Antioxidant Defense Research Articles

Hemoglobin in Submicron Particles (HbMPs) Is Stabilized Against Oxidation

Superoxide dismutase (SOD) and Catalase (CAT) play a crucial role as the first line of defense antioxidant enzymes in a living cell. These enzymes neutralize the superoxide anion from the autooxidation of oxyhemoglobin (Oxy-Hb) and convert hydrogen peroxides into water and molecular oxygen. In this study, we fabricated hemoglobin submicron particles (HbMPs) using the Coprecipitation Crosslinking Dissolution (CCD) technique and incorporating first-line antioxidant enzymes (CAT, SOD) and second-line antioxidant (ascorbic acid, Vit. C) to investigate a protective effect of modified HbMPs via cyclically oxygenation and deoxygenation. Thereafter, the total hemoglobin (Hb) content and Oxy-Hb content to HbMPs were determined. The results revealed that the HbMPs have a protective effect against oxidation from hydrogen peroxide and potentially neutralizing hydrogen peroxide to water over 16 times exposure cycles. No significant differences in total Hb content were found between normal HbMPs and enzyme-modified HbMPs in the absence of Vit. C. The Oxy-Hb of CAT-HbMPs showed significantly higher values than normal HbMPs. The functional Hb of normal HbMPs and enzyme-modified HbMPs was increased by 60–77% after a short time Vit. C (1:25) exposure. The co-immobilization of CAT and SOD in hemoglobin particles (CAT-SOD-HbMPs) in the presence of Vit. C provides protective effects against oxidation in cyclic Oxygenation and Deoxygenation and shows the lowest reduction of functional Hb. Our studies show that the CCD technique-modified HbMPs containing antioxidant enzymes and a reducing agent (ascorbic acid) demonstrate enhanced Hb functionality, providing protective effects and stability under oxidative conditions.

Assessment of Placental Antioxidant Defense Markers in Vaccinated and Unvaccinated COVID-19 Third-Trimester Pregnancies

Background: Pregnancy has been identified as a risk factor for severe COVID-19, leading to maternal and neonatal complications. The safety and effects of the SARS-CoV-2 vaccination during pregnancy, particularly on placental function and oxidative stress (OxS), remain underexplored. We investigated the impact of vaccination on third-trimester placental antioxidant defense markers. Methods: Ninety full-term pregnant women were divided into the following groups: vaccinated (n = 27) and unvaccinated (n = 25) COVID-19-positive pregnant women; control subgroups were composed of vaccinated (n = 19) or unvaccinated (n = 19) COVID-19-negative women with a healthy term singleton pregnancy with no signs of COVID-19. Placental samples were collected after delivery. Lipid peroxidation (TBARS), gene expression of HIF-1α, and catalase (CAT), superoxide dismutase-1 (SOD1) and CAT-SOD1 enzymatic activity were measured. Results: COVID-19-positive placentae exhibited significantly higher TBARS and HIF-1α levels compared to controls, regardless of vaccination status. Vaccination significantly increased placental CAT and SOD1 expression and activity in COVID-19-positive women, suggesting enhanced antioxidant defense. Unvaccinated women showed a higher incidence of COVID-19 symptoms and lower antioxidant enzyme activity. Conclusions: SARS-CoV-2 infection induced placental OxS, which is countered by a placental adaptive antioxidant response. Vaccination during pregnancy enhances placental defense, further supporting the safety and benefits of COVID-19 vaccination in preventing complications and protecting fetal development.

Oxidative Stress Biomarkers and Metabolic Parameters in Healthy Holstein Dairy Cows and Cows With Left Displacement Abomasum During the Transitional Period.

During the transitional period, dairy cows experience oxidative stress and are more susceptible to diseases, including left displacement of the abomasum (LDA). This study aimed to compare oxidative stress biomarker levels in cows with LDA to those in healthy conditions and investigate the associations between predictive metabolites linked to LDA and oxidative stress biomarkers. In this case-control study, 400 healthy multiparous Holstein cows were matched for lactation number, milk production and calving date. Blood samples were collected at four time points: 21 and 7 days before, as well as 7 and 21 days after parturition from all animals. During the observation period, seven cows diagnosed with LDA in the main population, and seven healthy cows were randomly selected as controls for the comparison of oxidative stress, liver enzymes and metabolic parameters. Analysis of blood parameters utilized repeated measures ANOVA, and the degree of relationship between oxidative stress biomarkers and other measured parameters was assessed using Pearson correlations. The LDA group exhibited significantly higher levels of urea, blood urea nitrogen (BUN), gamma-glutamyl transferase, glucose, cholesterol, triglyceride (TAG), β-hydroxybutyric acid (BHBA), aspartate aminotransferase (AST), sorbitol dehydrogenase, serum amyloid A (SAA), chloride, sodium, potassium, total antioxidant capacity (TAC) and malondialdehyde (MDA) compared to the control cows (p<0.05). Positive correlations were observed among BUN, glucose, TB, AST, SAA, BHBA, TAG and MDA. Conversely, these parameters displayed negative correlations with TAC. Negative correlations were found among chloride, sodium, potassium, calcium, phosphorus and MDA, whereas positive correlations were observed with TAC. These findings highlight the elevated level of oxidative stress and compromised antioxidant defence in cows with LDA and the intricate interplay among oxidative stress, metabolic parameters and liver enzymes.

Melittin - the main component of bee venom: a promising therapeutic agent for neuroprotection through keap1/Nrf2/HO-1 pathway activation

The Nuclear factor erythroid 2–related factor (Nrf2)/ Heme oxygenase-1 (HO-1) pathway, known for its significant role in regulating innate antioxidant defense mechanisms, is increasingly being recognized for its potential in neuroprotection studies. Derived from bee venom, melittin's neuroprotective effects have raised interest. This study confirmed that melittin specificity upregulated the weakened Nrf2/HO-1 signaling in mice brain. Interestingly, we also revealed melittin’s efficient tactic, as it was suggested to first restore redox balance and then gradually stabilized other regulations of the mouse hippocampus. Using a neuro-stress-induced scopolamine model, chromatography and mass spectrometry analysis revealed that melittin crossed the compromised blood–brain barrier and accumulated in the hippocampus, which provided the chance to interact directly to weakened neurons. A wide range of improvements of melittin action were observed from various tests from behavior Morris water maze, Y maze test to immune florescent staining, western blots. As we need to find out what is the focus of melittin action, we conducted a careful observation in mice which showed that: the first signs of changes, in the hippocampus, within 5 h after melittin administration were the restoration of the Nrf2/HO-1 system and suppression of oxidative stress. After this event, from 7 to 12.5 h after administration, a diversity of conditions was all ameliorated: inflammation, apoptosis, neurotrophic factors, cholinergic function, and tissue ATP level. This chain reaction underscores that melittin focus was on redox balance's role, which revived multiple neuronal functions. Evidence of enhancement in the mouse hippocampus led to further exploration with hippocampal cell line HT22 model. Immunofluorescence analysis showed melittin-induced Nrf2 translocation to the nucleus, which would initiating the translation of antioxidant genes like HO-1. Pathway inhibitors pinpointed melittin's direct influence on the Nrf2/HO-1 pathway. 3D docking models and pull-down assays suggested melittin's direct interaction with Keap1, the regulator of the Nrf2/HO-1 pathway. Overall, this study not only highlighted melittin specifically effect on Nrf2/HO-1, thus rebalancing cellular redox, and also showed that this is an effective multi-faceted therapeutic strategy against neurodegeneration.

Synergistic Protection of Vitamin B Complex and Alpha-Lipoic Acid Against Hepatic Ischemia–Reperfusion Injury: Boosting Antioxidant Defenses in Rats

This study aimed to investigate the protective effects of vitamin B complex and alpha-lipoic acid (ALA) pre-treatments on hepatic ischemia–reperfusion injury (IRI) in rats, focusing on their potential to enhance antioxidant defense mechanisms and reduce post-ischemic liver damage. Thirty male Wistar albino rats were divided into four groups: sham group (n = 10), IRI group (n = 10), vitamin B group (n = 10), vitamin B + ALA group (n = 10). In the IRI, vitamin B, and vitamin B + ALA groups, the rats underwent 45 min of hepatic ischemia followed by 60 min of reperfusion. Serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea, creatinine, and lactate dehydrogenase (LDH) were measured. Additionally, serum total antioxidant status (TAS) and total oxidant status (TOS) were assessed, and the oxidative stress index (OSI) was calculated. Liver tissue samples were collected for morphological evaluation. In the vitamin B and vitamin B + ALA groups, ALT, AST, urea, creatinine and LDH levels were better compared with the IRI group but the difference was statistically significant for only LDH levels in the vitamin B group and ALT, urea, and LDH levels in the vitamin B + ALA group (p &lt; 0.05). The lowest TOS and OSI levels were reported in the vitamin B and vitamin B + ALA groups and these groups had statistically significantly higher TAS compared with the sham and IRI groups (p &lt; 0.05). Our findings suggest that a vitamin B complex alone or a vitamin B complex + ALA combination reduces post-ischemic hepatic injury by enhancing the anti-oxidative status. The low dose of ALA may be a co-factor in these results and studies with larger doses of ALA are required to determine its effects on IRI of the liver.

Esculin, a Coumarin Glucoside Prevents Fluoride-Induced Oxidative Stress and Cardiotoxicity in Zebrafish Larvae.

Fluoride (F-) is a major groundwater contaminant spread across the world. In excess concentrations, F- can be detrimental to living beings. F- exposure is linked to cellular redox dyshomeostasis, leading to oxidative stress-mediated pathologies including heart dysfunction. Due to its potent antioxidant properties, various phytochemicals are found to alleviate the symptoms of F- toxicity. Hence, we explore the protective effect of esculin (Esc), a coumarin glucoside on F--induced oxidative stress and cardiotoxicity in zebrafish larvae. The experimental groups consisted of NaF (50 ppm) and Esc (100 μM) groups treated alone and in combination with a control group for 6 h. The groups were maintained till 78 hpf after which the level of oxidants (ROS, LPO, and PCC) and antioxidants (GST, GSH, GPx, SOD, and CAT) were assessed. The results revealed that Esc pretreatment restored the depleted antioxidant markers and reduced the levels of oxidant in the Esc+NaF group, exhibiting its antioxidant potential. In addition, analyses of the heartbeat rate and hemoglobin integrity using o-Dianisidine staining were conducted in the control and experimental groups. Esc treatment prevents F- induced cardiac changes including tachycardia and altered blood flow. Further, the mRNA expression level of antioxidant genes (nrf2, gstp1, hmox1a, prdx1, and nqo1) and cardiac developmental genes (bmp2b, nkx2.5, myh6, and myl7) confirmed that Esc acts as a potent free radical scavenger and antioxidant defense enhancer, protecting zebrafish larvae from NaF-induced oxidative stress and heart dysfunction.

Combined application of biochar and silicon nanoparticles enhance soil and wheat productivity under drought: Insights into physiological and antioxidant defense mechanisms

Agricultural drought periods are enhanced by the change in global climate that threatens food security. One of the main factors affecting soil and wheat productivity worldwide is drought stress. Especially, in semi-arid climates, studies are not found about adding organic amendments may be a viable way to reduce the adverse impacts of drought on crops while enhancing soil qualities and water use efficiency. In this study, we aimed to explore the effects of combined application of biochar (BC = 5 %) and silicon nanoparticles (SiNP = 900 mg/L) on soil and wheat drought resistance. Drought stress was applied at the three most critical growth stages of wheat, tillering (DTS), flowering (DFS), and grain filling (DGFS) stages, then evaluated the soil nutrients and wheat drought physiological resistance under applied treatments (BC, SiNP, BC+SiNP). Results showed that combined treatment of BC and SiNP greatly reduced the adverse effects of drought by enhancing plant height (9.36 %), spike length (25.63 %), number of fertile tillers (29.26 %), grains per spike (10.86 %), thousand-grain weight (18.25 %), and biological yield (16.34 %) with comparison to the control application. Additionally, physiological measures like water use efficiency (20.58 %), stomatal conductance (29.26 %), chlorophyll a (16.25 %), chlorophyll b (18.96 %), transpiration rate (21.65 %), photosynthetic rate (22.94 %), electrolyte leakage (-17.77 %), MDA (29.25 %), hydrogen peroxide (-19.88 %), superoxide dismutase (11.19 %), catalase (9.26 %), peroxidase (18.59 %), nitrogen (14.81 %), phosphorus (13.89 %), and potassium (17.61 %) were also meaningfully improved by treated application. BC and SiNP also significantly amended the organic carbon, nitrogen, and minerals percentage in soil. In conclusion, using the synergistic application of BC and SiNP could be a successful strategy to promote the biological soil properties, plant growth, yield, and quality of wheat crops as compared to all other treatments via dropping the dangerous effects of drought stress.

Effect of Ceramides Derivatives from the Peach on Skin Function Improvement in UV-Irradiated Hairless Mice

This study investigated the protective effects of a ceramides derivates from the peach (PF3) on photoaging by UV-irradiated hairless mice. Mice were randomly divided into seven groups: AIN93G without UVB exposure (normal control, NC), AIN93G with UVB exposure (control, C), AIN93G supplemented 100 mg/kg body weight (BW) of L-ascorbic acid with UVB exposure (AA), AIN93G supplemented 100 mg/kg BW of arbutin with UVB exposure (Arbutin), AIN93G supplemented 10 mg/kg BW of PF3 with UVB exposure (10PF3), AIN93G supplemented 20 mg/kg BW of PF3 with UVB exposure (20PF3), and AIN93G supplemented 40 mg/kg BW of PF3 with UVB exposure (40PF3). The study examined the impact of PF3 on skin hydration, wrinkle formation, and melanogenesis using enzyme-linked immunosorbent assay (ELISA), real-time polymerase chain reaction (real-time PCR), and Western blot analysis. The PF3 demonstrated significant protective effects against photoaging by reducing skin wrinkle formation, decreasing epidermal and dermal thickening, and improving skin hydration. It also enhanced the expression of moisture-related factors (hyaluronic acid synthase [HAS], long-chain ceramides [LCBs], dihydroceramide desaturase 1 [DEGS1], and type I collagen [COL1A]) and antioxidant enzyme activities while reducing pro-inflammatory cytokines and oxidative stress markers. The PF3 supplementation positively modulated skin wrinkle formation-related factors, increasing collagen-related gene expression and decreasing matrix metalloproteinases. Additionally, PF3 showed potential in regulating melanogenesis by reducing the nitric oxide and cAMP content, as well as the expression of melanogenesis-related proteins. These comprehensive findings suggest that PF3 supplementation may be an effective strategy for preventing and treating UVB-induced skin photoaging through multiple mechanisms, including improved skin structure, hydration, antioxidant defense, and reduced inflammation and pigmentation.

Green and chemical synthesis of iron oxide nanoparticles: Comparative study for antimicrobial activity and toxicity concerns

Infections caused by multidrug-resistant bacteria in healthcare settings highlight the critical need for robust infection control strategies and effective antimicrobial stewardship. This was further emphasized by the COVID-19 pandemic. This necessitates developing more effective antimicrobial agents. Iron oxide nanoparticles (IONPs) were reported to be biocompatible and possess a broad-spectrum antimicrobial activity with less susceptibilty of developing microbial resistance. Chemical synthesis is employed for industrial manufacture of IONPs. However, concerns about potential toxicity associated with chemical synthesis have led to a preference for green synthesis approaches, despite ongoing debates regarding their cytotoxicity. According to the authors' knowledge, the impact of synthesis methodology on the antimicrobial activity and cytotoxicity of IONPs has not been previously studied. Therefore, the current study is considered the first attempt to assess and compare the toxicity and antimicrobial effect of IONPs synthesized using different methodologies (green versus chemical synthesis) while correlating these effects to the physicochemical properties of IONPs. Additionally, this study addresses for the first time the disturbance of the microbial antioxidant defense system in multidrug-resistant bacteria after their exposure to IONPs. Both types of IONPs effectively eradicated SARS-CoV-2, and multidrug-resistant bacteria: S. aureus, and E. Coli, in addition, both disrupted the antioxidant bacterial defense mechanism, indicating that reactive oxygen species play a significant role in their antimicrobial activities. Chemically synthesized IONPs showed superior antimicrobial activity to those produced by green methods and in-vivo study demonstrated their greater compatibility with skin and eyes. Therefore, chemically synthesized IONPs might be used as a disinfectant in healthcare settings.

Sediment contamination in two German estuaries: A biomarker-based toxicity test with the ragworm Hediste diversicolor under intermittent oxygenation

Toxicity testing is an important tool for risk assessment of sediment contamination in estuaries. However, there has been a predominant focus on fitness parameters as toxic endpoints and on crustaceans as test organisms, while effects at the sub-organismal level and on other benthic taxa have received less attention. Also, interactions between sediment contamination and natural stressors such as oxygen are often neglected in traditional toxicity tests. Here we conducted a toxicity test of sediments from the Elbe and Oder (Odra) estuaries under three weeks of continuous and intermittent oxygenation, using biomarkers in an annelid, the ragworm Hediste diversicolor. Contaminated sediments affected worm survival and some biomarkers of antioxidant defense, electrophilic stress, and energy status with response ratios of above 20%. Toxic effects were most pronounced in sediments from the upper Elbe estuary, which contained high levels of heavy metals and organic chemicals. Oxygen regimes hardly changed the sediment effects, suggesting the robustness of the biomarker-based toxicity test with ragworms.

Effects of effluents from the Villa Victoria Reservoir (Mexico) on the development of Danio rerio at early life stages through apoptotic response and oxidative-induced state

As one of Mexico's most crucial water storage facilities, the Villa Victoria Reservoir (VVR) supplies water to over six million people residing in the Mexico City Metropolitan Area. In recent years, this water resource has been subjected to significant risks due to several factors, including human population growth, alterations in global climate patterns, excessive resource utilization, and insufficient protective regulations, thereby endangering not only the biocenosis itself, but also the water supply for numerous inhabitants. This study aimed to evaluate the current state of the reservoir through the determination of conventional and emerging pollutants present in the sampling points, as well as embryotoxicity and oxidative damage in Danio rerio embryos exposed to effluents from the VVR. Embryotoxicity was quantified using the General Morphology Score (GMS) and teratogenic index, whereas oxidative damage was assessed based on lipid peroxidation, hydroperoxide content, oxidized proteins, antioxidant enzyme activity, and gene expression. These results revealed the presence of heavy metals, diverse pharmaceutical compounds, and pesticides. In addition, elevated lipid, hydroperoxide, and protein oxidation accompanied by alterations in superoxide dismutase (SOD) and catalase (CAT) enzymatic activity were observed during exposure. GMS resulted in impaired embryo development and teratogenic effects, including pericardial, axial, and skeletal edema. Furthermore, the upregulation of genes associated with apoptotic processes and antioxidant defense reflects a comprehensive response to oxidative stress. The study concluded that pollutants in VVR water induced oxidative damage, modified antioxidant activity, elicited embryotoxicity, and upregulated oxidative damage-related genes. The findings underscore the necessity of undertaking restoration efforts for water sources, as pollution can potentially endanger aquatic organisms and human well-being.

Therapeutic Effects of Liposomal Resveratrol in the Mitigation of Diabetic Nephropathy via Modulating Inflammatory Response, Oxidative Stress, and Apoptosis.

An important factor in the development of diabetes and its associated consequences is prolonged chronic hyperglycemia, which weakens the antioxidant defense system and produces reactive oxygen species. Phytochemicals have been found to scavenge free radicals and exhibit antioxidant effects necessary to increase insulin sensitivity and reduce the development of diabetes-related complications. Current treatments for managing diabetes and diabetic nephropathy are often not very effective and come with several limitations and side effects. Resveratrol, for example, has shown therapeutic potential in mitigating kidney damage induced by high glucose levels, but its short bioavailability is a significant limitation. This accentuates the need for alternatives that not only improve the disease but also reduce the side effects associated with treatment. To enhance the therapeutic efficacy of resveratrol, we investigated the protective effects of liposomal resveratrol (LR) in a streptozotocin-induced diabetic rat model at doses of 20 and 40mg/kg. We compared the impact of LR to that of resveratrol alone (at a dose of 40mg/kg) on various parameters, including serum levels of biochemical markers, tissue levels of pro-inflammatory cytokines, nuclear transcription factor, oxidative stress indices, and apoptotic markers. LR, as a highly absorbable and metabolized form of resveratrol, has demonstrated beneficial effects in diabetic rats. Administered at both 20mg/kg and 40mg/kg dosages over a 5-week period, it demonstrated notable efficacy in alleviating inflammation. This was accomplished by diminishing the levels of pro-inflammatory mediators, TNF-α and IL-6, through the inhibition of NF-κB translocation. Additionally, LR influenced apoptotic markers, specifically caspase, BCL-2, and BAX. Furthermore, it enhanced the expression of key antioxidant enzymes such as catalase and glutathione peroxidase while significantly lowering malondialdehyde levels. These significant biochemical and immunological protective effects correlated with improved histological integrity and overall kidney architecture. Notably, resveratrol alone was not as effective as LR in restoring kidney function, highlighting its potential as a therapeutic candidate for the treatment of diabetic nephropathy. However, more in-depth studies are needed to explore its mechanism of action and improved bioavailability.

Physiological and ecological responses of flue-cured tobacco to field chilling stress: insights from metabolomics and proteomics

IntroductionCurrently, research on tobacco's response to chilling stress is mostly limited to laboratory simulations, where temperature is controlled to study physiological and molecular responses. However, laboratory conditions cannot fully replicate the complex environment of field chilling stress, so conducting research under field conditions is crucial for understanding the multi-level adaptive mechanisms of tobacco to chilling stress in natural environments. MethodsThis study aims to use field trials, starting from physiological responses, combined with proteomics and untargeted metabolomics, to systematically reveal the physiological and biochemical characteristics and key molecular mechanisms of tobacco leaves under chilling stress. It provides new insights into tobacco's adaptation strategies under chilling stress. ResultsThe results showed that (1) chilling stress damages the appearance of tobacco leaves, reduces the chlorophyll content, increases H2O2 and malondialdehyde (MDA) levels in cold-injured tobacco leaves, and damages the plasma membrane system. Although catalase (CAT) activity increases to cope with the accumulation of reactive oxygen species (ROS), the activities of key antioxidant enzymes superoxide dismutase (SOD) and peroxidase (POD) significantly decrease, indicating that the antioxidant system of tobacco leaves fails in environments with sudden temperature drops. (2) Proteomics analysis indicated that 410 differentially expressed proteins were identified in cold-stressed tobacco leaves, with 176 upregulated and 234 downregulated. Tobacco leaves under chilling stress attempt to maintain energy supply and physiological stability by enhancing glycolysis, starch, and sucrose metabolism pathways. Concurrently, chilling stress triggers the expression of proteins related to cell wall reinforcement and antioxidant defense. However, due to impaired ribosomal function, protein synthesis is significantly inhibited, which aggravates damage to photosynthesis and cellular functions. (3) Metabolomics analysis revealed that the differential metabolites in cold-stressed tobacco leaves were mainly enriched in tyrosine metabolism, isoquinoline alkaloid biosynthesis, and fatty acid degradation pathways. This indicates that under chilling stress, tobacco leaves enhance adaptability by regulating energy metabolism, increasing antioxidant capacity, and stabilizing cell membrane structure. ConclusionsTherefore, under chilling stress, tobacco leaves exhibit complex physiological adaptability through multiple regulatory mechanisms involving proteins and metabolites. The research results provide important insights into the metabolic regulatory mechanisms of tobacco in response to extreme environments and also enhance the theoretical foundation for addressing low-temperature stress in practical production.

Peculiarities of Plant Mineral Composition in Semi-Desert Conditions

Plant–soil interactions in semi-desert conditions elicit the development of plant-specific adaptation strategies, including selective accumulation of macro- and microelements. Using an ICP-MS analysis of 12 plant species belonging to Asteraceae, Fabaceae, Poaceae, Ephedraceae, Amarantaceae, and Lamiaceae families of the Baskunchak Nature Reserve, remarkable species differences in accumulation of 22 macro- and microelements were recorded. The most common Artemisia species and Poaceae representatives belong to two different groups of plants with high content of Na, K, Zn, Cu, V and high antioxidant status and low Si typical for the former group and the opposite characteristics for the latter one. The mentioned phenomenon indicates two diverse powerful adaptation mechanisms based on the antioxidant defense and Si protection, respectively. The high frequency of remarkable levels of Se in plants with BCF exceeding 1 (Glycyrrhiza aspera, Phlomis pungens, Tanacetum nullifolium, Helichrysum nogaicum, and Jurinea ewersmannii), Zn in all species except Poa angustifolia, and Cu in the Asteraceae plants Phlomis pungens and Krascheninnikovia ceratoides suggests the significance of these elements in plant tolerance to environmental stresses. Plant–soil positive correlations were recorded for Sr (r = 0.866; p &lt; 0.001); plant Sr, Fe, Co, Pb levels and soil salinity (r = 0.763, p &lt; 0.001; r = 0.606, p &lt; 0.03; r = 0.627, p &lt; 0.02; r = 0.548, p &lt; 0.05, respectively); and Cr only for Asteraceae species (r = 0.986, p &lt; 0.001). The results obtained in this research may be used in plant adaptability evaluation in conditions of environmental stress.

Construction and biological activity of metallothionein fused with ELP

Metallothionein (MT) plays a significant role in heavy metal removal, antioxidant defense, and immune regulation. The current predominant approach for obtaining natural MT is extraction from tissue, which often entails complex procedures resulting in limited yields. In recent years, researchers have adopted the strategy of fusing labels such as GST or His for the heterologous expression of MT. However, a challenge in industrial production arises from the subsequent removal of these labels, which often leads to a significant reduction in the yield. The fusion with elastin-like polypeptides (ELPs) offers a promising solution for achieving soluble expression of the target protein, while providing a simple and fast purification process. In this study, ELP was fused with MT, which significantly up-regulated the soluble expression of MT. The fusion protein ELP-MT with the purity above 97% was obtained efficiently and simply by inverse transition cycling (ITC). ELP-MT exhibited a remarkable 2,2'-azinobis(3-ethylbenzothiazoline-6- sulfonic acid) ammonium salt (ABTS) scavenging activity, with the half maximal inhibitory concentration (IC50) of 0.77 μmol/L, which was 53.7 times that of the vitamin E derivative Trolox. In addition, the fusion protein demonstrated strong 1,1-diphenyl-2-trinitrohydrazine (DPPH) scavenging ability. Furthermore, ELP-MT had no toxicity to the proliferation and promoted the adhesion and migration of NIH/3T3 cells. All these results indicated that ELP-MT had good biocompatibility. We constructed the fusion protein ELP-MT combining the unique properties of MT and elastin, laying a technical foundation for the large-scale production of recombinant MT and facilitating the applications in food, health supplement, and cosmetic industries.

Low-Dose Ozone as a Eustress Inducer: Experimental Evidence of the Molecular Mechanisms Accounting for Its Therapeutic Action

Ozone (O3) is an unstable, highly oxidative gas that rapidly decomposes into oxygen. The therapeutic use of O3 dates back to the beginning of 20th century and is currently based on the application of low doses, inducing moderate oxidative stress that stimulates the antioxidant cellular defences without causing cell damage. In recent decades, experimental investigations allowed the establishment of some basic mechanisms accounting for the therapeutic effects of eustress-inducing low-dose O3. In this review, special attention was given to the impact of O3 administration on the cell oxidant–antioxidant status, O3 anti-inflammatory and analgesic properties, efficacy in improving tissue regeneration, and potential anticancer action. Low O3 concentrations proved to drive the cell antioxidant response mainly by activating nuclear factor erythroid 2-related factor 2. The anti-inflammatory effect relies on the downregulation of pro-inflammatory factors and the modulation of cytokine secretion. The painkilling action is related to anti-inflammatory processes, inhibition of apoptosis and autophagy, and modulation of pain receptors. The regenerative potential depends on antioxidant, anti-inflammatory, anti-apoptotic, and pro-proliferative capabilities, as well as fibroblast activation. Finally, the anticancer potential is based on oxidant and anti-inflammatory properties, as well as the inhibition of cell proliferation, invasion, and migration and the induction of apoptosis.

Glyphosate and glyphosate-based herbicides induce Poecilia reticulata to maintain redox equilibrium during and after coexposure to iron oxide nanoparticles (y-Fe2O3)

Iron oxide nanoparticles (IONPs) are being increasingly recognized as viable materials for environmental remediation due to their capacity to adsorb contaminants such as glyphosate (GLY) on their surfaces. Nevertheless, the ecotoxicological implications of IONPs associated with GLY necessitate thorough evaluation to ascertain the safety of such remediation strategies. In this context, the present investigation was conducted to examine hepatic biomarkers pertinent to the redox system, as well as ultrastructural hepatic alterations in Poecilia reticulata, following a 21-day exposure to environmentally relevant concentrations of IONPs, iron ions (Fe), and glyphosate in its pure form (GLY) as well as a commercial glyphosate-based herbicide (GBH). After this exposure, the fish underwent a 21-day recovery in uncontaminated water. The results indicated an increase in the activity of catalase (CAT) and glutathione S-transferase (GST) and in the concentration of glutathione (GSH) in the animals subjected to IONP+GBH and IONP+GLY treatments. This biochemical response persisted for the duration of both the exposure and recovery phases. Concurrently, hepatocytes displayed mitochondria with increased electron density, augmented lipid droplet accumulation, and expanded necrotic areas within the hepatic tissue. In contrast, fish exposed solely to IONPs exhibited sustained redox homeostasis throughout the investigative timeline. These findings suggest that the coexposure toxicity of IONP+GLY and IONP+GBH is attributable to the agent adsorbed onto the IONPs and that P. reticulata could maintain an active antioxidant defense mechanism throughout the entire study period.

Luciferase-Based Reporter System for Investigating GPx4-Mediated Ferroptosis and Its Therapeutic Implications in Diabetes.

Ferroptosis, a distinct form of regulated cell death, is characterized by iron-dependent lipid peroxide accumulation in cell membranes from dysregulated cellular iron homeostasis and compromised antioxidant defense mechanisms. Glutathione peroxidase 4 (GPx4) is crucial in the regulation of ferroptosis by controlling lipid peroxide accumulation. Recent research established the association of ferroptosis with several diseases, prompting investigation toward ferroptosis-targeted therapeutic approaches. However, there is a lack of sensor systems designed to evaluate ferroptosis modulation in intact cells. In this study, we developed a highly sensitive luciferase-based reporter system to study GPx4-mediated ferroptosis in cells. We constructed a novel vector flanking the GPx4 promoter driving luciferase gene expression, demonstrating ferroptosis-specific luciferase activity in transfected HEK293T cells. We established stable cells expressing the construct and optimized its suitability for high-throughput screening using well-established ferroptosis modulators. We identified eugenol, a phenolic compound, as a potent ferroptosis inhibitor using the developed reporter system. Eugenol demonstrated dose-dependent protection against ferroptosis-induced damage in pancreatic beta cells, as assessed by the expression of the key markers such as GPx4, SLC7A11, NRF2, and HO1. Further, we showed the regulation of iron levels and total iron-binding capacity of beta cells by eugenol in streptozotocin (STZ) -induced diabetic mice. Additionally, the diabetes-induced downregulation of GPx4 and antioxidant Nrf2 in pancreatic tissue was significantly mitigated by eugenol, as evidenced by both immunohistochemistry and gene expression analysis. This research validates the functionality of the ferroptosis sensor and offers an approach to develop antidiabetic therapy by targeting ferroptosis to protect beta-cell viability and function.

Optimizing selenium-enriched yeast supplementation in laying hens: Enhancing egg quality, selenium concentration in eggs, antioxidant defense, and liver health

This study evaluated the effects of selenium-enriched yeast (SY) supplementation at various levels on health and production parameters in laying hens, including egg production, egg quality, selenium (Se) concentrations in eggs, liver health, serum biochemical markers, antioxidant function, and immune responses. A total of 360 Hy-Line Brown hens (28 weeks old) were randomly assigned to four dietary groups with six replicates of 15 birds each, monitored over a 12-week feeding trial after a two-week acclimatization period. The dietary groups included a control (basal diet without selenium) and three SY-supplemented groups with Se levels of 0.3 mg/kg (SY03), 1.5 mg/kg (SY15), and 6.0 mg/kg (SY60). The results showed no significant effects of dietary SY on laying performance or feed efficiency (P > 0.05). However, the SY15 group showed significant improvements in egg quality, particularly in albumen height, Haugh Unit and yolk color (P < 0.05). Selenium concentrations in eggs, albumen, and yolk increased dose-dependently, with significant differences in the SY-supplemented groups (P < 0.001). Increased activities of liver enzymes including alanine transaminase, alkaline phosphatase, and aspartate transaminase, alongside elevated levels of uric acid were notable in the SY60 group (P < 0.05). In addition, histological analysis revealed significant hepatocyte degeneration and a higher liver organ index (P < 0.05), in the SY60 group. All of which suggests potential liver toxicity at higher selenium levels. Antioxidant capacity of the birds were significantly enhanced due to dietary supplementation of SY as indicated by increased serum levels of total antioxidant capacity, and activities of catalase, glutathione peroxidase, and superoxide dismutase (P < 0.05). Analysis of hepatic genes expression revealed that SY15 supplementation significantly upregulated key antioxidant-related genes (Nrf2, HO-1, CAT, and NQO1) and downregulated Keap1 expression (P < 0.05), suggesting strong activation of the antioxidant defense system. In conclusion, SY supplementation at 1.5 mg/kg improved egg quality, increased Se concentrations in eggs, and enhanced antioxidant capacity without affecting laying performance or liver health. This makes it a balanced approach to improving egg quality and poultry health. However, higher supplementation levels (6.0 mg/kg) resulted in liver damage, underscoring the importance of careful dosage consideration.

Polymorphic variants of the genes for enzymes of the antioxidant system, apoptosis and inflammation as potential predictors of myocardial infarction

Myocardial infarction (MI) is a multifactorial polygenic disease that develops as a result of a complex interaction of numerous genetic factors and the external environment. Accordingly, the contribution of each of them separately is usually not large and may significantly depend on the state of other accompanying factors. The purpose of the study was to search for informative predictors of MI risk based on polygenic analysis of polymorphic variants of (1) the antioxidant defense enzyme genes PON1 (rs662), PON2 (rs7493), CAT (rs1001179), MSRA (rs10098474) and GSTP1 (rs1695); (2) the apoptosis genes CASP8 (rs3834129), TP53 (rs1042522) and BCL2 (rs12454712); and (3) the inflammation genes CRP (rs1205), CX3CR1 (rs3732378), IL6 (rs1800795) and CCL2 (rs1024611). 591 DNA samples were used in the study (280 patients with the onset at 30 to 60 years, with an average age of 46.02 ± 6.17, and 311 control subjects aged 30 to 62, with an average age of 44.65 ± 7.07). All the participants were male and Tatars by ethnicity. The logistic regression analysis with various models demonstrated associations with MI of polymorphic variants of the genes CX3CR1 (rs3732378) (overdominant model – G/G + A/A vs A/G P = 0.0002, OR = 1.9), MSRA (rs10098474) (dominant model – T/T vs T/C + C/C P = 0.015, OR = 1.51), CCL2 (rs1024611) (recessive model – P = 0.0007 – A/A + A/G vs G/G OR = 2.63), BCL2 (rs12454712) (log-additive model – *C allele, P = 0.005, OR = 1.38). Using the Monte Carlo method and Markov chains (APSampler), combinations of alleles/ genotypes of the studied polymorphic loci associated with a high risk of MI were obtained, which, in addition to those identified during single-locus analysis, contained polymorphic variants of the genes CASP8, TP53, CAT, PON2, CRP, IL6, GSTP1. Among the combinations obtained, a pairwise analysis of possible non-linear interactions between the identified combinations of alleles/genotypes was carried out, which showed synergistic interactions of the polymorphic variants CX3CR1*A/G and CASP8*I/I, MSRA*C and CRP*C, CAT*C/T and MSRA*C, CAT*C/T and CX3CR1*A contributing to the development of MI. Based on the results obtained using multivariate logistic regression analysis, a predictive model was built to assess the risk of developing MI, the predictive ability of which reached the value AUC = 0.71 (AUC – area under the curve in ROC analysis).