Zinc Superoxide Dismutase Research Articles

Protective effect of betaine supplementation in low-met diet against Aeromonas hydrophila-induced intestinal injury in grass carp (Ctenopharyngodon idella)

This study evaluated the effect of supplemental betaine in low-methionine (Met) diet concerning the regulatory role of bacteria-induced intestinal injury in fish. 540 grass carp (Ctenopharyngodon Idella) (210 ± 0.68 g) were selected and randomly divided into 6 groups, which contained a reference group fed with the diet supplemented with DL-Met (3.1 g kg−1 diet) and 5 groups fed on the diets added with increasing levels of betaine (0–6.4 g kg−1 diet) for 60 days. Then, a 14-day Aeromonas hydrophila challenge was conducted. Compared with BET0 group, results indicated that appropriate betaine supplementation (1) enhanced antioxidant enzymes activities (except glutathione S-transferase pi-1 (GSTP1) and copper zinc superoxide dismutase (CuZnSOD) and glutathione (GSH) contents, and NF-E2-related-factor 2 (Nrf2) protein nuclear translocation, while reduced reactive oxygen species (ROS), and representative oxidative damage markers; (2) up-regulated the transcription abundance of anti-apoptosis-related molecules while reduced DNA-ladder occurrence and the transcription levels of pro-apoptosis-related genes; (3) down-regulated the transcription of tight junction-related proteins (such as zonula occludens-1 (ZO-1) etc.) in fish intestines. Furthermore, correlation analysis revealed that feeding betaine ameliorating the oxidative damage, apoptosis and tight junction breakdown was possibly related to intestinal Nrf2, p38 mitogen-activated protein kinase (p38 MAPK) and c-Jun N-terminal kinases (JNK), and myosin light chain kinase (MLCK) signaling, respectively. According to the regression analysis for intestinal ROS abundance, claudin-b and caspase-2 transcription, the appropriate betaine supplementations were estimated to be 3.82, 3.98, and 3.62 g kg−1 diet, respectively. Notably, betaine has the potential to spare part Met and the efficacy of betaine relative to Met was 206.67 % based on intestinal ROS contents in grass carp (210–776 g).

Evolutionary Conserved and Divergent Responses to Copper Zinc Superoxide Dismutase Inhibition in Plants.

After an initial evolution in a reducing environment, life got successively challenged by reactive oxygen species (ROS), especially during the great oxidation event (GOE) that followed the development of photosynthesis. Therefore, ROS are deeply intertwined into the physiological, morphological and transcriptional responses of most present-day organisms. Copper-zinc superoxide dismutases (CuZnSODs) evolved during the GOE and are present in charophytes and extant land plants, but nearly absent from chlorophytes. The chemical inhibitor of CuZnSOD, lung cancer screen 1 (LCS-1), could greatly facilitate the study of SODs in diverse plants. Here, we determined the impact of chemical inhibition of plant CuZnSOD activity, on plant growth, transcription and metabolism. We followed a comparative approach by using different plant species, including Marchantia Polymorpha and Physcomitrium patens, representing bryophytes, the sister lineage to vascular plants, and Arabidopsis thaliana. We show that LCS-1 causes oxidative stress in plants and that the inhibition of CuZnSODs provoked a similar core response that mainly impacted glutathione homoeostasis in all plant species analysed. That said, Physcomitrium and Arabidopsis, which contain multiple CuZnSOD isoforms showed a more complex and exacerbated response. In addition, an untargeted metabolomics approach revealed a specific metabolic signature for each plant species. Our comparative analysis exposes a conserved core response at the physiological and transcriptional level towards LCS-1, while the metabolic response largely varies. These differences correlate with the number and localization of the CuZnSOD isoforms present in each species.

Elevated Renal Oxidative Stress and Na+ Transporters are Associated with Hypertension in Postpartum Preeclamptic Rats

Background: Approximately 5-10% of US pregnancies result in preeclampsia (PE). PE is characterized by new onset hypertension (HTN) during pregnancy and is usually accompanied by end-organ damage, especially in the kidneys. Postpartum (PP) women and dams that had PE have an increased risk of developing HTN and chronic kidney disease (CKD) later in life. However, mechanisms linking PE to the long-term development of HTN and CKD are unknown. One aspect that may contribute to renal injury in PP PE women and dams is oxidative stress. Elevated concentrations of oxidative stress have been shown to augment the abundance and activity of renal transporters to increase sodium (Na+) reabsorption and blood volume. These alterations in renal transporters can consequently increase Na+ and water to facilitate HTN. Thus, we hypothesize that at 6 weeks PP (~3 human years), PE dams will display increased oxidative stress, renal Na+ transporter abundance, and elevated blood pressure (BP). Methods: Pregnant Sprague Dawley rats were randomly assigned to two groups: normal (CON) and PE dams. On gestational day 14, the reduced uterine perfusion pressure surgery was performed to mimic placental ischemia and generate a model of PE. Dams gave birth naturally and weaned for 3 weeks. After 6 weeks PP, BP was measured via carotid catheterization, and kidneys were removed and sectioned. Western blots were used to quantify renal Na+ transporters: Na+ K+ 2Cl− transporter (NKCC2) in the kidney medulla (KM) and epithelial Na+ channel (ENaC) in both the kidney cortex (KC) and KM. Oxidative stress was evaluated by heat shock protein 1 (HSP-1), copper zinc superoxide dismutase (CuZnSOD), and manganese superoxide dismutase (MnSOD) via Western blots. Hydrogen peroxide (H2O2) and antioxidant capacity concentrations were assessed via colorimetric assays. Results: PP PE dams had increased BP (126.3±6.18vs105.7±3.74 mmHg, p<0.05) at 6 weeks after birth. KC HSP-1, H2O2, MnSOD, and antioxidant capacity were unchanged between groups. However, KC CuZnSOD protein abundance was decreased in PP PE dams (69.51±11.64vs100±5.73 IU/Protein/Control%, p<0.05). In the KM, HSP-1 abundance (113.7±3.3vs100±5.07 IU/Protein/Control%, p=0.06) and H2O2 concentrations (1.97±0.11vs1.31± 0.38 nM H2O2/mg Protein, p=0.08) were elevated in PP PE dams. MnSOD, CuZnSOD, and antioxidant capacity were unchanged between groups in the KM. No changes occurred in KC and KM ENaC protein abundance. However, NKCC2 protein abundance was elevated by ~50% in PP PE dams (151.71±22.17vs100±5.59 IU/Protein/Control%, p=0.06). Conclusion: In summary, BP, oxidative stress, and NKCC2 were elevated in PP PE dams at 6 weeks. Perhaps, the presence of oxidative stress in the KM may lead to increased NKCC2 abundance. However, more studies are warranted to make this conclusion. NKCC2 elevation may result in increased Na+ and water reabsorption, leading to an increase in BP. Future studies will assess the role of renal oxidative stress to regulate Na+ transporters in PP PE dams and determine the timeline after birth (PP) in which changes in oxidative stress, Na+ transporters, and BP occur. In summary, this study is clinically relevant because it indicates that both oxidative stress and NKCC2 in the KM, separately or together, may have a formative role in the pathogenesis of HTN and CKD in PP PE women later in life. This research was supported by the Neurobiology of Aging and Alzheimer’s Disease Training Program (funded by NIH training grant T32 AG020494). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Chitosan oligosaccharide improves intestinal function by promoting intestinal development, alleviating intestinal inflammatory response, and enhancing antioxidant capacity in broilers aged d 1 to 14

This study was conducted to investigate the effects of chitosan oligosaccharide (COS) supplementation on intestinal development and functions, inflammatory response, antioxidant capacity and the related signaling pathways in broilers aged d 1 to 14. A total of 240 one-day old male Arbor Acres broilers (40.47 ± 0.30 g) were randomly allotted to 4 groups, and each group consisted of 6 replicate pens with 10 broilers per replicate. Broilers fed a basal diet supplementation with COS at 0 (CON group), 200 (COS200 group), 400 (COS400 group), and 800 mg/kg (COS800 group) for 14 d, respectively. Broilers in the COS supplementation groups had no significant effects on growth performance. Compared to the CON group, dietary COS supplementation increased (P < 0.05) the relative weight of duodenum, jejunal lipase activity, duodenal and ileal villus surface area, and lower (P < 0.05) ileal amylase and alkaline phosphatase activity, and crypt depth. The expression level of duodenal glucose transporter 1 (GLUT1), Na+-glucose cotransporter 1 (SGLT1), peptide transporter 1 (PepT1), occludin, zonula occludens-1 (ZO-1), toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), and interleukin-10 (IL-10), jejunal SGLT1, PepT1, occludin, tumor necrosis factor-α (TNF-α), and ileal SGLT1, PepT1, and fatty acid binding protein 1 (FABP1) was upregulated by COS. However, the expression level of duodenal FABP1 and TNF-α, jejunal GLUT1, ZO-1, TLR4, MyD88, nuclear factor kappa-B p65 (NF-κB p65), and IL-1β, and ileal GLUT1, NF-κB p65, and IL-1β was downregulated by COS. Furthermore, dietary COS supplementation increased duodenal catalase (CAT), glutathione peroxidase (GSH-Px), and total superoxide dismutase (T-SOD) activity, jejunal CAT and T-SOD activity, upregulated the expression level of duodenal nuclear factor-erythroid 2-related factor 2 (Nrf2), CAT, glutathione peroxidase 1 (GPX1), and copper and zinc superoxide dismutase (Cu/Zn SOD), jejunal CAT, and ileal Nrf2, CAT, and GPX1. These results suggested that COS could promote intestinal development and functions in broilers aged d 1 to 14, which might be mediated by alleviating intestinal inflammatory response and enhancing antioxidant capacity.

Abstract P420: Changes In Cardiac Oxidative Stress Is Protected In Intrauterine Growth Restricted Females Vs Males Before The Development Of Hypertension And Cardiovascular Diseases

Preeclampsia (PE) is a pregnancy complication characterized by hypertension (HTN) typically during the third trimester. PE is the leading cause for intrauterine growth restricted (IUGR) offspring (OFS). Women with PE and their OFS (IUGR) are at a higher risk for developing cardiovascular diseases (CVD) later in life. Two major factors that contribute to the development of CVDs are HTN and oxidative stress (OS). Studies show that human and animals adult IUGR OFS have a higher risk of developing HTN and CVDs compared to females later in life. The rationale for these sex differences is unknown; and we hypothesize that male IUGR rats will have elevated cardiac oxidative stress, while female IUGR will have lower cardiac OS .To test, we will examine sex differences between 12 week IUGR and control (CON) OFS that are not hypertensive, to determine if there are any change in cardiac OS before the development of HTN and CVD. Methods: Sprague Dawley rats were divided into 4 groups: IUGR male, IUGR female, and control (CON) male and female. IUGR rats were derived from the OFS reduced uterine perfusion pressure (RUPP) PE rat model. CON OFS were from normal pregnant rats. At 12wk, BP was measured via carotid catheterization. Heart tissue was collected to assess oxidative stress, via examining reactive oxygen species (ROS) Heat shock protein 1 (HSP-1) and hydrogen peroxide (H2O2). Antioxidants were measured via manganese superoxide dismutase (MnSOD), copper zinc superoxide dismutase (CuZnSOD) and antioxidant capacity (AC). There were no significant changes in HSP-1, CuZnSOD, and H2O2 between male and female OFS. However, female IUGR OFS had a 20% increase in antioxidant MnSOD (119.9 ± 6.133 vs 93.98 ± 5.197 %MnSOD U/protein/CON, p &lt;0.05) compared to CON in the heart. Male OFS had no significant changes for MnSOD. IUGR female AC had a 2-fold increase in activity (22.13 ± 1.019 vs 14.15 ± 1.852 mM Trolox/mg protein, p &lt;0.05) compared to its CON. Furthermore, IUGR male OFS had a slight decrease in AC compared to CON male (11.00 ± 1.420 vs 13.24 ± 0.4676 mM Trolox/mg protein, ns). The significant increase of AC and MnSOD in IUGR female OFS suggests that females may be more protected from OS, via mitochondria-specific antioxidants compared to males.

Dietary zinc supplementation in breeding pigeons improves the carcass traits of squabs through regulating antioxidant capacity and myogenic regulatory factor expression

The purpose of this experiment was to explore the effects of zinc supplementation in breeding pigeons diet on carcass traits, meat quality, antioxidant capacity and mRNA expressions of myogenic regulatory factors of squabs. A total of 120 healthy White King pigeons were randomly assigned to 5 treatments, each involving 8 replicates. The experiment lasted for 46 d (18-d incubation period of eggs and 28-d growth period of squabs). The 5 groups were 0, 30, 60, 90, and 120 mg/kg zinc addition. Results showed that the 28-d body weight, breast muscle yield, zinc content in crop milk and myogenic factor 6 (MyF6) abundance of breast muscle were linearly increased (P < 0.050), but the abdominal fat yield linearly decreased (P=0.040) with increasing dietary zinc supplementation. Both the linear (P < 0.050) and quadratic responses (P < 0.001) were observed in copper zinc superoxide dismutase (Cu-Zn SOD), total antioxidant capacity (T-AOC) and malondialdehyde (MDA) contents in liver and breast muscle. The 28-d body weight was increased by 90 mg/kg zinc supplementation (P < 0.05), and there is no significant difference between 90 and 120 mg/kg zinc addition. The breast muscle yield, Cu-Zn SOD and T-AOC contents in breast muscle and liver, zinc contents in crop milk and breast muscle, MyF6 mRNA expression in breast muscle were higher (P < 0.05) in the group supplemented with 120 mg/kg zinc than the control. The abdominal fat yield was numerically lowest, and MDA contents in breast muscle and liver were significantly lowest in the group fed 120 mg/kg zinc (P < 0.05). However, the meat quality traits were not affected (P > 0.05) by zinc supplementation, except for shear force. It should be stated dietary zinc supplementation at the level of 120 mg/kg for breeding pigeons increased body weight and breast muscle yield of squabs, which may be associated with the up-regulating MyF6 mRNA expression and antioxidant capacity in liver and breast muscle.

Screening and comparative studies of conducting polymers for developing effective molecular imprinted sensors for copper, zinc superoxide dismutase

This work describes the design, development, and screening of conducting polymers based molecularly imprinting sensors (MIP) for copper, zinc superoxide dismutase (SOD1). It is clinically significant for a wide variety of cardiovascular, neurodegenerative, Covid-19, and chronic immune illness. The SOD1 MIP sensors were undertaken by electropolymerization of various monomers on Screen Printed carbon electrode (SPCE) using cyclic voltammetry (CV) to examine the molecular recognition capability. The MIP receptors film binding towards SOD1 was studied by fitting experimental CV data to the Langmuir and Freundlich isotherms. Among the various monomers EDOT (3,4-ethylenedioxythiophene), Py (Pyrrole), and DA (Dopamine), the binding affinity (KL) of the poly(3-amionphenylboronic acid) (P3APBA) imprinted MIP system was considerably higher than the other conducting polymer MIP systems. Based on the above studies, 3APBA was chosen to develop a molecularly imprinted poly(3-aminophenylboronic acid) (MIP3APBA) sensor for sensitive and selective detection of SOD1. This MIP3APBA sensor’s behavior and analytical ability were characterized by Cyclic Voltammetry (CV), Differential Pulse Voltammetry (DPV) and Electrochemical Impedance Spectroscopy (EIS). It showed a lowest detection limit of 0.4 μM and a linear range of 1 μM to 500 μM. Further, this electrochemical MIP3APBA sensor was also used to quantify SOD1 levels in plasma samples.

Lack of Sod1 induces disruption of innervation accompanied by Schwann cell proliferation in skeletal muscle

The neuromuscular junction (NMJ) is an excitatory synapse that constitutes the primary site of communication between the nervous system and skeletal muscle and therefore any degenerative changes at the NMJ have the potential to impair muscle function. The primary cellular components of the NMJ are the motor neuron, the muscle fiber and terminal Schwann cells (tSC) that surround the NMJ. The importance of redox homeostasis for the maintenance of NMJ integrity is supported by work from our group showing degenerative changes at NMJs in mice lacking copper zinc superoxide dismutase (CuZnSOD; Sod1 -/- ) as early as 4 months of age. Prior work from others suggests that tSCs contribute trophic support for the NMJ and facilitate motor unit remodeling, but changes in tSC structure and function under conditions of elevated oxidative stress have not been thoroughly explored. Using in vitro assays, we previously showed that primary Schwann cells treated with a sublethal dose of the reactive oxygen species inducing agent Paraquat (PQ) resulted in increased Schwann cell number, ki67+ staining, and gene expression of trophic factors and the critical NMJ organizing protein Agrin. Furthermore, differentiated C2C12 myotubes treated with conditioned media from PQ treated Schwann cell cultures showed increased number of acetylcholine receptor (AChR) plaques. The goal of the current study was to characterize NMJs in young Sod1 -/- mice (2-months) to determine the early effects of elevated oxidative stress on NMJ structure in vivo with a specific focus on tSCs. We hypothesized that loss of Sod1 would lead to aberrant tSC and NMJ structure associated with decreased muscle function. We assessed maximum isometric force, NMJ morphology, and number of Schwann cells in gastrocnemius muscles of WT (n = 3) and Sod1 -/- (n = 3) mice. In Sod1 -/- mice, muscle force was reduced by 26% ( p = 2e-4) with nerve stimulation compared to direct muscle stimulation, and overlap between motor nerve terminals and AChRs was reduced by nearly 60% compared to WT controls. Post-synaptic AChR area was increased by 37% in Sod1 -/- mice, and tSC area and numbers per NMJ were increased by 60% and 155%, respectively in Sod1 -/- mice. Finally, Schwann cells proximal to NMJs showed increased Ki67+ labeling in Sod1 -/- mice compared to WT mice. This analysis demonstrates that early changes in both pre- and post-synaptic NMJ components and reduced neurotransmission due to loss of Sod1 are associated with marked increases in tSC proliferation. Our data implicate a protective role of muscle-resident Schwann cells in preserving NMJ integrity in response to alterations in redox homeostasis that may be mediated in part through activation and proliferation of these cells. These findings indicate that further investigation of the dynamics between Schwann cells and NMJ function is important to increase understanding of neuromuscular degenerativeconditions that involve altered redox homeostasis such as sarcopenia. Supported by the University of Michigan Medical School Office of Research, the American Physiological Society, and NIH AR069620. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

ALS drug receives advisory panel’s thumbs-up

A panel of external advisers to the US Food and Drug Administration has given Biogen’s amyotrophic lateral sclerosis (ALS) treatment, tofersen, its unanimous support for accelerated approval. The recommendation, made March 22, came despite the drug’s failed Phase 3 trial; instead, it is based on biomarker data. The copper zinc superoxide dismutase 1 ( SOD1 ) mutation  is the second-most-common genetic mutation that causes familial ALS. Though the mutation affects only about 330 people in the US, they have heartbreaking stories to tell: during the hearing, one patient shared that her family has recorded 33 deaths from ALS across seven generations and counting. The first drug candidate for SOD1 -ALS, Biogen’s tofersen is an antisense oligonucleotide that binds to messenger RNA to silence the expression of the faulty SOD1 protein. But a reduced protein level doesn’t necessarily translate to improvement among patients. In a 6-month double-anonymous Phase 3 trial

Bone Marrow Mesenchymal Stem-Cell-Derived Exosomes Ameliorate Deoxynivalenol-Induced Mice Liver Damage.

Deoxynivalenol (DON) is a kind of Fusarium toxin that can cause a variety of toxic effects. DON is mainly metabolized and detoxified by the liver. When the concentration of DON exceeds the metabolic capacity of the liver, it will trigger acute or chronic damage to the liver tissue. Previous studies demonstrated that bone marrow mesenchymal stem-cell-secreted exosomes (BMSC-exos) reduce liver injury. Therefore, we issue a hypothesis that in vitro-cultured rat BMSC-secreted exos could ameliorate liver damage after 2 mg/kg bw/day of DON exposure. In total, 144 lipids were identified in BMEC-exos, including high polyunsaturated fatty acid (PUFA) levels. BMSC-exos treatment alleviated liver pathological changes and decreased levels of alanine aminotransferase, aspartate aminotransferase, inflammatory factors interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and lipid peroxidation. Otherwise, low or high BMSC-exos treatment obviously changes DON-induced hepatic oxylipin patterns. According to the results from our correlation network analysis, Pearson correlation coefficient analysis, and hierarchical clustering analysis, the top 10% oxidized lipids can be classified into two categories: one that was positively correlated with copper-zinc superoxide dismutase (Cu/Zn SOD) and another that was positively correlated with liver injury indicators. Altogether, BMSC-exos administration maintained normal liver function and reduced oxidative damage in liver tissue. Moreover, it could also significantly change the oxylipin profiles under DON conditions.

Evidence and Metabolic Implications for a New Non-Canonical Role of Cu-Zn Superoxide Dismutase

Copper-zinc superoxide dismutase 1 (SOD1) has long been recognized as a major redox enzyme in scavenging superoxide radicals. However, there is little information on its non-canonical role and metabolic implications. Using a protein complementation assay (PCA) and pull-down assay, we revealed novel protein-protein interactions (PPIs) between SOD1 and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (YWHAZ) or epsilon (YWHAE) in this research. Through site-directed mutagenesis of SOD1, we studied the binding conditions of the two PPIs. Forming the SOD1 and YWHAE or YWHAZ protein complex enhanced enzyme activity of purified SOD1 in vitro by 40% (p < 0.05) and protein stability of over-expressed intracellular YWHAE (18%, p < 0.01) and YWHAZ (14%, p < 0.05). Functionally, these PPIs were associated with lipolysis, cell growth, and cell survival in HEK293T or HepG2 cells. In conclusion, our findings reveal two new PPIs between SOD1 and YWHAE or YWHAZ and their structural dependences, responses to redox status, mutual impacts on the enzyme function and protein degradation, and metabolic implications. Overall, our finding revealed a new unorthodox role of SOD1 and will provide novel perspectives and insights for diagnosing and treating diseases related to the protein.

Tobacco smoke induces oxidative stress and alters pro-inflammatory cytokines and some trace elements in healthy indoor cats.

This study was aimed to assess oxidative stress, pro-inflammatory cytokines and some trace elements in healthy pet cats exposed to environmental tobacco smoke. Forty healthy cats were included in this study. Cats were divided in two groups: Exposed to tobacco smoke (ETS; n = 20) and non-exposed to tobacco smoke (NETS; n = 20). Blood levels of cotinine, total oxidant status (TOS), oxidative stress index (OSI), lipid hydroperoxide (LOOH), protein carbonyl (PCO), advanced oxidative protein products (AOPP), total antioxidant status (TAS), copper, zinc-superoxide dismutase (Cu, Zn-SOD), catalase (CAT), total thiol (T-SH), interferon gamma (INF-γ), tumor necrosis factor (TNF-α), interleukin β (IL-1β), interleukin 6 (IL-6), interleukin-8 (IL-8), inter-leukin 2 (IL-2) and iron (Fe), zinc (Zn), copper (Cu), selenium (Se) levels were measured. Hematological and biochemical parameters were also measured. Serum cotinine, TOS, OSI, PCO, AOPP and LOOH levels were higher, whereas TAS and Cu, Zn-SOD levels were lower in ETS group. In ETS group INF-γ, IL-1β, IL-2, and IL-6 levels were higher. The Cu level was higher in ETS group. Blood reticulocyte number, serum creatinine and glucose were higher in ETS group. It could be concluded that exposure to tobacco smoke in cats impaired the oxidant/antioxidant balance and potentially triggered the release of pro-inflammatory cytokines.

Alterations in the Expression of Antioxidant Enzyme Genes in Response to Pesticide Exposure During Embryonic Development in the Native Reptile Species Caiman latirostris

The aim of this study was to quantify the expression levels of Catalase (cat) and copper, zinc Superoxide dismutase (Cu, Zn-sod) genes involved in the antioxidant response in Caiman latirostris (broad-snouted caiman) blood, after embryonic exposure to the formulations cypermethrin (CYP), chlorpyrifos (CPF), glyphosate (GLY), and their binary and ternary mixtures. Experimental groups were: negative control (NC-distilled water), vehicle control (VC-ethanol), GLY-2%, CYP- 0.12%, CPF- 0.8%, a ternary mixture of them (TM), and three binary mixtures. The applications were made on the nest material in contact with the eggs at the beginning of the incubation period. After hatching, RNA was isolated from blood and expression levels analyzed through qPCR. The results showed downregulation in the expression of sod and cat genes in the three binary mixtures studied, compared to the controls. In addition, we found a possible antagonistic effect between different pesticides in the TM on the expression of both genes.

Organic copper promoted copper accumulation and transport, enhanced low temperature tolerance and physiological health of white shrimp (Litopenaeus vannamei Boone, 1931)

This study was conducted to assess the effects of dietary copper source and level on hematological parameters, copper accumulation and transport, resistance to low temperature, antioxidant capacity and immune response of white shrimp (Litopenaeus vannamei Boone, 1931). Seven experimental diets with different copper sources and levels were formulated: C, no copper supplementation; S, 30 mg/kg copper in the form of CuSO4·5H2O; SO, 15 mg/kg copper in CuSO4·5H2O + 7.5 mg/kg copper in Cu-proteinate; O1, O2, O3 and O4, 10, 20, 30 and 40 mg/kg copper in the form of Cu-proteinate, respectively. A total of 840 shrimp (5.30 ± 0.04 g) were randomly distributed to 21 tanks (3 tanks/diet, 40 shrimp/tank). An 8-week feeding trial was conducted. The results showed that there was no significant difference in growth performance and whole shrimp chemical compositions among all groups. Compared with inorganic copper, dietary organic copper (O2 and O3) increased total protein, albumin, and glucose content of plasma, while decreased triglyceride and total cholesterol of plasma. Copper concentration in plasma and muscle and gene expression of metallothionein and copper-transporting ATPase 2 like in hepatopancreas were higher in shrimp fed organic copper (SO, O2, O3 and O4). The lowest mortality after low temperature (10 °C) challenge test was observed in the O2 and O3 groups. Organic copper (SO, O2, O3 and O4) significantly enhanced the antioxidant capacity (in terms of higher activities of total superoxide dismutase, copper zinc superoxide dismutase, catalase, glutathione peroxidase and total antioxidant capacity, lower malondialdehyde concentration of plasma, and up-regulated gene expression of superoxide dismutase, copper zinc superoxide dismutase, catalase and glutathione peroxidase of hepatopancreas). Organic copper (SO, O2, O3 and O4) enhanced the immune response (in terms of higher number of total hemocytes, higher activities of acid phosphatase, alkaline phosphatase, phenoloxidase, hemocyanin and lysozyme in plasma, and higher gene expressions of alkaline phosphatase, lysozyme and hemocyanin in hepatopancreas). Inorganic copper (Diet S) also had positive effects on white shrimp compared with the C diet, but the SO, O2, O3 and O4 diets resulted in better results, among which the O2 diet appeared to be the best one. In conclusion, organic copper was more beneficial to shrimp health than copper sulfate.

Resveratrol ameliorates oxidative stress, inflammatory response and lipid metabolism in common carp (Cyprinus carpio) fed with high-fat diet

High-fat diet is regarded as crucial inducers of oxidative stress, inflammation, and metabolic imbalance. In order to investigate the ameliorative potential of resveratrol against the progression of liver injury towards steatohepatitis, common carp (Cyprinus carpio) were distributed into six experimental groups and were fed with a normal-fat diet, a high-fat diet, and supplemented with resveratrol (0.8, 1.6, 2.4, and 3.2 g/kg diet) for 8 weeks. The high-fat diet decreased the antioxidant capacities, as well as causing the inflammatory response and lipid deposition of common carp. Resveratrol induced a marked elevation in the final body weight, weight gain rate, condition factor and significant decrease in the feed conversion ratio. Moreover, dietary resveratrol showed a significant decrease in the alanine aminotransferase, aspartate aminotransferase, triglyceride and low-density lipoprotein levels, which was accompanied by an increase in high-density lipoprotein concentration in serum. A significant elevation in total superoxide dismutase, catalase, glutathione peroxidase and a decreased malondialdehyde content were observed, along with a substantial elevation in antioxidant activities were found. Additionally, fish fed with resveratrol had an up-regulation of hepatic catalase, copper, zinc superoxide dismutase, glutathione peroxidase 1a, and glutathione peroxidase 1b gene expression via Nrf2 signaling pathway. Expectedly, our results also demonstrated that resveratrol regulates hepatic lipid metabolism in fish by inhibiting the expression of hepatic lipogenesis genes (acetyl-CoA carboxylase 1, fatty acid synthase, and sterol regulatory element binding protein 1), fatty acid uptake-related genes of lipoprotein lipase, and β-oxidation-related genes via PPAR-γ signaling pathway. Furthermore, dietary resveratrol reduced inflammation, as evident by down-regulating the interleukin-1β, interleukin-6, interleukin-8, and tumor necrosis factor-α expression levels and upregulating the interleukin-10 and transforming growth factor-β2 expression levels via NF-κB signaling pathway. As a whole, our results demonstrated that resveratrol defensed the impacts against high-fat diet on the serum biochemical, hepatic antioxidants, inflammation, and lipid metabolism.

Effect of replacing inorganic minerals with small peptide chelated minerals on production performance, some biochemical parameters and antioxidant status in broiler chickens.

Due to the low bio-availability of inorganic trace minerals, its application in poultry production has been causing many problems such as environment pollution and waste of resources. The current study was designed to evaluate if replacing inorganic trace minerals (ITM) with small peptide chelate trace minerals (SPM) affects production performance, some biochemical parameters and antioxidant status, tibia mineral deposition, and fecal mineral content in 817 white-feathered broilers. A total of 432 broilers (21-day-old) were randomly divided into four groups with six replicates of 18 chicks each. The four groups included inorganic trace minerals group (addition of 1,000mg/kg ITM; common practice by commercial poultry farms), three organic trace minerals groups with supplementation of 150, 300, and 500mg/kg SPM, respectively. The experiment lasted for 30days. The results showed that there was no significant difference in growth performance and slaughter performance among the four groups (p > 0.05). Total cholesterol in the SPM group was significantly lower than those in the ITM groups (p < 0.01). Compared with the ITM group, the serum urea nitrogen in 150 and 300mg/kg SPM groups decreased significantly (p < 0.01). Among all SPM treatments, 300mg/kg SPM groups had the highest serum glutathione peroxidase (GSH-Px) activity (p < 0.01). The activity of copper and zinc superoxide dismutase (Cu/Zn SOD) of liver in ITM group was the lowest among the four groups (p < 0.01). The catalase (CAT) activity of liver in the 150mg/kg SPM group was significantly higher than the ITM group and 300mg/kg SPM group (p < 0.05). Compared to the ITM group, the iron content of the tibia was significantly increased in 300mg/kg SPM group (p < 0.05) and 500mg/kg SPM group (p < 0.01). Compared to the ITM group, dietary supplementation with SPM significantly reduced fecal content of zinc and manganese (p < 0.01). The 150mg/kg SPM and 300mg/kg SPM group had significantly reduced content of iron (p < 0.05). This study demonstrated that replacing inorganic minerals with low doses of SPM (300 and 500mg/kg) did not negatively affect growth and slaughter performance, as well as the antioxidant status of broiler chickens. In addition, SPM can also promote mineral content in the tibia and reduce mineral content in the feces.

Heat Shock Proteins and Antioxidant Genes Involved in Heat Combined with Drought Stress Responses in Perennial Rye Grass.

The frequent occurrence of heat and drought stress can severely reduce agricultural production of field crops. In comparison to a single stress, the combination of both heat (H) and drought (D) further reduce plant growth, survival and yield. This study aimed to explore the transcriptional responses of heat shock protein (HSP) and antioxidant genes under H combined D stress in perennial rye grass (PRG). The results demonstrated that oxidative stress indicators (hydrogen peroxide, lipid peroxidation) significantly increased, particularly in the case of combined H and D treatment, suggesting that oxidative stress-induced damage occurred in plants under the combined stresses. Transcriptional responses of heat shock protein 70 (HSP70), heat shock protein 90-6 (HSP90-6), and the mitochondrial small heat shock protein HSP26.2 (HSP26.2) occurred rapidly, and showed high level of expression particularly under H and D stress. Antioxidant genes including ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), catalase (CAT), copper–zinc superoxide dismutase (Cu/ZnSOD), peroxidase (POD), ferredoxin–thioredoxin (FTR), thioredoxin (Trx), 2-cysteine peroxiredoxin (2-Cys Prx) showed response to combined H and D, followed by either D or H stress alone in rye grass. An interactome map revealed the close partnership of these heat shock protein genes and antioxidant genes, respectively. These candidate genes were predominantly linked to stress responses and antioxidant defense in plants. These findings may advance our understanding about the HSP and the antioxidant genes underlying combined abiotic stress response and tolerance in perennial rye grass.

Glycine-serine-rich effector PstGSRE4 in Puccinia striiformis f. sp. tritici inhibits the activity of copper zinc superoxide dismutase to modulate immunity in wheat.

Puccinia striiformis f. sp. tritici (Pst) secretes an array of specific effector proteins to manipulate host immunity and promote pathogen colonization. In a previous study, we functionally characterized a glycine-serine-rich effector PstGSRE1 with a glycine-serine-rich motif (m9). However, the mechanisms of glycine-serine-rich effectors (GSREs) remain obscure. Here we report a new glycine-serine-rich effector, PstGSRE4, which has no m9-like motif but inhibits the enzyme activity of wheat copper zinc superoxide dismutase TaCZSOD2, which acts as a positive regulator of wheat resistance to Pst. By inhibiting the enzyme activity of TaCZSOD2, PstGSRE4 reduces H2O2 accumulation and HR areas to facilitate Pst infection. These findings provide new insights into the molecular mechanisms of GSREs of rust fungi in regulating plant immunity.

Oxidative stress and impaired insulin secretion in cystic fibrosis pig pancreas

Cystic fibrosis-related diabetes (CFRD) is one the most common comorbidities in cystic fibrosis (CF). Pancreatic oxidative stress has been postulated in the pathogenesis of CFRD, but no studies have been done to show an association. The main obstacle is the lack of suitable animal models and no immediate availability of pancreas tissue in humans. In the CF porcine model, we found increased pancreatic total glutathione (GSH), glutathione disulfide (GSSG), 3-nitrotyrosine- and 4-hydroxynonenal-modified proteins, and decreased copper zinc superoxide dismutase (CuZnSOD) activity, all indicative of oxidative stress. CF pig pancreas demonstrated increased DHE oxidation (as a surrogate marker of superoxide) in situ compared to non-CF and this was inhibited by a SOD-mimetic (GC4401). Catalase and glutathione peroxidase activities were not different between CF and non-CF pancreas. Isolated CF pig islets had significantly increased DHE oxidation, peroxide production, reduced insulin secretion in response to high glucose and diminished secretory index compared to non-CF islets. Acute treatment with apocynin or an SOD mimetic failed to restore insulin secretion. These results are consistent with the hypothesis that CF pig pancreas is under significant oxidative stress as a result of increased O2●− and peroxides combined with reduced antioxidant defenses against reactive oxygen species (ROS). We speculate that insulin secretory defects in CF may be due to oxidative stress.

Molecular characterization of two CuZn-SOD family proteins in the Pacific oyster Crassostrea gigas.

Superoxide dismutases (SOD) are multifamily antioxidant enzymes, playing an important role in the defense against oxidative stress in all organisms. Genomic information indicated the presence of genetic diversification of the copper and zinc SOD (CuZn-SOD) family in oysters. In the present research, we characterized two CuZn-SOD family proteins, Cg-CuZn-SOD and Cg-dominin3, in the Pacific oyster Crassostrea gigas using comprehensive sequence analyses, recombinant proteins and site-directed mutagenesis, and observations of gene expression in larval and adult oysters. We found that Cg-CuZn-SOD possessed sequence and structural elements conserved in a CuZn-SOD molecule and the recombinant protein was confirmed empirically to have the SOD enzyme activity. In contrast, Cg-dominin3 lacked five of the seven residues essential for the conformation of SOD active center and the recombinant protein did not have the enzyme activity. However, recombinant Cg-dominin3 showed strong binding activities toward zinc and copper ions. Substitutions of five conserved His residues in the active center demolished the SOD activity but enhanced the metal binding capacity in Cg-CuZn-SOD. On the other hand, reinstallation of the five His residues that were assumed to be activity essential and lost in evolution did not restore the SOD enzyme activity in Cg-dominin3. Additionally, the coding genes of the two proteins exhibited different patterns of expression during larval development and in adult oyster in response to zinc challenges. These results have led to the discovery of the first cytoplasmic CuZn-SOD molecule and the confirmation of molecular diversification of extracellular CuZn-SOD homologs in oysters.