Effect of Hormone Complexes on Reactive Oxygen Species and Membrane Lipid Metabolism in Postharvest Kiwifruit Fruit

BackgroundEnergy production is a necessary process to continue physical activities, and exercise is associated with more oxygen consumption and increase of oxidative stress. what seems important is the numerical relationship between antioxidant and free radicals. Although the activity of some enzymes increases with physical activities, but it is possible that gene expression of this enzyme is not changed during exercise.ObjectivesThe aim of the present study is to investigate the antioxidant enzymes gene expression and changes in malondialdehyde (MDA) and total antioxidant capacity (TAC) levels in men and women affected by a session of incremental exercise and to carefully and numerically assess the relationship between MDA changes and gene expression and activity of antioxidant enzymes.Materials and Methods12 active men and 12 active women (21 - 24 years old) participated voluntarily in this study. Peripheral blood samples were taken from the subjects in three phases, before and after graduated exercise test (GXT) and 3 hours later (recovery).ResultsThe gene expression of manganese superoxide dismutase (MnSOD) enzyme increased significantly in women in the recovery phase (P < 0.05). Catalase gene expression significantly increased in men in both phases (immediately & recovery) (P < 0.05). But the changes in active women were only significant immediately after the exercise. TAC levels increased significantly in men in the recovery phase and in active women immediately after the exercise (P < 0.05). MDA activity also increased significantly in men in both phases (P < 0.05). However, in women the increase was significant only in the recovery phase (P < 0.05). There was a reverse relationship between changes in MnSOD and copper- and zinc-containing superoxide dismutase (Cu/ZnSOD) levels and MDA in men (P < 0.05). In active women there was also a significant relationship between changes in MDA and gene expression of Cu/ZnSOD and TAC (P < 0.05).ConclusionsThe increase in free radicals during incremental exercises challenges gene expression and activity of antioxidant enzymes. However, despite the negative effects of free radicals, in women, activity and gene expression of antioxidant enzymes respond appropriately to free radicals.

The mechanisms underlying long-term (30days) intermittent hypoxia (LT-IH)-evoked DNA methylation of anti-oxidant enzyme (AOE) gene repression in the carotid body (CB) reflex pathway were examined. LT-IH-treated rats showed increased reactive oxygen species (ROS) levels in the CB reflex pathway. Administration of a ROS scavenger or CB ablation blocked LT-IH-evoked DNA methylation and AOE gene repression in the central and efferent limbs of the CB reflex. LT-IH increased DNA methyltransferase (Dnmt) activity through upregulation of Dnmt1 and 3b proteins by ROS-dependent inactivation of glycogen synthase kinase 3β (GSK3β) by Akt. A pan-Akt inhibitor prevented LT-IH-induced GSK3β inactivation, elevated Dnmt protein expression and activity, AOE gene methylation, sympathetic activation and hypertension. Long-term exposure to intermittent hypoxia (LT-IH; 30days), simulating blood O2 profiles during sleep apnoea, has been shown to repress anti-oxidant enzyme (AOE) gene expression by DNA methylation in the carotid body (CB) reflex pathway, resulting in persistent elevation of plasma catecholamine levels and blood pressure. The present study examined the mechanisms by which LT-IH induces DNA methylation. Adult rats exposed to LT-IH showed elevated reactive oxygen species (ROS) in the CB, nucleus tractus solitarius (nTS) and rostroventrolateral medulla (RVLM) and adrenal medulla (AM), which represent the central and efferent limbs of the CB reflex, respectively. ROS scavenger treatment during the first ten days of IH exposure prevented ROS accumulation, blocked DNA methylation, and normalized AOE gene expression, suggesting that ROS generated during the early stages of IH activate DNA methylation. CB ablation prevented the ROS accumulation, normalized AOE gene expression in the nTS, RVLM, and AM and blocked DNA methylation, suggesting that LT-IH-induced DNA methylation in the central and efferent limbs of the CB reflex is indirect and requires CB neural activity. LT-IH increased DNA methyl transferase (Dnmt) activity through upregulation of Dnmt1 and 3b protein expression due to ROS-dependent inactivation of glycogen synthase kinase 3β (GSK3β) by protein kinase B (Akt). Treating rats with the pan-Akt inhibitor GSK690693 blocked the induction of Dnmt activity, Dnmt protein expression, and DNA methylation, leading to normalization of AOE gene expression as well as plasma catecholamine levels and blood pressure.

The infection of maize and peanut with Aspergillus flavus and subsequent contamination with aflatoxin pose a threat to global food safety and human health, and is exacerbated by drought stress. Drought stress-responding compounds such as reactive oxygen species (ROS) are associated with fungal stress responsive signaling and secondary metabolite production, and can stimulate the production of aflatoxin by A. flavus in vitro. These secondary metabolites have been shown to possess diverse functions in soil-borne fungi including antibiosis, competitive inhibition of other microbes, and abiotic stress alleviation. Previously, we observed that isolates of A. flavus showed differences in oxidative stress tolerance which correlated with their aflatoxin production capabilities. In order to better understand these isolate-specific oxidative stress responses, we examined the transcriptional responses of field isolates of A. flavus with varying levels of aflatoxin production (NRRL3357, AF13, and Tox4) to H2O2-induced oxidative stress using an RNA sequencing approach. These isolates were cultured in an aflatoxin-production conducive medium amended with various levels of H2O2. Whole transcriptomes were sequenced using an Illumina HiSeq platform with an average of 40.43 million filtered paired-end reads generated for each sample. The obtained transcriptomes were then used for differential expression, gene ontology, pathway, and co-expression analyses. Isolates which produced higher levels of aflatoxin tended to exhibit fewer differentially expressed genes than isolates with lower levels of production. Genes found to be differentially expressed in response to increasing oxidative stress included antioxidant enzymes, primary metabolism components, antibiosis-related genes, and secondary metabolite biosynthetic components specifically for aflatoxin, aflatrem, and kojic acid. The expression of fungal development-related genes including aminobenzoate degradation genes and conidiation regulators were found to be regulated in response to increasing stress. Aflatoxin biosynthetic genes and antioxidant enzyme genes were also found to be co-expressed and highly correlated with fungal biomass under stress. This suggests that these secondary metabolites may be produced as part of coordinated oxidative stress responses in A. flavus along with antioxidant enzyme gene expression and developmental regulation.

Drought stress severely impacts the growth, yield and quality of apple (Malus domestica). Abscisic acid (ABA) and basic helix-loop-helix (bHLH) transcription factors play crucial roles in regulating the drought response in many plants, but the potential interactions between bHLH and ABA in response to drought in apple still need to be discovered. Herein, we identified a bHLH transcription factor, ORG2 (OBP3-responsive gene 2), from M. hupehensis, and the expression of which is induced by drought and ABA. Apple plants that overexpressed MhORG2 were more sensitive to drought stress, while silencing MhORG2 caused the opposite phenotype. Specifically, we found that MhORG2 could directly bind to the DRE element in the MhAAO3 promoter and repress its expression, thereby ultimately reducing drought tolerance. Furthermore, MhORG2 represses the expression of antioxidant enzyme genes (MhSOD, MhAPX1 and MhCAT), leading to the accumulation of reactive oxygen species (ROS) and consequently reducing the drought tolerance of apple plants. Our findings uncover a novel mechanism by which MhORG2 negatively regulates drought tolerance in apple plants, offering a potential target for the development of drought-tolerant crops via biotechnological approaches.

Reactive oxygen intermediates play a role in chronic renal injury and glomerulosclerosis. We investigate changes in renal cortex antioxidant enzyme gene expression in the rat remnant-kidney model of chronic renal failure and compare the new data to enzyme activities published earlier. Antioxidant enzyme gene expression is evaluated by Northern blot analysis of cortex mRNA, using cDNA probes for catalase, copper/zinc-containing superoxide dismutase, and glutathione peroxidase. Catalase gene expression decreases during development of renal failure; this decrease is accompanied by decreased catalase activity during the glomerulosclerosis phase of the remnant-kidney model. Copper/zinc superoxide dismutase and glutathione peroxidase gene expression remain at a normal level during progression of the model, whereas their activities show a temporary decrease in the early remnant kidney. In the remnant-kidney model, catalase seems to be more vulnerable to reactive oxygen intermediates than superoxide dismutase and glutathione peroxidase. Our results show that antioxidant enzyme activity and gene expression do not change in the same direction at all times during disease development and that all antioxidant enzymes do not respond in the same way.

The inflammatory process in Crohn's disease (CD) is closely associated with the formation of reactive oxygen species. Antioxidant enzymes can play an important role in the outcome of CD and may influence postoperative recurrence in these patients. The aim of our study was to evaluate gene expression of intracellular antioxidant enzymes in surgically resected intestinal specimens of patients with CD, both in macroscopically normal and in inflamed tissue. A total of 28 patients referred for elective bowel resection were enrolled in the study. Full-thickness small intestinal specimens were investigated. Gene expression of antioxidant enzymes - superoxide dismutase (SOD), glutathione peroxidase (GPX), glutathione reductase (GSR) - was evaluated both in macroscopically normal and inflamed samples. There were significantly lower levels of SOD1 mRNA (p = 0.007) and GSR mRNA (p = 0.027) in inflamed tissue compared to macroscopically normal areas. No significant differences were found between affected and non-affected intestinal segments in mRNA for SOD2, SOD3 and GPX. Our pilot data clearly showed that the gene expression of major antioxidant enzymes is not a uniform mechanism in the pathogenesis of Crohn's disease. Topically decreased gene expression of SOD1 and GSR might facilitate the segmental tissue injury caused by reactive oxygen species.

Twenty-eight rats were examined in a 5-week experiment to investigate the effect of curcumin on gene expression and activities of hepatic antioxidant enzymes in rats intoxicated with aflatoxin B1 (AFB1 ). The rats were divided into four groups. Rats in 1-4 groups served as control, oral curcumin treated (15 mg/kg body weight), single i.p. dose of AFB1 (3 mg/kg body weight) and combination of single i.p. dose of AFB1 with oral curcumin treated, respectively. AFB1 Liver damage and oxidative stress were evident in untreated AFB1 -intoxicated rats as indicated by a significant elevation in hepatic transaminases, elevation in lipid peroxide biomarkers (thiobarbituric acid reactive substances; TBARS), reduction of reduced glutathione (GSH) concentration, reduction in the activities of antioxidant enzymes namely catalase (CAT), total superoxide dismutase (SOD), glutathione peroxidase (GPX) and glutathione-S-transferase (GST) and down-regulation of gene expression of these antioxidant enzymes compared to control. Liver sections of rats intoxicated with AFB1 showed a disrupted lobular architecture, scattered necrotic cells and biliary proliferation. Administration of curcumin with AFB1 resulted in amelioration of AFB1 -induced effects compared to untreated AFB1 -intoxicated rats via an up-regulation of antioxidant enzyme gene expression, activation of the expressed genes and increase in the availability of GSH.

Toxoplasmosis is a widespread parasitic infection; it affects about 30% of the global population, either through acute toxoplasmosis or its sequels. Our aim was to determine how curcumin affected testicular infection in mice with chronic toxoplasmosis using toxoplasma gondii strain ME49. Forty male BALB/c mice (6-8 weeks old) weighing between 20 and 25g were randomly divided into four groups. The control group, uninfected animals, received 1cc of normal saline (vehicle) for 2 weeks. Toxo infection in the Toxo and Toxo + CUR groups continued for 4 weeks. After infection, animals in the Toxo and Toxo + CUR groups were treated orally for 2 weeks with 1cc of normal saline (vehicle) or curcumin (CUR) (200mg/kg) respectively [1]. Levels of oxidative stress markers, antioxidant enzyme activities and gene expression, sperm parameters, and histopathological changes were measured and evaluated [1]. Levels of oxidative stress indicators, antioxidant enzyme activity and gene expression, sperm parameters and histopathological changes were measured and evaluated. Toxoplasma gondii infection decreased the activity and gene expression of testosterone and serum antioxidant enzymes (SOD, GPx, and CAT), while elevating FSH and LH levels. Histological alterations, including maturational anomalies, intratubular necrosis, and inflammatory infiltration, were noted in mice infected with Toxoplasma gondii. Curcumin decreased FSH and LH levels while enhancing sperm parameters, histological alterations, and the activity and gene expression of antioxidant enzymes (SOD, GPx, and CAT), as well as testosterone levels. Curcumin treatment mitigated testicular infection induced by Toxoplasma gondii by enhancing antioxidant enzymes, improving sperm parameters, and decreasing pathological alterations in testicular tissue.

The effect of acute nitrite stress on gene expression of antioxidant enzymes, Hsp70 and metabolic enzymes in gill tissue of adult Procambarus clarkii (Girard, 1852) was evaluated. A 48 hours nitrite exposure was conducted, using four test concentrations |$({\rm{N}}{{\rm{O}}_{{2^ - }}} - {\rm{N}} = 1,2,3\,{\rm{and}}\,4\;{\rm{mg}}\;{{\rm{l}}^{ - 1}})$| plus a control group. Significantly increased mRNA expression was observed for all the antioxidant enzymes. A significant increase of Hsp70 mRNA expression was observed when exposed to |$2\;{\rm{mg}}\;{{\rm{l}}^{ - 1}}$| nitrite-N at 12 h. An elevation in AST mRNA expression occurred in the |$3\;{\rm{mg}}\;{{\rm{l}}^{ - 1}}$| nitrite-N group at 12 h. No significant variation was observed for ALT. Elevations in SDH and |${\rm{N}}{{\rm{a}}^ + }/{{\rm{K}}^ + }$|-ATPase |$\alpha $|-subunit mRNA expression were found throughout the experiment. This study shows that gene expression of antioxidant and metabolic enzymes could be helpful in predicting sublethal nitrite toxicity and it will also improve our understanding of the toxicological mechanisms of nitrite on crustacean.

The purposes of this study were to 1) examine the immune and oxidative stress responses following high-intensity interval training (HIIT); 2) determine changes in antioxidant enzyme gene expression and enzyme activity in lymphocytes following HIIT; and 3) assess pre-HIIT, 3-h post-HIIT, and 24-h post-HIIT lymphocyte cell viability following hydrogen peroxide exposure in vitro. Eight recreationally active males completed three identical HIIT protocols. Blood samples were obtained at preexercise, immediately postexercise, 3 h postexercise, and 24 h postexercise. Total number of circulating leukocytes, lymphocytes, and neutrophils, as well as lymphocyte antioxidant enzyme activities, gene expression, cell viability (CV), and plasma thiobarbituric acid-reactive substance (TBARS) levels, were measured. Analytes were compared using a three (day) × four (time) ANOVA with repeated measures on both day and time. The a priori significance level for all analyses was P < 0.05. Significant increases in superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) activities were observed in lymphocytes following HIIT. No significant increases in lymphocyte SOD, CAT, or GPX gene expression were found. A significant increase in TBARS was found immediately post-HIIT on days 1 and 2. Lymphocyte CV in vitro significantly increased on days 2 and 3 compared with day 1. Additionally, there was a significant decrease in CV at 3 h compared with pre- and 24 h postexercise. These findings indicate lymphocytes respond to oxidative stress by increasing antioxidant enzyme activity. Additionally, HIIT causes oxidative stress but did not induce a significant postexercise lymphocytopenia. Analyses in vitro suggest that lymphocytes may become more resistant to subsequent episodes of oxidative stress. Furthermore, the analysis in vitro confirms that lymphocytes are more vulnerable to cytotoxic molecules during recovery from exercise.

The response of lymphocyte and plasma antioxidant defences to a prolonged exercise as a cycling stage in a professional race was analysed. Antioxidant enzyme activities and gene expression, carbonyl derivative and MDA levels were determined in lymphocytes. Plasma levels of vitamin E, carotenes, protein carbonyl derivatives and the test d-Roms were measured. Significant increases in plasmatic carbonyls and in the test d-Roms were observed after the cycling stage. No significant differences were found in the lymphocyte MDA and carbonyl derivative levels. A significant increase was found in plasma vitamin E concentration after the cycling stage; however, the lymphocyte vitamin E concentration did not change. Significant increases were observed in lymphocyte total superoxide dismutase (SOD) activity and in the levels of CuZn-SOD and Mn-SOD isoenzymes. The moderate levels of oxidative stress in the lymphocyte induced a cellular adaptation to exercise enough to counteract the negative effects of oxidative stress.

Hydroxypropyl cellulose reduces chilling injury in green bell pepper (Capsisum annuum L.) by regulating the activity and gene expression of enzymes involved in antioxidant and membrane lipid metabolism

Root-associated symbiotic fungi can enhance the host plant’s ability to tolerate adverse environmental conditions, but it’s unclear whether and how they enhance waterlogged peach plants. This study aimed to investigate the effects of Funneliformis mosseae (Fm), an arbuscular mycorrhizal fungus, and Serendipita indica (Si), an endophytic fungus, on plant growth, root development, reactive oxygen species (ROS) levels, total flavonoids, ROS scavenging activity of flavonoids, and activities and gene expression of antioxidant enzymes in roots of waterlogged peach seedlings. Although waterlogging treatment had little effect on root development, Fm and Si, particularly Fm, improved root development variables to varying degrees. Waterlogging significantly promoted superoxide anion radicals (O2•−) and hydrogen peroxide (H2O2) levels, whereas Fm and Si distinctly reduced O2•− and H2O2 levels under waterlogging. Waterlogging triggered an increase in total flavonoids in fungi-inoculated seedlings, and the two fungi also significantly increased total flavonoid concentrations and the scavenging activity of total flavonoids to hydroxyl radical, O2•−, and 2,2-diphenyl-1-picrylhydrazyl radical. In addition, root fungal colonization rate was negatively and significantly correlated with O2•− and H2O2, but positively with total flavonoids. Under waterlogging conditions, Fm significantly raised root superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) activities, and Si likewise elevated root SOD, CAT, GPX, and peroxidase (POD) activities, compared with uninoculated treatment. Although the expression of various antioxidant enzyme genes in roots remained mostly unchanged or were inhibited by the waterlogging, Fm up-regulated the expression of PpSOD3 and PpSOD7, whereas Si up-regulated the expression of PpSOD3, PpCAT1, PpGPX6, PpGPX8, and PpPOD1. In conclusion, low oxidative bursts in Fm- and Si-inoculated peach plants exposed to waterlogging were associated with an increase in total flavonoids as well as activities and gene expression of antioxidant enzymes.Graphical

Açai (Euterpe oleracea Mart.) has recently emerged as a promising source of natural antioxidants. Because increased oxidative stress and impaired antioxidant defense mechanisms are important factors in the development of diabetic complications and many health claims have been reported for açai, the present study was undertaken to evaluate the possible protective effects of açai on the production of reactive oxygen species by neutrophils and on the liver antioxidant defense system in control and streptozotocin-induced diabetic rats. Diet supplementation with 2% açai was found to increase mRNA levels for gamma-glutamylcysteine synthetase and glutathione peroxidase in liver tissue and to decrease reactive oxygen species production by neutrophils. Compared to control animals, diabetic rats exhibited lower levels of mRNA coding for Zn-superoxide dismutase, glutathione peroxidase and gamma-glutamylcysteine synthetase and higher levels of reactive oxygen species production by neutrophils, thiobarbituric acid-reactive substances and carbonyl proteins in hepatic tissues. Although açai supplementation was not effective in restore gene expression of antioxidant enzymes in diabetic rats, it showed a protective effect, decreasing thiobarbituric acid-reactive substances levels and increasing reduced glutathione content in the liver. These findings suggest that açai can modulate reactive oxygen species production by neutrophils and that it has a significant favorable effect on the liver antioxidant defense system under fisiological conditions of oxidative stress and partially revert deleterious effects of diabetes in the liver.

Thioredoxin reductase (TrxR) is a conserved protein that is involved in protecting organisms against various oxidative stresses. In this study, a thioredoxin reductase gene was cloned from the mud crab Scylla paramamosain (SpTrxR). The full-length cDNA of SpTrxR is comprised of 2724 bp with a 1791 bp open reading frame that encodes a putative protein of 596 amino acids. The deduced amino acid sequence of SpTrxR contains the typical TrxR domain. Quantitative real-time PCR analysis revealed that the SpTrxR mRNA was distributed abundantly in mud crabs, while strong expression was observed mainly in the gills. The expression of antioxidant enzyme genes (SpTrxR, SpTrx, SpSOD, and SpCAT) was measured using quantitative real-time PCR after acute ammonia and nitrite exposure. The results show that antioxidant enzyme genes (SpTrxR, SpTrx, SpSOD, and SpCAT) were modulated by acute ammonia and nitrite exposure. These results suggest that antioxidant enzyme genes play an important role in protecting organisms against oxidative stress