Radical Oxygen Species Research Articles

Expression Pattern of Ngb in Astrocytes after Spinal Cord Injury and the Clinical Significance

In the current study, we evaluated the expression pattern of neuroglobin (Ngb) in the astrocytes after spinal cord injury (SCI) and explore the clinical significance. For this purpose, a total of 48 Sprague-Dawley rats were divided into the SCI group (n = 40) and Sham group (n = 8). Rats in the SCI group were used to prepare the SCI models by using the modified Allen’s method, followed by the HE staining to observe the post-SCI pathological changes and immunofluorescent staining to observe the dynamic changes of Ngb in astrocytes after SCI. Then, oxidative stress injury models were constructed on the astrocytes in the spinal cord of rats by using peroxide in different concentrations (0, 50, 100, 150, 200 and 400μmol/L), and at 6 and 12 h after treatment, the vitality of astrocytes that were treated by peroxide in different concentrations was determined using the MTT method, while the ability of astrocytes to generate radical oxygen species (ROS) was determined by using the flow cytometry. The mRNA expression of Ngb after the oxidative stress injury in astrocytes was measured by using the real-time quantitative PCR. Results of HE staining demonstrated that rats with SCI presented with the gradual transition from acute injury into the glial scar, a natural repair, while the results of immunofluorescent staining indicated that after SCI, expression of Ngb in the astrocytes experienced an increase followed by a decrease, and the peak level was attained at 14 d after SCI. Following the treatment of H2O2 at different concentrations (50, 100, 150, 200 and 400μmol/L) for 6 and 12 h, the vitality of astrocytes in the model groups was significantly lower than that in the control groups (all P < 0.05). As the concentration of H2O2 increased (50, 100, 150, 200, 400μmol/L) and exposure to H2O2 prolonged (6, 12 h), mRNA expression was firstly increased but then decreased in astrocytes in a time-dose dependent pattern (all P < 0.05). After SCI, the expression of Ngb in the astrocytes of spine was upregulated, suggesting that Ngb may be involved in the anti-oxidative stress injury in astrocytes after SCI, thereby playing as an endogenous protector of cells.

Identification of genes involved in drought tolerance in seedlings of the desert grass, Psammochloa villosa (Poaceae), based on full-length isoform sequencing and de novo assembly from short reads

Psammochloa villosa is a perennial herbaceous plant that is dominant within arid regions of the Inner Mongolian Plateau and the Qinghai-Tibet Plateau in China, where it is an endemic species and exhibits strong drought tolerance and wind resistance. To study drought tolerance in P. villosa and determine its molecular basis, we simulated high and moderate drought stress in a controlled environment and then analyzed transcriptome sequences by combining long-read sequences from a representative, wild-grown individual with short reads from the treatment groups. We obtained 184,076 high-quality isoforms as a reference and 168,650 genes (91.6%), which we were able to annotate according to public databases. Ultimately, we obtained 119,005 unigenes representing the transcriptome of P. villosa under drought stress and, among these, we identified 3089 differentially expressed genes and 1484 transcription factors. Physiologically, P. villosa that was exposed to high and moderate drought stress had reduced germination rates and shorter buds but generated more chlorophyll, which is atypical under drought stress and possibly reflects an adaptation of these plants to their arid environment. We inferred that significantly upregulated genes were annotated as ‘Chlorophyll a-b binding protein’ and ‘Light-harvesting chlorophyll-protein’ among drought and control groups. Broadly, our analyses revealed that drought stress triggered many genome-level responses, especially related to mitigation of radical oxygen species (ROS), which increase in concentration under drought stress. In particular, in the high drought stress group compared with the control, GO enrichment analysis revealed a significant enrichment of upregulated genes (n = 10) involved in mitigation of oxidative stress. Similarly, using KEGG we found significant enrichment of genes in the phenylpropanoid biosynthesis pathway (11 genes), which yields phenols that scavenge ROS. We also inferred that many genes involved in metabolism of arginine and proline, which may serve as both scavengers of ROS and osmoprotectants that interact with stress response genes based on our protein–protein interaction network analysis. We verified the relative expression levels of eight genes associated with mitigation of ROS, DNA repair, and transmembrane transporter activity using qRT-PCR, and the results were consistent with our inferences from transcriptomes. This study provides insights into the genomic and physiological basis of drought tolerance in P. villosa and represents a resource for development of the species as a forage crop via molecular breeding within arid lands.

Photodynamic action of photosensitizers based on polycationic derivatives of synthetic bacteriochlorin against human lung cancer cells A549

In view of their potential in the treatment of oncological diseases, photosensitizers (PS) have been the object of active research for the last years. The applied therapeutic photodynamic method involves the activation of a PS by light at a specific wavelength. PS interacts with oxygen and catalyzes the production of singlet oxygen molecules or radical oxygen species which leads to tumor cell death. This work is devoted to the study of the effectiveness of the photodynamic action of new PSs based on polycationic derivatives of synthetic bacteriochlorin on human lung cancer cells A549. The results obtained show that these PS effectively bind to these cells, have high phototoxicity and low ‘dark’ cytotoxicity for them. Morphological and immunohistochemical studies show that photodynamic effect leads to induction of cell necrosis and apoptosis, as well as a sharp decrease in mitotic activity.

Coffee-Derived Phenolic Compounds Activate Nrf2 Antioxidant Pathway in I/R Injury In Vitro Model: A Nutritional Approach Preventing Age Related-Damages.

Age-related injuries are often connected to alterations in redox homeostasis. The imbalance between free radical oxygen species and endogenous antioxidants defenses could be associated with a growing risk of transient ischemic attack and stroke. In this context, a daily supply of dietary antioxidants could counteract oxidative stress occurring during ischemia/reperfusion injury (I/R), preventing brain damage. Here we investigated the potential antioxidant properties of coffee-derived circulating metabolites and a coffee pulp phytoextract, testing their efficacy as ROS scavengers in an in vitro model of ischemia. Indeed, the coffee fruit is an important source of phenolic compounds, such as chlorogenic acids, present both in the brewed seed and in the discarded pulp. Therefore, rat brain endothelial cells, subjected to oxygen and glucose deprivation (OGD) and recovery (ogR) to mimic reperfusion, were pretreated or not with coffee by-products. The results indicate that, under OGD/ogR, the ROS accumulation was reduced by coffee by-product. Additionally, the coffee extract activated the Nrf2 antioxidant pathway via Erk and Akt kinases phosphorylation, as shown by increased Nrf2 and HO-1 protein levels. The data indicate that the daily intake of coffee by-products as a dietary food supplement represents a potential nutritional strategy to counteract aging.

Kv7.4 channels regulate potassium permeability in neuronal mitochondria

Mitochondrial K+ permeability regulates neuronal apoptosis, energy metabolism, autophagy, and protection against ischemia–reperfusion injury. Kv7.4 channels have been recently shown to regulate K+ permeability in cardiac mitochondria and exert cardioprotective effects. Here, the possible expression and functional role of Kv7.4 channels in regulating membrane potential, radical oxygen species (ROS) production, and Ca2+ uptake in neuronal mitochondria was investigated in both clonal (F11 cells) and native brain neurons.In coupled mitochondria isolated from F11 cells, K+-dependent changes of mitochondrial membrane potential (ΔΨ) were unaffected by the selective mitoBKCa channel blocker iberiotoxin and only partially inhibited by the mitoKATP blockers glyburide or ATP. Interestingly, K+-dependent ΔΨ decrease was significantly reduced by the Kv7 blocker XE991 and enhanced by the Kv7 activator retigabine. Among Kv7s, western blot experiments showed the expression of only Kv7.4 subunits in F11 mitochondrial fractions; immunocytochemistry experiments showed a strong overlap between the Kv7.4 fluorescent signal and that of the mitochondrial marker Mitotracker. Silencing of Kv7.4 expression significantly suppressed retigabine-dependent decrease in ΔΨ in intact F11 cells. Expression of Kv7.4 subunits was also detected by western blot in isolated mitochondria from total mouse brain and by immunofluorescence in mouse primary cortical neurons. Pharmacological experiments revealed a relevant functional role for Kv7.4 channels in regulating membrane potential and Ca2+ uptake in isolated neuronal mitochondria, as well as ΔΨ and ROS production in intact cortical neurons. In conclusion, these findings provide the first experimental evidence for the expression of Kv7.4 channels and their contribution in regulating K+ permeability of neuronal mitochondria.

Antioxidant and Anti-Inflammatory Activity of Combined Phycocyanin and Palmitoylethanolamide in Human Lung and Prostate Epithelial Cells

Inflammation involving the innate and adaptive immune systems is a normal response to infection; however, when allowed to continue unchecked, inflammation may result in several pathologies. Natural molecules with antioxidant properties can target the key players of inflammation and exert beneficial health effects. In this study, human normal bronchial (Beas-2B) and prostate (HPrEpiC) epithelial cell lines were exposed to infectious stimulation and treated with phycocyanin (PC) and palmitoylethanolamide (PEA), with the aim of demonstrating the enhanced antioxidant and anti-inflammatory properties of the combination. The cotreatment protected from cytotoxicity and greatly abated both the production of radical oxygen species (ROS) and the transcription of several inflammatory cytokines. Oxidative stress and inflammation were curtailed by affecting three main pathways: (1) inhibition of cyclooxygenase-2 enzyme and consequent decrease of signaling generating ROS; (2) increased synthesis of glutathione and therefore strengthening of the natural antioxidant defenses of the cells; (3) decreased infection-driven mitochondrial respiratory burst which generates oxidative stress. Based on the mounting interest in using nutraceuticals as adjuvants in the clinical practice, the present study unveils new mechanisms of action and enhanced efficacy of PC and PEA, supporting the possible exploitation of this combination in human disorders.

Vanadium (V)-doped ZnFe layered double hydroxide for enhanced sonocatalytic degradation of pymetrozine

In ultrasonic (US) processes, the development of environmentally friendly, effective, low-cost, and durable catalysts is needed to degrade pollutants. Here, ZnFe layered double hydroxide (LDH) was doped with vanadium (V) for the sonocatalytic degradation of a pesticide pymetrozine. The resulting catalyst had an average thickness of 25 nm, a specific surface area of 125.38 m2/g, and a bandgap value of 2.20 eV. In 90 min ultrasonic treatment, V-doped ZnFe LDH had 73% pymetrozine removal efficiency, which was 32% more than that of undoped ZnFe LDH. The US/V-doped ZnFe LDH process had a strong synergistic effect (synergy factor 7.17), which resulted in 57% and 68% greater efficiencies than the US alone and V-doped ZnFe LDH alone, respectively. The role of radical oxygen species was confirmed by carrying out radical trapping experiments using different scavengers and electron paramagnetic resonance analyses. Due to its high stability, the catalyst had good reuse potential with only an 8% performance reduction after 5 reuse cycles. Besides, the leaching of heavy metals was insignificant owing to the high integrity of the catalyst as confirmed by SEM and X-ray diffraction analysis. According to the GC–MS analysis, pymetrozine was first transformed into cyclic compounds then into aliphatic compounds such as animated products and carboxylic acids.

Evolutionary impacts of purine metabolism genes on mammalian oxidative stress adaptation.

Many mammals risk damage from oxidative stress stemming from frequent dives (i.e., cycles of ischemia/reperfusion and hypoxia/reoxygenation), high altitude and subterranean environments, or powered flight. Purine metabolism is an essential response to oxidative stress, and an imbalance between purine salvage and de novo biosynthesis pathways can generate damaging reactive oxygen species (ROS). Here, we examined the evolution of 117 purine metabolism-related genes to explore the accompanying molecular mechanisms of enhanced purine metabolism in mammals under high oxidative stress. We found that positively selected genes, convergent changes, and nonparallel amino acid substitutions are possibly associated with adaptation to oxidative stress in mammals. In particular, the evolution of convergent genes with cAMP and cGMP regulation roles may protect mammals from oxidative damage. Additionally, 32 genes were identified as under positive selection in cetaceans, including key purine salvage enzymes (i.e., HPRT1), suggesting improved re-utilization of non-recyclable purines avoid hypoxanthine accumulation and reduce oxidative stress. Most intriguingly, we found that six unique substitutions in cetacean xanthine dehydrogenase (XDH), an enzyme that regulates the generation of the ROS precursor xanthine oxidase (XO) during ischemic/hypoxic conditions, show enhanced enzyme activity and thermal stability and diminished XO conversion activity. These functional adaptations are likely beneficial for cetaceans by reducing radical oxygen species production during diving. In summary, our findings offer insights into the molecular and functional evolution of purine metabolism genes in mammalian oxidative stress adaptations.

Preparation of magnetic copper ferrite nanoparticle as peroxymonosulfate activating catalyst for effective degradation of levofloxacin.

Magnetic CuFe2O4 nanoparticles were successfully synthesized with a coprecipitation method at 500 °C calcination temperature, and were utilized to degrade levofloxacin (LEV) as a peroxymonosulfate (PMS) activator. The structure and composition of the nanocatalyst were characterized by a series of methods, including scanning electron microscopy, X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy, X-ray diffraction, vibrating sample magnetometer and thermogravimetric analysis. The effects of the PMS concentration, the catalyst dosage, the LEV initial concentration, the pH value and the inorganic anions on the LEV degradation were also explored. The results revealed that the designed CuFe2O4/PMS system had high activity and excellent stability in the complex conditions. The degradation efficiency of LEV still reached above 80% after four recycles of CuFe2O4 catalyst. The reactive species quenching experiments and electron paramagnetic resonance analysis suggested the existence of superoxide radicals, single oxygen, hydroxy radicals and sulfate radicals, and the first two were dominant radical oxygen species. Based on the mechanism analyses, the efficient degradation of LEV was probably due to the continuous generation of reactive species under the condition of Fe(III)/Fe(II) and Cu(II)/Cu(I) redox cycles. The research provided a reasonable reference for the PMS activation mechanism-based spinel-type ferrite catalysis.

Redox Properties of Human Erythrocytes Are Adapted for Vitamin C Recycling.

Ascorbic acid (AA; or vitamin C) is an important physiological antioxidant and radical scavenger. Some mammalian species, including homo sapiens, have lost the ability to synthetize AA and depend on its nutritional uptake. Erythrocytes from AA-auxotroph mammals express high amounts of the glucose transporter GLUT1. This isoform enables rapid uptake of glucose as well as dehydroascorbate (DHA), the fully oxidized form of AA. Here, we explored the effects of DHA uptake on the redox metabolism of human erythrocytes. DHA uptake enhanced plasma membrane electron transport (PMET) activity. This process is mediated by DCytb, a membrane bound cytochrome catalyzing extracellular reduction of Fe3+ and ascorbate free radical (AFR), the first oxidized form of AA. DHA uptake also decreased cellular radical oxygen species (ROS) levels. Both effects were massively enhanced in the presence of physiological glucose concentrations. Reduction of DHA to AA largely depleted intracellular glutathione (GSH) and induced the efflux of its oxidized form, GSSG. GSSG efflux could be inhibited by MK-571 (IC50 = 5 μM), indicating involvement of multidrug resistance associated protein (MRP1/4). DHA-dependent GSH depletion and GSSG efflux were completely rescued in the presence of 5 mM glucose and, partially, by 2-deoxy-glucose (2-DG), respectively. These findings indicate that human erythrocytes are physiologically adapted to recycle AA both intracellularly via GLUT1-mediated DHA uptake and reduction and extracellularly via DCytb-mediated AFR reduction. We discuss the possibility that this improved erythrocyte-mediated AA recycling was a prerequisite for the emergence of AA auxotrophy which independently occurred at least twice during mammalian evolution.

Ferroptosis in Lung Cancer: From Molecular Mechanisms to Prognostic and Therapeutic Opportunities.

Lung cancer is the second commonly diagnosed malignancy worldwide and has the highest mortality rate among all cancers. Tremendous efforts have been made to develop novel strategies against lung cancer; however, the overall survival of patients still is low. Uncovering underlying molecular mechanisms of this disease can open up new horizons for its treatment. Ferroptosis is a newly discovered type of programmed cell death that, in an iron-dependent manner, peroxidizes unsaturated phospholipids and results in the accumulation of radical oxygen species. Subsequent oxidative damage caused by ferroptosis contributes to cell death in tumor cells. Therefore, understanding its molecular mechanisms in lung cancer appears as a promising strategy to induce ferroptosis selectively. According to evidence published up to now, significant numbers of research have been done to identify ferroptosis regulators in lung cancer. Therefore, this review aims to provide a comprehensive standpoint of molecular mechanisms of ferroptosis in lung cancer and address these molecules’ prognostic and therapeutic values, hoping that the road for future studies in this field will be paved more efficiently.

Ascorbic Acid in Health and Disease: A Review

: Ascorbic acid is an essential nutrient, and required for various metabolic activities in humans. Typically citrus fruits, vegetables and organ meat are good source of vitamin C. It acts as strong antioxidant and act as a scavenger in defence against free radical oxygen species. It has also contributed to rejuvenate photo aged skin. It has ability to control the pigmentation of melanin by inhibiting the enzyme tyrosinase by interacting with copper ions. It serves as a co-antioxidant with vitamin E to regenerate alpha tocopherol, thereby retards cellular damage. Ascorbic acid is deprotonated to form ascorbate anion, contributes to its prooxidant properties and act as a potential anticancer agent. It reduces the mutation rate in mismatch-repair deficient human colon cancer cells. Ascorbic acid is a phytochemical has micronutrients that act against the inflammation in arthritis. Currently, challenges lies finding most stable formulation for achieving optimum results.

Attenuation of postharvest peel browning and chilling injury of banana fruit by Astragalus polysaccharides

The effect of Astragalus polysaccharides (APS) on peel browning and chilling injury (CI) of banana fruit was investigated. The results showed that APS induced the activities of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and reactive oxygen species (ROS) scavenging, reduced the H2O2 accumulation and the membrane lipid peroxidation and electrolyte leakage, by which delayed fruit CI, alleviated peel browning and the loss of nutrients in the flesh. It also improved the antioxidant capacity of fruit, enhanced the expression of MaSOD, MaCAT, MaAPX and led to the increase of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) activities. Moreover, APS up-regulated the expression of MaPOD, MaPAL, MaCHS and down-regulated the expression of MaPPO, resulting in the composition changes of the phenolics and flavonoids in banana fruit. Higher levels of hesperidin, ferulic acid, acacetin and myricetin were detected in APS treated fruit and were involved in alleviating fruit CI and peel browning. The present results for the first time demonstrated that APS treatment alleviated the CI of banana fruit, and provided a promising strategy for enhancing postharvest chilling tolerance.

Chlamydomonas Responses to Salinity Stress and Possible Biotechnological Exploitation

Salinity is among the main drivers affecting growth and distribution of photosynthetic organisms as Chlamydomonas spp. These species can live in multiple environments, including polar regions, and have been frequently studied for their adaptation to live at different salinity gradients. Upon salinity stress (hypersalinity is the most studied), Chlamydomonas spp. were found to alter their metabolism, reduce biomass production (growth), chlorophyll content, photosynthetic activity, and simultaneously increasing radical oxygen species production as well as lipid and carotenoid contents. This review summarizes the current literature on salt stress related studies on the green algae from the genus Chlamydomonas considering physiological and molecular aspects. The overall picture emerging from the data suggests the existence of common features of the genus in response to salinity stress, as well as some differences peculiar to single Chlamydomonas species. These differences were probably linked to the different morphological characteristics of the studied algae (e.g., with or without cell wall) or different sampling locations and adaptations. On the other hand, molecular data suggest the presence of common reactions, key genes, and metabolic pathways that can be used as biomarkers of salt stress in Chlamydomonas spp., with implications for future physiological and biotechnological studies on microalgae and plants.

The enhanced photocatalytic sterilization of MOF-Based nanohybrid for rapid and portable therapy of bacteria-infected open wounds

Open wounds are prone to infection and difficult to heal, which even threatens the life of patients because bacterial infections can induce other lethal complications without prompt treatment. The commonly used antibiotics treatment for bacterial infections has been reported to cause globally bacterial resistance and even the occurrence of superbacteria. The highly effective and antibiotic-independent therapeutic strategies are urgently needed for treating various kinds of bacteria-infected diseases. In this work, we synthesized an eco-friendly nanohybrid material (ZnDMZ) consisting of a kind of biodegradable metal organic framework (MOF, ZIF-8) combined with Zn-doped MoS2 (Zn–MoS2) nanosheets, which exhibited great ability to kill bacteria and promote the healing of bacteria-infected wounds under 660 nm light irradiation. The underlying mechanism is that besides the local hyperthermia, the nanohybrid material exhibits enhanced photocatalytic performance than single component in it, i.e., it can also be excited by 660 nm light to produce more oxygen radical species (ROS) due to the following factors. On one hand, the Zn doping can reduce the work function and the band gap of MoS2, which promotes the movement of photoexcited electrons to the surface of the material. On the other hand, the combination between Zn–MoS2 and MOF induces the formation of a built-in electric field due to their work function difference, thus accelerating the separation of photoexcited electron-hole pairs. Because of the synergy of photocatalytic effect, photothermal effect and the released Zn ions, the synthesized ZnDMZ possessed a highly effective antibacterial efficacy of 99.9% against Staphylococcus aureus under 660 nm light irradiation for 20 min without cytotoxicity. In vivo tests showed that this nanohybrid material promoted the wound healing due to the released Zn ions. This nanohybrid will be promising for rapid and portable treatment of bacteria-infected open wounds in pathogenic bacteria contaminated environments.

An updated overview on metal nanoparticles toxicity.

Although thousands of different nanoparticles (NPs) have been identified and synthesized to date, well-defined, consistent guidelines to control their exposure and evaluate their potential toxicity have yet to be fully established. As potential applications of nanotechnology in numerous fields multiply, there is an increased awareness of the issue of nanomaterials' toxicity among scientists and producers managing them. An updated inventory of customer products containing NPs estimates that they currently number over 5.000; ten years ago, they were one fifth of this. More often than not, products bear no information regarding the presence of NPs in the indicated list of ingredients or components. Consumers are therefore largely unaware of the extent to which nanomaterials have entered our lives, let alone their potential risks. Moreover, the lack of certainties with regard to the safe use of NPs is curbing their applications in the biomedical field, especially in the diagnosis and treatment of cancer, where they are performing outstandingly but are not yet being exploited as much as they could. The production of radical oxygen species is a predominant mechanism leading to metal NPs-driven carcinogenesis. The release of particularly reactive metal ions capable of crossing cell membranes has also been implicated in NPs toxicity. In this review we discuss the origin, behavior and biological toxicity of different metal NPs with the aim of rationalizing related health hazards and calling attention to toxicological concerns involved in their increasingly widespread use.

Development of bioactive catechol functionalized nanoparticles applicable for 3D bioprinting

Efficient wound treatments to target specific events in the healing process of chronic wounds constitute a significant aim in regenerative medicine. In this sense, nanomedicine can offer new opportunities to improve the effectiveness of existing wound therapies. The aim of this study was to develop catechol bearing polymeric nanoparticles (NPs) and to evaluate their potential in the field of wound healing. Thus, NPs wound healing promoting activities, potential for drug encapsulation and controlled release, and further incorporation in a hydrogel bioink formulation to fabricate cell-laden 3D scaffolds are studied. NPs with 2 and 29 M % catechol contents (named NP2 and NP29) were obtained by nanoprecipitation and presented hydrodynamic diameters of 100 and 75 nm respectively. These nanocarriers encapsulated the hydrophobic compound coumarin-6 with 70% encapsulation efficiency values. In cell culture studies, the NPs had a protective effect in RAW 264.7 macrophages against oxidative stress damage induced by radical oxygen species (ROS). They also presented a regulatory effect on the inflammatory response of stimulated macrophages and promoted upregulation of the vascular endothelial growth factor (VEGF) in fibroblasts and endothelial cells. In particular, NP29 were used in a hydrogel bioink formulation using carboxymethyl chitosan and hyaluronic acid as polymeric matrices. Using a reactive mixing bioprinting approach, NP-loaded hydrogel scaffolds with good structural integrity, shape fidelity and homogeneous NPs dispersion, were obtained. The in vitro catechol NPs release profile of the printed scaffolds revealed a sustained delivery. The bioprinted scaffolds supported viability and proliferation of encapsulated L929 fibroblasts over 14 days. We envision that the catechol functionalized NPs and resulting bioactive bioink presented in this work offer promising advantages for wound healing applications, as they: 1) support controlled release of bioactive catechol NPs to the wound site; 2) can incorporate additional therapeutic functions by co-encapsulating drugs; 3) can be printed into 3D scaffolds with tailored geometries based on patient requirements.

Role of TLR4 in Neutrophil Dynamics and Functions: Contribution to Stroke Pathophysiology.

Background and PurposeThe immune response subsequent to an ischemic stroke is a crucial factor in its physiopathology and outcome. It is known that TLR4 is implicated in brain damage and inflammation after stroke and that TLR4 absence induces neutrophil reprogramming toward a protective phenotype in brain ischemia, but the mechanisms remain unknown. We therefore asked how the lack of TLR4 modifies neutrophil function and their contribution to the inflammatory process.MethodsIn order to assess the role of the neutrophilic TLR4 after stroke, mice that do not express TLR4 in myeloid cells (TLR4loxP/Lyz-cre) and its respective controls (TLR4loxP/loxP) were used. Focal cerebral ischemia was induced by occlusion of the middle cerebral artery and infarct size was measured by MRI. A combination of flow cytometry and confocal microscopy was used to assess different neutrophil characteristics (circadian fluctuation, cell surface markers, cell complexity) and functions (apoptosis, microglia engulfment, phagocytosis, NETosis, oxidative burst) in both genotypes.ResultsAs previously demonstrated, mice with TLR4 lacking-neutrophils had smaller infarct volumes than control mice. Our results show that the absence of TLR4 keeps neutrophils in a steady youth status that is dysregulated, at least in part, after an ischemic insult, preventing neutrophils from their normal circadian fluctuation. TLR4-lacking neutrophils showed a higher phagocytic activity in the basal state, they were preferentially engulfed by the microglia after stroke, and they produced less radical oxygen species (ROS) in the first stage of the inflammatory process.ConclusionsTLR4 is specifically involved in neutrophil dynamics under physiological conditions as well as in stroke-induced tissue damage. This research contributes to the idea that TLR4, especially when targeted in specific cell types, is a potential target for neuroprotective strategies.

A Versatile Light‐Triggered Radical‐Releasing Surface Coating Technology

AbstractOrganic based redox active materials and coatings are increasingly being investigated as solutions for more efficient energy devices, catalysts, sensors, and biomedical technologies. Their advantage is negating the necessity for expensive and unsustainable rare earth metals seen in other redox active materials. Challenges to their wide‐spread usage remain, including versatility of coatings on a wide range of substrates, and creating smart devices that can respond to environmental stimuli. A new light responsive radical releasing redox coating has been developed, that can be rapidly and stably applied to a diverse range of substrates including silicon dioxide, plastics, and microparticles. These coatings rapidly cleave upon irradiation with UV‐A light to release stable nitroxides from the surface coating. These coatings are developed into microparticle sensors for radical oxygen species, which are able to detect the generation of free radicals in a model system for particulate matter pollution. These new responsive organic redox coatings also offer future potential to function as switchable organic electrodes, and biomedical surface coatings to prevent biofouling and reduce inflammation.

Attenuation of Hg(II)-induced cellular and DNA damage in human blood cells by uric acid.

Mercury (Hg) is a widespread environmental pollutant and toxicant that induces multiple organ damage in humans and animals. Hg toxicity is mediated by the induction of oxidative stress in the target cells. We used uric acid (UA), a potent antioxidant found in biological fluids, to protect human red blood cells (RBC) and lymphocytes against Hg-mediated cell, organelle, and genotoxicity. RBCs were incubated with mercuric chloride (HgCl2), an Hg(II) compound, either alone or in the presence of UA. Incubation of RBCs with only HgCl2 increased the production of nitrogen and oxygen radical species, enhanced methemoglobin levels, heme degradation, free ferrous iron, oxidation of proteins and membrane lipids, and reduced the antioxidant capacity of cells. UA enhanced the antioxidant capacity of RBCs and restored metabolic, plasma membrane-bound, and antioxidant enzyme activities. Scanning electron microscopy showed that UA prevented HgCl2-mediated morphological changes in RBCs. HgCl2 dissipated the mitochondrial membrane potential and increased lysosomal membrane damage in lymphocytes, but UA pre-treatment attenuated these effects. Genotoxicity analysis by comet assay showed that UA protected lymphocyte DNA from HgCl2-induced damage. Importantly, UA itself did not exhibit any deleterious effects on RBCs or lymphocytes. Thus, UA protects human blood cells from Hg(II)-mediated oxidative damage, reducing the harmful effects of this extremely toxic metal. We suggest that UA has a similar protective role in plasma against heavy metal toxicity.