were identified in the fraction. Full article

Selenium compounds influence cell growth and are highly interesting candidate compounds for cancer chemotherapy. Over decades an extensive number of publications have reported highly efficient growth inhibitory effects with a number of suggested mechanisms f especially for redox-active selenium compounds. However, the studies are difficult to compare due to a high degree of variations in half-maximal inhibitor concentration (IC₅₀) dependent on cultivation conditions and methods to assess cell viability. Among other factors, the variability in culture conditions may affect the experimental outcome. To address this, we have compared the maintenance effects of four commonly used cell culture media on two cell lines, A549 and HepG2, evaluated by the toxic response to selenite and seleno-methylselenocysteine, cell growth and redox homeostasis. We found that the composition of the cell culture media greatly affected cell growth and sensitivity to selenium cytotoxicity. We also provided evidence for change of phenotype in A549 cells when maintained under different culture conditions, demonstrated by changes in cytokeratin 18 (CK18) and vimentin expression. In conclusion, our results have shown the importance of defining the cell culture medium used when comparing results from different studies. Full article

The use of green marine seaweed Ulva spp. as foods, feed supplements, and functional ingredients has gained increasing interest. Microwave-assisted extraction technology was employed to improve the extraction yield and composition of Ulva pertusa polysaccharides. The antioxidant activity of ulvan was also evaluated. The impacts of four independent variables, i.e., extraction time (X₁, 30 to 60 min), power (X₂, 500 to 700 W), water-to-raw-material ratio (X₃, 40 to 70), and pH (X₄, 5 to 7) were evaluated. The chemical structure of different polysaccharides fractions was investigated via FT-IR and the determination of their antioxidant activities. A response surface methodology based on a Box–Behnken design (BBD) was used to optimize the extraction conditions as follows: extraction time of 43.63 min, power level of 600 W, water-to-raw-material ratio of 55.45, pH of 6.57, and maximum yield of 41.91%, with a desired value of 0.381. Ulvan exerted a strong antioxidant effect against 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and showed reducing power in vitro. Ulvan protected RAW 264.7 cells against H₂O₂-induced oxidative stress by upregulating the expression and enhancing the activity of oxidative enzymes such as superoxide dismutase (SOD) and superoxide dismutase (CAT). The results suggest that the polysaccharides from U. pertusa might be promising bioactive compounds for commercial use. Full article