Redox Defense Research Articles

ER-GUARD: an evolutionarily conserved antioxidant defense system at ER membranes.

Oxidative protein folding in the endoplasmic reticulum (ER) is essential for all eukaryotic cells yet generates hydrogen peroxide (H2O2), a reactive oxygen species (ROS). The ER-transmembrane protein that provides reducing equivalents to ER and guards the cytosol for antioxidant defense remains unidentified. Here we combine AlphaFold2-based and functional reporter screens in C. elegans to identify a previously uncharacterized and evolutionarily conserved protein ERGU-1 that fulfills these roles. Deleting C. elegans ERGU-1 causes excessive H2O2 and transcriptional gene up-regulation through SKN-1, homolog of mammalian antioxidant master regulator NRF2. ERGU-1 deficiency also impairs organismal reproduction and behaviors. Both C. elegans and human ERGU-1 proteins localize to ER membranes and form network reticulum structures. We name this system ER-GUARD, Endoplasmic Reticulum Guardian Aegis of Redox Defense. Human and Drosophila homologs of ERGU-1 can rescue C. elegans mutant phenotypes, demonstrating evolutionarily ancient and conserved functions. Together, our results reveal an ER-membrane-specific protein machinery and defense-net system ER-GUARD for peroxide detoxification and suggest a previously unknown but conserved pathway for antioxidant defense in animal cells.

The Role of One-Carbon Metabolism and Methyl Donors in Medically Assisted Reproduction: A Narrative Review of the Literature.

One-carbon (1-C) metabolic deficiency impairs homeostasis, driving disease development, including infertility. It is of importance to summarize the current evidence regarding the clinical utility of 1-C metabolism-related biomolecules and methyl donors, namely, folate, betaine, choline, vitamin B12, homocysteine (Hcy), and zinc, as potential biomarkers, dietary supplements, and culture media supplements in the context of medically assisted reproduction (MAR). A narrative review of the literature was conducted in the PubMed/Medline database. Diet, ageing, and the endocrine milieu of individuals affect both 1-C metabolism and fertility status. In vitro fertilization (IVF) techniques, and culture conditions in particular, have a direct impact on 1-C metabolic activity in gametes and embryos. Critical analysis indicated that zinc supplementation in cryopreservation media may be a promising approach to reducing oxidative damage, while female serum homocysteine levels may be employed as a possible biomarker for predicting IVF outcomes. Nonetheless, the level of evidence is low, and future studies are needed to verify these data. One-carbon metabolism-related processes, including redox defense and epigenetic regulation, may be compromised in IVF-derived embryos. The study of 1-C metabolism may lead the way towards improving MAR efficiency and safety and ensuring the lifelong health of MAR infants.

Improving effect of N-acetylcysteine on growth and intestinal health in juvenile Cyprinus carpio involve the Nrf2/MAPK/NF-κB pathways

N-acetylcysteine (NAC) plays a beneficial role in improving animal growth and health. The current study evaluated the impact of dietary supplementation with NAC on the growth performance, physiological status and intestinal health of juvenile Cyprinus carpio. The experiment involved 5 groups, with 3 replicates in each group, and each replicate group containing 30 juvenile Cyprinus carpio (average weight 2.06 ± 0.01 g). Five experimental diets (36% crude protein and 9% crude lipid) were prepared by supplementing graded levels (0, 0.15%, 0.30%, 0.45% and 0.60%) of NAC in the basal diet. Subsequently, these 5 diets were utilized for an 56 d feeding trial with Cyprinus carpio. During the feeding trial, the water temperature was maintained between 24 and 28 °C, and the feeding rate was 3%–5% of body weight. After the feeding trial, fish were fasted for 24 h, anesthetized, and dissected on ice trays to collect samples for subsequent analysis. Results showed that NAC (0.30%, 0.45% and 0.60% diets) significantly improved the growth performance and intestinal mucosa development in Cyprinus carpio, and inhibited apoptosis. Similarly, NAC (0.30%, 0.45% and 0.60% diets) could enhance the homeostasis of the redox defense system in intestine of Cyprinus carpio, which is linked to enhanced glutathione metabolism, scavenging of reactive oxygen species and activation of Nrf2 pathway. Furthermore, dietary NAC improved anti-inflammatory and immunomodulatory functions, and involved in the regulation of MAPK/NF-κB signaling pathway. Summary, NAC (0.30%, 0.45% and 0.60% diets) could significantly promote the growth and intestinal health in Cyprinus carpio. These findings provide partial available information for promoting the utilization of NAC in aquafeed and improving the ecological and healthy cultivation of Cyprinus carpio.

The Effectiveness of Four Nicotinamide Adenine Dinucleotide (NAD+) Precursors in Alleviating the High-Glucose-Induced Damage to Hepatocytes in Megalobrama amblycephala: Evidence in NAD+ Homeostasis, Sirt1/3 Activation, Redox Defense, Inflammatory Response, Apoptosis, and Glucose Metabolism.

The administration of NAD+ precursors is a potential approach to protect against liver damage and metabolic dysfunction. However, the effectiveness of different NAD+ precursors in alleviating metabolic disorders is still poorly elucidated. The current study was performed to compare the effectiveness of four different NAD+ precursors, including nicotinic acid (NA), niacinamide (NAM), nicotinamide riboside (NR), and nicotinamide mononucleotide (NMN) in alleviating high-glucose-induced injury to hepatocytes in a fish model, Megalobrama amblycephala. An in vitro high-glucose model was successfully established to mimic hyperglycemia-induced damage to the liver, which was evidenced by the reduced cell viability, the increased transaminase activity, and the depletion of cellular NAD+ concentration. The NAD+ precursors all improved cell viability, with the maximal effect observed in NR, which also had the most potent NAD+ boosting capacity and a significant Sirt1/3 activation effect. Meanwhile, NR presented distinct and superior effects in terms of anti-oxidative stress, inflammation inhibition, and anti-apoptosis compared with NA, NAM, and NMN. Furthermore, NR could effectively benefit glucose metabolism by activating glucose transportation, glycolysis, glycogen synthesis and the pentose phosphate pathway, as well as inhibiting gluconeogenesis. Moreover, an oral gavage test confirmed that NR presented the most potent effect in increasing hepatic NAD+ content and the NAD+/NADH ratio among four NAD+ precursors. Together, the present study results demonstrated that NR is most effective in attenuating the high-glucose-induced injury to hepatocytes in fish compared to other NAD+ precursors.

The P4' Peptide-Carrying Bacillus subtilis in Cottonseed Meal Improves the Chinese Mitten Crab Eriocheir sinensis Innate Immunity, Redox Defense, and Growth Performance.

This study developed a recombinant Bacillus subtilis to carry the LGSPDVIVIR peptide (cmP4) isolated from the hydrolyzed products of cottonseed meal with excellent antioxidant and immune-enhancing properties in vitro. It was carried as a tandem of five cmP4 peptides (cmP4') to be stably expressed on a large scale. Then, its effectiveness was evaluated in Chinese mitten crab (Eriocheir sinensis) based on growth performance, redox defense, and innate immunity. A total of 280 crabs (mean body weight: 41.40 ± 0.14) were randomly assigned to seven diets including a control one (without B. subtilis) and six experimental ones with different doses (107,108, and 109 CFU/kg) of unmodified and recombinant B. subtilis, respectively, for 12 weeks. Each diet was tested in four tanks of crabs (10/tank). In terms of bacterial dosages, the final weight (FW), weight gain (WG), hemolymph and hepatopancreatic activities of superoxide dismutase (SOD), catalase (CAT), lysosome (LZM), acid phosphatase (ACP) and alkaline phosphatase (AKP), and hepatopancreatic transcriptions of cat, mitochondrial manganese superoxide dismutase (mtmnsod), thioredoxin-1 (trx1), and prophenoloxidase (propo) all increased significantly with increasing B. subtilis dosages, while hemolymph and hepatopancreatic malondialdehyde (MDA) content and the transcriptions of toll like receptors (tlrs), NF-κB-like transcription factor (relish), and lipopolysaccharide-induced TNF-α factor (litaf) all decreased remarkably. In terms of bacterial species, the recombinant B. subtilis group obtained significantly high values of FW, WG, hemolymph, and hepatopancreatic activities of SOD, CAT, LZM, ACP, and AKP, and the transcriptions of mtmnsod, peroxiredoxin 6 (prx6), and propo compared with the unmodified B. subtilis, while opposite results were noted in hemolymph and hepatopancreatic MDA content and the transcriptions of tlrs, relish, and litaf. These results indicated that dietary supplementation with 109 CFU/kg of recombinant B. subtilis can improve the growth performance, redox defense, and nonspecific immunity of E. sinensis.

Targeting one-carbon metabolism for cancer immunotherapy.

One-carbon (1C) metabolism is a metabolic network that plays essential roles in biological reactions. In 1C metabolism, a series of nutrients are used to fuel metabolic pathways, including nucleotide metabolism, amino acid metabolism, cellular redox defence and epigenetic maintenance. At present, 1C metabolism is considered the hallmark of cancer. The 1C units obtained from the metabolic pathways increase the proliferation rate of cancer cells. In addition, anticancer drugs, such as methotrexate, which target 1C metabolism, have long been used in the clinic. In terms of immunotherapy, 1C metabolism has been used to explore biomarkers connected with immunotherapy response and immune-related adverse events in patients. We collected numerous literatures to explain the roles of one-carbon metabolism in cancer immunotherapy. In this review, we focus on the important pathways in 1C metabolism and the function of 1C metabolism enzymes in cancer immunotherapy. Then, we summarise the inhibitors acting on 1C metabolism and their potential application on cancer immunotherapy. Finally, we provide a viewpoint and conclusion regarding the opportunities and challenges of targeting 1C metabolism for cancer immunotherapy in clinical practicability in the future. Targeting one-carbon metabolism is useful for cancer immunotherapy.

Tumor necrosis factor α1 (TNFα1) administration can disrupt barrier function and attenuate redox defense in midgut of red crucian carp (Carassius auratus red var)

TNFα belongs to superfamily of tumor necrosis factor that can exert the pleiotropic effort in a large quantity of biological processes, including immune homeostasis, intracellular modulation and etiopathogenesis, but its regulatory role in mucosal immune regulation of fish is unclear. Currently, this study aimed to evaluate the immunoregulatory function of TNFα1 on gut barrier in teleost fish. In this study, TNFα1 sequences were identified from red crucian carp (RCC, Carassius auratus red var). The high-level expression of RCC-TNFα1 mRNA was detected in gill among all the isolated samples. Then, RCC-TNFα1 expression increased dramatically in immune-related tissues after A. hydrophila infection and in cultured fish cells after lipopolysaccharide (LPS) treatment. RCC-TNFα1 fusion protein was generated and purified in vitro. RCC receiving RCC-TNFα1 perfusion showed an increased levels of villi fusion and edema in injured midgut with the fuzzy appearance. In addition, The mRNA expressions of tight junction (TJ) genes, mucin genes and redox sensitive genes decreased sharply in TNFα1-treated midgut in comparison with those of the control (P < 0.05), whereas the expression levels of apoptotic genes involved in caspase signals and unfolded protein response (UPR) attained the dramatic increase. These results demonstrated that RCC-TNFα1 stimulation could impair midgut structural integrity and immune function by induction of antioxidant collapse and apoptotic activation.

New insights into searching patulin degrading enzymes in Saccharomyces cerevisiae through proteomic and molecular docking analysis

Global warming has increased the contamination of mycotoxins. Patulin (PAT) is a harmful contaminant that poses a serious threat to food safety and human health. Saccharomyces cerevisiae biodegrades PAT by its enzymes during fermentation, which is a safe and efficient method of detoxification. However, the key degradation enzymes remain unclear. In this study, the proteomic differences of Saccharomyces cerevisiae under PAT stress were investigated. The results showed that the proteins involved in redox reactions and defense mechanisms were significantly up-regulated to resist PAT stress. Subsequently, molecular docking was used to virtual screen for degrading enzymes. Among 18 proteins, YKL069W showed the highest binding affinity to PAT and was then expressed in Escherichia coli, where the purified YKL069W completely degraded 10 μg/mL PAT at 48 h. YKL069W was demonstrated to be able to degrade PAT into E-ascladiol. Molecular dynamics simulations confirmed that YKL069W was stable in catalyzing PAT degradation with a binding free energy of − 7.5 kcal/mol. Furthermore, it was hypothesized that CYS125 and CYS101 were the key amino acid residues for degradation. This study offers new insights for the rapid screening and development of PAT degrading enzymes and provides a theoretical basis for the detoxification of mycotoxins.

Proteomic analysis of the faba bean-wheat intercropping system in controlling the occurrence of faba bean fusarium wilt due to stress caused by Fusarium oxysporum f. sp. fabae and benzoic acid

BackgroundIn faba bean, continuous cropping severely affects plant growth and increases the incidence of fusarium wilt due to the accumulation of pathogens and autotoxic substances. The intercropping of faba bean and wheat is commonly used to alleviate the occurrence of fusarium wilt in the faba bean.ObjectiveTo investigate the role of Fusarium oxysporum f. sp. Fabae(FOF) and benzoic acid in the occurrence of faba bean fusarium wilt and unravel the potential mechanism of intercropping in alleviating its occurrence.MethodsHydroponic experiment was carried out using monocropping faba bean (M) and intercropping faba bean and wheat (I) patterns under FOF alone stress (M + F, I + F), FOF and benzoic acid double stress (M + F + B, I + F + B). The growth of faba bean seedlings under FOF and benzoic acid dual stresses were analyzed as well as the protein expression profile of monocropping and intercropping faba bean roots.ResultUnder FOF stress, the growth of faba bean seedlings was inhibited, and the inhibitory effect was enhanced under the dual stress of FOF and benzoic acid. However, faba bean-wheat intercropping alleviated the inhibitory effect of FOF and benzoic acid on faba bean growth. In faba bean, the up-regulated protein was involved in different functions, such as redox, hydrogen peroxide decomposition, and metabolic processes under FOF stress (M + F, I + F) compared to the control. Compared with FOF stress (M + F, I + F), under the dual stress of FOF and benzoic acid (M + F + B, I + F + B), the up-regulated protein in faba bean were involved in intracellular redox balance, defense, and maintenance of cell integrity. Compared with monocropping (M, M + F, M + F + B), the up-regulated protein function of intercropping(I, I + F, I + F + B) was mainly involved in the biosynthesis of secondary metabolites, redox balance, biological carbon fixation of photosynthesis, and so on. KEGG enrichment analysis results showed that intercropping increased ethylene and jasmonic acid synthesis and other related pathways to improve resistance against fusarium wilt in the faba bean.ConclusionThe growth of faba bean was inhibited under FOF stress and the inhibitory effect was enhanced under the dual stress of FOF and benzoic acid, which promoted the occurrence of faba bean fusarium wilt. This might be due to the down-regulation of energy and cytoplasmic matrix proteins under FOF and benzoic acid stress. The faba bean wheat intercropping alleviated the inhibition of FOF and benzoic acid stress by up-regulating the biosynthesis of secondary metabolites, redox homeostasis, photosynthetic carbon fixation, and other related proteins. Besides, it also promoted the biosynthesis of ethylene, and jasmonic acid, improved the resistance of faba bean plants, and alleviated the occurrence of faba bean fusarium wilt. This provides a theoretical basis for the determination of jasmonic acid and ethylene content.

Thiamine-induced resistance in carnation against Fusarium oxysporum f. sp dianthi and mode of action studies based on the proteomics analysis of root tissue

Colombia is the main producer of carnations worldwide. However, its production is affected by vascular wilting caused by Fusarium oxysporum f. sp dianthi (Fod). Previous studies have proposed thiamine as a potential inducer of resistance against this pathogen, but the biochemical mechanisms involved in its action have not been elucidated. The objective of the present work was to investigate the underlying mechanism of thiamine–induced resistance by analyzing changes in the root protein profile. A greenhouse experiment was performed with two cultivars, evaluating the following treatments: control, Fod–inoculated plants, foliar application of thiamine, and application of thiamine and subsequent Fod inoculation. Incidence and severity were determined for each treatment. Applicating thiamine reduced the incidence (22%) and severity (62%) of vascular wilting in a susceptible cultivar confirming the effect as Resistance Inducer. In addition, a shotgun (nLC–MS/MS) proteomic analysis of root tissues was carried out at 96 h post–inoculation. The application of thiamine and subsequent inoculation with Fod, increased the abundance of 44 proteins in the susceptible cultivar and 70 in the resistant one. These proteins belonged to different functional groups, including protein synthesis and degradation, redox homeostasis, transport, cell organization, stress, and defense. In conclusion, the results obtained suggest that the treatment with thiamine previously to Fod inoculation allows the early activation of defense mechanisms that protect the plant against the pathogen. These results increase the knowledge of thiamine use as a biostimulant agent in plants and its protein–level mode of action against pathogens.

Effects of Curcumin Treatment on Cell Energy Status, Levels of Mitochondrial Enzymes, and Gene Expression of Glucose-related Mechanism in Pancreatic Cancer Cell Lines

Background and Purpose:. Curcumin is an active component of turmeric, has antitumor, immunomodulatory, anti-inflammatory effects. It was aimed to investigate the effects of the administration of curcumin on the energy metabolism, the abnormal redox defense mechanism profile, the malignant transformation indicator of Panc-1 and BxPC-3 pancreatic cancer cells. &#x0D; Methods: BxPC-3 and Panc-1 cells were incubated, were replaced with containing various concentrations of curcumin (10-125 μM) for 24 h. Cell lysate Adenosine triphosphate (ATP), Adenosine diphosphate (ADP), Adenosine monophosphate (AMP), Manganese superoxidase (MnSOD), and cytochrome p450 reductase (CPR) concentrations were analyzed with HPLC and ELISA methods. Genes expression of Lactate dehydrogenase (LDH), mitochondrially encoded ATP synthase membrane subunit 6 (MTATP6), Glucose transporter 1 (GLUT1), and cytochrome p450 were analyzed. &#x0D; Results and Conclusion: IC50 values for 24 hours were found as 47,26 μM in BxPC-3 and 45,84 μM in Panc-1 cells. Treatment with curcumin inhibits oxidative stress by increasing MnSOD enzyme levels. ATP levels did not change in BxPC-3 cells, but it showed an increase in Panc-1 supplemented with curcumin. The effects of curcumin on GLUT-1 are significantly important at a dose of curcumin of 45 μM concentration and affect glucose consumption in both cells. Curcumin showed anti-proliferative, and antioxidant effects.

MicroRNAs-associated with FOXO3 in cellular senescence and other stress responses.

FOXO3 is a member of the FOXO transcription factor family and is known for regulating cellular survival in response to stress caused by various external and biological stimuli. FOXO3 decides cell fate by modulating cellular senescence, apoptosis and autophagy by transcriptional regulation of genes involved in DNA damage response and oxidative stress resistance. These cellular processes are tightly regulated physiologically, with FOXO3 acting as the hub that integrates signalling networks controlling them. The activity of FOXO3 is influenced by post-translational modifications, altering its subcellular localisation. In addition, FOXO3 can also be regulated directly or indirectly by microRNAs (miRNAs) or vice versa. This review discusses the involvement of various miRNAs in FOXO3-driven cellular responses such as senescence, apoptosis, autophagy, redox and inflammation defence. Given that these responses are linked and influence cell fate, a thorough understanding of the complex regulation by miRNAs would provide key information for developing therapeutic strategy and avoid unintended consequences caused by off-site targeting of FOXO3.

Role of mitochondrial disruption and oxidative stress in plasticizer phthalate-induced cytotoxicity to human bone osteoblasts.

Phthalates are frequently utilized in a wide range of products such as plasticizers with reported negative effects on bones. The current study evaluated the effect of butyl cyclohexyl phthalate on the human osteoblasts via different assays. MTT and lactate dehydrogenase assays were used to examine the in-vitro cytotoxic effect of butyl cyclohexyl phthalate on human bone osteoblasts in concentrations 0.1, 1, 10, and 100μM for 12 to 72h postexposures. Incubation of osteoblasts with butyl cyclohexyl phthalate significantly reduced cell viability based on its concentrations and durations of exposure. In parallel, osteoblast secretion of procollagen type 1, osteocalcin, as well as alkaline phosphatase was significantly decreased by butyl cyclohexyl phthalate in concentrations (1 or 2μM). Butyl cyclohexyl phthalate decreased ATP synthesis and mitochondrial complexes I and III activities, with increased lactate production, all of which were detrimental to cellular bioenergetics. The cellular redox defense systems were significantly depleted by increased lipid peroxidation, elevated reactive oxygen species, decreased catalase and superoxide dismutase enzymes activities, and decreased intracellular reduced glutathione (GSH). Redox stress was also induced. Interestingly, preincubating osteoblasts with reduced GSH before exposing them to butyl cyclohexyl phthalate significantly lowered the cytotoxicity of the butyl cyclohexyl phthalate, suggesting that antioxidants may play a helpful protective effect.

The Benefits of Nanosized Magnesium Oxide in Fish Megalobrama amblycephala: Evidence in Growth Performance, Redox Defense, Glucose Metabolism, and Magnesium Homeostasis.

This study evaluated the effects of dietary magnesium oxide nanoparticles (MgO NPs) on the growth, redox defense, glucose metabolism, and magnesium homeostasis in blunt snout bream. Fish (12.42 ± 0.33 g) were fed seven diets containing graded levels of MgO NPs (0, 60, 120, 240, 480, 960, and 1920 mg/kg) for 12 weeks. Whole-body Mg retention decreased significantly as the dietary Mg increased. As dietary MgO NPs levels reached 120 mg/kg, the growth performance and feed utilization remarkably improved. When added at 240 mg/kg, oxidative stress was significantly reduced evidenced by the increased Mn-sod transcription and the decreased CAT and GSH-Px activities and the MDA content. Meanwhile, it enhanced glucose transport, glycolysis, and glycogen synthesis, while inhibiting gluconeogenesis, as was characterized by the increased transcriptions of glut2, gk, and pk, and the decreased transcriptions of fbpase and g6pase. In addition, the supplementation of 120 mg/kg MgO NPs promoted Mg transport marked by a significant increase in the protein expressions of TRMP7, S41A3, and CNNM1. In conclusion, the moderate supplementation of MgO NPs improved the growth performance, reduced hepatic oxidative stress, and promoted glucose transport, glycolysis, glycogen synthesis, and magnesium homeostasis in fish while inhibiting glu.

Each Cellular Compartment Has a Characteristic Protein Reactive Cysteine Ratio Determining Its Sensitivity to Oxidation.

Signaling and detoxification of Reactive Oxygen Species (ROS) are important patho-physiologcal processes. Despite this, we lack comprehensive information on individual cells and cellular structures and functions affected by ROS, which is essential to build quantitative models of the effects of ROS. The thiol groups from cysteines (Cys) in proteins play a major role in redox defense, signaling, and protein function. In this study, we show that the proteins in each subcellular compartment contain a characteristic Cys amount. Using a fluorescent assay for -SH in thiolate form and amino groups in proteins, we show that the thiolate content correlates with ROS sensitivity and signaling properties of each compartment. The highest absolute thiolate concentration was found in the nucleolus, followed by the nucleoplasm and cytoplasm whereas protein thiolate groups per protein showed an inverse pattern. In the nucleoplasm, protein reactive thiols concentrated in SC35 speckles, SMN, and the IBODY that accumulated oxidized RNA. Our findings have important functional consequences, and explain differential sensitivity to ROS.

Radical cation scavenging activity of berberine bridge enzyme-like oligosaccharide oxidases acting on short cell wall fragments

Oligogalacturonide-oxidases (OGOXs) and cellodextrin-oxidase (CELLOX) are plant berberine bridge enzyme-like oligosaccharide-oxidases (OSOXs) that oxidize, respectively, oligogalacturonides (OGs) and cellodextrins (CDs), thereby inactivating their elicitor nature and concomitantly releasing H2O2. Little is known about the physiological role of OSOX activity. By using an ABTS·+-reduction assay, we identified a novel reaction mechanism through which the activity of OSOXs on cell wall oligosaccharides scavenged the radical cation ABTS·+ with an efficiency dependent on the type and length of the oxidized oligosaccharide. In contrast to the oxidation of longer oligomers such as OGs (degree of polymerization from 10 to 15), the activity of OSOXs on short galacturonan- and cellulose-oligomers (degree of polymerization ≤ 4) successfully counteracted the radical cation-generating activity of a fungal laccase, suggesting that OSOXs can generate radical cation scavenging activity in the apoplast with a power proportional to the extent of degradation of the plant cell wall, with possible implications for redox homeostasis and defense against oxidative stress.

Dietary-derived vitamin B12 protects Caenorhabditis elegans from thiol-reducing agents

BackgroundOne-carbon metabolism, which includes the folate and methionine cycles, involves the transfer of methyl groups which are then utilised as a part of multiple physiological processes including redox defence. During the methionine cycle, the vitamin B12-dependent enzyme methionine synthetase converts homocysteine to methionine. The enzyme S-adenosylmethionine (SAM) synthetase then uses methionine in the production of the reactive methyl carrier SAM. SAM-binding methyltransferases then utilise SAM as a cofactor to methylate proteins, small molecules, lipids, and nucleic acids.ResultsWe describe a novel SAM methyltransferase, RIPS-1, which was the single gene identified from forward genetic screens in Caenorhabditis elegans looking for resistance to lethal concentrations of the thiol-reducing agent dithiothreitol (DTT). As well as RIPS-1 mutation, we show that in wild-type worms, DTT toxicity can be overcome by modulating vitamin B12 levels, either by using growth media and/or bacterial food that provide higher levels of vitamin B12 or by vitamin B12 supplementation. We show that active methionine synthetase is required for vitamin B12-mediated DTT resistance in wild types but is not required for resistance resulting from RIPS-1 mutation and that susceptibility to DTT is partially suppressed by methionine supplementation. A targeted RNAi modifier screen identified the mitochondrial enzyme methylmalonyl-CoA epimerase as a strong genetic enhancer of DTT resistance in a RIPS-1 mutant. We show that RIPS-1 is expressed in the intestinal and hypodermal tissues of the nematode and that treating with DTT, β-mercaptoethanol, or hydrogen sulfide induces RIPS-1 expression. We demonstrate that RIPS-1 expression is controlled by the hypoxia-inducible factor pathway and that homologues of RIPS-1 are found in a small subset of eukaryotes and bacteria, many of which can adapt to fluctuations in environmental oxygen levels.ConclusionsThis work highlights the central importance of dietary vitamin B12 in normal metabolic processes in C. elegans, defines a new role for this vitamin in countering reductive stress, and identifies RIPS-1 as a novel methyltransferase in the methionine cycle.

Nanobiotechnology-based strategies for enhanced crop stress resilience.

Nanobiotechnology approaches to engineering crops with enhanced stress tolerance may be a safe and sustainable strategy to increase crop yield. Under stress conditions, cellular redox homeostasis is disturbed, resulting in the over-accumulation of reactive oxygen species (ROS) that damage biomolecules (lipids, proteins and DNA) and inhibit crop growth and yield. Delivering ROS-scavenging nanomaterials to plants has been shown to alleviate abiotic stress. Here we review the current state of knowledge of using ROS-scavenging nanomaterials to enhance plant stress tolerance. When present below a threshold level, ROS can mediate redox signalling and defence pathways that foster plant acclimatization against stress. We find that ROS-triggering nanomaterials, such as nanoparticulate silver and copper oxide, have the potential to be judiciously applied to crop species to stimulate the defence system, prime stress responses and subsequently increase the stress resistance of crops.

Evaluation of erythrocyte viability, antioxidant capacity and antiplasmodial activity induced by alkaloid extract of Phyllanthus amarus

Introduction: The production of large amounts of reactive oxygen species in severe malarial infection is due to parasite invasion to erythrocytes. Malaria resistance to medication has left malaria-endemic countries with no alternate source of medications but traditional medicine. One of such plants utilized by traditional healers is Phyllanthus amarus. Therefore, this study aims at ascertaining the antiplasmodial and cytotoxic activities of P. amarus and its specific actions on retaining erythrocyte viability and antioxidant activity. Methods: Antiplasmodial and erythrocyte viability activities were determined in vitro via parasite suppression and tetrazolium-based colorimetric assays, respectively. Antioxidant capacity was determined by measuring extract’s ability to inhibit lipid peroxidation, scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide ions, reduce iron(III) ions, and chelate iron in vitro using documented methods. Results: Alkaloid extracts of P. amarus showed great antiplasmodial activity (IC50=0.52μg/mL) and low cytotoxic activity (CC50=54.95 μg/mL). Erythrocyte viability assay showed the minimal impact of the extract on the uninfected erythrocytes but improved viability of the infected RBC in a dose dependent manner, and antioxidant activity manifested mainly in its iron chelating activity (EC50=0.34 μg/mL). Conclusion: This study suggests that the alkaloid extract of P. amarus has significant antiplasmodial and antioxidant activities. These activities promoted the repair of parasite-induced free radical damage to the erythrocyte membrane but distorted the parasites redox balance and defense mechanism, and hence survival rate as indicated by the parasite suppression associated with alkaloid extract treatment of malarial infection.

Selenoprotein: Potential Player in Redox Regulation in Chlamydomonas reinhardtii.

Selenium (Se) is an essential micro-element for many organisms, including Chlamydomonas reinhardtii, and is required in trace amounts. It is obtained from the 21st amino acid selenocysteine (Sec, U), genetically encoded by the UGA codon. Proteins containing Sec are known as selenoproteins. In eukaryotes, selenoproteins are present in animals and algae, whereas fungi and higher plants lack them. The human genome contains 25 selenoproteins, most of which are involved in antioxidant defense activity, redox regulation, and redox signaling. In algae, 42 selenoprotein families were identified using various bioinformatics approaches, out of which C. reinhardtii is known to have 10 selenoprotein genes. However, the role of selenoproteins in Chlamydomonas is yet to be reported. Chlamydomonas selenoproteins contain conserved domains such as CVNVGC and GCUG, in the case of thioredoxin reductase, and CXXU in other selenoproteins. Interestingly, Sec amino acid residue is present in a catalytically active domain in Chlamydomonas selenoproteins, similar to human selenoproteins. Based on catalytical active sites and conserved domains present in Chlamydomonas selenoproteins, we suggest that Chlamydomonas selenoproteins could have a role in redox regulation and defense by acting as antioxidants in various physiological conditions.