Antioxidant Enzyme Genes Research Articles

Pectic oligosaccharide inhibits postharvest decay and maintains kiwifruit quality by inducing host defense response

Pectin-derived bioactive molecules have been used as elicitors of defense response and other physiological processes in plants, including harvested fruit. In the present study, the postharvest use of a pectic oligosaccharide (POS) was investigated for its ability to inhibit blue mold (Penicillium expansum) and black rot (Alternaria alternata) in artificially-inoculated kiwifruit, as well as naturally-occurring infection. Results demonstrated that POS markedly inhibited blue mold and black rot, as well as natural infection in kiwifruit, relative to untreated, control fruit. POS upregulated the expression of defense-related and antioxidant enzyme genes, and corresponding enzyme activity, including chitinase, β-1,3-glucanase and superoxide dismutase. Our data indicate that POS induces host defense responses in kiwifruit that increase resistance to postharvest decay pathogens. POS-treated fruit also maintained a higher level of fruit firmness and total phenolic compounds than untreated, control fruit, and exhibited a reduced level of weight loss. Collectively, our study indicates that the use of POS may represent a potential strategy for prolonging the postharvest quality of kiwifruit.

Foliar Spraying of Glycine Betaine Alleviated Growth Inhibition, Photoinhibition, and Oxidative Stress in Pepper (Capsicum annuum L.) Seedlings under Low Temperatures Combined with Low Light.

Low temperature combined with low light (LL stress) is a typical environmental stress that limits peppers' productivity, yield, and quality in northwestern China. Glycine betaine (GB), an osmoregulatory substance, has increasingly valuable effects on plant stress resistance. In this study, pepper seedlings were treated with different concentrations of GB under LL stress, and 20 mM of GB was the best treatment. To further explore the mechanism of GB in response to LL stress, four treatments, including CK (normal temperature and light, 28/18 °C, 300 μmol m-2 s-1), CB (normal temperature and light + 20 mM GB), LL (10/5 °C, 100 μmol m-2 s-1), and LB (10/5 °C, 100 μmol m-2 s-1 + 20 mM GB), were investigated in terms of pepper growth, biomass accumulation, photosynthetic capacity, expression levels of encoded proteins Capsb, cell membrane permeability, antioxidant enzyme gene expression and activity, and subcellular localization. The results showed that the pre-spraying of GB under LL stress significantly alleviated the growth inhibition of pepper seedlings; increased plant height by 4.64%; increased root activity by 63.53%; and decreased photoinhibition by increasing the chlorophyll content; upregulating the expression levels of encoded proteins Capsb A, Capsb B, Capsb C, Capsb D, Capsb S, Capsb P1, and Capsb P2 by 30.29%, 36.69%, 18.81%, 30.05%, 9.01%, 6.21%, and 16.45%, respectively; enhancing the fluorescence intensity (OJIP curves), the photochemical efficiency (Fv/Fm, Fv'/Fm'), qP, and NPQ; improving the light energy distribution of PSΠ (Y(II), Y(NPQ), and Y(NO)); and increasing the photochemical reaction fraction and reduced heat dissipation, thereby increasing plant height by 4.64% and shoot bioaccumulation by 13.55%. The pre-spraying of GB under LL stress also upregulated the gene expression of CaSOD, CaPOD, and CaCAT; increased the activity of the ROS-scavenging ability in the pepper leaves; and coordinately increased the SOD activity in the mitochondria, the POD activity in the mitochondria, chloroplasts, and cytosol, and the CAT activity in the cytosol, which improved the LL resistance of the pepper plants by reducing excess H2O2, O2-, MDA, and soluble protein levels in the leaf cells, leading to reduced biological membrane damage. Overall, pre-spraying with GB effectively alleviated the negative effects of LL stress in pepper seedlings.

Temporal correlations of ferroptosis, inflammation and oxidative stress under acute ammonia exposure in brain tissue of yellow catfish (Pelteobagrus fulvidraco)

Ammonia is one of the most serious environmental stressors which severely affect fishery production. Ammonia toxicity to fish has a tight relationship with oxidative stress, inflammation and ferroptosis (a type of programmed cell death characterized by iron-dependent lipid peroxidation), but the temporal response of the above three in brain remains unclear. In the present study, yellow catfish were exposed to three concentrations of ammonia: low concentration (TA-N ˂ 0.01 mg L−1, LA), middle concentration (TA-N 5.70 mg L−1, MA), high concentration (TA-N 28.50 mg L−1, HA) for 96 h. Brain was selected as target tissues for analysis. Results showed that ammonia stress resulted in firstly increased contents of hydroxyl radical at 1 h, total iron at 12 h, malondialdehyde at 48 h, respectively, and decreased contents of GSH at 3 h. The initial high expression levels of ferroptosis (GPX4, system xc−, TFR1) and inflammatory-related factors (NF-ƙB p65, TNF, COX-2, and LOX-15B), antioxidant enzymes genes (SOD and CAT) were observed at first hour upon MA or HA stress. Combining all, it suggested that brain ferroptosis and inflammation were the first to be activated at the initial stage of ammonia stress, and then that provoked oxidative stress.

Variation in antioxidant enzyme activity and key gene expression during fruit development of blackberry and blackberry–raspberry hybrids

Blackberry (Rubus spp.) belongs to the Rubus genus in the Rosaceae family, and its fruits are rich in nutrients and function in exerting antioxidant effects. Based on this, the fruits of 'Chester' blackberry, 'Hull' blackberry and the hybrid cultivars 'Boysen' and 'Young' at different developmental stages were selected to evaluate antioxidant-related indexes, antioxidant enzyme activity and the expression patterns of genes encoding antioxidant enzymes. The results showed that the total antioxidant capacity of the hybrid fruits was higher than that of the pure blackberry fruits. PCA (principal component analysis) and correlation analysis showed that the antioxidant capacity was related to anthocyanin, ASA (ascorbic acid), GSH (glutathione), MDA contents and O2- production rate. However, the correlations between antioxidant capacity and metabolites were different among the cultivars. The O2- production rate and MDA (malondialdehyde) content increased with fruit development. The activities of CAT (catalase) and POD (peroxidase) were maintained at a relatively low level, while SOD (superoxide) activity was maintained at a relatively high level. This study was the first to analyze the content of antioxidant substances in different developmental stages of blackberry fruits and blackberry-raspberry hybrids fruits, which provided some suggestion for the extraction of specific antioxidant substances in different developmental stages. Based on the transcriptome data, the role of antioxidant enzyme genes in antioxidant system was concerned, providing a practical basis for the expression characteristics of key genes.

Riluzole, a Derivative of Benzothiazole as a Potential Anti-Amoebic Agent against Entamoeba histolytica.

Amoebiasis is produced by the parasite Entamoeba histolytica; this disease affects millions of people throughout the world who may suffer from amoebic colitis or amoebic liver abscess. Metronidazole is used to treat this protozoan, but it causes important adverse effects that limit its use. Studies have shown that riluzole has demonstrated activity against some parasites. Thus, the present study aimed, for the first time, to demonstrate the in vitro and in silico anti-amoebic activity of riluzole. In vitro, the results of Entamoeba histolytica trophozoites treated with IC50 (319.5 μM) of riluzole for 5 h showed (i) a decrease of 48.1% in amoeba viability, (ii) ultrastructural changes such as a loss of plasma membrane continuity and alterations in the nuclei followed by lysis, (iii) apoptosis-like cell death, (iv) the triggering of the production of reactive oxygen species and nitric oxide, and (v) the downregulation of amoebic antioxidant enzyme gene expression. Interestingly, docking studies have indicated that riluzole presented a higher affinity than metronidazole for the antioxidant enzymes thioredoxin, thioredoxin reductase, rubrerythrin, and peroxiredoxin of Entamoeba histolytica, which are considered as possible candidates of molecular targets. Our results suggest that riluzole could be an alternative treatment against Entamoeba histolytica. Future studies should be conducted to analyze the in vivo riluzole anti-amoebic effect on the resolution of amebic liver abscess in a susceptible model, as this will contribute to developing new therapeutic agents with anti-amoebic activity.

Epicatechin promotes oocyte quality in mice during repeated superovulation

The negative impacts of repeated superovulation on mitochondrial function and oocyte quality remain unresolved. Epicatechin (EC), a polyphenolic compound found in the human diet with strong antioxidant activity, was investigated for its effects and underlying mechanism on embryonic development after repeated superovulation. The results showed that as the number of superovulation cycles increased, the number of 2-cell embryos decreased, the development of embryos in subsequent in vitro culture was delayed, the apoptosis rate of blastocyst cells increased and the number of blastocyst cells decreased. However, intraperitoneal injection of EC (10 mg/kg body-weight) for two consecutive days during repeated superovulation increased mitochondrial DNA copies in 2-cell embryos of mice. It also promoted the expression of antioxidant enzyme genes in ovaries, increased the content of glutathione (GSH) content and improved the antioxidant capacity of ovaries. Altogether, these results revealed that intraperitoneal injection of EC could increase the embryonic mitochondrial DNA copy number (mtDNA-CN) and enhance the ovary's antioxidant capacity and GSH content, ultimately promoting the quality of mouse embryos in the process of repeated superovulation.

In vivo and in silico investigations on the efficacy of albendazole against Enterocytozoon hepatopenaei (EHP) infecting Penaeus vannamei

The microsporidian Enterocytozoon hepatopenaei (EHP) is an intercellular spore former that causes hepatopancreatic microsporidiosis (HPM), a commercially important disease in Penaeus vannamei (white leg shrimp) farming. In this study, albendazole an inhibitor of β-tubulin polymerization in microsporidians was evaluated for its efficacy in reducing EHP infection in P. vannamei. EHP-infected shrimps were exposed to 1, 5, 10, 25, 50 and 75 mg L−1 of albendazole and evaluated for the reduction in EHP DNA copy numbers at 3, 6, 9, 12, 15, 18, 21, 24 days post-exposure (dpe) by absolute quantitative real-time PCR (RT-qPCR) assay. The expressions of antioxidant enzyme genes viz., cytosolic manganese superoxide dismutase (cMnSOD), catalase (CAT) and glutathione peroxidase (GPx); haemolymph aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) were assessed at 24 dpe. A maximum of about 98% reduction in EHP DNA copies was observed in the shrimps treated with 75 mg L−1 per day for 24 days. A significant upregulation in relative expressions of GPx genes and a downregulation cMnSOD and CAT genes were observed in shrimps exposed to 25, 50 and 75 mg L‐1 at 24 dpe. A significant reduction in haemolymph AST and ALT levels was observed upon treatment with 25, 50 and 75 mg L−1 at 24 dpe and there are no significant changes in ALP. A virtual molecular docking between albendazole and EHP encoded β-tubulin revealed a best binding interaction with the presence of 11 hydrogens bonds, multiple non-covalent bonds and a very much lower solvation-free energy gain of −15.6 kcal mol−1 in the active sites of protein interactions. Thus, this study identified albendazole as a potential drug to control EHP infection in shrimp, which might help to combat the production losses caused by EHP.

Transcriptomic and metabolomic analyses reveal the potential mechanism of waterlogging resistance in cotton (Gossypium hirsutum L.).

Cotton (Gossypium hirsutum L.) is susceptible to long-term waterlogging stress; however, genomic information of cotton response mechanisms toward long days of waterlogging is quite elusive. Here, we combined the transcriptome and metabolome expression level changes in cotton roots after 10 and 20 days of waterlogging stress treatment pertaining to potential resistance mechanisms in two cotton genotypes. Numerous adventitious roots and hypertrophic lenticels were induced in CJ1831056 and CJ1831072. Transcriptome analysis revealed 101,599 differentially expressed genes in cotton roots with higher gene expression after 20 days of stress. Reactive oxygen species (ROS) generating genes, antioxidant enzyme genes, and transcription factor genes (AP2, MYB, WRKY, and bZIP) were highly responsive to waterlogging stress among the two genotypes. Metabolomics results showed higher expressions of stress-resistant metabolites sinapyl alcohol, L-glutamic acid, galactaric acid, glucose 1-phosphate, L-valine, L-asparagine, and melibiose in CJ1831056 than CJ1831072. Differentially expressed metabolites (adenosine, galactaric acid, sinapyl alcohol, L-valine, L-asparagine, and melibiose) significantly correlated with the differentially expressed PRX52, PER1, PER64, and BGLU11 transcripts. This investigation reveals genes for targeted genetic engineering to improve waterlogging stress resistance to enhance abiotic stress regulatory mechanisms in cotton at the transcript and metabolic levels of study.

Oxidative damage contributes to bisphenol S-induced development block at 2-cell stage preimplantation embryos in mice through inhibiting of embryonic genome activation

Although bisphenol S (BPS), as a bisphenol A (BPA) substitute, has been widely used in the commodity, it is embryotoxic in recent experiments. Nowadays, it remains unclear how BPS affects preimplantation embryos. Here, my team investigated the effects of BPS on preimplantation embryos and the possible molecular mechanisms in mice. The results showed that 10–6 mol/L BPS exposure delayed the blastocysts stage, and exposure to 10–4 mol/L BPS induced 2-cell block in mice preimplantation embryos. A significant increase in reactive oxygen species (ROS) level and antioxidant enzyme genes Sod1, Gpx1, Gpx6, and Prdx2 expression were shown, but the level of apoptosis was normal in 2-cell blocked embryos. Further experiments demonstrated that embryonic genome activation (EGA) specific genes Hsp70.1 and Hsc70 were significantly decreased, which implied that ROS and EGA activation have the potential to block 2-cell development. Antioxidant enzymes, including superoxide dismutase (SOD), coenzyme Q10 (CoQ10), and folic acid (FA) were used to further explore the roles of ROS and EGA in 2-cell block. Only 1200 U/mL SOD was found to alleviate the phenomenon of 2-cell block, reduce oxidative damage, and restore the expression of EGA-specific genes Hsp70.1 and Hsc70. Conclusively, this study demonstrates for the first time that BPS can induce 2-cell block, which is mainly mediated by ROS aggregation and results in the failure of EGA activation.

Lipid nanoparticle delivery of siRNA targeting Cyp2e1 gene attenuates subacute alcoholic liver injury in mice.

To investigate the effect and mechanism of lipid nanoparticle (LNP) delivery of small interfering RNA (siRNA) targeting Cyp2e1 gene on subacute alcoholic liver injury in mice. siRNA targeting Cyp2e1 gene was encapsulated in LNP (si-Cyp2e1 LNP) by microfluidic technique and the resulting LNPs were characterized. The optimal dose of si-Cyp2e1 LNP administration was screened. Forty female C57BL/6N mice were randomly divided into blank control group, model control group, si-Cyp2e1 LNP group, LNP control group and metadoxine group. The subacute alcoholic liver injury mouse model was induced by ethanol feeding for 10 d plus ethanol gavage for the last 3 d. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, and the superoxide dismutase (SOD) activity as well as malondialdehyde, reactive oxygen species, glutathione, triacylglycerol, total cholesterol contents in liver tissue were measured in each group, and liver index was calculated. The expression of genes related to oxidative stress, lipid synthesis and inflammation in each group of mice were measured by realtime RT-PCR. Compared with the model control group, the levels of liver index, serum ALT, AST activities, malondialdehyde, reactive oxygen species, triacylglycerol, total cholesterol contents in liver tissue decreased, but the SOD activity as well as glutathione increased in the si-Cyp2e1 LNP group (all P<0.01). Hematoxylin-eosin staining result showed disorganized hepatocytes with sparse cytoplasm and a large number of fat vacuoles and necrosis in the model control group, while the si-Cyp2e1 LNP group had uniformly sized and arranged hepatocytes with normal liver tissue morphology and structure. Oil red O staining result showed si-Cyp2e1 LNP group had lower fat content of the liver compared to the model control group (P<0.01), and no fat droplets accumulated. Anti-F4/80 monoclonal antibody fluorescence immunohistochemistry showed that the si-Cyp2e1 LNP group had lower cumulative optical density values compared to the model control group (P<0.01) and no significant inflammatory reaction. Compared with the model control group, the expression of catalytic genes P47phox, P67phox and Gp91phox were reduced (all P<0.01), while the expression of the antioxidant enzyme genes Sod1, Gsh-rd and Gsh-px were increased (all P<0.01). The mRNA expression of the lipid metabolism genes Pgc-1α and Cpt1 were increased (all P<0.01) and the lipid synthesis-related genes Srebp1c, Acc and Fasn were decreased (all P<0.01); the expression of liver inflammation-related genes Tgf-β, Tnf-α and Il-6 were decreased (all P<0.01). The si-Cyp2e1 LNP may attenuate subacute alcoholic liver injury in mice mainly by reducing reactive oxygen levels, increasing antioxidant activity, blocking oxidative stress pathways and reducing ethanol-induced steatosis and inflammation.

Polymorphism of antioxidant defense enzyme genes as the risk &#x0D; for formation of chronic dust bronchitis in long-term labour experience coal mine workers

Introduction. The antioxidant defense system is involved in protecting the body from long-term exposure to harmful damaging factors. Polymorphisms in the genes of antioxidant defense enzymes change the response to oxidative stress in occupational respiratory diseases such as asbestosis, silicosis, occupational asthma, and others.&#x0D; The aim was to study the polymorphism of the MnSOD (rs4880) and GPX1 (rs1050450) genes to predict the risk of developing chronic dust bronchitis in miners of the main occupations of coal enterprises in the South of Kuzbass.&#x0D; Materials and methods. One hundred eighty two coal mine workers with long-term exposure to high concentrations of coal-rock dust (exceeding the maximum permissible concentrations by up to 35 times), including 116 people with a previously proven diagnosis of chronic dust bronchitis, were examined. The comparison group (66 workers) consisted of the persons without a proven diagnosis, working in the same sanitary and hygienic conditions. Genomic DNA was isolated from peripheral blood leukocytes using phenol-chloroform extraction method. Typing of the MnSOD (rs4880) and GPX1 (rs1050450) genes was performed using the Real-Time method.&#x0D; Results. The chance of detecting the AA genotype of the MnSOD gene and the GG genotype of the GPX1 gene in long-term labour experience miners with chronic dust bronchitis was found to be 2 times and 6 times higher than in the comparison group, respectively. The AA GPX1 genotype had been shown to be a factor in resistance to the development of this disease. The combination of AA/GG genotypes of the MnSOD/GPX1 genes was statistically significantly associated with a 1.5-fold risk of developing chronic dust bronchitis. Combinations of genotypes of the MnSOD/GPX1 GG/AA, AA/AA, and AG/AA genes were associated with the resistance to the development of chronic dust bronchitis. The data obtained can be used for personalized prediction of the risk of developing chronic dust bronchitis in long-term labour experience miners of the main occupations.&#x0D; Limitations. The study was limited to the number of miners who underwent periodic medical examinations and were hospitalized at the Research Institute for Complex Problems of Hygiene and Occupational Diseases.&#x0D; Conclusion. The results obtained indicate to the contribution of the polymorphism of the MnSOD (rs4880) and GPX1 (rs1050450) genes to the development of chronic dust bronchitis.

Benzophenone-3 causes oxidative stress in the brain and impairs aversive memory in adult zebrafish

Oxybenzone (BP-3) is an ultraviolet (UV) filter widely used in industries that is directly or indirectly released into the aquatic environment. However, little is known about its effects on brain performance. Here, we investigated whether BP-3 exposure affects the redox imbalance in zebrafish and how they respond to a task that requires memory of an aversive situation. Fish were exposed to BP-3 10 and 50 μg L−1 for 15 days and then tested using an associative learning protocol with electric shock as a stimulus. Brains were extracted for reactive oxygen species (ROS) measurement and qPCR analysis of antioxidant enzyme genes. ROS production increased for exposed animals, and catalase (cat) and superoxide dismutase 2 (sod 2) were upregulated. Furthermore, learning and memory were reduced in zebrafish exposed to BP-3. These results suggested that BP-3 may lead to a redox status imbalance, causing impaired cognition and reinforcing the need to replace the toxic UV filters with filters that minimize environmental effects.

Chronic dietary exposure to arsenic at environmentally relevant concentrations impairs cognitive performance in adult zebrafish (Danio rerio) via oxidative stress and dopaminergic dysfunction

The current study was designed to evaluate the effects of chronic dietary arsenic exposure on the cognitive performance of adult zebrafish and uncover probable pathways by which arsenic mediates such neurotoxic effects. Adult zebrafish were treated with 3 different dietary arsenic concentrations (30, 60, and 100 μg/g dry weight (dw), as arsenite) in addition to control for 60 days. A latent learning paradigm, which employs a complex maze, was used to assess the cognitive performance of fish. Our results demonstrated that dietary treatment with arsenic, especially at medium (60 μg/g dw) and high (100 μg/g dw) exposure dose levels, significantly impaired the performance of fish in various latent learning tasks evaluated in the present study. Concomitant with cognitive dysfunction, chronic dietary exposure to arsenic was also found to increase arsenic accumulation and dopamine levels, and induce oxidative stress (reduced thiol redox, increased lipid peroxidation and expression of antioxidant enzyme genes) in the brain of zebrafish in a dose-dependent manner. Dopaminergic system in the brain is known to play a critical role in regulating cognitive behaviours in fish, and our observations suggested that chronic dietary treatment with medium and high arsenic doses leads to significant alterations in the expression of genes involved in dopamine signaling (dopamine receptors), synthesis (thyroxine hydroxylase) and metabolism (monoamine oxidase) in the zebrafish brain. Moreover, we also recorded significant downregulation of genes such as the brain-derived neurotrophic factor (BDNF) and ectonucleotidases (entpd2_mg, entpd2_mq, and 5′-nucleotidase), which are critical for learning and memory functions, in the zebrafish brain following chronic dietary exposure to arsenic. Overall, the present study suggests that chronic environmentally relevant dietary exposure to arsenic can impair the cognitive performance in zebrafish, essentially by inducing oxidative stress and disrupting the dopaminergic neurotransmission in the brain.

Boosting NAD blunts T H17 inflammation via arginine biosynthesis and redox regulatory control in healthy control and psoriasis human subjects

Abstract Although NAD +levels modulate metabolism, how this regulates immunometabolism/inflammation remains unclear. Employing Nicotinamide Riboside (NR) to boost NAD, we explored T H17-linked inflammatory in Psoriasis. Primary Psoriasis CD4 +T cells exposed to NR reduced IL-17 secretion and RNA-seq/pathway analysis implicated NR in modulating sequestosome 1 (SQSTM1/p62)-coupled oxidative stress. In parallel, NR reduced CD4 +T cell reactive oxygen species (ROS) levels and activated the NRF2 transcription factor. Depletion of NRF2 or SQSTM1 diminished NR effects on ROS and T H17 activation. Metabolomic analysis implicated NR in arginine biosynthesis and L-arginine replicated NR CD4 +T cell effects. Genetic disruption of the argininosuccinate lyase synthesis enzyme ameliorated NR antiinflammatory effects, supporting that NR, via arginine biosynthesis orchestrates NRF2 activation, CD4 +T cell antioxidant defenses and blunted T H17 responsiveness. A pilot in-vivo human study similarly showed that NR increased arginine, antioxidant enzyme gene expression and blunted T H17 regulation, implicating NAD metabolism in CD4 +T cell-linked inflammation. NHLBI Division of Intramural Research (MNS – ZIA-HL005102), NIH Bench to Bedside award (MNS and RT, HL-129510-04S1) and the NIH Office of Dietary Supplementation (JT) and the UK MRC (JLG – MR/P011705/2; UKDRI-5002; MAP UK).

Stress memory in seedlings of primed seed chickpea (Cicer arietinum L.) using sodium nitroprusside under cold stress

Pea cultivation has been developed as a rich source of protein in many countries. The spring cultivation of chickpeas has low performance and efficiency due to the short length of the growth period and the confrontation of the reproductive period with drought and high temperature stress. Autumn cultivation of chickpeas is introduced as an important solution in reducing the yield caused by high temperature and drought stress during the flowering time of spring cultivation. However, in autumn cultivation, sensitivity to cold is the main limitation. So, the present study was performed to investigate the effect of nitric oxide on stress memory due to seed priming with sodium nitroprusside for cold tolerance of chickpea seedlings under simulated autumn temperatures. This experiment was conducted as a factorial in a completely randomized design with nine replicates at the Research Center for Plant Sciences of Ferdowsi University, Mashhad Iran, in 2020. Experimental factors included seed priming at two levels (control and sodium nitroprusside) and chickpea cultivars, including a susceptible genotype (MCC505) and a cold-tolerant cultivar (Saral). This experiment evaluated the gene expression of antioxidant enzymes (superoxide dismutase, catalase, peroxidase, and ascorbate peroxidase) in the seedling stage. The results revealed that superoxide dismutase, catalase, peroxidase, and ascorbate peroxidase genes were expressed in sodium nitroprusside and control treatments. In the Saral cultivar, sodium nitroprusside increased the gene expression of superoxide dismutase, catalase, peroxidase, and ascorbate peroxidase enzymes by 29, 88, 54, and 63%, respectively. On the other hand, the gene expression of these enzymes in the control treatment may be due to the effect of temperature on them. There was a significant increase in the expression of superoxide dismutase, catalase, peroxidase, and ascorbate peroxidase genes in the Saral cultivar compared to the MCC505 genotype. In general, sodium nitroprusside increased the activity of antioxidant enzymes in the Saral cultivar through the expression of antioxidant enzyme genes.

Effects of Hermetia illucens larvae meal on the Pacific white shrimp (Litopenaeus vannamei) revealed by innate immunity and 16S rRNA gene sequencing analysis

The larvae of the black soldier fly, Hermetia illucens, are now attracting attention and becoming promising sources for aquafeed ingredient due to the nutritious substance. However, the introduction of a novel ingredient into the recipe may have unpredictable effects on the innate immune function and gut bacteria composition of crustaceans. Therefore, the present study aimed to evaluate how dietary black soldier fly larvae meal (BSFLM) affected the antioxidant ability, innate immunity and gut microbiome of shrimp (Litopenaeus vannamei) fed with a practical diet, including the gene expression of Toll and immunodeficiency (IMD) pathways. Six experimental diets were formulated by replacing gradient levels of fish meal (0 %, 10 %, 20 %, 30 %, 40 % and 50 %) based on a commercial shrimp diet. Four replicates of shrimp were fed different diets three times daily for 60 days. Growth performance linearly decreased with increasing BSFLM inclusion. Results of antioxidative enzyme activities and gene expression suggested that low dietary BSFLM levels activated the antioxidant capacity of shrimp, while dietary BSFLM levels up to 100 g/kg may induce oxidative stress and inhibit glutathione peroxidase activity. Although traf6, toll1, dorsal and relish were significantly upregulated in different BSFLM groups, the expression of tak1 was significantly downregulated in groups containing BSFLM, implying the immune susceptibility may be weakened. Gut flora analysis indicated dietary BSFLM altered both beneficial and opportunistic pathogenic bacterial abundance, with low levels of dietary BSFLM increased the abundance of bacteria that may contribute to carbohydrate utilization, while high levels of dietary BSFLM may cause intestinal disease and low intestinal immune response. To conclude, 60–80 g/kg of dietary BSFLM showed no adverse effects on the growth, antioxidant capacity and gut flora of shrimp, which was the adequate level in shrimp diet. While 100 g/kg dietary BSFLM may induce oxidative stress and potentially weaken the innate immunity of shrimp.

Antioxidant enzyme and DNA base repair genetic risk scores' associations with systemic oxidative stress biomarker in pooled cross-sectional studies.

Background: Oxidative stress is hypothesized to contribute to the pathogenesis of several chronic diseases. Numerous dietary and lifestyle factors are associated with oxidative stress; however, little is known about associations of genetic factors, individually or jointly with dietary and lifestyle factors, with oxidative stress in humans. Methods: We genotyped 22 haplotype-tagging single nucleotide polymorphisms (SNPs) in 3 antioxidant enzyme (AE) genes and 79 SNPs in 14 DNA base excision repair (BER) genes to develop oxidative stress-specific AE and BER genetic risk scores (GRS) in two pooled cross-sectional studies (n = 245) of 30-74-year-old, White, cancer- and inflammatory bowel disease-free adults. Of the genotypes, based on their associations with a systemic oxidative stress biomarker, plasma F2-isoprostanes (FiP) concentrations, we selected 4 GSTP1 SNPs for an AE GRS, and 12 SNPs of 5 genes (XRCC1, TDG, PNKP, MUTYH, and FEN1) for a BER GRS. We also calculated a previously-reported, validated, questionnaire-based, oxidative stress biomarker-weighted oxidative balance score (OBS) comprising 17 anti- and pro-oxidant dietary and lifestyle exposures, with higher scores representing a higher predominance of antioxidant exposures. We used general linear regression to assess adjusted mean FiP concentrations across GRS and OBS tertiles, separately and jointly. Results: The adjusted mean FiP concentrations among those in the highest relative to the lowest oxidative stress-specific AE and BER GRS tertiles were, proportionately, 11.8% (p = 0.12) and 21.2% (p = 0.002) higher, respectively. In the joint AE/BER GRS analysis, the highest estimated mean FiP concentration was among those with jointly high AE/BER GRS. Mean FiP concentrations across OBS tertiles were similar across AE and BER GRS strata. Conclusion: Our pilot study findings suggest that DNA BER, and possibly AE, genotypes collectively may be associated with systemic oxidative stress in humans, and support further research in larger, general populations.

Effects of Dietary Vegetable Oil Mixtures including Soybean Oil on Intestinal Oxidative Stress in Gilthead Sea Bream (Sparus aurata).

Fish oil is commonly replaced by vegetable oils in sea bream diets, but little is known about their effects on intestinal health regarding oxidative stress biomarkers. The negative effects of lipid peroxidation on digestive mucosa could have consequences in animal nutrition and welfare. In this study, five isonitrogenous (46%) and isolipidic (22%) diets with 75% of vegetable oils inclusion were evaluated: soybean oil (S) alone or different mixtures containing soybean oil with linseed (SL), linseed and rapeseed (SLR), linseed and palm (SLP), and linseed, rapeseed, and palm (SLRP). Gilthead sea bream juveniles were fed twice a day for 18 weeks. Pyloric caeca and proximal intestine samples were collected 24 h post feeding for lipid peroxidation (LPO), antioxidant enzyme activities (SOD, CAT, GPx, GST, and GR) and gene expression analyses. Pyloric caeca presented larger unhealthy changes in oxidative status than proximal intestine. Although SL-fed fish showed the highest antioxidant activities, they were unable to cope with LPO that in pyloric caeca was 31.4 times higher than in the other groups. Instead, SLP fish presented the best oxidative status, with low LPO levels, antioxidant enzyme activities, and gene expression. In summary, between the vegetable oils dietary mixtures tested, SPL would maintain better intestinal health.

Effect of redroot pigweed interference on antioxidant enzyme and light response of common bean (Phaseolus vulgaris L.) depends on cultivars and growth stages

Redroot Pigweed (Amaranthus retroflexus L.) is an important weed that is highly competitive with common bean. Photosynthetic pigments, the activity of antioxidant enzymes, the relative expression of a number of antioxidant enzyme and light response genes, were studied in three of common bean cultivars and in V4 and R7 stages under Redroot Pigweed free and infested. The presence of weeds reduced the content of chlorophyll, relative chlorophyll and anthocyanin of common bean leaves. With the increase of weed competition, the expression of antioxidant genes and enzymes increased, which indicates the increase of their activity in order to reduce the amount of reactive oxygen species. Among the studied antioxidant enzymes, the activity of catalase and ascorbate peroxidase produced in the leaves was higher than that of superoxide dismutase. With the increase of weed interference, the expression of phytochrome interacting factor 3 (PIF3) gene as a positive regulator of light signals is increased and the expression of phytochrome rapidly regulated1 (PAR1) gene as a negative regulator is decreased. Chlorophyll a/b-binding protein (CAB1) and auxin-responsive protein IAA8 (IAA8) genes also down-regulated with increasing competition. Along with the decrease of CAB expression in the conditions of competition with weeds, the chlorophyll a, b content also decreased. Correlation between gene expression and physiological traits related to them highlights the prominent role of CWCP in maintaining yield potential.

DNA damage, alterations in the expression of antioxidant enzyme genes and in the histoarchitecture of gill cells of zebrafish exposed to 17-α-ethinylestradiol

Endocrine disruptors, such as estrogen, are chemical substances with the potential to alter the hormonal balance of organisms. Their origin can be natural or artificial, and they can act at very low doses. The estrogen 17α-ethinylestradiol (EE2) is used worldwide as an oral contraceptive and is a potential contaminant in aquatic ecosystems. It is well documented that these environmental pollutants can act directly or indirectly on the reproductive system, impairing development and fertility. However, little is known about the alteration of the cell oxidative status induced by EE2. The main objective of this study was to evaluate the effect on the gill cells of adult zebrafish exposed in vivo to EE2, analyzing cell histology, DNA damage and the expression levels of genes encoding the main enzymes involved in oxidative stress pathways. The histological study showed that EE2 produces moderate to high damage to the gill tissue, an increase in gill cell DNA damage and the mRNA levels of the genes corresponding to the manganese superoxide dismutase (Mn-sod) and catalase (cat) after exposure to 5 ng/L EE2. The results indicate that EE2 causes tissue alterations, DNA damage and oxidative stress. EE2 produced important alterations in the gills, a fundamental organ for the survival of fish. There is a clear need for further research on the ecological consequences of EDCs on non-target organisms.