Non-protein Thiol Content Research Articles

Multiple Biomarker Responses in Aegla castro Exposed to Copper: A Laboratory Approach.

Although some biomarkers have already been determined in aeglids collected in the field, data from laboratory exposures are scarce. To our knowledge, no studies have investigated oxidative stress biomarkers in aeglids exposed to metals in the laboratory, or performed hemocyte counts and the comet assay using gill and hepatopancreas of aeglids. Thus, we investigated the effects of acute Cu exposure on intermolt males of Aegla castro, collected from a reference stream, acclimated for 6days in the laboratory, and then exposed to 11μgL-1 of dissolved Cu (Cu 11) or only to water (CTR), for 24h. Gill and hepatopancreas samples were used to determine Cu accumulation, DNA damage, and metallothionein content (MT), while hemolymph samples were used to determine Cu accumulation, DNA damage, and hemocyte counts. Muscle samples were used to determine Cu accumulation and acetylcholinesterase activity (AChE). Non-protein thiol content (NPSH), catalase (CAT), glutathione S-transferase activities (GST), lipoperoxidation (LPO), and protein carbonylation content (PCC) were measured only in the hepatopancreas. Aegla castro exposed to Cu accumulated this metal in gills and activated detoxification mechanisms, through increased MT content in the gill, and showed an immune response, evidenced by an increase in hyaline hemocytes. Therefore, gill and hemocytes appear to have a protective role in preventing the transport and bioavailability of Cu through the body. On the other hand, we observed a decrease in MT content in the hepatopancreas of crabs exposed to Cu, suggesting the excretion of MT in association with Cu bound to the sulfhydryl groups of this protein.

Hormetic responses to cadmium exposure in wheat seedlings: insights into morphological, physiological, and biochemical adaptations.

Cadmium is commonly recognized as toxic to plant growth, low-level Cd has promoting effects on growth performance, which is so-called hormesis. Although Cd toxicity in wheat has been widely investigated, knowledge of growth response to a broad range of Cd concentrations, especially extremely low concentrations, is still unknown. In this study, the morphological, physiological, and biochemical performance of wheat seedlings to a wide range of Cd concentrations (0-100 µΜ) were explored. Low Cd treatment (0.1-0.5µM) improved wheat biomass and root development by enhancing the photosynthetic system and antioxidant system ability. Photosynthetic rate (Pn) was improved by 5.72% under lower Cd treatment (1 µΜ), but inhibited by 6.05-49.85% from 5 to 100 µΜ. Excessive Cd accumulation induced oxidative injury manifesting higher MDA content, resulting in lower photosynthetic efficiency, stunted growth, and reduction of biomass. Further, the contents of ascorbate, glutathione, non-protein thiols, and phytochelatins were improved under 5-100 µΜ Cd treatment. The ascorbate peroxidase activity in the leaf showed a hormetic dose-response characteristic. Correlation analysis and partial least squares (PLS) results indicated that antioxidant enzymes and metabolites were closely correlated with Cd tolerance and accumulation. The results of the element network, correlation analysis, and PLS showed a crucial role for exogenous Cd levels in K, Fe, Cu, and Mn uptake and accumulation. These results provided a deeper understanding of the hormetic effect of Cd in wheat, which would be beneficial for improving the quality of hazard and risk assessments.

Role of SbNRT1.1B in cadmium accumulation is attributed to nitrate uptake and glutathione-dependent phytochelatins biosynthesis

Phytoremediation of cadmium (Cd)-polluted soil by using sweet sorghum displays a tremendous potential as it is a fast-growing, high biomass and Cd tolerant energy plant. Previous study has demonstrated SbNRT1.1B expression change is in accordance with enhanced Cd accumulation by external nitrate supply in sweet sorghum. Nevertheless, underlying mechanism of SbNRT1.1B response to Cd stress is still elusive. SbNRT1.1B exhibited a positive response to Cd stress in sweet sorghum. Overexpressing SbNRT1.1B increased primary root length, shoot fresh weight, nitrate and chlorophyll concentrations compared with Col-0 under Cd stress, while complementary SbNRT1.1B rescued these decreased values in mutant chl1–5. Cd concentrations in overexpressing SbNRT1.1B, complementary SbNRT1.1B and Col-0 lines were 3.2–4.1, 2.5–3.1 and 1.2–2.1 folds of that in chl1–5. Consistent with Cd concentrations, non-protein thiol (NPT), reduced glutathione (GSH) and phytochelatins (PCs) concentrations as well as the related genes expression levels showed the same trends under Cd stress. GSH biosynthesis inhibitor failed to reverse the patterns of GSH-dependent PCs concentrations changes in different lines, suggesting that SbNRT1.1B plays an upstream role in GSH-dependent PCs biosynthesis under Cd treatment. Altogether, SbNRT1.1B enhances nitrate concentrations contributing to increased chlorophyll concentrations and GSH-dependent PCs metabolites biosynthesis, thereby improving growth and Cd concentrations in plants.

Protective Role of Polyethylene Glycol Towards the Damaging Effects of Cadmium.

This study aimed to evaluate the role of drought-induced changes in the effects of cadmium (Cd) in plants. Cd is the most hazardous and important environmental pollutant. Water deficit is the most common environmental stress encountered by plants and affects most of the plant functions. The present study assessed the effect of Cd and water deficit on Capsicum frutescens seedlings in single and combined treatments. The seedlings of Capsicum were grown in a hydroponic solution and treated with Cd. The seedlings were subjected to water deficit with the help of polyethylene glycol (PEG). The other set of seedlings was treated with combined Cd + PEG. In the absence of PEG, maximum Cd accumulation was observed. The root and shoot growth of the seedlings were affected under all treatments with maximum inhibition in Cd. Pigment, protein and sugar contents and nitrate reductase activity decreased significantly in all treatments, while proline content increased. Induction of oxidative damage occurred through the formation of free radicals which caused alteration in electrolyte leakage, lipid peroxidation and activities of antioxidant enzymes, viz. superoxide dismutase, catalase, ascorbate peroxidase, guaiacol peroxidase and non-enzymatic non-protein thiol content and ascorbic acid in the stressed seedlings. Water deficit buttressed the toxic effect of Cd on chilli seedlings.

Photosynthetic variation and detoxification strategies based on cadmium uptake, non-protein thiols, and secondary metabolites in Miscanthus sacchariflorus under cadmium exposure.

Miscanthus sacchariflorus is previously demonstrated to be a potential candidate for remediation of cadmium (Cd) pollution. To explore its resistance strategy to Cd, a hydroponic experiment was conducted to determine the variations of photosynthetic activity in leaves and physiological response in roots of this plant. Results showed that the root of M. sacchariflorus was the primary location for Cd accumulation. The bioconcentration factor in the roots and rhizomes was >1, and the translocation factor from underground to aboveground was <1. Throughout the experimental period, treatment with 0.06 mM Cd2+ did not significantly alter the contents of chlorophyll a, chlorophyll b, or carotenoid. By contrast, treatment with 0.15 and 0.30 mM Cd2+ decreased the contents of chlorophyll a, chlorophyll b, and carotenoid; caused the deformation of the chlorophyll fluorescence transient curve; reduced the photochemical efficiency of photosystem II; and increased the contents of non-protein thiols, total flavone, and total phenol. These results indicate that M. sacchariflorus has good adaptability to 0.06 mM Cd2+. Moreover, the accumulation of the non-protein thiols, total flavone, and total phenol in roots may promote the chelation of Cd2+, thus alleviating Cd toxicity. This study provides theoretical support for using M. sacchariflorus to remediate Cd-polluted wetlands.

Emerging contaminants at trace levels of pesticides perturbs biomolecules in different organs in mice: Role of peroxisome proliferator-activated receptor-alpha

Objectives: Information is lacking on the consequences of chronic exposure to emerging contaminants at environmentally relevant (trace concentrations) on biomolecules. Environmental exposure to these chemical mixtures happens at trace concentrations and at multiple molecular interactions. The consequences of trace concentrations of multiple pesticides (MPs) on the regulation of selected biomolecules nitric oxide (NO), thiols, superoxide dismutase (SOD), and glutathione S-transferase (GST) in the tissues from wild type (WT) and genetically deficient- peroxisome proliferator-activated receptor-alpha (PPARα) knockout (Null) mice were investigated. Material and Methods: Mice were exposed to trace concentrations of MPs: Atrazine, dieldrin, endrin, endosulfan, and anthracene (1–100 ng/L) in drinking water for 6 weeks. Organs were collected and homogenized; NO, protein and non-protein thiol levels, as well as SOD and GST activities were determined. Results: Differential and organ selective effects of the treatments were observed in the WT and PPARα knockout. Increased NO levels were observed in the organs from WT with limited increase in the kidney (Null). SOD activity was decreased in the organs from the WT and was increased in the PPARα knockout when compared to the control. Thiol level was significantly increased in the heart and spleen in the WT and in the heart of the PPARα knockout mice when compared to the control. Non-protein thiol concentration was reduced in the heart and kidney (WT) and reduced in the liver of the PPARα knockout when compared to the control. GST activity was significantly decreased in the liver and spleen (WT) and was significantly elevated in all organs in the PPARα knockout mice when compared to the WT. Conclusion: The low concentrations of MPs may have caused selective dysregulation of biomolecules in different organs of the body. These effects observed may be influenced by genetic status such as in PPARα deficiency. These results present a scenario that implicates nanoconcentrations of series of organic contaminants that can cause cellular and molecular dysregulations of biomolecules precipitating toxicity and pathology that can be a threat to human health. Further, investigation into the molecular mechanism(s) and signaling pathway(s) implicated in these dysregulations is warranted.

Nitrogen-deficient leaves and roots can keep high abilities to scavenge reactive oxygen species and methylglyoxal, and protect them against oxidative damage in Citrus sinensis seedlings

Nitrogen-deficiency (ND) usually occurs in some citrus orchard soils in China. The roles of reactive oxygen species (ROS) and methylglyoxal (MG) generation and their detoxification systems in ND tolerance of horticultural woody plants still need to be revealed. For the first time, we examined the effects of ND on ROS and MG generation and their detoxification systems in leaves and roots of Citrus sinensis seedlings. The objectives are to test the hypotheses that N-deficient leaves and roots can keep high abilities to scavenge ROS and MG, thereby protecting them from oxidative damage, and that ND-induced alterations of ROS and MG formation and their detoxification systems in leaves and roots are different. ND augmented superoxide anion production rate and MG concentrations, but it decreased malondialdehyde (MDA) concentrations and electrolyte leakage in leaves and roots. ND increased the activities of most enzymes involved in ROS (ascorbate-glutathione cycle-related enzymes, antioxidant enzymes, and sulfur metabolism-related enzymes) and MG (glyoxalases) detoxification expressed on a protein basis with a few exceptions, and the concentrations of ascorbate, phytochelatins, and total non-protein thiols in leaves and roots. These results suggested that nitrogen-deficient leaves and roots could keep high abilities to detoxify ROS and MG, and protect them from oxidative damage. Generally viewed, ND affected the production and removal of ROS and MG more in roots than in leaves.

Toxicological assessment of sodium benzoate in Drosophila melanogaster.

Sodium benzoate (SB), the sodium salt of benzoic acid, is a food preservative with wide applications in the food, cosmetic and pharmaceutical industries due to its ability to kill many microorganisms effectively. Experimental evidence however suggests that excessive intake of SB poses detrimental health risks among consumers in the population. The present study investigated the toxic effects of various concentrations of SB using Drosophila melanogaster as a model. Adult wild-type flies of Canton S strain (1- to 3-daysold) was orally exposed to SB (0, 0.5, 1.0, 2.0 and 5.0 mg/5 g diet) to evaluate survival rates for 21days. Thereafter, we evaluated markers of oxidative stress, antioxidant status and behavioral activity in D. melanogaster exposed to SB for seven (7) days. We observed that SB (2.0 and 5.0 mg/5 g diet) decreased the survival of D. melanogaster. Also, SB inhibited glutathione-S-transferase activity and depleted total thiols and nonprotein thiols contents. Moreover, SB (5 mg/5 g diet) increased nitric oxide (nitrite/nitrate) level and reduced flies' emergence rate. Conclusively, findings from this study revealed that exposure to high concentrations of SB reduced survival rate and induced toxicity via the induction of oxidative stress and inhibition of antioxidant enzymes in D. melanogaster.

Physiological variations in hypovirus-infected wild and model long-term laboratory strains of Cryphonectria parasitica.

Forest ecosystems are highly threatened by the simultaneous effects of climate change and invasive pathogens. Chestnut blight, caused by the invasive phytopathogenic fungus Cryphonectria parasitica, has caused severe damage to European chestnut groves and catastrophic dieback of American chestnut in North America. Within Europe, the impacts of the fungus are widely mitigated through biological control that utilizes the RNA mycovirus: Cryphonectria hypovirus 1 (CHV1). Viral infections, similarly to abiotic factors, can cause oxidative stress in their hosts leading to physiological attrition through stimulating ROS (reactive oxygen species) and NOx production. To fully understand the interactions leading to the biocontrol of chestnut blight, it is vital to determine oxidative stress damage arising during CHV1 infection, especially considering that other abiotic factors, like long-term cultivation of model fungal strains, can also impact oxidative stress. Our study compared CHV1-infected C. parasitica isolates from two Croatian wild populations with CHV1-infected model strains (EP713, Euro7 and CR23) that have experienced long-term laboratory cultivation. We determined the level of oxidative stress in the samples by measuring stress enzymes' activity and oxidative stress biomarkers. Furthermore, for the wild populations, we studied the activity of fungal laccases, expression of the laccase gene lac1, and a possible effect of CHV1 intra-host diversity on the observed biochemical responses. Relative to the wild isolates, the long-term model strains had lower enzymatic activities of superoxide dismutase (SOD) and glutathione S-transferase (GST), and higher content of malondialdehyde (MDA) and total non-protein thiols. This indicated generally higher oxidative stress, likely arising from their decades-long history of subculturing and freeze-thaw cycles. When comparing the two wild populations, differences between them in stress resilience and levels of oxidative stress were also observed, as evident from the different MDA content. The intra-host genetic diversity of the CHV1 had no discernible effect on the stress levels of the virus-infected fungal cultures. Our research indicated that an important determinant modulating both lac1 expression and laccase enzyme activity is intrinsic to the fungus itself, possibly related to the vc type of the fungus, i.e., vegetative incompatibility genotype.

Synergistic Reduction of Arsenic Uptake and Alleviation of Leaf Arsenic Toxicity in Maize (Zea mays L.) by Arbuscular Mycorrhizal Fungi (AMF) and Exogenous Iron through Antioxidant Activity.

Arbuscular mycorrhizal fungi (AMF) play key roles in enhancing plant tolerance to heavy metals, and iron (Fe) compounds can reduce the bioavailability of arsenic (As) in soil, thereby alleviating As toxicity. However, there have been limited studies of the synergistic antioxidant mechanisms of AMF (Funneliformis mosseae) and Fe compounds in the alleviation of As toxicity on leaves of maize (Zea mays L.) with low and moderate As contamination. In this study, a pot experiment was conducted with different concentrations of As (0, 25, 50 mgꞏkg-1) and Fe (0, 50 mgꞏkg-1) and AMF treatments. Results showed that under low and moderate As concentrations (As25 and As50), the co-inoculation of AMF and Fe compound significantly increased the biomass of maize stems and roots, phosphorus (P) concentration, and P-to-As uptake ratio. Moreover, the co-inoculation of AMF and Fe compound addition significantly reduced the As concentration in stem and root, malondialdehyde (MDA) content in leaf, and soluble protein and non-protein thiol (NPT) contents in leaf of maize under As25 and As50 treatments. In addition, co-inoculation with AMF and Fe compound addition significantly increased the activities of catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) in the leaves of maize under As25 treatment. Correlation analysis showed that stem biomass and leaf MDA content were very significantly negatively correlated with stem As content, respectively. In conclusion, the results indicated that the co-inoculation of AMF and Fe compound addition can inhibit As uptake and promote P uptake by maize under low and moderate As contamination, thereby mitigating the lipid peroxidation on maize leaves and reducing As toxicity by enhancing the activities of antioxidant enzymes under low As contamination. These findings provide a theoretical basis for the application of AMF and Fe compounds in the restoration of cropland soil contaminated with low and moderate As.

Silicon as an attenuator of the toxic effects of aluminum in Schinus terebinthifolius plants.

Aluminum (Al) is highly toxic to plants, since it causes stress and inhibits plant growth. Silicon (Si) is known to mitigate the stress caused by Al in several plant species. Thus, the current study aims to investigate the soothing effects of Si on morphophysiological and photosynthetic variables, and the attributes associated with oxidative stress in Schinus terebinthifolius plants exposed to Al. Treatments have followed a completely randomized design, with three repetitions based on the following Al/Si combinations (in mM): Treatment 1: 0 Al + 0 Si; Treatment 2: 0 Al + 2.5 Si; Treatment 3: 1.85 Al + 0 Si; Treatment 4: 1.85 Al + 2.5 Si; Treatment 5: 3.71 Al + 0 Si; Treatment 6: 3.71 Al + 2.5 Si. Each sampling unit consisted of a tray with 15 plants, totaling forty-five per treatment. Shoot and root morphological variables, photosynthetic variables, photosynthetic pigments, hydrogen peroxide concentration, lipid peroxidation (MDA), guaiacol peroxidase (POD) and superoxide dismutase (SOD) enzymes, and non-enzymatic antioxidant such as Ascorbic acid (AsA) and non-protein thiol (NPSH) concentration were assessed. Root growth inhibition followed by changes in root morphological variables have negatively affected root and shoot biomass production in plants only subjected to Al. However, adding 2.5 mM Si to the treatment has mitigated the toxic effects caused by 1.85 mM of aluminum on S. terebinthifolius plants.

Protective effects of 2-aminoethylthiosulfuric acid and structurally analogous organosulfur compounds against ionizing radiation.

High efficacy and minimal toxicity radioprotectors are desirable options for the hazards posed by nuclear medical and energy technologies and the dangers presented by nuclear weapons in an unstable global situation. Although cysteamine is an effective radioprotector, it has considerable toxicity. In this study, the protective effects of the less toxic organosulfur compounds 2-aminoethylthiosulfate (AETS), thiotaurine (TTAU), and hypotaurine (HTAU) against X-ray damage in mice were compared with that of cysteamine. Intraperitoneal injection of either AETS or cysteamine (2.2 mmol/kg body weight) 30 min before X-ray irradiation (7.0 Gy) provided 100% survival for 30 days, limited the decrease in erythrocytes and neutrophils over 9 days, and reduced damage to bone marrow and spleen over 9 days. Neither TTAU nor HTAU provided any protection. In mice, 30 min after AETS administration, non-protein thiol content increased in the spleen, indicating cysteamine generation by AETS hydrolysis, the active protective species of AETS. All examined compounds scavenged •OH under diffusion control in aqueous solution, which is inconsistent with the difference in the protective effects among the compounds. The results indicate that AETS protects animals from ionizing radiation by several mechanisms, including scavenging •OH as cysteamine.

Uma abordagem de multibiomarcadores para investigar os efeitos do paracetamol no eixo de regulação da reprodução de machos do bagre Neotropical Rhamdia quelen

Abstract: Aim Paracetamol (PCM), or acetaminophen, is one of the most used drugs for human treatment of pain and fever. Since it has often been found in the aquatic environment, the aim of this study was to investigate the effects of PCM on the reproductive axis of male Rhamdia quelen catfish. Methods Different biomarkers were evaluated in hypothalamus, liver and gonads, as well as the plasma sexual hormone quantification. The fish were exposed to three PCM concentrations: 0.25, 2.5 and 25 µg.L-1 and to a control group (solvent acetone 0.0003%). After 14 days of exposure, fish were anesthetized, for blood sampling and biometrics, and after euthanasia, the tissues were sampled. Plasma was used for 11- keto testosterone and 17β - estradiol quantification. The hypothalamus was collected for brain aromatase (cyp19a1b) gene expression; the liver for the vitellogenin (vtg) gene expression and biochemical biomarkers; and gonad for the biochemical and histological biomarkers analyses. Results No alterations were observed in the hormone’s levels, sexual maturation level or in cyp19a1b and vtg gene expression. In the liver the non-protein thiols concentration increased at 2.5 µg.L-1 of PCM, and the superoxide dismutase (SOD) activity was reduced at 0.25 µg.L-1 of PCM. In gonads, glutathione S-transferase (GST) activity decreased and SOD activity increased at 25 µg.L-1 of PCM, while the glutathione peroxidase (GPx) activity reduced after exposure to both PCM concentrations. Conclusion The results showed that environmental concentrations of PCM can cause alterations in the antioxidant system, mainly in the gonads of R. quelen males. However, without significant change in the hormones levels or in the expression of genes related to the reproduction axis. These alterations occurred at concentrations already found in aquatic environment, including in Brazil.

Nitrate enhances cadmium accumulation through modulating sulfur metabolism in sweet sorghum

Sweet sorghum deploys tremendous potential for phytoremediation of cadmium (Cd)-polluted soils. Nitrate increases Cd accumulation in sweet sorghum, but the mechanism underlying this is still elusive. Sulfur-containing metabolites have been corroborated to play important roles in Cd tolerance in plants. Thus, whether sulfur metabolism contributed to nitrate-increased Cd accumulation in sweet sorghum was investigated in the present study. Two-way ANOVA analysis showed that most sulfur-containing metabolites concentrations and relevant enzymes activities were regulated by nitrate, Cd and interplay of nitrate and Cd. By using grey correlation analysis and Pearson correlation coefficient, Cd accumulation in shoots as affected by nitrate was also mainly ascribed to sulfur metabolism. ATP sulfurylase (ATPS) activities and non-protein thiol (NPT) concentrations in leaves were the two prominent factors that positively correlated with Cd accumulation in shoots. Excess nitrate elevated ATPS activities in leaves which contributed to increased NPT and phytochelatins (PCs) concentrations in leaves. Nitrate enhanced Cd accumulation in shoots of sweet sorghum under a low level of Cd treatment. Intriguingly, Cd accumulation in shoots of sweet sorghum was similar between a low level and a high level of Cd treatment. Principal Components Analysis (PCA) based on 34 parameters failed to separate the low Cd treatment from the high Cd treatment either, suggesting sweet sorghum is exclusively suitable for phytoremediation of slight Cd-polluted arable lands. Taken together, enhanced Cd accumulation in shoots of sweet sorghum by excess nitrate application is closely correlated with sulfur metabolism containing elevated ATPS activities, NPT and PCs concentrations in leaves.

Thiol, volatile and semi-volatile compounds alleviate the stress of zinc oxide nanoparticles of the pomegranate callus

Pomegranate trees are tropical and subtropical shrubs with nutritional benefits and pharmaceutical and therapeutic uses. Antioxidative systems protect the structure and function of cellular membranes. This study demonstrated the connection between oxidative stress generated by excess nanoparticles ZnO (ZnO-NPs) accumulation in pomegranate calli and the involvement of thiol groups and volatile and semi-volatile compounds in alleviating this stress. The effect of the non-enzymatic antioxidant system was studied using callus treated with three levels of ZnO-NPs or bulk particles (ZnO-BPs). With rising ZnO levels in the media, callus growth was gradually decreased by ZnO in both forms (NPs and BPs). Malondialdehyde (MDA) measurements revealed that different concentrations of both forms promoted lipid peroxidation. The supply of both forms had a considerable stimulatory influence on the cysteine (Cys) content in calli. Raised ZnO-NP concentrations increased glutathione (GSH) and non-protein thiols (NPTs) content in calli, but higher ZnO-BP concentrations lowered their content. Conversely, ZnO-NP levels reduced the protein thiols (PTs) content in calli, but ZnO-BP concentrations increased their content. GC-MS analysis was employed to investigate the volatile and semi-volatile chemical profiles within calli following exposure to 0 and 150 μg mL−1 of ZnO in both forms. GC-MS analysis detected 77, 67, and 83 compounds in ZnO-treated calli, of which 14, 16, and 20 with a similarity value greater than 70%, based on a NIST library, were recognized as metabolites for ZnO untreated and NPs- and BPs-treated calli, respectively. Six substances, including five alkanes and one morphinan, showed similarities in metabolite composition between control and NPs- or BPs-treated calli. ZnO-NPs-treated calli contained two alkane compounds only similar to the control, but ZnO-BPs-treated calli had six metabolites, including four alkanes, one carboxylic acid, and one ester. However, eight alkanes were similar within the callus treated with NPs and BPs.

Canavalia ensiformis lectin induced oxidative stress mediate both toxicity and genotoxicity in Drosophila melanogaster

Mannose/glucose-binding lectin from Canavalia ensiformis seeds (Concanavalin A - ConA) has several biological applications, such as mitogenic and antitumor activity. However, most of the mechanisms involved in the in vivo toxicity of ConA are not well known. In this study, the Drosophila melanogaster model was used to assess the toxicity and genotoxicity of different concentrations of native ConA (4.4, 17.5 and 70 μg/mL) in inhibited and denatured forms of ConA. The data show that native ConA affected: the survival, in the order of 30.6 %, and the locomotor performance of the flies; reduced cell viability to levels below 50 % (4.4 and 17.5 μg/mL); reduced nitric oxide levels; caused lipid peroxidation and increased protein and non-protein thiol content. In the Comet assay, native ConA (17.5 e 70 μg/mL) caused DNA damage higher than 50 %. In contrast, treatments with inhibited and denatured ConA did not affect oxidative stress markers and did not cause DNA damage. We believe that protein-carbohydrate interactions between ConA and carbohydrates of the plasma membrane are probably the major events involved in these activities, suggesting that native ConA activates mechanisms that induce oxidative stress and consequently DNA damage.

Modulatory effects of selected compounds on oxidative stress in hydrogen peroxide-induced Drosophila melanogaster

The balance between oxidant production and their conversion to harmless products is vital to the survival of living organisms. Hence the comparative free radical scavenging activity of quercetin, caffeic acid, ascorbic acid and gallic acid was studied using in vitro, in silico and in vivo model of Drosophila melanogaster. The FRAP and DPPH scavenging activities was performed using standard protocol, the molecular docking of the compounds against the active site of glutathione-s-transferase-2 was carried out using autodoc vina (v11-1). Hydrogen peroxide-induced D. melanogaster was use for evaluation of antioxidant parameters using standard procedures. The results revealed that, ascorbic acid has the highest percentage DPPH inhibition across the concentration with no significant difference in the ability of the compounds to reduce Fe3+ to Fe2+. The antioxidant compounds ameliorate weight loss and decrease percentage negative geotaxis observed in H2O2-induced flies without treatment. Data from this study showed that, the compounds mitigated against H2O2-induced accumulation of nitric acid, hydrogen peroxide and malondialdehyde. Moreover, the compounds improved total thiol and non-protein thiol concentration compared with H2O2 induced flies. The in silico study showed that, quercetin has the highest binding affinity more than the standard drug (methyldopa). All the compounds showed better reactivity potential compared with methyldopa. In conclusion, all the compounds offer protection against H2O2-induced oxidative stress in D. melanogaster.

Impact of Ferrous Sulfate on Thylakoidal Multiprotein Complexes, Metabolism and Defence of Brassica juncea L. under Arsenic Stress.

Forty-day-old Brassica juncea (var. Pusa Jai Kisan) plants were exposed to arsenic (As, 250 µM Na2HAsO4·7H2O) stress. The ameliorative role of ferrous sulfate (2 mM, FeSO4·7H2O, herein FeSO4) was evaluated at 7 days after treatment (7 DAT) and 14 DAT. Whereas, As induced high magnitude oxidative stress, FeSO4 limited it. In general, As decreased the growth and photosynthetic parameters less when in the presence of FeSO4. Furthermore, components of the antioxidant system operated in better coordination with FeSO4. Contents of non-protein thiols and phytochelatins were higher with the supply of FeSO4. Blue-Native polyacrylamide gel electrophoresis revealed an As-induced decrease in almost every multi-protein-pigment complex (MPC), and an increase in PSII subcomplex, LHCII monomers and free proteins. FeSO4 supplication helped in the retention of a better stoichiometry of light-harvesting complexes and stabilized every MPC, including supra-molecular complexes, PSI/PSII core dimer/ATP Synthase, Cytochrome b6/f dimer and LHCII dimer. FeSO4 strengthened the plant defence, perhaps by channelizing iron (Fe) and sulfur (S) to biosynthetic and anabolic pathways. Such metabolism could improve levels of antioxidant enzymes, and the contents of glutathione, and phytochelatins. Important key support might be extended to the chloroplast through better supply of Fe-S clusters. Therefore, our results suggest the importance of both iron and sulfur to combat As-induced stress in the Indian mustard plant at biochemical and molecular levels through enhanced antioxidant potential and proteomic adjustments in the photosynthetic apparatus.

Amelioration of cadmium toxicity by enhancing nitrogen assimilation and photosynthetic activity by two different nitrogen supplements in rice (Oryza sativa L.) cv. Lalat

Rapid industrialization and over-use of agrochemicals may lead to cadmium (Cd) contamination in agricultural soil, posing serious threat to human health. In recent times, application of plant nutrients and other growth promoting substances, like, nanoparticles (nZVI), silicon, salicylic acid were reported to have ameliorated Cd toxicity, in this regard, nitrogen (N) amendment can be a promising technique to ameliorate Cd toxicity as, N is an essential macronutrient responsible for plant growth and development. In the present study, seven days old seedlings of Oryza sativa L. cv. Lalat were co-treated with 10 µM CdCl2 and two different nitrogen supplements (Calcium nitrate and Ammonium acetate) separately to evaluate its ameliorating potential. It was observed that, Cd stress impaired plant vigor, biomass, RWC, nitrogen-containing components and non-enzymatic antioxidants causing significant reduction in photosynthesis and N-assimilation, indicating sensitivity of the cultivar to Cd stress. Cd stress also enhanced lipid peroxidation, enzymatic-antioxidant activity, proline and non-protein thiol accumulation. N-supplementation to Cd stressed plants rescued plant growth, biomass, protein content and nitrogen containing compounds leading to the upregulation of photosynthesis and nitrogen assimilation. N amendment was found to downregulate lipid peroxidation, enzymatic antioxidant activity, proline and non-protein thiol content and upregulate non-enzymatic antioxidant. Agronomical parameters and yield also improved significantly by N supplementation as compared to Cd stressed plants. Supplementation with N salts also improved grain quality by enhancing grain nutrients and reduced grain Cd by decreasing bioavailable Cd in soil, thereby reducing daily Cd intake and cancer risk. Among the two N-supplements, ammonium acetate showed better ameliorating effect compared to calcium nitrate.

Protective capacity of carotenoid trans-astaxanthin in rotenone-induced toxicity in Drosophila melanogaster

Trans-astaxanthin (TA), a keto-carotenoid found in aquatic invertebrates, possesses anti-oxidative and anti-inflammatory activities. Rotenone is used to induce oxidative stress-mediated Parkinson’s disease (PD) in animals. We probed if TA would protect against rotenone-induced toxicity in Drosophila melanogaster. Trans-astaxanthin (0, 0.1, 0.5, 1.0, 2.5, 10, and 20 mg/10 g diet) and rotenone (0, 250 and 500 μM) were separately orally exposed to flies in the diet to evaluate longevity and survival rates, respectively. Consequently, we evaluated the ameliorative actions of TA (1.0 mg/10 g diet) on rotenone (500 μM)-induced toxicity in Drosophila after 7 days’ exposure. Additionally, we performed molecular docking of TA against selected pro-inflammatory protein targets. We observed that TA (0.5 and 1.0 mg/10 g diet) increased the lifespan of D. melanogaster by 36.36%. Moreover, TA (1.0 mg/10 g diet) ameliorated rotenone-mediated inhibition of Catalase, Glutathione-S-transferase and Acetylcholinesterase activities, and depletion of Total Thiols and Non-Protein Thiols contents. Trans-astaxanthin prevented behavioural dysfunction and accumulation of Hydrogen Peroxide, Malondialdehyde, Protein Carbonyls and Nitric Oxide in D. melanogaster (p < 0.05). Trans-astaxanthin showed higher docking scores against the pro-inflammatory protein targets evaluated than the standard inhibitors. Conclusively, the structural features of TA might have contributed to its protective actions against rotenone-induced toxicity.