The oxidative damage induced by abiotic stress factors such as salinity, drought, extreme temperatures, heavy metals, pollution, and high irradiance has been studied in Arabidopsis thaliana. Ultra-weak photon emission (UPE) is presented as a signature reflecting the extent of the oxidation process and/or damage. It can be used to predict the physiological state and general health of plants. This study presents an overview of a potential research platform where the technique can be applied. The results presented can aid in providing invaluable information for developing strategies to mitigate abiotic stress in crops by improving plant breeding programs with a focus on enhancing tolerance. This study evaluates the applicability of charged couple device (CCD) imaging in evaluating plant stress and degree of damage and to discuss the advantages and limitations of the claimed non-invasive label-free tool.
- Klíčová slova
- Antioxidants, Reactive oxygen species, Stress imaging, Two-dimensional photon emission imaging, Wounding,
- MeSH
- Arabidopsis * fyziologie MeSH
- fotony * MeSH
- fyziologický stres * MeSH
- oxidační stres MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Neutrophils mediate the early innate immune response through extracellular traps comprising intracellular protein and DNA. These traps play a pivotal role in both immunity against invading pathogens and the development of immunopathological reactions through the production of reactive oxygen species (ROS). Proteins serve as the main target for ROS, resulting in the formation of protein adducts. Herein, we report that the superoxide anion radical (O2˙-) plays a vital role in neutrophil function through sequential events involving 5-lipoxygenase (5-LOX) and NADPH oxidase (NOX). More specifically, differences in NOX homologs expression were observed post-stimulation with PMA and LPS. Differentiation conditions and O2˙- generation were confirmed using flow cytometry. Immunoblotting analysis confirmed the time-dependent expression of NOX underlying its requirement and 5-LOX-mediated lipid peroxidation events in neutrophil function. Protein-malondialdehyde (MDA) adducts formed were detected using immunoblotting, and quercetin was evaluated for its ability to scavenge free radicals through electron paramagnetic resonance (EPR) spin-trapping spectroscopy and results were confirmed with blotting analysis. Free radical-mediated protein oxidation events influence neutrophil function and protein adducts formed serve as markers of neutrophil activation upon infection and inflammation. The study warrants further corroboration and the study of specific proteins involved in neutrophil activation and their role in inflammation.
- Publikační typ
- časopisecké články MeSH
Extracellular vesicles (EVs) are a type of cytoplasmic vesicles secreted by a variety of cells. EVs originating from cells have been known to participate in cell communication, antigen presentation, immune cell activation, tolerance induction, etc. These EVs can also carry the active form of Nicotinamide Adenine Dinucleotide Phosphate Oxidase Hydrogen (NADPH) oxidase, which is very essential for the production of reactive oxygen species (ROS) and that can then modulate processes such as cell regeneration. The aim of this study is to characterize the EVs isolated from U-937 and THP-1 cells, identify the NADPH oxidase (NOX) isoforms, and to determine whether EVs can modulate NOX4 and NOX2 in monocytes and macrophages. In our study, isolated EVs of U-937 were characterized using dynamic light scattering (DLS) spectroscopy and immunoblotting. The results showed that the exogenous addition of differentiation agents (either phorbol 12-myristate 13-acetate (PMA) or ascorbic acid) or the supplementation of EVs used in the study did not cause any stress leading to alterations in cell proliferation and viability. In cells co-cultured with EVs for 72 h, strong suppression of NOX4 and NOX2 is evident when monocytes transform into macrophagic cells. We also observed lower levels of oxidative stress measured using immunoblotting and electron paramagnetic resonance spectroscopy under the EVs co-cultured condition, which also indicates that EVs might contribute significantly by acting as an antioxidant source, which agrees with previous studies that hypothesized the role of EVs in therapeutics. Therefore, our results provide evidence for NOX regulation by EVs in addition to its role as an antioxidant cargo.
- Klíčová slova
- NADPH oxidase, NOX2, NOX4, extracellular vesicles, free radicals, macrophages, monocytes, reactive oxygen species,
- Publikační typ
- časopisecké články MeSH
The innate immune response represents the first-line of defense against invading pathogens. Reactive oxygen species (ROS) and reactive nitrogen species (RNS) have been implicated in various aspects of innate immune function, which involves respiratory bursts and inflammasome activation. These reactive species widely distributed within the cellular environment are short-lived intermediates that play a vital role in cellular signaling and proliferation and are likely to depend on their subcellular site of formation. NADPH oxidase complex of phagocytes is known to generate superoxide anion radical (O2 •-) that functions as a precursor for antimicrobial hydrogen peroxide (H2O2) production, and H2O2 is utilized by myeloperoxidase (MPO) to generate hypochlorous acid (HOCl) that mediates pathogen killing. H2O2 modulates the expression of redox-responsive transcriptional factors, namely NF-kB, NRF2, and HIF-1, thereby mediating redox-based epigenetic modification. Survival and function of immune cells are under redox control and depend on intracellular and extracellular levels of ROS/RNS. The current review focuses on redox factors involved in the activation of immune response and the role of ROS in oxidative modification of proteins in macrophage polarization and neutrophil function.
- Klíčová slova
- inflammation, innate immune response, macrophage, neutrophils, oxidative stress, protein oxidation, reactive oxygen species,
- MeSH
- kyselina chlorná MeSH
- oxidace-redukce MeSH
- oxidační stres MeSH
- peroxid vodíku * MeSH
- přirozená imunita MeSH
- superoxidy * MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- kyselina chlorná MeSH
- peroxid vodíku * MeSH
- superoxidy * MeSH
In this study, we investigated the properties of ascorbic acid (vitamin C), which is a naturally occurring water-soluble vitamin. Our goal is to evaluate its pro-oxidative and/or antioxidant capabilities. To do this, we initially used a confocal laser scanning microscope (CLSM) to visualize the differentiation pattern in U-937 cells under the treatment of variable concentrations of ascorbic acid. Prior to induction, U-937 cells showed a spherical morphology. After treatment, significant morphological changes were observed in the form of prominent pseudopodia and amoeboid structures. Interestingly, pseudopodia incidences increased with an increase in ascorbic acid concentrations. In addition, our analysis of protein modification using anti-malondialdehyde antibodies showed changes in more than one protein. The findings reveal the link between the differentiation of U-937 cells into macrophages and the protein modifications triggered by the production of reactive oxygen species when U-937 cells are exposed to ascorbic acid. Furthermore, the transformation of ascorbic acid from a pro-oxidative to an antioxidant property is also demonstrated.
- Klíčová slova
- Antioxidants, Human cells, Pro-oxidant, Reactive oxygen species, Vitamin C,
- Publikační typ
- časopisecké články MeSH
Human skin is exposed to various physical and chemical stress factors, which commonly cause the oxidation of lipids and proteins. In this study, azo initiator AAPH [2,2' -azobis(2-methylpropionamidine) dihydrochloride] was employed to initiate lipid peroxidation in porcine skin as an ex vivo model for human skin. We demonstrate that malondialdehyde (MDA), a secondary product of lipid peroxidation, is covalently bound to collagen in the dermis, forming MDA-collagen adducts. The binding of MDA to collagen results in an unfolding of the collagen triple helix, formation of the dimer of α-chains of collagen, and fragmentation of the collagen α-chain. It is proposed here that the MDA is bound to the lysine residues of α-chain collagen, which are involved in electrostatic interaction and hydrogen bonding with the glutamate and aspartate of other α-chains of the triple helix. Our data provide crucial information about the MDA binding topology in the skin, which is necessary to understand better the various types of skin-related diseases and the aging process in the skin under stress.
- Klíčová slova
- Collagen, Lysine, Malondialdehyde, Reactive oxygen species, Skin,
- MeSH
- kolagen * metabolismus MeSH
- lidé MeSH
- malondialdehyd metabolismus MeSH
- oxidace-redukce MeSH
- oxidační stres * MeSH
- peroxidace lipidů MeSH
- prasata MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- kolagen * MeSH
- malondialdehyd MeSH
Acetaldehyde can be found in human cells as a byproduct of various metabolic pathways, including oxidative processes such as lipid peroxidation. This secondary product of lipid peroxidation plays a role in various pathological processes, leading to various types of civilization diseases. In this study, the formation of free acetaldehyde induced by oxygen-centred radicals was studied in monocyte-like cell line U937. Exposure of U937 cells to peroxyl/alkoxyl radicals induced by azocompound resulted in the formation of free acetaldehyde. Acetaldehyde is formed by the cleavage of fatty acids, which represents the breakdown of fatty acids into smaller fragments initiated by the cyclization of lipid peroxyl radical and β-scission of lipid alkoxyl radical. The cleavage of fatty acids alters the integrity of the plasma and nuclear membrane, leading to the loss of cell viability. Understanding the pathological processes of acetaldehyde formation is an active area of research with potential implications for preventing and treating various diseases associated with oxidative stress.
- Klíčová slova
- Aldehyde, Azocompound, Cell viability, Lipid peroxidation, Membrane integrity, Peroxyl radical,
- MeSH
- acetaldehyd * MeSH
- lidé MeSH
- mastné kyseliny metabolismus MeSH
- monocyty * metabolismus MeSH
- peroxidace lipidů MeSH
- reaktivní formy kyslíku MeSH
- U937 buňky MeSH
- volné radikály metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- acetaldehyd * MeSH
- mastné kyseliny MeSH
- perhydroxyl radical MeSH Prohlížeč
- reaktivní formy kyslíku MeSH
- volné radikály MeSH
Oxidative processes in all types of organisms cause the chemical formation of electronically excited species, with subsequent ultraweak photon emission termed biological auto(chemi)luminescence (BAL). Imaging this luminescence phenomenon using ultrasensitive devices could potentially enable monitoring of oxidative stress in optically accessible areas of the human body, such as skin. Although oxidative stress induced by UV light has been explored, for chemically induced stress, there is no in vivo-quantified imaging of oxidative processes in human skin using BAL under the controlled extent of oxidative stress conditions. Furthermore, the mechanisms and dynamics of BAL from the skin have not been fully explored. Here, we demonstrate that different degrees of chemically induced oxidative stress on the skin can be spatially resolved quantitatively through noninvasive label-free BAL imaging. Additionally, to gain insight into the underlying mechanisms, a minimal chemical model of skin based on a mixture of lipid, melanin, and water was developed and used to show that it can be used to reproduce essential features of the response of real skin to oxidative stress. Our results contribute to novel, noninvasive photonic label-free methods for quantitative sensing of oxidative processes and oxidative stress.
- MeSH
- fotony MeSH
- kůže * metabolismus MeSH
- lidé MeSH
- luminiscence * MeSH
- oxidační stres MeSH
- ultrafialové záření MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Under environmental conditions, plants are exposed to various abiotic and biotic stress factors, which commonly cause the oxidation of lipids and proteins. Lipid peroxidation constantly produces malondialdehyde (MDA), a secondary product of lipid peroxidation, which is covalently bound to proteins forming MDA-protein adducts. The spatial distribution of MDA-protein adducts in Arabidopsis leaves shows that MDA-protein adducts are located in the chloroplasts, uniformly spread out over the thylakoid membrane. At the lumenal side of thylakoid membrane, MDA interacts with PsbP, an extrinsic subunit of the photosystem II (PSII), which is in electrostatic interaction with the PSII core proteins. Under heat stress, when MDA is moderately enhanced, the electrostatic interaction between PsbP and PSII core proteins is weakened, and PsbP with bound MDA is released in the lumen. It is proposed here that the electrophilic MDA is bound to the nucleophilic lysine residues of PsbP, which are involved in electrostatic interactions with the negatively charged glutamate of the PSII core protein. Our data provide crucial information about the MDA binding topology in the higher plant PSII complex, which is necessary to understand better the physiological functions of MDA for plant survival under stress.
- Klíčová slova
- Heat stress, Lysine, Malondialdehyde, Photosystem II, Reactive oxygen species,
- MeSH
- Arabidopsis * MeSH
- kyselina glutamová MeSH
- lysin MeSH
- malondialdehyd MeSH
- reakce na tepelný šok MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- kyselina glutamová MeSH
- lysin MeSH
- malondialdehyd MeSH
The heart of oxygenic photosynthesis is the water-splitting photosystem II (PSII), which forms supercomplexes with a variable amount of peripheral trimeric light-harvesting complexes (LHCII). Our knowledge of the structure of green plant PSII supercomplex is based on findings obtained from several representatives of green algae and flowering plants; however, data from a non-flowering plant are currently missing. Here we report a cryo-electron microscopy structure of PSII supercomplex from spruce, a representative of non-flowering land plants, at 2.8 Å resolution. Compared with flowering plants, PSII supercomplex in spruce contains an additional Ycf12 subunit, Lhcb4 protein is replaced by Lhcb8, and trimeric LHCII is present as a homotrimer of Lhcb1. Unexpectedly, we have found α-tocopherol (α-Toc)/α-tocopherolquinone (α-TQ) at the boundary between the LHCII trimer and the inner antenna CP43. The molecule of α-Toc/α-TQ is located close to chlorophyll a614 of one of the Lhcb1 proteins and its chromanol/quinone head is exposed to the thylakoid lumen. The position of α-Toc in PSII supercomplex makes it an ideal candidate for the sensor of excessive light, as α-Toc can be oxidized to α-TQ by high-light-induced singlet oxygen at low lumenal pH. The molecule of α-TQ appears to shift slightly into the PSII supercomplex, which could trigger important structure-functional modifications in PSII supercomplex. Inspection of the previously reported cryo-electron microscopy maps of PSII supercomplexes indicates that α-Toc/α-TQ can be present at the same site also in PSII supercomplexes from flowering plants, but its identification in the previous studies has been hindered by insufficient resolution.
- MeSH
- alfa-tokoferol * analýza metabolismus MeSH
- elektronová kryomikroskopie MeSH
- fotosyntéza MeSH
- fotosystém II (proteinový komplex) * metabolismus MeSH
- rostliny metabolismus MeSH
- tylakoidy metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- alfa-tokoferol * MeSH
- fotosystém II (proteinový komplex) * MeSH
