Reactive Oxygen Species Formation Research Articles

Chlorogenic Acid Alleviates High Glucose-induced HK-2 Cell Oxidative Damage through Activation of KEAP1/NRF2/ARE Signaling Pathway.

To investigate the alleviating effect of chlorogenic acid (CGA) on oxidative damage in high glucose (HG)-induced HK-2 cells and to explore its potential mechanisms. We cultured the human proximal tubular cell line HK-2 and divided them into the control group and different concentrations of CGA groups (0, 5, 10, 25, 50, 100, 200 μM). The trypan blue dye test was used to detect CGA's potential cytotoxicity on HK-2 cells. Then, we treated HK-2 with HG and CGA; the Cell Counting Kit-8 (CCK-8) method was used to detect the cell viability of HK-2 cells in each group. Flow cytometry was employed to measure the apoptosis rate of cells. Western blot was performed to detect the expression of apoptosis proteins B-cell lymphoma-2 (BCL-2), BCL-2-associated X protein (BAX), cysteinyl aspartate specific proteinase (CASPASE)-9, and CASPASE-3. In addition, enzymatic activities, including superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), and lipid peroxide (LPO), were measured with the corresponding detection kits. 2',7'-Dichlorodihydrofluorescein diacetate (DCFH-DA) assay and flow cytometry were performed to detect reactive oxygen species (ROS) production. Western blot analysis and Reverse Transcription-Polymerase Chain Reaction (RT-PCR) were conducted to evaluate protein and mRNA expressions of the Kelch-like ECH-associated protein-1 (KEAP1)/Nuclear factor erythroid 2-related factor 2 (NRF2)/Antioxidant Response Elements (ARE) signaling pathway. The outcomes showed that, in a dose-dependent way, CGA dramatically increased the vitality of HK-2 induced by HG. Furthermore, CGA significantly reduced the HG-stimulated HK-2 cell apoptosis, which may be linked to the promotion of BCL-2 and the suppression of BAX, cleaved-CASPASE-3, and cleaved-CASPASE-9 expression. In HK-2 cells, CGA reduced the formation of ROS generated by HG levels and markedly boosted the activity of the antioxidant enzymes SOD, GSH-Px, and CAT. Furthermore, compared with the HG group, CGA significantly raised NRF2 nuclear expression and downregulated NRF2 cytosolic expression and increased the mRNA expression of NRF2 and its target genes, heme oxygenase-1 (HO-1), KEAP1, and NAD(P)H dehydrogenase quinone 1 (NQO1). These results show that CGA might be useful in managing oxidative damage in HG-induced HK-2 cells.

Protective Role of Resveratrol Against Toluene-Induced Oxidative Stress in Rats

Toluene is a toxic substance commonly used in the industry that leads to oxidative stress in the body and causes in the formation of reactive oxygen species. In addition to the antioxidants produced by our body, we also obtain antioxidants through foods. Resveratrol is a polyphenol found in many plant species and possesses antioxidant properties. The effects of oxidative stress induced by toluene and the protective effect of resveratrol were investigated through changes in malondialdehyde (MDA) and glutathione (GSH) levels. Male rats were administered different doses of toluene and resveratrol (5 mg/kg, 10 mg/kg, 20 mg/kg), while control groups were given physiological saline and ethanol. At the end of the experiment, blood and lung tissues of the animals were examined, and MDA and GSH levels were measured. According to the obtained data, a significant increase in GSH activity was observed after the administration of toluene. However, in groups treated with combinations of toluene and various doses of resveratrol, a significant decrease in GSH levels was detected. While a significant increase in MDA levels in lung tissues was observed between the control groups and the toluene group, no significant change was observed in blood samples.

Superoxide Photoproduction from Wetland Plant-Derived Dissolved Organic Matter: Implications for Biogeochemical Impacts of Plant Invasion.

Although the impacts of exotic wetland plant invasions on native biodiversity, landscape features, and carbon-nitrogen cycles are well appreciated, biogeochemical consequences posed by ecological competition, such as the heterogeneity of dissolved organic matter (DOM) from plant detritus and its impact on the formation of reactive oxygen species, are poorly understood. Thus, this study delves into O2•- photogeneration potential of DOM derived from three different parts (stem, leaf, and panicle) of invasive Spartina alterniflora (SA) and native Phragmites australis (PA). It is found that DOM from the leaves of SA and the panicles of PA has a superior ability to produce O2•-. With more stable aromatic structures and a higher proportion of sulfur-containing organic compounds, SA-derived DOM generally yields more O2•- than that derived from PA. UVA exposure enhances the leaching of diverse DOM molecules from plant detritus. Based on the reported monitoring data and our findings, the invasion of SA is estimated to approximately double the concentration of O2•- in the surrounding water bodies. This study can help to predict the underlying biogeochemical impacts from the perspective of aquatic photochemistry in future scenarios of plant invasion, seawater intrusion, wetland degradation, and elevated solar UV radiation.

The potency of yellow pigment extract from the marine bacterium Pseudomonas oryzihabitans SAB E-3 as an antioxidant agent

Abstract. Putri DA, Astuti RI, Wahyudi AT. 2024. The potency of yellow pigment extract from the marine bacterium Pseudomonas oryzihabitans SAB E-3 as an antioxidant agent. Biodiversitas 25: 2565-2575. Metabolic processes in cells can produce byproducts in the form of Reactive Oxygen Species (ROS). Excessive amounts of ROS are one of the causes of oxidative stress. Antioxidants are known to act as free radical-scavenging compounds. Pigment-producing bacteria associated with sponges have been widely reported to have antioxidant compounds. In this study, the marine bacterium Pseudomonas oryzihabitans SAB E-3 produced intracellular yellow pigments with potential antioxidant activity. Therefore, this study aimed to assess the in vitro antioxidant and mitochondrial activity at the cellular level and identify chemical compounds. The intracellular pigments produced by P. oryzihabitans SAB E-3 were extracted using methanol, acetone, chloroform, and ethyl acetate. The methanol pigment extract contained the highest number of phenolic and flavonoid compounds, which may have contributed to its antioxidant activity. Furthermore, the active fraction obtained from the bioautographic TLC and harvested using preparative TLC had stronger antioxidant activity against DPPH radicals (75.71 µg/mL) than the crude pigment extract. However, the ABTS radical-reducing activity was lower (226.59 µg/mL) than the crude pigment extract. The maximum wavelength of the pigment extract was 453 nm, indicating the presence of carotenoids. In addition, the lowest concentration of the crude pigment extract (50 µg/mL) and selected active fractions (18.75 µg/mL) increased yeast Schizosaccharomyces pombe ARC039 tolerance to oxidative stress and induced mitochondrial activity. Active fraction one extract analyzed using LC-MS/MS contained piceatannol, resveratrol, isorhapontigenin, isoliquiritigenin, liquiritin, and 2-Omethylisohemigossylic acid lactone, which may contribute to its antioxidant activity. Therefore, the yellow pigment extract and the active fraction produced by P. oryzihabitans SAB E-3 are potential candidates for natural antioxidant sources.

Recent Progress in Thermally Activated Delayed Fluorescence Photosensitizers for Photodynamic Therapy.

Photodynamic therapy (PDT) is a rapidly growing discipline that is expected to become an encouraging noninvasive therapeutic strategy for cancer treatment. In the PDT process, an efficient intersystem crossing (ISC) process for photosensitizers from the singlet excited state (S1) to the triplet excited state (T1) is critical for the formation of cytotoxic reactive oxygen species and improvement of PDT performance. Thermally activated delayed fluorescence (TADF) molecules featuring an extremely small singlet-triplet energy gap and an efficient ISC process represent an enormous breakthrough for the PDT process. Consequently, the development of advanced TADF photosensitizers has become increasingly crucial and pressing. The most recent developments in TADF photosensitizers aimed at enhancing PDT efficiency for bio-applications are presented in this review. TADF photosensitizers with water dispersibility, targeting ability, activatable ability, and two-photon excitation properties are highlighted. Furthermore, the future challenges and perspectives of TADF photosensitizers in PDT are proposed.

Visible light triggers the formation of reactive oxygen species in monoclonal antibody formulations

Most monoclonal antibody formulations require the presence of a surfactant, such as polysorbate, to ensure protein stability. The presence of high concentrations of polysorbate have been shown to enhance photooxidation of certain protein drug products when exposed to visible light. The current literature, however, suggest that photooxidation of polysorbate only occurs when exposed to visible light in combination with UVA light. This is probable as peroxides present in polysorbate solutions can be cleaved homolytically in the UVA region. In the visible region, photooxidation is not expected to occur as cleavage of peroxides is not expected at these wavelengths.This report presents findings suggesting that the presence of one or more photosensitiser(s) in polysorbate must be a cause and is required to catalyse the aerobic oxidation of polysorbate solutions upon exposure to visible light. Our investigation aimed to clarify the mechanism(s) of polysorbate photooxidation and explore the kinetics and the identity of the generated radicals and their impact on monoclonal antibody (mAb) degradation. Our study reveals that when polysorbate solutions are exposed to visible light between 400 - 800 nm in the absence of proteins, discolouration, radical formation, and oxygen depletion occur. We discuss the initial formation of reactive species, most likely occurring directly after reaction of molecular oxygen, with the presence of a triplet state photosensitiser, which is generated by intersystem crossing of the excited singlet state. When comparing the photooxidation of PS20 and PS80 in varying quality grades, we propose that singlet oxygen possesses potential for reacting with unsaturated fatty acids in PS80HP, however, PS20HP itself exhibited no measurable oxidation under the tested conditions. The study’s final part delves into the photooxidation behaviour of different PS grades, examining its influence on the integrity of a mAb in the formulation. Finally, we examined the effect of photooxidation on the integrity of monoclonal antibodies. Our findings show that the exposure to visible light in polysorbate-containing mAb solutions at high PS concentrations of 4 mg·ml−1 results in increased monoclonal antibody degradation, highlighting the need for cautious evaluation of the correct PS concentration to stabilise protein therapeutics.

Oxidative and Nitrous Stress Underlies Vascular Malfunction Induced by Ionizing Radiation and Diabetes.

Oxidative stress results from the production of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in quantities exceeding the potential activity of the body's antioxidant system and is one of the risk factors for the development of vascular dysfunction in diabetes and exposure to ionizing radiation. Being the secondary products of normal aerobic metabolism in living organisms, ROS and RNS act as signaling molecules that play an important role in the regulation of vital organism functions. Meanwhile, in high concentrations, these compounds are toxic and disrupt various metabolic pathways. The various stress factors (hyperglycemia, gamma-irradiation, etc.) trigger free oxygen and nitrogen radicals accumulation in cells that are capable to damage almost all cellular components including ion channels and transporters such as Na+/K+-ATPase, BKCa, and TRP channels. Vascular dysfunctions are governed by interaction of ROS and RNS. For example, the reaction of ROS with NO produces peroxynitrite (ONOO-), which not only oxidizes DNA, cellular proteins, and lipids, but also disrupts important signaling pathways that regulate the cation channel functions in the vascular endothelium. Further increasing in ROS levels and formation of ONOO- leads to reduced NO bioavailability and causes endothelial dysfunction. Thus, imbalance of ROS and RNS and their affect on membrane ion channels plays an important role in the pathogenesis of vascular dysfunction associated with various disorders.

The sensitivity of acute myeloid leukemia cells to cytarabine is increased by suppressing the expression of Heme oxygenase-1 and hypoxia-inducible factor 1-alpha

BackgroundAcute myeloid leukemia (AML), a malignancy Often resistant to common chemotherapy regimens (Cytarabine (Ara-c) + Daunorubicin (DNR)), is accompanied by frequent relapses. Many factors are involved in causing chemoresistance. Heme Oxygenase-1 (HO-1) and Hypoxia-Inducible Factor 1-alpha (HIF-1α) are two of the most well-known genes, reported to be overexpressed in AML and promote resistance against chemotherapy according to several studies. The main chemotherapy agent used for AML treatment is Ara-c. We hypothesized that simultaneous targeting of HO-1 and HIF-1α could sensitize AML cells to Ara-c.MethodIn this study, we used our recently developed, Trans-Activator of Transcription (TAT) - Chitosan-Carboxymethyl Dextran (CCMD) - Poly Ethylene Glycol (PEG) - Nanoparticles (NPs), to deliver Ara-c along with siRNA molecules against the HO-1 and HIF-1α genes to AML primary cells (ex vivo) and cell lines including THP-1, KG-1, and HL-60 (in vitro). Subsequently, the effect of the single or combinational treatment on the growth, proliferation, apoptosis, and Reactive Oxygen Species (ROS) formation was evaluated.ResultsThe designed NPs had a high potential in transfecting cells with siRNAs and drug. The results demonstrated that treatment of cells with Ara-c elevated the generation of ROS in the cells while decreasing the proliferation potential. Following the silencing of HO-1, the rate of apoptosis and ROS generation in response to Ara-c increased significantly. While proliferation and growth inhibition were considerably evident in HIF-1α-siRNA-transfected-AML cells compared to cells treated with free Ara-c. We found that the co-inhibition of genes could further sensitize AML cells to Ara-c treatment.ConclusionsAs far as we are aware, this study is the first to simultaneously inhibit the HO-1 and HIF-1α genes in AML using NPs. It can be concluded that HO-1 causes chemoresistance by protecting cells from ROS damage. Whereas, HIF-1α mostly exerts prolific and direct anti-apoptotic effects. These findings imply that simultaneous inhibition of HO-1 and HIF-1α can overcome Ara-c resistance and help improve the prognosis of AML patients.

Liraglutide alleviated alpha-pyrrolidinovalerophenone (α-PVP) induced cognitive deficits in rats by modifying brain mitochondrial impairment

The use of NPS compounds is increasing, and impairment in spatial learning and memory is a growing concern. Alpha-pyrrolidinovalerophenone (α-PVP) consumption, as a commonly used NPS, can impair spatial learning and memory via the brain mitochondrial dysfunction mechanism. Liraglutide isone of the most well-known Glucagon-Like Peptide 1 (GLP-1) agonists that is used as an anti-diabetic and anti-obesity drug. According to current research, Liraglutide likely ameliorates cognitive impairment in neurodegenerative conditions and substance use disorders. Hence, the purpose of this study is examining the effect of Liraglutide on α-PVP-induced spatial learning and memory problems due to brain mitochondrial dysfunction. Wistar rats (8 in each group) received α-PVP (20 mg/kg/d for 10 consecutive days, intraperitoneally (I.P.)). Then, Liraglutide was administered at 47 and 94 μg/kg/d, I.P., for 4 weeks following the α-PVP administration. The Morris Water Maze (MWM) task evaluated spatial learning and memory 24 h after Liraglutide treatment. Bedside, brain mitochondrial activity parameters, including reactive oxygen species (ROS) level, mitochondrial membrane potential (MMP), cytochrome c release, mitochondrial outer membrane damage and swelling, and brain ADP/ATP ratio, were studied. Our results showed that Liraglutide ameliorated α-PVP-induced spatial learning and memory impairments through alleviating brain mitochondrial dysfunction (which is indicated by increasing ROS formation, collapsed MMP, mitochondrial outer membrane damage, cytochrome c release, mitochondrial swelling, and increased brain ADP/ATP ratio). This study could be used as a starting point for future studies about the possible role of Liraglutide in ameliorating mitochondrial dysfunction leading to substance use disorder- induced cognitive impairment.

Water-Catalyzed Formation of Reactive Oxygen Species from NO2 on a Weakly Hydrated Calcite Surface.

The interfaces of weakly hydrated mineral substrates have been shown to serve as catalytic sites for chemical reactions that may not be accessible in the gas phase or under bulk conditions. Currently known mechanisms for the formation of reactive oxygen species (ROS) from nitrogen dioxide (NO2) involve NO2 dimerization. Here, we report the formation of the ROS HONO via a mechanism involving simple adsorption of a single NO2 molecule on a weakly hydrated calcite substrate. First-principles molecular dynamics simulations coupled with enhanced sampling techniques show how an adsorbed water sublayer can enhance NO2 adsorption on calcite compared to adsorption on a bare dry substrate. On the weakly hydrated calcite surface, an interfacial electric field facilitates proton extraction from water, thus allowing HONO formation from a single adsorbed NO2, i.e., without the need for the formation of a NO2 dimer precomplex. HONO formation on calcite is kinetically more favorable than that in the gas phase, with a reaction barrier of 14 kcal/mol on the weakly hydrated calcite surface compared to 27 kcal/mol in the gas phase. Further photocatalysed HONO production by visible light and HONO dissociation are hampered on calcite, unlike the process on silica. NO2 is a significant anthropogenic pollutant, and understanding its chemistry is crucial for explaining the high ROS levels and haze formation in polluted areas or prebiotic ROS generation. These findings emphasize how mineral substrates under water-restricted hydration conditions can trigger chemical pathways that are unexpected in the gas phase or under bulk conditions.

Atomic Confinement Empowered CoZn Dual-Single-Atom Nanotubes for H2O2 Production in Sequential Dual-Cathode Electro-Fenton Process.

Single-atom catalysts (SACs) are flourishing in various fields because of their 100% atomic utilization. However, their uncontrollable selectivity, poor stability and vulnerable inactivation remain critical challenges. According to theoretical predictions and experiments, a heteronuclear CoZn dual-single-atom confined in N/O-doped hollow carbon nanotube reactors (CoZnSA@CNTs) is synthesized via spatial confinement growth. CoZnSA@CNTs exhibit superior performance for H2O2 electrosynthesis over the entire pH range due to dual-confinement of atomic sites and O2 molecule. CoZnSA@CNTs is favorable for H2O2 production mainly because the synergy of adjacent atomic sites, defect-rich feature and nanotube reactor promoted O2 enrichment and enhanced H2O2 reactivity/selectivity. The H2O2 selectivity reaches ∼100% in a range of 0.2-0.65V versus RHE and the yield achieves 7.50M gcat -1 with CoZnSA@CNTs/carbon fiber felt, exceeding most of the reported SACs in H-type cells. The obtained H2O2 is converted directly to sodium percarbonate and sodium perborate in a safe way for H2O2 storage/transportation. The sequential dual-cathode electron-Fenton process promotes the formation of reactive oxygen species (•OH, 1O2 and •O2 -) by activating the generated H2O2, enabling accelerated degradation of various pollutants and Cr(VI) detoxification in actual wastewater. This work proposes a promising confinement strategy for catalyst design and selectivity regulation of complex reactions.

Performance, mechanism, and ecotoxicity assessment of sulfamethoxazole degradation by coupling visible-light photocatalysis with persulphate activation over Fe3O4/ZnO composites

ABSTRACT To probe the efficient and ecofriendly treatment of emerging pollutants, a magnetically separable Fe3O4/ZnO composite was facilely synthesised and utilised to establish a combined oxidation process by coupling photocatalysis with persulphate activation (Fe3O4/ZnO-PS). As proved by various characterisations, the Fe3O4/ZnO composite was constituted of highly dispersed Fe3O4 nanoparticles with ZnO nanorods as the support, and it showed the increased visible-light harvesting and facilitated charge transfer. The Fe3O4/ZnO-PS process significantly enhanced sulfamethoxazole (SMX) degradation under visible light irradiation, and the degradation rate constant and synergy coefficient approached 0.222 h−1 and 1.98, respectively. The enhancement mechanism of Fe3O4/ZnO-PS process for SMX degradation was resulted from the promotion of reactive oxygen species formation. Electron spin resonance and fluorescence measurements indicated that •OH and SO4•‒ majorly contribute to the SMX degradation. In addition, the effects of catalyst dosage, PS dosage, solution pH and inorganic anions were systematically investigated. Based on liquid chromatography – mass spectrometry (LC/MS) identification, there were totally 12 products of SMX degradation found in the Fe3O4/ZnO-PS process, the degradation pathways of which involve hydroxylation, S‒N bond cleavage, and SO2 abstraction. The ecotoxicities of these degradation products were predicted by quantitative structure-activity relationship software and were verified by actual experiments using Vibrio fischeri. The ecotoxicity assessment results indicated that although the Fe3O4/ZnO-PS process degraded SMX effectively, more toxic intermediates appeared and accordingly increased the ecological risk, which should be deeply concerned.

Silicon Dioxide Nanoparticles-Based Amelioration of Cd Toxicity by Regulating Antioxidant Activity and Photosynthetic Parameters in a Line Developed from Wild Rice.

An extremely hazardous heavy metal called cadmium (Cd) is frequently released into the soil, causing a considerable reduction in plant productivity and safety. In an effort to reduce the toxicity of Cd, silicon dioxide nanoparticles were chosen because of their capability to react with metallic substances and decrease their adsorption. This study examines the processes that underlie the stress caused by Cd and how SiO2NPs may be able to lessen it through modifying antioxidant defense, oxidative stress, and photosynthesis. A 100 μM concentration of Cd stress was applied to the hydroponically grown wild rice line, and 50 μM of silicon dioxide nanoparticles (SiO2NPs) was given. The study depicted that when 50 μM SiO2NPs was applied, there was a significant decrease in Cd uptake in both roots and shoots by 30.2% and 15.8% under 100 μM Cd stress, respectively. The results illustrated that Cd had a detrimental effect on carotenoid and chlorophyll levels and other growth-related traits. Additionally, it increased the levels of ROS in plants, which reduced the antioxidant capability by 18.8% (SOD), 39.2% (POD), 32.6% (CAT), and 25.01% (GR) in wild rice. Nevertheless, the addition of silicon dioxide nanoparticles reduced oxidative damage and the overall amount of Cd uptake, which lessened the toxicity caused by Cd. Reduced formation of reactive oxygen species (ROS), including MDA and H2O2, and an increased defense system of antioxidants in the plants provided evidence for this. Moreover, SiO2NPs enhanced the Cd resistance, upregulated the genes related to antioxidants and silicon, and reduced metal transporters' expression levels.

How water acidification influences the organism antioxidant capacity and gill structure of Mediterranean mussel (Mytilus galloprovincialis, Lamarck, 1819) at normoxia and hypoxia

The effect of water acidification in combination with normoxia or hypoxia on the antioxidant capacity and oxidative stress markers in gills and hemolymph of the Mediterranean mussel (Mytilus galloprovincialis), as well as on gill microstructure, has been evaluated through an in vivo experiment. Mussels were exposed to a low pH (7.3) under normal dissolved oxygen (DO) conditions (8 mg/L), and hypoxia (2 mg/L) for 8 days, and samples were collected on days 1, 3, 6, and 8 to evaluate dynamic changes of physiological responses. Cytoplasmic concentrations of reactive oxygen species (ROS) and levels of DNA damage were measured in hemocytes, while the activity of catalase (CAT) and superoxide dismutase (SOD) and histopathological changes were assessed in gills. The results revealed that while water acidification did not significantly affect the activity of SOD and CAT in gills under normoxic and hypoxic conditions, there was a trend towards suppression of CAT activity at the end of the experimental period (day 8). Similarly, we did not observe increased formation of ROS in hemocytes or changes in the levels of DNA damage during the experimental period. These results strongly suggest that the oxidative stress response system in mussels is relatively stable to experimental conditions of acidification and hypoxia. Experimental acidification under normoxia and hypoxia caused changes to the structure of the gills, leading to various histopathological alterations, including dilation, hemocyte infiltration into the hemal sinuses, intercellular edema, vacuolization of epithelial cells in gill filaments, lipofuscin accumulation, changes in the shape and adjacent gill filaments, hyperplasia, exfoliation of the epithelial layer, necrosis, swelling, and destruction of chitinous layers (chitinous rods). Most of these alterations were reversible, non-specific changes that represent a general inflammatory response and changes in the morphology of the gill filaments. The dynamics of histopathological alterations suggests an active adaptive response of gills to environmental stresses. Taken together, our data indicate that Mediterranean mussels have a relative tolerance to water acidification and hypoxia at tissue and cellular levels.

N6-methyladenosine demethylase fat mass and obesity-associated protein suppresses hyperglycemia-induced endothelial cell injury by inhibiting reactive oxygen species formation via autophagy promotion

IntroductionHyperglycemia-induced endothelial cell injury is one of the main causes of diabetic vasculopathy. Fat mass and obesity-associated protein (FTO) was the first RNA N6-methyladenosine (m6A) demethylase identified; it participates in the pathogenesis of diabetes. However, the role of FTO in hyperglycemia-induced vascular endothelial cell injury remains unclear. Materials and methodsThe effects of FTO on cellular m6A, autophagy, oxidative stress, proliferation, and cytotoxicity were explored in human umbilical vein endothelial cells (HUVECs) treated with high glucose (33.3 mmol/mL) after overexpression or pharmacological inhibition of FTO. MeRIP-qPCR and RNA stability assays were used to explore the molecular mechanisms by which FTO regulates autophagy. ResultsHigh glucose treatment increased m6A levels and reduced FTO protein expression in HUVECs. Wild-type overexpression of FTO markedly inhibited reactive oxygen species generation by promoting autophagy, increasing endothelial cell proliferation, and decreasing the cytotoxicity of high glucose concentrations. The pharmacological inhibition of FTO showed the opposite results. Mechanistically, we identified Unc-51-like kinase 1 (ULK1), a gene responsible for autophagosome formation, as a downstream target of FTO-mediated m6A modification. FTO overexpression demethylated ULK1 mRNA and inhibited its degradation in an m6A-YTHDF2-dependent manner, leading to autophagy activation. ConclusionsOur study demonstrates the functional importance of FTO-mediated m6A modification in alleviating endothelial cell injury under high glucose conditions and indicates that FTO may be a novel therapeutic target for diabetic vascular complications.

Low energy availability increases immune cell formation of reactive oxygen species and impairs exercise performance in female endurance athletes

The effects of low energy availability (LEA) on the immune system are poorly understood. This study examined the effects of 14 days of LEA on immune cell redox balance and inflammation at rest and in response to acute exercise, and exercise performance in female athletes. Twelve female endurance athletes (age: 26.8±3.4yrs, maximum oxygen uptake (V˙O2max): 55.2±5.1 mL×min-1×kg-1) were included in a randomized, single-blinded crossover study. They were allocated to begin with either 14 days of optimal energy availability diet (OEA, 52±2kcal×kg fat free mass (FFM)-1×day-1) or LEA diet (22±2kcal×kg FFM-1×day-1), followed by 3 days of refueling (OEA) with maintained training volume. Peripheral blood mononuclear cells (PBMCs) were isolated, and plasma obtained at rest before and after each dietary period. The PBMCs were used for analysis of mitochondrial respiration and H2O2 emission and specific proteins. Exercise performance was assessed on cycle by a 20-min time trial and time to exhaustion at an intensity corresponding to ∼110% V˙O2max). LEA was associated with a 94% (P=0.003) increase in PBMC NADPH oxidase 2 protein content, and a 22% (P=0.013) increase in systemic cortisol. LEA also caused an alteration of several inflammatory related proteins (P<0.05). Acute exercise augmented H2O2 emission in PBMCs (P<0.001) following both OEA and LEA, but to a greater extent following LEA. LEA also reduced the mobilization of white blood cells with acute exercise. After LEA, performance was reduced in both exercise tests (P<0.001), and the reduced time trial performance remained after the 3 days of refueling (P<0.001). 14 days of LEA in female athletes increased cortisol levels and had a pronounced effect on the immune system, including increased capacity for ROS production, altered plasma inflammatory proteome and lowered exercise induced mobilization of leukocytes. Furthermore, LEA resulted in a sustained impairment in exercise performance.

Regulating Reactive Oxygen Intermediates of Fe–N–C SAzyme via Second‐Shell Coordination for Selective Aerobic Oxidation Reactions

Reactive oxygen species (ROS) regulation for single‐atom nanozymes (SAzymes), e.g., Fe–N–C, is a key scientific issue that determines the activity, selectivity, and stability of aerobic reaction. However, the poor understanding of ROS formation mechanism on SAzymes greatly hampers their wider deployment. Herein, inspired by cytochromes P450 affording bound ROS intermediates in O2 activation, we report Fe–N–C containing the same FeN4 but with tunable second‐shell coordination can effectively regulate ROS production pathways. Remarkably, compared to the control Fe–N–C sample, the second‐shell sulfur functionalized Fe–N–C delivered a·2.4‐fold increase of oxidase‐like activity via the bound Fe=O intermediate. Conversely, free ROS (•O2‐) release was significantly reduced after functionalization, down to only 17% of that observed for Fe–N–C. The detailed characterizations and theoretical calculations revealed that the second‐shell sulfur functionalization significantly altered the electronic structure of FeN4 sites, leading to an increase of electron density at Fermi level. It enhanced the electron transfer from active sites to the key intermediate *OOH, thereby ultimately determining the type of ROS in aerobic oxidation process. The proposed Fe–N–Cs with different second‐shell anion were further applied to three aerobic oxidation reactions with enhanced activity, selectivity, and stability.

Exposure to O3 and NO2 on the interfacial chemistry of the pulmonary surfactant and the mechanism of lung oxidative damage

Exposure to ozone (O3) and nitrogen dioxide (NO2) are related to pulmonary dysfunctions and various lung diseases, but the underlying biochemical mechanisms remain uncertain. Herein, the effect of inhalable oxidizing gas pollutants on the pulmonary surfactant (PS, extracted from porcine lungs), a mixture of active lipids and proteins that plays an important role in maintaining normal respiratory mechanics, is investigated in terms of the interfacial chemistry using in-vitro experiments; and the oxidative stress induced by oxidizing gases in the simulated lung fluid (SLF) supplemented with the PS is explored. The results showed that O3 and NO2 individually increased the surface tension of the PS and reduced its foaming ability; this was accompanied by the surface pressure-area isotherms of the PS monolayers shifting toward lower molecular areas, with O3 exhibiting more severe effects than NO2. Moreover, both O3 and NO2 produced reactive oxygen species (ROS) resulting in lipid peroxidation and protein damage to the PS. The formation of superoxide radicals (O2•–) was correlated with the decomposition of O3 and the reactions of O3 and NO2 with antioxidants in the SLF. These radicals, in the presence of antioxidants, led to the formation of hydrogen peroxide and hydroxyl radicals (•OH). Additionally, the direct oxidation of unsaturated lipids by O3 and NO2 further caused an increase in the ROS content. This change in the ROS chemistry and increased •OH production tentatively explain how inhalable oxidizing gases lead to oxidative stress and adverse health effects. In summary, our results indicated that inhaled O3 and NO2 exposure can significantly alter the interfacial properties of the PS, oxidize its active ingredients, and induce ROS formation in the SLF. The results of this study provide a basis for the elucidation of the potential hazards of inhaled oxidizing gas pollutants in the human respiratory system.

Regulating Reactive Oxygen Intermediates of Fe-N-C SAzyme via Second-Shell Coordination for Selective Aerobic Oxidation Reactions.

Reactive oxygen species (ROS) regulation for single-atom nanozymes (SAzymes), e.g., Fe-N-C, is a key scientific issue that determines the activity, selectivity, and stability of aerobic reaction. However, the poor understanding of ROS formation mechanism on SAzymes greatly hampers their wider deployment. Herein, inspired by cytochromes P450 affording bound ROS intermediates in O2 activation, we report Fe-N-C containing the same FeN4 but with tunable second-shell coordination can effectively regulate ROS production pathways. Remarkably, compared to the control Fe-N-C sample, the second-shell sulfur functionalized Fe-N-C delivered a 2.4-fold increase of oxidase-like activity via the bound Fe=O intermediate. Conversely, free ROS (⋅O2 -) release was significantly reduced after functionalization, down to only 17 % of that observed for Fe-N-C. The detailed characterizations and theoretical calculations revealed that the second-shell sulfur functionalization significantly altered the electronic structure of FeN4 sites, leading to an increase of electron density at Fermi level. It enhanced the electron transfer from active sites to the key intermediate *OOH, thereby ultimately determining the type of ROS in aerobic oxidation process. The proposed Fe-N-Cs with different second-shell anion were further applied to three aerobic oxidation reactions with enhanced activity, selectivity, and stability.

Construction of biomimetic camouflaged neutrophil membrane nanoparticles for precise delivery and augmented glioma cancer treatment

Developing novel nanomedicine formulations that successfully penetrate the blood-brain barrier (BBB) to treat and diagnose glioma effectively is still a significant obstacle. This study presents the development of polymer nanogels camouflaged with macrophage membranes. The nanogels are loaded with manganese dioxide (MnO2) and carboplatin. They are designed for the treatment of glioma using a combination of chemotherapy and chemodynamic therapy (CDT). The precipitation polymerization process was used to create redox-responsive poly(caprolactone) (PCL) nanogels (NGs). These NGs were then loaded with MnO2 and physically encapsulated with carboplatin. The resulting nanogels had an average diameter of 102.52 nm and were covered with macrophage membranes to provide excellent stability. The developed NGs exhibit redox and pH responsiveness, releasing carboplatin and Mn(II) in a controlled manner. NGs may effectively deplete glutathione (GSH) in the tumour milieu. The Mn(II) generated in this process enhances the effectiveness of the loaded carboplatin by exerting its chemotherapeutic action and promoting the formation of reactive oxygen species to improve the efficiency of CDT. The results of our study show a very efficient and promising nanomedicine platform for the treatment and diagnosis of glioma, a type of cancer. This platform is self-adaptive, cooperative, and based on NG technology. Furthermore, this platform can potentially be used for other challenging types of cancer.