Elevated Catalase Activities Research Articles

Antioxidant System Activity in Roots and Shoots of Bean Cultivars in Response to Seed Treatment with Auxin as a Potential Model of Interaction with Endophytic Bacteria

Plant growth-promoting endophytic bacteria (PGPEB), producing auxins, are offered for a promising eco-friendly crop production. Precise bacterial strain selection is essential to ensure consistent and effective plant growth and resilience. Creating a model for the optimal dose-dependent interactions between PGPEB and hosts is necessary for understanding the mechanisms of high-precision selection of the inoculant composition to enhance bacterial preparations’ efficacy. This study investigated the impact of pre-sowing treatment with exogenous auxin indole-3-acetic acid (IAA) at various concentrations (0, 10, 1, 0.1, 0.01, 0.001, 0.0001, 0.00001 mg L−1) on the growth and antioxidant responses of three cultivars (cvs) of Phaseolus vulgaris L. (bean): Ufimskaya, Elsa, and Zolotistaya. The findings showed dose-dependent and cultivar-specific responses of 7-day-old bean seedlings to exogenous IAA. Ufimskaya cv exhibited significant increases in shoot, main root, and total root lengths at 0.001 mg L−1 IAA, while higher and lower concentrations inhibited growth. The reduced catalase (CAT) activity in roots and the elevated CAT activity in shoots correlated with shoot length and total root length of Ufimskaya cv. Importantly, the growth parameters exhibited weak or no correlations with malondialdehyde (MDA) and H2O2 content in roots and shoots, which is a peculiarity of the Ufimskaya cv response to exogenic IAA in contrast to the shown earlier response to inoculation with endophytes. The growth of only the main root of Elsa cv peaked at 0.1 mg L−1 IAA, and there were neutral or inhibitory effects with other concentrations. The positive correlation between CAT activity in shoots and the main root length and total root length as well as positive correlation between MDA content in roots and the total root length of Elsa cultivar were revealed. The shoot length and total root length of Zolotistaya cv were neutral or negatively responded to all concentration IAA, but the number of roots increased by 2–4 times. For Zolotistaya cv, positive correlations were observed between CAT activity in roots and the length of the main root and the total root length. Overall, these cultivar-specific antioxidant responses to exogenous IAA may help create models for optimal dose-dependent interactions between auxin-producing PGPEB and plants, enhancing the effectiveness of microbial preparations for consistent bean growth promotion.

Antifungal Effect of Oregano Essential Oil Against Penicillium expansum on Pyrus sinkiangensis.

Given the increasing demand for fruit safety assurance and environmental protection, plant essential oils have gained significant attention as natural alternatives for control of postharvest decay caused by various pathogens. In postharvest management, it is particularly important to effectively control postharvest decay without compromising the quality attributes of fruits. Although oregano essential oil (OEO) has been extensively studied for its antimicrobial properties against various postharvest pathogens, few studies have focused on its interactions with postharvest fruits. In this study, OEO was applied for management of postharvest decay of Pyrus sinkiangensis caused by Penicillium expansum, and its antifungal mechanisms and impacts on the quality attributes of pears were investigated. The OEO exhibited notable inhibitory effects, with determined MIC (0.02%) and MFC (0.04%) against P. expansum, which highlighted its potential as a viable alternative to synthetic fungicides. Our findings revealed that OEO disrupted P. expansum by compromising the integrity of the fungal plasma membrane, as evidenced by changes in plasma membrane permeability and the leakage of cellular components. The OEO treatment significantly reduced weight loss, maintained firmness, and preserved soluble-solid content in the treated pears. In addition, OEO treatment stimulated the intrinsic antioxidant mechanisms of pears, as demonstrated by elevated activities of superoxide dismutase and catalase during storage. This study provides compelling evidence for the antifungal and quality-preserving properties of OEO in the postharvest management of pears, underscoring its potential as an alternative to synthetic fungicides for controlling postharvest decay. The elucidation of the interaction between OEO and P. sinkiangensis would greatly enhance our comprehensive understanding of the biological activities of OEO and facilitate its practical application in the postharvest management of pears.

Effect of exogenous melatonin on growth and antioxidant system of pumpkin seedlings under waterlogging stress.

Melatonin regulates defense responses in plants under environmental stress. This study aimed to explore the impact of exogenous melatonin on the phenotype and physiology of 'BM1' pumpkin seedlings subjected to waterlogging stress. Waterlogging stress was induced following foliar spraying of melatonin at various concentrations (CK, 0, 10, 100, 200, and 300 μmol·L-1). The growth parameters, malondialdehyde (MDA) content, antioxidant enzyme activity, osmoregulatory substance levels, and other physiological indicators were assessed to elucidate the physiological mechanisms underlying the role of exogenous melatonin in mitigating waterlogging stress in pumpkin seedlings. The results indicate that pumpkin seedlings exhibit waterlogging symptoms, such as leaf wilting, water loss, edge chlorosis, and fading, under waterlogging stress conditions. Various growth indicators of the seedlings, including plant height, stem diameter, root length, fresh and dry weight, and leaf chlorophyll content, were significantly reduced. Moreover, the MDA content in leaves and roots increased significantly, along with elevated activities of superoxide dismutase, catalase, peroxidase, and soluble protein contents. When different concentrations of melatonin were sprayed on the leaves post waterlogging stress treatment, pumpkin seedlings showed varying degrees of recovery, with the 100 μmol·L-1 treatment displaying the best growth status and plant morphological phenotypes. There were no significant differences compared to the control group. Seedling growth indicators, chlorophyll content, root activity, antioxidant enzyme activities, soluble protein content, and osmotic adjustment substance content all increased to varying degrees with increasing melatonin concentration, peaking at 100 μmol·L-1. Melatonin also reduced membrane damage caused by oxidative stress and alleviated osmotic imbalance. Exogenous melatonin enhanced the activities of antioxidant enzymes and systems involved in scavenging reactive oxygen species, with 100 μmol·L-1 as the optimal concentration. These findings underscore the crucial role of exogenous melatonin in alleviating waterlogging stress in pumpkins. The findings of this study offer a theoretical framework and technical assistance for cultivating waterlogging-resistant pumpkins in practical settings. Additionally, it establishes a theoretical groundwork for the molecular breeding of pumpkins with increased tolerance to waterlogging.

Metagenomic and transcriptome insights into the disease resistance mechanisms in fresh-cut apple under pyridoxine treatment

Microbial spoilage poses a significant challenge to fresh-cut apples during storage, impacting quality and consumer health. This study explores pyridoxine’s (vitamin B6; VB6) potential mode of action to mitigate microbial spoilage in fresh-cut apples through physiological, transcriptomic, and metagenomic analysis. VB6 notably reduces microbial spoilage at room temperature by suppressing bacterial genera (Pseudomonas, Curtobacterium, Frigoribacterium) and fungal genera (Alternaria, Penicillium, Aureobasidium, Pseudogymnoascus) proliferation. Spoilage-causing identified biomarkers include species related to Pseudomonas, Curtobacterium, Frigoribacterium, and Penicillium. KEGG analysis suggests correlations of induced disease resistance with tropane alkaloids biosynthesis, plant-pathogen interaction, and phenylpropanoid biosynthesis pathways in fresh-cut apples. Moreover, VB6 treatment enhanced the antioxidant system via maintaining higher total phenolic contents, total flavonoid contents, and ascorbic acid under elevated catalase, superoxide dismutase, and ascorbate peroxidase enzymes activities, induced disease resistance under higher enzymes activities including phenylalanine ammonialyase, chitinase, cinnamate 4-hydroxylase, β-1,3-glucanase, and 4-coumarate: co-enzyme A ligase enzymes activities, maintained elevated VB6 contents, and reduced H2O2 accumulation. Notably, 6 elevated transcription factors involving WRKY, NAC and MYB might regulate disease resistance pathways in fresh-cut apples under VB6 treatment. This research establishes a theoretical and practical foundation for preserving post-processing quality of fresh-cut apples by restraining microbial proliferation through the proposed VB6-induced disease resistance regulatory mechanism.

Mitigating Effects of Tanacetum balsamita L. on Metabolic Dysfunction-Associated Fatty Liver Disease (MAFLD).

The metabolic syndrome and its associated co-morbidities have been recognized as predisposing risk factors for the development of metabolic-associated fatty liver disease (MAFLD). The present study reports on the beneficial effect of the Tanacetum balsamita methanol-aqueous extract (ETB) at 150 and 300 mg/kg bw on biochemical parameters related to oxidative stress, metabolic syndrome, and liver function in rat animal models with induced MAFLD. ETB was found to be non-toxic with LD50 > 3000 mg/kg and did not affect cell viability of hepatic HEP-G2 cells in a concentration up to 800 μg/mL. The pathology was established by a high-calorie diet and streptozotocin. Acarbose and atorvastatin were used as positive controls. At the higher dose, ETB reduced significantly (p < 0.05) the blood glucose levels by about 20%, decreased lipase activity by 52%, total cholesterol and triglycerides by 50% and 57%, respectively, and restored the amylase activity and leukocytes compared to the MAFLD group. ETB ameliorated oxidative stress biomarkers reduced glutathione and malondialdehyde in a dose-dependent manner. At 300 mg/kg, the beneficial effect of the extract on antioxidant enzymes was evidenced by the elevated catalase, glutathione peroxidase, and superoxide dismutase activity by 70%, 29%, and 44%, accordingly, compared to the MAFLD rats. ETB prevents the histopathological changes related to MAFLD. ETB, rich in 3,5-dicafeoylquinic, chlorogenic, and rosmarinic acids together with the isorhamnetin- and luteolin-glucoside provides a prominent amelioration of MAFLD.

Potential Involvement of MnCYP710A11 in Botrytis cinerea Resistance in Arabidopsis thaliana and Morus notabilis.

Cytochrome P450 (CYP) is a crucial oxidoreductase enzyme that plays a significant role in plant defense mechanisms. In this study, a specific cytochrome P450 gene (MnCYP710A11) was discovered in mulberry (Morus notabilis). Bioinformatic analysis and expression pattern analysis were conducted to elucidate the involvement of MnCYP710A11 in combating Botrytis cinerea infection. After the infection of B. cinerea, there was a notable increase in the expression of MnCYP710A11. MnCYP710A11 is overexpressed in Arabidopsis and mulberry and strongly reacts to B. cinerea. The overexpression of the MnCYP710A11 gene in Arabidopsis and mulberry led to a substantial enhancement in resistance against B. cinerea, elevated catalase (CAT) activity, increased proline content, and reduced malondialdehyde (MDA) levels. At the same time, H2O2 and O2- levels in MnCYP710A11 transgenic Arabidopsis were decreased, which reduced the damage of ROS accumulation to plants. Furthermore, our research indicates the potential involvement of MnCYP710A11 in B. cinerea resistance through the modulation of other resistance-related genes. These findings establish a crucial foundation for gaining deeper insights into the role of cytochrome P450 in mulberry plants.

Evaluation of dietary Fe on juvenile mirror carp (Cyprinus carpio var. specularis) and lettuce (Lactuca sativa var. ramosa Hort) in recirculating aquaponic system

In this study, we assessed the effects of dietary Fe level on the growth performance, nutritional status and health condition of mirror carp (Cyprinus carpio var. specularis) (12.59 ± 1.73 g), production and quality of lettuce (Lactuca sativa var. ramosa Hort) in recirculating aquaponic system for a 60-day feeding trial. Five semi-purified diets with ferrous sulfate as the iron source were used in the feeding trail, which were: T1 (53.81 mg Fe/kg); T2 (118.83 mg Fe/kg); T3 (209.24 mg Fe/kg); T4 (328.44 mg Fe/kg); T5 (498.74 mg Fe/kg). The findings indicated the following: (1) The growth results showed that compare to other groups, the weight gain of mirror carp in Group T2 was the highest, and the feed conversion ratio was the lowest (P < 0.05). (2) The body composition results showed that muscle crude fat and crude protein of the mirror carp significantly increase in the T2 group compared to the T1 group (P < 0.05). Additionally, the C20:5n-3 levels in both the T2 and T3 groups were significantly higher than that of T1 group (P < 0.05). (3) Group T2 exhibited the highest liver superoxide dismutase activity, with significantly elevated catalase activity compared to the other groups (P < 0.05), and the liver malondialdehyde content in group T2 was significantly lower than the T4 and T5 groups (P < 0.05). (4) The morphological measurements of intestinal tissues in mirror carp showed that compared to Group T1 and Group T5, Group T2 exhibited the highest villus length and muscular layer thickness (P < 0.05). (5) The lowest yield of lettuce is in the T1group (3.35 kg/m2/month), and the highest yield (3.96 kg/m2/month) is in the T5 group, along with the highest chlorophyll and soluble sugar content (P < 0.05). (6) The T5 group had the highest nitrogen utilization efficiency (69.44%), while the T1 group had the lowest nitrogen utilization efficiency (58.37%). In addition, the different dietary iron levels did not adversely affect the water quality of the aquaponic systems. Based on the line regression analysis of weight gain and feed conversion ratio with feed iron content, it was found that the most suitable iron content in feed is 118.22–120.44 mg/kg, and the production and quality of lettuce will increase with the increase of feed iron content. According to the results of fish and vegetable growth, as well as nitrogen utilization efficiency, it is suggested that the dietary iron level in the feed of juvenile mirror carp utilized within aquaponic systems should ideally fall within the range of 120–200 mg/kg.

How does water stress affect the bioaccumulation of galanthamine and lycorine, growth performance, phenolic content and defense enzyme activities in summer snowflake (Leucojum aestivum L.)?

Leucojum aestivum L. is an Amaryllidaceae bulbous plant with two alkaloids that have remarkable medicinal potential: galanthamine and lycorine. Although the presence of galanthamine in L. aestivum has commercial value for the pharmaceutical industry and the effect of water stress (WS) applications on secondary metabolite enhancement is well established in a variety of plants, no studies have been carried out to reveal the effectiveness of WS on this beneficial medicinal plant. Objective of the study was to investigate the effects of eight different WS treatments [Control, waterlogging (WL) condition, and drought stress conditions (water deficiency generated by water deficit irrigation-WDI 25%, 50%, and 75%- and polyethylene glycol-PEG 6000 15%, 30%, and 45%-)] on growth parameters, alkaloid levels (galanthamine and lycorine), non-enzymatic antioxidant activities (total phenol-flavonoid content and free radical scavenging activity), and enzymatic antioxidant activities [superoxide dismutase (SOD) and catalase (CAT)] of L. aestivum in a pot experiment. Based on the findings, maximum increases in growth parameters were obtained with PEG-induced WS treatments. Moderate water deficiency (50% WDI) produced the highest levels of galanthamine and lycorine, total phenol-flavonoid content, and antioxidant capacity, along with moderately elevated CAT activity in the bulbs. All WS treatments resulted in increased CAT activity in the bulbs. It was observed that bulbs had higher SOD and CAT activities under WL conditions had lower fresh weights and were close to control in terms of alkaloid levels, total phenol-flavonoid content, and free radical scavenging activity. When all of the outcomes were taken into account, it can be concluded that moderate water-deficit stress (50% WDI) was regarded as the most effective treatment for increasing the pharmaceutical value of L. aestivum.Graphical abstract

Cobalt protoporphyrin modulates antioxidant enzyme activity in the hypothalamus and motor cortex of female rats.

Cobalt protoporphyrin (CoPP) is a synthetic heme analog that has been observed to reduce food intake and promote sustained weight loss. While the precise mechanisms responsible for these effects remain elusive, earlier research has hinted at the potential involvement of nitric oxide synthase in the hypothalamus. This study aimed to delve into CoPP's impact on the activities of crucial antioxidant enzymes: superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione-S-transferase (GST) across seven distinct brain regions (hippocampus, hypothalamus, prefrontal cortex, motor cortex, striatum, midbrain, and cerebellum), as well as in the liver and kidneys. Female Wistar rats weighing 180 to 200 grams received a single subcutaneous dose of 25 μmol/kg CoPP. After six days, brain tissue was extracted to assess the activities of antioxidant enzymes and quantify malondialdehyde levels. Our findings confirm that CoPP administration triggers the characteristic effects of decreased food intake and reduced body weight. Moreover, it led to an increase in SOD activity in the hypothalamus, a pivotal brain region associated with food intake regulation. Notably, CoPP-treated rats exhibited elevated enzymatic activity of catalase, GR, and GST in the motor cortex without concurrent signs of heightened oxidative stress. These results underscore a strong connection between the antioxidant system and food intake regulation. They also emphasize the need for further investigation into the roles of antioxidant enzymes in modulating food intake and the ensuing weight loss, using CoPP as a valuable research tool.

Enhancing Postharvest Quality and Shelf Life of Strawberries through Advanced Coating Technologies: A Comprehensive Investigation of Chitosan and Glycine Betaine Nanoparticle Treatments.

The application of natural polymer-based coatings presents a viable approach to prolong the longevity of fruits and tissue damage. This study investigates the impact of treatments involving glycine betaine (GB), chitosan (CTS), and chitosan-coated glycine betaine nanoparticles (CTS-GB NPs) on preserving the quality and reducing decay in strawberry fruits. The fruits were subjected to treatments with GB (1 mM), CTS (0.1%), CTS-GB NPs (0.1%), or distilled water at 20 °C for 5 min, followed by storage at 4 °C for 12 days. The results indicate that CTS and CTS-GB NPs treatments resulted in the highest tissue firmness, total anthocyanin content, and ascorbate peroxidase activity, while exhibiting the lowest decay percentage and weight loss, as well as reduced malondialdehyde levels at the end of storage. GB, CTS, and CTS-GB NPs treatments demonstrated elevated catalase activity and antioxidant capacity, coupled with lower electrolyte leakage and hydrogen peroxide levels. These treatments did not significantly differ from each other but were markedly different from the control. The results substantiate that CTS and CTS-GB NPs treatments effectively preserve strawberry quality and extend storage life by bolstering antioxidant capacity and mitigating free radical damage.

Evaluation of Resistance Resources and Analysis of Resistance Mechanisms of Maize to Stalk Rot Caused by Fusarium graminearum.

Stalk rot is one of the most destructive and widely distributed diseases in maize plants worldwide. Research on the performance and resistance mechanisms of maize against stem rot is constantly improving. In this study, among 120 inbred maize lines infected by Fusarium graminearum using the injection method, 4 lines (3.33%) were highly resistant to stalk rot, 28 lines (23.33%) were resistant, 57 lines (47.50%) were susceptible, and 31 lines (25.84%) were highly susceptible. The inbred lines 18N10118 and 18N10370 were the most resistant and susceptible with disease indices of 7.5 and 75.6, respectively. Treatment of resistant and susceptible maize inbred seedlings with F. graminearum showed that root hair growth of the susceptible inbred lines was significantly inhibited, and a large number of hyphae attached and adsorbed multiple conidia near the root system. However, the resistant inbred lines were delayed and inconspicuous, with only a few hyphae and spores appearing near the root system. Compared with susceptible inbred lines, resistant maize inbred line seedlings treated with F. graminearum exhibited elevated activities of catalase, phenylalanine ammonia-lyase, polyphenol oxidase, and superoxide dismutase. We identified 153 genes related to disease resistance by transcriptome analysis. The mitogen-activated protein kinase signaling and peroxisome pathways mainly regulated the resistance mechanism of maize inbred lines to F. graminearum infection. These two pathways might play an important role in the disease resistance mechanism, and the function of genes in the two pathways must be further studied, which might provide a theoretical basis for further understanding the molecular resistance mechanism of stalk rot and resistance gene mining.

Exogenous Ascorbate Administration Elevates Testicular Oxidative Damage and Histological Injuries in Rats after Busulfan Treatment

Several anticancer drugs are coadministered with ascorbate (ASCB) to complement their cytotoxic effects. However, it is not known if the treatment regimen prevents collateral oxidative damage to non-target sites. The current study evaluated the effect of ASCB cotreatment on the testes of young adult rats treated with an anticancer drug, busulfan (BUS). About 40 Wistar rats were arbitrarily assigned into four groups (N = 10), namely, control (&lt;0.2% dimethyl sulfoxide vehicle), BUS (4 mg/kg b.w.; intraperitoneally for 4 days), BUS + ASCB (4 mg BUS/kg b.w. for 4 days + 100 mg ASCB/kg b.w.; intraperitoneally for 14 days and 7 days prior to start of BUS injection), and ASCB (100 mg/kg b.w.; intraperitoneally for 21 days). At the end of study, ASCB + BUS cotreatment reduced spermatogenesis score index, superoxide dismutase activity, total ASCB and decreased hydrogen peroxide level, and elevated catalase activity and nitrite concentration much more than treatment with BUS alone ( P &lt; 0.05 ). Other observations included elevated malondialdehyde level, DNA damage, and diminished glutathione concentration in the testes of BUS + ASCB animals. Interestingly, ASCB administration raised testicular ASCB concentration beyond the control values ( P &lt; 0.05 ) and the antioxidant status of the testes. Histological aberrations included many single layers of germ cells, shrinked tubules, and vacuolated structures in the epithelium of BUS + ASCB, similar to those of BUS-treated animals. In conclusion, BUS treatment deregulated the redox status of the testes and caused a dramatic consumption of ASCB which were enhanced by exogenous ASCB resulting to testicular damage.

Nano-TiO2 aggravates the adverse effect of pentachlorophenol on antioxidant and immune response in anti-predatory mussels

Nano-TiO2 can act as a vector to organic compounds, such as pentachlorophenol (PCP) posing a potential threat to the marine ecosystems. Studies showed that nano pollutant toxicity can be modulated by abiotic factors, but little is known about the potential influence of biotic stressors (such as predators) on the physiological responses to pollutants in marine organisms. We explored the effects of n-TiO2 and PCP on the mussel Mytilus coruscus in the presence of its natural predator, the swimming crab Portunus trituberculatus. Exposure to n-TiO2, PCP, and predation risk showed interactive effects on antioxidant and immune parameters of the mussels. Elevated activities of catalase (CAT), glutathione peroxidase (GPX), acid phosphatase (ACP) and alkaline phosphatase (AKP), suppressed activity of superoxide dismutase (SOD), lower levels of glutathione (GSH) and increased malondialdehyde (MDA) levels indicated dysregulation of the antioxidant system and immune stress induced by single PCP or n-TiO2 exposure. Integrated biomarker (IBR) response values showed the effect of PCP was concentration dependent. Of the two used n-TiO2 sizes (25 and 100 nm), larger particles induced higher antioxidant and immune disturbances indicating higher toxicity possibly due to higher bioavailability. Compared to single PCP exposure, the combination of n-TiO2 and PCP enhanced the imbalance of SOD/CAT and GSH/GPX and led to elevated oxidative lesions and activation of immune-related enzymes. Overall, the combined impacts of pollutants and biotic stress exhibited a greater magnitude of adverse effects on antioxidant defense and immune parameters in mussels. The toxicological effects of PCP were exacerbated in the presence of n-TiO2, and the deleterious impact of these stressors was further amplified under predator-induced risk after prolonged (28 days) exposure. However, the underlying physiological regulatory mechanisms governing the interplay of these stressors and predatory cues on mussels remain elusive, warranting further investigation.

Iridoids from Morinda lucida, (Benth.) Rubiaceae, produced analgesic and anti-inflammatory activities via agonism at the kappa and delta opioid receptors, inhibition of COX-2 besides elevation of CAT and SOD activities.

Pain and inflammation are the major symptoms of almost every human disease. Herbal preparations from Morinda lucida are used to treat pain and inflammation in traditional medicine. However, the analgesic and anti-inflammatory activities of some of its chemical constituents are not known. The aim of this study is to evaluate the analgesic and anti-inflammatory activities and possible mechanisms of these activities of iridoids from Morinda lucida. The compounds were isolated using column chromatography and characterized by NMR spectroscopy and LC-MS. Anti-inflammatory activity was evaluated using carrageenan-induced paw edema. Whereas, the analgesic activity was assessed in the hot plate and acetic acid-induced writhing assays. Mechanistic studies were conducted using pharmacological blockers, determination of antioxidant enzymes, lipid peroxidation, and docking studies. The iridoid, ML2-2 exhibited inverse dose-dependent anti-inflammatory activity (42.62% maximum at 2 mg/kg p. o). ML2-3 produced dose-dependent anti-inflammatory activity (64.52% maximum at 10 mg/kg p. o.). Anti-inflammatory activity of diclofenac sodium was 58.60% at 10 mg/kg p. o. Furthermore, ML2-2 and ML2-3 produced analgesic activity (P < 0.01) of 44.44 ± 5.84 and 54.18 ± 19.01%. at 10 mg/kg p. o. respectively in the hot plate assay and 64.88 and 67.44% in the writhing assay. ML2-2 significantly elevated catalase activity. However, ML2-3 elevated SOD and catalase activity significantly. In the docking studies, both iridoids formed stable crystal complexes with delta and kappa opioid receptors, and the COX-2 enzyme with very low free binding energies (ΔG) from -11.2 to -14.0 kcal/mol. However, they did not bind with the mu opioid receptor. The lower bound RMSD of most of the poses were found to be ≤ 2. Several amino acids were involved in the interactions through various inter molecular forces. These results indicate that ML2-2 and ML2-3 possessed very significant analgesic and anti-inflammatory activities via acting as both delta and kappa opioid receptor agonist, elevation of anti-oxidant activity and inhibition of COX-2.

Potential negative effect of long-term exposure to nitrofurans on bacteria isolated from wastewater

Nitrofurans are broad-spectrum bactericidal agents used in a large quantity for veterinary and human therapy. This study reports the long-term impact of two nitrofuran representatives, nitrofurantoin (NFT) and furaltadone (FTD) on the bacterial strains Sphingobacterium siyangense FTD2, Achromobacter pulmonis NFZ2, and Stenotrophomonas maltophilia FZD2, isolated from a full-scale wastewater treatment plant. Bacterial whole genome sequencing was used for preliminary strains characterization. The metabolomic, electrochemical, and culture methods were applied to understand changes in the bacterial strains after 12-month exposure to nitrofurans. The most significantly altered metabolic pathways were observed in amino acid and sugar metabolism, and aminoacyl-tRNA biosynthesis. Disrupted protein biosynthesis was measured in all strains treated with antibiotics. Prolonged exposure to NFT and FTD also triggered mutagenic effects, affected metabolic activity, and facilitated oxidative stress within the cells. Nitrofuran-induced oxidative stress was evidenced from an elevated activity of catalase and glutathione S-transferases. NFT and FTD elicited similar but not identical responses in all analyzed strains. The results obtained in this study provide new insights into the potential risks of the prolonged presence of antimicrobial compounds in the environment and contribute to a better understanding of the possible impacts of nitrofuran antibiotics on the bacterial cells.

Phytotoxic Effects of Polystyrene and Polymethyl Methacrylate Microplastics on Allium cepa Roots

Plastic contamination has become one of the most pressing environmental issues due to rapidly increasing production of disposable plastic products, their fragmentation into smaller pieces, and long persistence in the environment, which affects all living organisms, including plants. In this study, Allium cepa roots were exposed to 0.01, 0.1, and 1 g L-1 of commercial polystyrene (PS-MPs) and polymethyl methacrylate microparticles (PMMA-MPs) for 72 h. Dynamic light scattering (DLS) analyses showed high stability of both types of MPs in ultrapure water used for A. cepa treatment. Morphometric analysis revealed no significant change in root length compared to control. Pyrolysis hyphenated to gas chromatography and mass spectrometry (Py-GC-MS) has proven PS-MPs uptake by onion roots in all treatments, while PMMA-MPs were recorded only upon exposure to the highest concentration. Neither MPs induced any (cyto)toxic effect on root growth and PMMA-MPs even had a stimulating effect on root growth. ROS production as well as lipid and protein oxidation were somewhat higher in PS-MP treatments compared to the corresponding concentrations of PMMA-MP, while neither of the applied MPs induced significant damage to the DNA molecule assayed with a Comet test. Significantly elevated activity of H2O2 scavenging enzymes, catalase, and peroxidases was measured after exposure to both types of MPs. Obtained results suggest that onion roots take up PS-MPs more readily in comparison to PMMA-MPs, while both types of MPs induce a successful activation of antioxidant machinery in root cells that prevented the occurrence of toxic effects.

Comparison of Oxygen Electrode Chronoamperometry and Spectrophotometry for Determination of Catalase Activity

Catalase is a key antioxidative enzyme, and a deficiency or malfunction of catalase is hypothesized to be related to various diseases. To investigate catalase activity, it is important to use reliable methods and experimental protocols enabling consistent fallouts. One major problem, however, is that the activity values obtained with different techniques and procedures can vary to a large extent. The aim of this work was to identify experimental conditions that provide similar catalase activity values with two different methods based on either spectrophotometry or chronoamperometry. The investigated parameters include the concentration of catalase and its substrate (H2O2), as well as the effect of deoxygenation of the catalase medium by nitrogen (N2). Within the frame of investigated conditions, we show that spectrophotometry is strongly affected by the catalase concentration, whereas chronoamperometry is shown to be more dependent on the substrate concentration. Deoxygenation leads to elevated catalase activity values in the case of chronoamperometry, whereas it shows no influence on the results obtained with spectrophotometry. In particular, in the case of low substrate concentrations (i.e., low catalase reaction rates), higher and more accurate results are obtained with deoxygenation in the case of chronoamperometry measurements due to minimized oxygen escape. The effect of deoxygenation, giving rise to elevated catalase activity values, however, is not statistically significant at high substrate concentrations, implying that the protocol can be simplified by excluding this step as long as the other parameters are optimized. Finally, by comparing the two methods at different experimental conditions, we identified protocols resulting in similar results, i.e., 10 mM H2O2 and catalase activity of 4–5 U/mL. Based on this work, improved consistency of catalase activity data obtained with different methodologies and in different labs is expected.

Assessing the hepatotoxicity of PFOA, PFOS, and 6:2 Cl-PFESA in black-spotted frogs (Rana nigromaculata) and elucidating potential association with gut microbiota

Pollution caused by per- and polyfluoroalkyl substances (PFASs) has become a major global concern. The association between PFAS-induced hepatotoxicity and gut microbiota in amphibians, particularly at environmentally relevant concentrations, remains elusive. Herein we exposed male black-spotted frogs (Rana nigromaculata) to 1 and 10 μg/L waterborne perfluorooctanoic acid (PFOA), perfluorooctanesulfonic acid (PFOS), and 6:2 chlorinated polyfluorinated ether sulfonate (6:2 Cl-PFESA) for 21 days; subsequently, liver histopathological, oxidative stress, molecular docking, gene/protein expression, and gut microbiome analyses were conducted. PFOS and 6:2 Cl-PFESA exposure enhanced serum alanine aminotransferase and aspartate aminotransferase activities, and markedly increased hepatic area of vacuoles and inflammatory cell infiltration, while PFOA exposure increased serum alanine aminotransferase but not aspartate aminotransferase activities and affected hepatic area of vacuoles and inflammatory cell infiltration to a lesser extent. All three PFASs elevated catalase, glutathione S-transferase, and glutathione peroxidase activities and glutathione and malondialdehyde contents in the liver, suggesting the induction of oxidative stress. Further, PFASs could bind to mitogen-activated protein kinases (p38, ERK, and JNK), upregulating not only their expression but also the expression of downstream oxidative stress-related genes and that of P-p38, P-ERK, and Nrf2 proteins. In addition, PFAS exposure significantly increased the relative abundance of Proteobacteria and Delftia and decreased that of Firmicutes and Dietzia, Mycoplasma, and Methylobacterium–Methylorubrum in the order of PFOS ≈ 6:2 Cl-PFESA > PFOA. Altogether, it appears that PFOS and 6:2 Cl-PFESA are more toxic than PFOA. Finally, microbiota function prediction, microbiota co-occurrence network, and correlation analysis between gut microbiota and liver indices suggested that PFAS-induced hepatotoxicity was associated with gut microbiota dysbiosis. Our data provide new insights into the role of gut microbiota in PFAS-induced hepatotoxicity in frogs.

Overexpression of PgCBF3 and PgCBF7 Transcription Factors from Pomegranate Enhances Freezing Tolerance in Arabidopsis under the Promoter Activity Positively Regulated by PgICE1

Cold stress limits plant growth, development and yields, and the C-repeat binding factors (CBFs) function in the cold resistance in plants. However, how pomegranate CBF transcription factors respond to cold signal remains unclear. Considering the significantly up-regulated expression of PgCBF3 and PgCBF7 in cold-tolerant Punica granatum ‘Yudazi’ in comparison with cold-sensitive ‘Tunisia’ under 4 °C, the present study focused on the two CBF genes. PgCBF3 was localized in the nucleus, while PgCBF7 was localized in the cell membrane, cytoplasm, and nucleus, both owning transcriptional activation activity in yeast. Yeast one-hybrid and dual-luciferase reporter assay further confirmed that PgICE1 could specifically bind to and significantly enhance the activation activity of the promoters of PgCBF3 and PgCBF7. Compared with the wild-type plants, the PgCBF3 and PgCBF7 transgenic Arabidopsis thaliana lines had the higher survival rate after cold treatment; exhibited increased the contents of soluble sugar and proline, while lower electrolyte leakage, malondialdehyde content, and reactive oxygen species production, accompanying with elevated enzyme activity of catalase, peroxidase, and superoxide dismutase; and upregulated the expression of AtCOR15A, AtCOR47, AtRD29A, and AtKIN1. Collectively, PgCBFs were positively regulated by the upstream PgICE1 and mediated the downstream COR genes expression, thereby enhancing freezing tolerance.

Improved Hydrogen Peroxide Stress Resistance of Zymomonas mobilis NADH Dehydrogenase (ndh) and Alcohol Dehydrogenase (adhB) Mutants

Unintended shifts in stress resistance of microbial strains with engineered central metabolism may impact their growth and production performance under oxidative, lignocellulosic, solvent, and other stress conditions, and as such, must be taken into account in bioprocess design. In the present work, we studied oxidative stress resistance in mutant strains of the facultatively anaerobic, ethanologenic bacterium Zymomonas mobilis with modified respiratory (inactivated NADH dehydrogenase Ndh, by disruption of ndh) and ethanologenic (inactivated iron-containing alcohol dehydrogenase isoenzyme ADH II, by disruption of adhB) catabolism, using exogenously added H2O2 in the concentration range of 2–6 mM as the oxidative stressor. Both mutations improved H2O2 resistance and enhanced catalase activity by a factor of 2–5, while the overexpression of Ndh had an opposite effect. Strains with a catalase-negative background were unable to grow already at 1 mM hydrogen peroxide, and their H2O2 resistance did not depend on AdhB or Ndh expression levels. Hence, the improved resistance of the ndh and adhB mutants to H2O2 resulted from their elevated catalase activity. The interrelation between these mutations, the catabolic redox balance, catalase activity, and oxidative stress defense in Z. mobilis is discussed.