Oxidative Alterations Research Articles

Insights into the potential mechanism of liquid nitrogen spray freezing's influence on volatile compounds in surimi gels with different cross-linking degrees: Focus on oxidation, protein structure, intermolecular force and free amino acid alterations

This study revealed the variations in odor characteristics and underlying mechanisms of different cross-linked surimi gels under liquid nitrogen (LN) spray freezing. The results demonstrated that LN spray freezing had an essential effect on the gels′ odor. The odor changes in the −80 °C LN spray freezing group were closer to the control group, while −35 °C LN spray freezing treatment had the greatest impact on the aroma quality of gels. Freezing reduced gels′ texture properties, intensified lipid and protein oxidation, altered protein conformation, increased surface hydrophobicity and hydrophobic interactions. These changes affected the gels' odor characteristics, leading to a reduction in fish aroma and an increase in fishy and oil odors after freezing. These tendencies were more pronounced at −35 °C LN spray freezing with lower cross-linking degrees, and reducing the freezing temperature to −80 °C and increasing the cross-linking degree to 62.99% mitigated the extent of deterioration in gel flavor quality.

DOP88 Lipids drive myofibroblast activation and the epithelial-mesenchymal transition process in Crohn’s Disease

Abstract Background Fibrostenosis represents one of the major clinical complications of Crohn’s Disease (CD). However, the molecular mechanisms underlying intestinal fibrogenesis remain poorly understood and anti-fibrotic therapy has not yet been developed. Growing evidences supports that oxidative stress and the epithelial-mesenchymal transition (EMT) process contribute to the development and progression of fibrosis. Here, we assessed the role of lipids in fibrogenesis and whether mitochondria dysfunction can support fibrosis formation. Methods Spatial metabolomics analysis and adipophilin (ADFP) staining were performed on surgical specimens from fibrotic ileum and the surrounding region (healthy) derived from CD patients. Fibroblasts isolated from fibrotic and healthy ileum and human induced pluripotent stem cell-derived intestinal organoids (iHOs) were treated for 1-3 weeks with lipid mix (2%) and analyzed by RNA sequencing. Pathways significantly modulated were validated by RT-PCR, WB, FACS analysis, and proliferative, migratory and contraction assays, and mitochondrial functions by Seahorse analyzer. Results Spatial metabolomics revealed a significant accumulation of lipids characterized by very long-chain fatty acids in the fibrotic area compared to surrounding healthy regions, which was further confirmed by ADFP staining. RNA-seq analysis showed overlapping dysregulated pathways between healthy fibroblasts stimulated with lipids and fibrotic fibroblasts, in particular genes related to cell cycle, cytoskeletal reorganization, migration, contractility and cell metabolism (oxidative stress) which were further confirmed by functional assays. Among the genes related to oxidative stress, lipid stimulation significantly decreased NFE2L2 and promoted an increase in CPT1A, gene encoding for a fatty oxidation mediator. Moreover, lipid-stimulated fibroblasts showed a reduction in mitochondrial respiration, particularly basal and maximal respiration, and spare respiratory capacity by Seahorse analysis. On the other hand, long exposure of iHOs to lipids promoted the EMT process through the upregulation of TWIST, ZEB, VIM, and S100a4. Conclusion Lipid stimulation induced myofibroblast activation through oxidative stress and alteration of mitochondrial metabolism, and increased the expression of EMT transcription factors in intestinal epithelial cells (iHOs) that may support fibroblast-mediated pro-fibrotic activity. These data indicate that regulators of fatty acid oxidation involved in the fibrotic process may represent new potential targets for the treatment of fibrostenotic CD patients.

Behavioral, Biometric and Oxidative Stress-Induced Alterations in Clarias gariepinus Juveniles Exposed to Chlorpyrifos

Chlorpyrifos is an organophosphate pesticide used to kill wide range of pest including insect and worms. The present study investigates the behavioral, biometric and oxidative stress alterations in Clarias gariepinus juveniles exposed to chlorpyrifos. The 96 h LC50 values of the pesticide estimated by probit analysis was 4.67 mg/L while the toxic unit was 21.41 indicating that the pesticide is very toxic. Fish were exposed to chlorpyrifos sublethal concentrations of 0.43 mg/L and 0.93 mg/L and the behavioral, biometric and oxidative stress parameters were determined on day 1, 5, 10, and 15. The results indicate changes in the behavior and hepatosomatic indices in C. gariepinus at different chlorpyrifos concentrations and time. There were concentration and duration dependent increase in lipid peroxidation, protein and glucose but mixed trends in catalase values. The use of chlopyrifos in the environment especially near aquatic ecosystem should be strictly monitored to avoid the hazards associated with its application.

Human umbilical cord-derived mesenchymal stem cells ameliorate liver fibrosis by improving mitochondrial function via Slc25a47-Sirt3 signaling pathway

Chronic Liver fibrosis may progress to liver cirrhosis and hepatocellular carcinoma (HCC), hence cause a substantial global burden. However, effective therapies for blocking fibrosis are still lacking. Although mesenchymal stem cells (MSCs) have been proven beneficial to liver regeneration after damage, the underlying mechanism of their therapeutic effects are not fully understood. Oxidative stress and mitochondrial functionality alteration directly contributes to the hepatocyte apoptosis and development of liver fibrosis. This study aims to elucidate the mechanism by which hUC-MSC alleviates liver fibrosis and mitochondrial dysfunction. RNA-sequencing was performed to characterize the transcriptomic changes after implantation of hUC-MSCs in mice with liver fibrosis. Next, western blot, RT-PCR, immunohistochemical and immunofluorescence staining were used to evaluate the expression of different genes in vitro and in vivo. Additionally, mitochondrial morphological and dynamic changes, ROS content, and ATP production were examined. Slc25a47, a newly identified liver-specific mitochondrial NAD+ transporter, was notably reduced in CCl4-treated mice and H2O2-stimulated hepatocytes. Conversely, hUC-MSCs increased the Slc25a47 expression and NAD+ level within mitochondria, thereby enhanced Sirt3 protein activity and alleviated mitochondrial dysfunction in the liver. Furthermore, Slc25a47 knockdown could partially abrogate the protective effects of hUC-MSCs on H2O2-induced mitochondrial fission and oxidative stress in hepatocytes. Our study illustrates that Slc25a47 is a key molecular for hUC-MSCs to improve liver fibrosis and regulates mitochondrial function through Sirt3 for the first time, and providing a theoretical basis for the clinical translation of hUC-MSCs transplantation in the treatment of patients with liver fibrosis/cirrhosis.

Lipopeptide Biosurfactant Produced by Acinetobacter junii B6 Attenuates Lung Inflammatory, Oxidative, and Histopathological Alterations due to Asthma in Rats

Lipopeptide Biosurfactant Produced by Acinetobacter junii B6 Attenuates Lung Inflammatory, Oxidative, and Histopathological Alterations due to Asthma in Rats

Dietary supplementation with natural antioxidants: assessment of growth performance and meat quality in broiler chickens

Exposure to air and light lowers the meat quality of chickens, which is mainly determined by the color alteration and accumulation of detrimental products from lipid oxidation. This study tests the effects of two supplements rich in polyphenols, Oxilem (OX) and OxiGem (OG), on broiler Ross 308 performance and meat quality in comparison to a control group (C). A total of 105 one-day-old Ross 308 male chicks were allocated to 21 pens and randomly assigned to one of the three treatments. The trial lasted 42 days. Individual live weight and feed intake per pen were recorded. Proximate analysis, color, cholesterol content and cholesterol oxidation products (COPs), fatty acids (FAs), volatile aldehydes profile, and secondary oxidation products of meat FAs were determined, with analyses for detecting oxidative alterations conducted on breast burgers preserved for seven days at 4°C. Birds fed OG grew 7 g/day more than those receiving OX, reaching a higher final weight. After slaughtering, meat from the OX group had a higher yellow index compared to C. After seven days of air and light exposure, the influence on the color parameters of the OG and OX burgers was significantly less pronounced than that of C. Secondary oxidation products of the FAs of the burgers were not significantly affected by the diet regimens. In addition, OX and OG burgers exhibited lower amounts of volatile aldehydes, triol and COPs. These results confirm the effectiveness of OX and OG supplementation against lipid oxidation at the inclusion level used in this trial.

The Multifaceted Roles of NRF2 in Cancer: Friend or Foe?

Physiological concentrations of reactive oxygen species (ROS) play vital roles in various normal cellular processes, whereas excessive ROS generation is central to disease pathogenesis. The nuclear factor erythroid 2-related factor 2 (NRF2) is a critical transcription factor that regulates the cellular antioxidant systems in response to oxidative stress by governing the expression of genes encoding antioxidant enzymes that shield cells from diverse oxidative alterations. NRF2 and its negative regulator Kelch-like ECH-associated protein 1 (KEAP1) have been the focus of numerous investigations in elucidating whether NRF2 suppresses tumor promotion or conversely exerts pro-oncogenic effects. NRF2 has been found to participate in various pathological processes, including dysregulated cell proliferation, metabolic remodeling, and resistance to apoptosis. Herein, this review article will examine the intriguing role of phase separation in activating the NRF2 transcriptional activity and explore the NRF2 dual impacts on tumor immunology, cancer stem cells, metastasis, and long non-coding RNAs (LncRNAs). Taken together, this review aims to discuss the NRF2 multifaceted roles in both cancer prevention and promotion while also addressing the advantages, disadvantages, and limitations associated with modulating NRF2 therapeutically in cancer treatment.

Bisphenol-A induced cyto-genotoxicity on retinal pigment epithelial cells is differentially modulated by a multi-supplement containing guarana, selenium, and L-carnitine.

Bisphenol A (BPA) may adversely affect human health by inducing oxidative stress and irreversible damage to cells. Bioactive compounds found in some functional foods, individually or in combination, can attenuate the negative effects of BPA exposure; an example is the multi-supplement containing guarana (Gua), selenium (Se), and L-carnitine (LC) -GSC- which has already demonstrated antioxidant, genoprotective, and immunomodulatory activities. This study aimed to determine the effect of GSC and its constituents on oxidative and genotoxic alterations triggered by BPA exposure in the retinal epithelial cell line. The cells exposed to BPA (0.001, 0.01, 0.1, 1, 3, and 10 µM) to determine the lowest concentration required to induce cyto-genotoxicity. ARPE-19 cells were then concomitantly exposed to the selected BPA concentration, GSC, and its components (Gua, 1.07 mg/mL; Se, 0.178 µg/mL; and LC, 1.43 mg/mL). Flow cytometry, biochemical assays, qRT-PCR, genotoxicity, apoptosis, and cellular proliferation. Based on our results, 10 µM of BPA could induce cyto-genotoxic and oxidative alterations. BPA did not alter the Bcl-2/BAX expression ratio but induced Casp3 and Casp8 overexpression, suggesting that apoptosis was induced mainly via the extrinsic pathway. GSC partially reversed the alterations triggered by BPA in ARPE-19 cells. However, Se had unexpected negative effects on ARPE-19 cells. The multi-supplement GSC may attenuate changes in oxidative and genotoxic markers related to exposure of ARPE-19 cells to BPA. our results revealed that the antioxidant, anti-apoptotic, and genoprotective properties of GSC were not universally shared by its individual, once Se did not exhibit any positive impact.

The Efficacy of Methanolic Extract of Calotropis procera Leaf in Treating Eimeriosis-Induced Inflammation and Oxidative Status Alteration of the Jejunum in Experimental Mice

Calotropis procera leaf extract was utilized to evaluate the anti-inflammatory and oxidative status of Eimeria papillate infection in mice. The mice were divided into five groups: Group I served as the control. In Group II, considered uninfected, a dose of 200 mg/kg of the extract was administered to evaluate toxicity. Groups III, IV, and V were infected with 1 x 103 sporulated E. papillate oocysts. Group III received no treatment. Group IV was administered 200 mg/kg extractants daily, while Group V received 25 mg/kg of toltrazuril daily via gavage. On day 5 p.i., mice were slaughtered and jejunum tissues were prepared for examination. GC-MS analysis revealed nine biologically active chemical compounds. Also, quantitative results showed phenolics, flavonoids, and tannin 160.22±0.652 GAE, 34.046±0.545 QE, and 65.869±0.263 TAE/(mg/g DW), respectively. Moreover, IC50 was obtained at 44.2±0.007 g/mL for Hep-G2 cell lines. Results showed statistically significant differences in mice that received a dose of 200 mg/kg of the extract, compared to the infected group (P ≤ 0.05), as the tissue injury within the jejunum decreased by 61%. The extract successfully reverses the effects of the E. papillate infection on glutathione, malondialdehyde, and total protein levels. It has been demonstrated that extract possesses anti-inflammatory activities, and improves oxidative status.

The Effect of Propofol on the Hippocampus in Chronic Cerebral Hypoxia in a Rat Model Through Klotho Regulation.

Chronic cerebral hypoxia often leads to brain damage and inflammation. Propofol is suggested to have neuroprotective effects under anaesthesia. This study used rat models with carotid artery coarctation or closure. Four groups of rats were compared: a control group, a propofol-treated group, a group with bilateral common carotid artery blockage (BCAO), and a BCAO group treated with propofol post-surgery. The Morris water maze test indicated cognitive impairment in BCAO rats, which also showed hippocampal structure changes, oxidative stress markers alteration, and reduced Klotho expression. Propofol treatment post-BCAO surgery improved these outcomes, suggesting its potential in mitigating chronic cerebral hypoxia effects. Propofol may increase klotho levels and reduce apoptosis and inflammation linked to oxidative stress in cognitively impaired mice.

Potential applications of biosurfactants in animal production and meat research

<abstract> <p>Muscle foods are perishable products that are subject to several contaminations such as microbial and/or chemical (lipid and protein oxidation) alterations, which result in their deterioration and quality loss. Several processing strategies are used to preserve and improve the stability, shelf-life and quality of meat and meat products, from which natural preservative agents are gaining interest from both industrials and consumers as green and eco-friendly strategies. Among these natural preservatives, biosurfactants are emerging molecules. Their natural origin and biodegradability make them appealing for use in the food industry. In meat research, biosurfactants are of great interest as antimicrobial and antioxidant agents to reduce meat spoilage and wastage as well as for improving the shelf-life of the products. We aimed to discuss the potential applications of biosurfactants with a focus on their antimicrobial and antioxidant activity within the objectives of reducing meat quality deterioration and improving the image quality (acceptability by consumers) of meat and meat products. Additionally, further perspectives under the context of practical applications of biosurfactants in meat emulsification have been discussed, serving as a reference to feed knowledge gaps in this emerging topic of research. Further studies and evaluations of biosurfactants in meat research are needed to establish more evidence of their potential benefits, applicability and feasibility at a larger scale.</p> </abstract>

Science and Applications of Plasma-Activated Solutions: Current Trends and Future Directions

Recent research into plasma-activated solutions has unveiled their considerable potential across a range of applications, including cancer treatment, industrial processes, and environmental management. This review summarizes key findings from recent studies on the effects of plasma-activated solutions, focusing on their ability to induce cell death in glioblastoma cells, enhance ethanol production in yeast, and convert organic compounds into environmentally friendly substances. The review highlights the mechanisms by which plasma-activated solutions exert their effects, such as oxidative stress induction and cellular metabolic alterations. Additionally, it discusses the implications of these findings for overcoming challenges in cancer therapy, improving agricultural practices, and developing novel environmental technologies. The ongoing advancements in plasma technology offer promising avenues for future research and applications, underscoring its transformative potential in both scientific and practical domains.

Exploring Metabolic and Gut Microbiome Responses to Paraquat Administration in Male Wistar Rats: Implications for Oxidative Stress.

In this study, we examined the metabolic and gut microbiome responses to paraquat (PQ) in male Wistar rats, focusing on oxidative stress effects. Rats received a single intraperitoneal injection of PQ at 15 and 30 mg/kg, and various oxidative stress parameters (i.e., MDA, SOD, ROS, 8-isoprostanes) were assessed after three days. To explore the omic profile, GC-qTOF and UHPLC-qTOF were performed to assess the plasma metabolome; 1H-NMR was used to assess the urine metabolome; and shotgun metagenomics sequencing was performed to study the gut microbiome. Our results revealed reductions in body weight and tissue changes, particularly in the liver, were observed, suggesting a systemic effect of PQ. Elevated lipid peroxidation and reactive oxygen species levels in the liver and plasma indicated the induction of oxidative stress. Metabolic profiling revealed changes in the tricarboxylic acid cycle, accumulation of ketone body, and altered levels of key metabolites, such as 3-hydroxybutyric acid and serine, suggesting intricate links between energy metabolism and redox reactions. Plasma metabolomic analysis revealed alterations in mitochondrial metabolism, nicotinamide metabolism, and tryptophan degradation. The gut microbiome showed shifts, with higher PQ doses influencing microbial populations (e.g., Escherichia coli and Akkermansia muciniphila) and metagenomic functions (pyruvate metabolism, fermentation, nucleotide and amino acid biosynthesis). Overall, this study provides comprehensive insights into the complex interplay between PQ exposure, metabolic responses, and gut microbiome dynamics. These findings enhance our understanding of the mechanisms behind oxidative stress-induced metabolic alterations and underscore the connections between xenobiotic exposure, gut microbiota, and host metabolism.

Micro(nano)plastics: an Emerging Burden for Human Health.

The escalation of plastic pollution represents a global environmental and health problem. Important toxic effects have been attributed to the increasing diffusion of microplastics (MPs) and nanoplastics (NPs) derived from the degradation of plastics. These particles have been ubiquitously observed in the environment, with humans being continuously exposed via ingestion, inhalation and skin contact. Nonetheless, the cellular homeostasis imbalance induced by micro- and nano- plastics (MNPs) in human health has been only recently shown, while most evidence and molecular mechanisms derived from studies in vitro and in vivo models. To date, the majority of available results testified the accumulation of MNPs in the cardiovascular, nervous, reproductive and digestive systems, and recently clear evidence about cardiovascular toxic effects of MNPs has been provided in humans. In this context, this review aims to provide a comprehensive update about the most recent studies reporting the effects of MNPs in different models, focusing on the available evidence in the main areas of study related to human health. Hopefully, this review will contribute to raise awareness about the toxicity and oxidative alteration exerted by MNPs, supporting the elaboration of new strategies to counteract plastic pandemics.

Investigating the Mitoprotective Effects of S1P Receptor Modulators Ex Vivo Using a Novel Semi-Automated Live Imaging Set-Up.

In multiple sclerosis (MS), mitochondrial alterations appear to contribute to disease progression. The sphingosine-1-phosphate receptor modulator siponimod is approved for treating secondary progressive MS. Its preceding compound fingolimod was shown to prevent oxidative stress-induced alterations in mitochondrial morphology. Here, we assessed the effects of siponimod, compared to fingolimod, on neuronal mitochondria in oxidatively stressed hippocampal slices. We have also advanced the model of chronic organotypic hippocampal slices for live imaging, enabling semi-automated monitoring of mitochondrial alterations. The slices were prepared from B6.Cg-Tg(Thy1-CFP/COX8A)S2Lich/J mice that display fluorescent neuronal mitochondria. They were treated with hydrogen peroxide (oxidative stress paradigm) ± 1 nM siponimod or fingolimod for 24 h. Afterwards, mitochondrial dynamics were investigated. Under oxidative stress, the fraction of motile mitochondria decreased and mitochondria were shorter, smaller, and covered smaller distances. Siponimod partly prevented oxidatively induced alterations in mitochondrial morphology; for fingolimod, a similar trend was observed. Siponimod reduced the decrease in mitochondrial track displacement, while both compounds significantly increased track speed and preserved motility. The novel established imaging and analysis tools are suitable for assessing the dynamics of neuronal mitochondria ex vivo. Using these approaches, we showed that siponimod at 1 nM partially prevented oxidatively induced mitochondrial alterations in chronic brain slices.

Regulation of MMP-2 by IL-8 in Vascular Endothelial Cells: Probable Mechanism for Endothelial Dysfunction in Women with Preeclampsia.

Endothelial dysfunction (ED) in preeclampsia (PE) results from the convergence of oxidative stress, inflammation, and alterations in extracellular matrix components, affecting vascular tone and permeability. The molecular network leading to ED includes IL-8 and MMP-2. In vitro, IL-8 regulates the concentration and activity of MMP-2 in the trophoblast; this interaction has not been studied in endothelial cells during PE. We isolated human umbilical vein endothelial cells (HUVECs) from women with healthy pregnancies (NP, n = 15) and PE (n = 15). We quantified the intracellular concentration of nitric oxide and reactive oxygen species with colorimetric assays, IL-8 with ELISA, and MMP-2 with zymography and using an ELISA-type system. An IL-8 inhibition assay was used to study the influence of this cytokine on MMP-2 concentration and activity. HUVECs from women with PE showed significantly higher oxidative stress than NP. IL-8 and MMP-2 were found to be significantly elevated in PE HUVECs compared to NP. Inhibition of IL-8 in HUVECs from women with PE significantly decreased the concentration of MMP-2. We demonstrate that IL-8 is involved in the mechanisms of MMP-2 expression in HUVECs from women with PE. Our findings provide new insights into the molecular mechanisms regulating the ED distinctive of PE.

Sophora interrupta bedd., ameliorates doxorubicin induced myocardial necrosis by attenuation of oxidative stress and structural cardiomyocyte alterations in rats

The present study aimed at investigating the cardioprotective activity of Sophora interrupta Bedd against doxorubicin induced myocardial necrosis by attenuation of oxidative stress and structural cardiomyocyte alterations in rats.The experimental procedure was carried out with male Wistar rats inducing cardiotoxicity by administrating Doxorubicin in normal saline 3.750 mg/kg i.p on day 7, 14, 21 and 28 and treated with test extract of Sophora interrupta Bedd at a dose of 200 & 400 mg/kg, b.wt., per orally once daily for 28 days. The heart weight to body weight ratio was measured to evaluate the effect of extract on MI. The plasma was evaluated for the biochemical parameters and cardiac markers. Pro-oxidant & anti-oxidant levels were measured along with the histopathological examination of the heart. The test extractsSophora interrupta Bedd showed a significant (p< 0.001) restoration of the altered plasma biochemical parameters and cardiac markers reducing the extent of damage in heart musculature. The anti-oxidant effect of the test extract plays a prominent role in curative effect of cardiomyopathy associated with doxorubicin.The test extract of Sophora interrupta Bedd was beneficial for doxorubicin induced cardiotoxicity by ameliorating oxidative stress and restoring the abnormal structural changes in the heart tissue.

Mitochondrial DNA oxidation, methylation, and copy number alterations in major and bipolar depression.

Mood disorders are common disabling psychiatric disorders caused by both genetic and environmental factors. Mitochondrial DNA (mtDNA) modifications and epigenetics are promising areas of research in depression since mitochondrial dysfunction has been associated with depression. In this study we aimed to investigate the mtDNA changes in depressive disorder (MDD) and bipolar disorder (BD). Displacement loop methylation (D-loop-met), relative mtDNA copy number (mtDNA-cn) and mtDNA oxidation (mtDNA-oxi) were investigated in DNA samples of individuals with MDD (n = 34), BD (n = 23), and healthy controls (HC; n = 40) using the Real-Time Polymerase Chain Reaction (RT-PCR). Blood samples were obtained from a subset of individuals with MDD (n = 15) during a depressive episode (baseline) and after remission (8th week). The study groups exhibited significant differences in D-loop-met (p = 0.020), while relative mtDNA-cn and mtDNA-oxi showed comparable results. During the remission phase (8th week), there were lower levels of relative mtDNA-cn (Z = -2.783, p = 0.005) and D-loop-met (Z = -3.180, p = 0.001) compared to the acute MDD baseline, with no significant change in mtDNA-oxi levels (Z = -1.193, p = 0.233). Our findings indicate significantly increased D-loop methylation in MDD compared to BD and HCs, suggesting distinct mtDNA modifications in these conditions. Moreover, the observed alterations in relative mtDNA-cn and D-loop-met during remission suggest a potential role of mtDNA alterations in the pathophysiology of MDD. Future studies may provide valuable insights into the dynamics of mtDNA modifications in both disorders and their response to treatment.

Toxic effects of lead on plants: integrating multi-omics with bioinformatics to develop Pb-tolerant crops.

Lead disrupts plant metabolic homeostasis and key structural elements. Utilizing modern biotechnology tools, it's feasible to develop Pb-tolerant varieties by discovering biological players regulating plant metabolic pathways under stress. Lead (Pb) has been used for a variety of purposes since antiquity despite its toxic nature. After arsenic, lead is the most hazardous heavy metal without any known beneficial role in the biological system. It is a crucial inorganic pollutant that affects plant biochemical and morpho-physiological attributes. Lead toxicity harms plants throughout their life cycle and the extent of damage depends on the concentration and duration of exposure. Higher levels of lead exposure disrupt numerous key metabolic activities of plants including oxygen-evolving complex, organelles integrity, photosystem II connectivity, and electron transport chain. This review summarizes the detrimental effects of lead toxicity on seed germination, crop growth, and yield, oxidative and ultra-structural alterations, as well as nutrient absorption, transport, and assimilation. Further, it discusses the Pb-induced toxic modulation of stomatal conductance, photosynthesis, respiration, metabolic-enzymatic activity, osmolytes accumulation, and antioxidant activity. It is a comprehensive review that reports on omics-based studies along with morpho-physiological and biochemical modifications caused by lead stress. With advances in DNA sequencing technologies, genomics and transcriptomics are gradually becoming popular for studying Pb stress effects in plants. Proteomics and metabolomics are still underrated and there is a scarcity of published data, and this review highlights both their technical and research gaps. Besides, there is also a discussion on how the integration of omics with bioinformatics and the use of the latest biotechnological tools can aid in developing Pb-tolerant crops. The review concludes with core challenges and research directions that need to be addressed soon.

Analysis of critical residues for peroxygenation and improved peroxygenase activity via in situ H2O2 generation in CYP105D18.

Limited numbers of CYPs have been reported to work naturally as peroxygenases. The peroxide shunt pathway can be efficiently used as an alternative for the NAD(P)H and reductase systems, particularly in high hydrogen peroxide (H2O2) resistance CYPs. We reported the structural and biochemical features of CYP105D18 peroxygenase for its high H2O2 tolerance capacity. Q348 was a crucial residue for the stability of CYP105D18 during the exposure to H2O2. In addition, the role of the hydrophilic amino acid T239 from the I helix for peroxygenation and regiospecificity toward testosterone was investigated. Interestingly, T239E differs in product formation from wild type, catalyzing testosterone to androstenedione in the presence of H2O2. The other variant, T239A, worked with the Pdx/Pdr system and was unable to catalyze testosterone conversion in the presence of H2O2, suggesting the transformation of peroxygenase into monooxygenase. CYP105D18 supported the alternative method of H2O2 used for the catalysis of testosterone. The use of the same concentration of urea hydrogen peroxide adducts in place of direct H2O2 was more efficient for 2β-hydroxytestosterone conversion. Furthermore, in situ H2O2 generation using GOx/glucose system enhanced the catalytic efficiency (kcat/Km) for wild type and F184A by 1.3- and 1.9-fold, respectively, compared to direct use of H2O2 The engineering of CYP105D18, its improved peroxygenase activity, and alteration in the product oxidation facilitate CYP105D18 as a potential candidate for biotechnological applications.