Free Radical Research最新文献

Plasma is the fourth physical state of matter, characterized by an ionized gaseous mixture, after solid, liquid, and gas phases, and contains a wide array of components such as ions, electrons, radicals, and ultraviolet ray. Whereas the sun and thunder are typical natural plasma, recent progress in the electronics enabled the generation of body-temperature plasma, designated as low-temperature plasma (LTP) or non-thermal plasma since the 1990s. LTP has attracted the attention of researchers for possible biological and medical applications. All the living species on earth utilize water as essential media for solvents and molecular transport. Thus, biological application of LTP naturally intervenes water whether LTP is exposed directly or indirectly, where plasma-activated lactate (PAL) is a standard, containing H₂O₂, NO₂^- and other identified molecules. Electron spin resonance and immunohistochemical studies demonstrated that LTP exposure is a handy method to load local oxidative stress. Cancer cells are characterized by persistent self-replication and high cytosolic catalytic Fe(II). Therefore, both direct exposure of LTP and PAL can provide higher damage to cancer cells in comparison to non-tumorous cells, which has been demonstrated in a variety of cancer types. The cell death mode is either apoptosis or ferroptosis, depending on the cancer-type. Thus, LTP and PAL are expected to work as an additional cancer therapy to the established guideline protocols, especially for use in somatic cavities or surgical margins.

COVID-19 has been pandemic since 2020 with persistent generation of new variants. Cellular receptor for SARS-CoV-2 is angiotensin-converting enzyme 2 (ACE2), where transmembrane serine protease-2 (TMPRSS2) is essential for viral internalization. We recently reported abundant expression of ACE2 and TMPRSS2 in the oral cavity of humans and mice. Therefore, oral cavity may work for COVID-19 infection gates. Here we undertook to evaluate whether vaccination in the tongue harbors any merit in comparison to subcutaneous injection. Low-temperature plasma (LTP) is the fourth physical state of matters with ionization above gas but at body temperature. LTP provides complex chemistry, eventually supplying oxidative and/or nitrosative stress on the interface. LTP-associated cellular death has been reported to cause apoptosis and/or ferroptosis. However, there is few data available on immunogenicity retention after LTP exposure. We therefore studied the effect of LTP exposure after the injection of keyhole limpet hemocyanin (KLH) or spike 2 protein of SARS-CoV-2 to the tongue of six-week-old male BALB/c mice, compared to subcutaneous vaccination. Whereas LTP did not change the expression of ACE2 and TMPRSS2 in the tongue, repeated LTP exposure after tongue vaccination significantly promoted systemic and specific IgM production at day 11. In contrast, repeated LTP exposure after subcutaneous vaccination of KLH decreased systemic IgM production. Of note, tongue injection produced significantly higher titer of IgM and IgG in the case of KLH. In conclusion, LTP significantly reinforced humoral immunity by IgM after tongue injection. Vaccination to the tongue can be a novel strategy to acquire immediate immunity.

Extracellular vesicles (EVs) are identified as a non-classical way to mediate iron efflux except ferroportin. Interestingly, recent studies indicated that EVs pathway is a novel way involved in iron efflux. Mitochondria-derived vesicles (MDVs) are the potential mediator to load mitochondrial iron into EVs. Additionally, iron-replete cells resist excess iron-induced damage by secreting iron-loaded EVs, and the uptake of these EVs induces oxidative damage in the recipient cell. Importantly, iron-loaded EVs play a key role in aberrant iron distribution, which drives the progress of diseases like nonalcoholic fatty liver disease (NAFLD) and neurodegenerative diseases. Herein, we summarize extant research on intracellular iron export with an emphasis on EVs and put our eyes on the relationship between iron-loaded EVs with both parent and target cells. Iron-loaded EVs will be an important avenue for later research on their vital role in iron redistribution.

Oxidative Stress (OS) is involved in the pathogenesis of COVID-19 and in the mechanisms by which SARS-CoV-2 causes injuries to tissues, leading to cytopathic hypoxia and ultimately multiple organ failure. The measurement of blood glutathione (GSH), H₂O₂, and catalase activity may help clarify the pathophysiology pathways of this disease. We developed and standardized a sensitive and specific chemiluminescence technique for H₂O₂ and GSH measurement in plasma and red blood cells of COVID-19 patients admitted to the intensive care unit (ICU). Contrary to what was expected, the plasma concentration of H₂O₂ was substantially reduced (10-fold) in COVID-19 patients compared to the healthy control group. From the cohort of patients discharged from the hospital and those who were deceased, the former showed a 3.6-fold and the later 16-fold H₂O₂ reduction compared to the healthy control. There was a 4.4 reduction of H₂O₂ concentration in the deceased group compared to the discharged group. Interestingly, there was no variation in GSH levels between groups, and reduced catalase activity was found in discharged and deceased patients compared to control. These data represent strong evidence that H₂O₂ is converted into highly reactive oxygen species (ROS), leading to the worst prognosis and death outcome in COVID-19 patients admitted to ICU. Considering the difference in the levels of H₂O₂ between the control group and the deceased patients, it is proposed the quantification of plasma H₂O₂ as a marker of disease progression and the induction of the synthesis of antioxidant enzymes as a strategy to reduce the production of oxidative stress during severe COVID-19.HighlightsH₂O₂ plasma levels is dramatically reduced in patients who deceased compared to those discharged and to the control group.Plasmatic quantification of H₂O₂ can be possibly used as a predictor of disease progression.Catalase activity is reduced in COVID-19.GSH levels remain unchanged in COVID-19 compared to the control group.

Heart ischemia/reperfusion (I/R) injury is related to iron content. However, the occurrence and mechanism of changes in labile iron pool (LIP) during I/R is debatable. Moreover, the identity of the iron form dominant in LIP during I/R is unclear. Herein, we measured changes of LIP during simulated ischemia (SI) and reperfusion (SR), in which ischemia was simulated in vitro with lactic acidosis and hypoxia. Total LIP did not change in lactic acidosis, whereas LIP, especially Fe³⁺, increased in hypoxia. Under SI, accompanied by hypoxia with acidosis, both Fe²⁺ and Fe³⁺ were significantly increased. Increased total LIP was maintained at 1 h post-SR. However, the Fe²⁺ and Fe³⁺ portion was changed. The increased Fe²⁺ was decreased, and conversely the Fe³⁺ was increased. BODIPY oxidized signal increased and through the time-course these changes correlated with blebbing of cell membrane and SR-induced LDH release. These data suggested lipid peroxidation occurred via Fenton's reaction. The experiments using bafilomycin A1 and zinc protoporphyrin suggested no role of ferritinophagy or heme oxidation in the increase of LIP during SI. The extracellular source, transferrin assessed using serum transferrin bound iron (TBI) saturation showed that the depletion of TBI reduced SR-induced cell damages and additive saturation of TBI accelerated SR-induced lipid peroxidation. Furthermore, Apo-Tf dramatically blocked the increase of LIP and SR-induced damages. In conclusion, Tf-mediated iron induces the increase of LIP during SI, and it causes Fenton reaction-mediated lipid peroxidation during the early phase of SR.

Urinary 8-oxo-7,8-dihydroguanosine (8-oxoGuo) and 8-oxo-7,8-dihydro-2'- deoxyguanosine (8-oxodGuo) are considered biomarkers of oxidative stress, and patients with nephrotic syndrome have been reported to have increased oxidative stress levels. In this study, we aimed to assess the value of 8-oxoGuo and 8-oxodGuo as novel biomarkers to evaluate the severity of nephrotic syndrome. In total, 107 patients with nephrotic syndrome and 116 healthy controls were recruited for this study. The concentrations of urinary 8-oxoGuo and 8-oxodGuo were measured using isotope-labeled liquid chromatography with tandem mass spectrometry. Urinary creatinine was used to regulate 8-oxoGuo and 8-oxodGuo concentrations. Urinary 8-oxoGuo and 8-oxoGuo/Cr levels in patients with nephrotic syndrome were significantly higher than those in healthy control participants. 8-oxoGuo/Cr showed a positive correlation with the 24 h urinary total protein (UTP) and UTP levels and negative correlations with serum total protein and albumin levels. After treatment, urinary 8-oxoGuo and 8-oxoGuo/Cr levels were significantly lower in the group with a low 24 h-UTP value (<3.5 g/d) than in the high value group. 8-oxoGuo can be used as a feasible and reliable biomarker for the assessment of nephrotic syndrome.HighlightsUrinary 8-oxoGuo level was significantly increased in patients with nephrotic syndrome.Urinary 8-oxoGuo level increased with an increase in plasma protein and a decrease in urine protein.Urinary 8-oxoGuo level decreased with nephrotic syndrome remission when urinary microalbumin showed no significant change.Urinary 8-oxoGuo level can be used as novel biomarkers of nephrotic syndrome.

Guaiacol (2-methoxyphenol) is naturally occurring phenolic compound essential in various research areas. Oxidative transformation of guaiacol can lead to the formation of various products, including 1,3-benzodioxole or ortho-quinone. Therefore, this study is focused on the investigation of the reaction enthalpies of experimentally observed guaiacol oxidation pathways in gas-phase, as well as in non-polar environment and aqueous solution. Corresponding Density Functional Theory (DFT) calculations were carried out using two hybrid functionals (M06-2X and B3LYP-D3). All reaction enthalpies, as well as Gibbs free energies, were also calculated using composite ab initio G4 method. M06-2X and G4 results show mutual agreement and the best accordance with available experimentally determined reaction enthalpies. Obtained Gibbs free reaction energies indicate that formation of ortho-quinone is thermodynamically preferred to formation of 1,3-benzodioxole at 298 K in studied environments. Moreover, all computational methods confirm that the reaction enthalpy of methoxy group demethylation, i.e. O-C bond dissociation enthalpy (BDE), is substantially lower in comparison to the enthalpy of hydrogen atom transfer from phenolic OH group. In the case of phenoxide anion of guaiacol, which can be formed in ionization supporting solvents, O-C BDE shows further significant decrease, exceeding 50 kJ mol^-1, in comparison to parent molecule.HIGHLIGHTSReaction enthalpies and Gibbs free energies of individual steps of guaiacol transformation to 1,3-benzodioxole or ortho-quinone are studied in three environments.M06-2X functional and composite ab initio G4 methods provide reliable O-H and O-C bond dissociation enthalpies.Dissociation enthalpy of methoxy group O-C bond is lower by ca. 100 kJ mol^-1 in comparison to phenolic O-H bond.Phenoxide anion of guaiacol shows substantially lower O-C BDE than parent molecule.

Betulinic acid (BA), a pentacyclic triterpenoid found in certain plant species, has been reported to have several health benefits including antioxidant and anti-apoptotic properties. However, the mechanism by which BA confers these properties is currently unknown. Saccharomyces cerevisiae, a budding yeast with a short life cycle and conserved cellular mechanism with high homology to humans, was used as a model for determining the role of BA in aging and programmed cell death (PCD). Treatment with hydrogen peroxide (H₂O₂) exhibited significantly increased (30-35%) survivability of antioxidant (sod1Δ, sod2Δ, cta1Δ, ctt1Δ, and tsa1Δ) and anti-apoptotic (pep4Δ and fis1Δ) mutant strains when cells were pretreated with BA (30 µM) as demonstrated in spot and CFU (Colony forming units) assays. Measurement of intracellular oxidation level using the ROS-specific dye H₂DCF-DA showed that all tested BA-pretreated mutants exhibited decreased ROS than the control when exposed to H₂O₂. Similarly, when mutant strains were pretreated with BA and then exposed to H₂O₂, there was reduced lipid peroxidation as revealed by the reduced malondialdehyde content. Furthermore, BA-pretreated mutant cells showed significantly lower apoptotic activity by decreasing DNA/nuclear fragmentation and chromatin condensation under H₂O₂-induced stress as determined by DAPI and acridine orange/ethidium bromide staining. In addition, BA treatment also extended the life span of antioxidant and anti-apoptotic mutants by ∼10-25% by scavenging ROS and preventing apoptotic cell death. Our overall results suggest that BA extends the chronological life span of mutant strains lacking antioxidant and anti-apoptotic genes by lowering the impact of oxidative stress, ROS levels, and apoptotic activity. These properties of BA could be further explored for its use as a valuable nutraceutical.

Oxidative stress is related to health problems including neurological and neurodegenerativedisturbs, such as Parkinson's disease. Natural compounds are reported as source of antioxidant molecules. Therefore, this study aimed to analyze the antioxidant and neuroprotective potential of a new diterpene isolated from C. argyrophylloides (MP-1). Male Wistar rats (250-300 g) were used to evaluate MP-1 antiparkinsonian potential through neurodegenerative model induced by the neurotoxin 6-hydroxydopamine (21 μg). On the 14th day, animals were submitted to behavioral tests and on the 15th day, brain areas were dissected to neurochemical analyzes. MP-1 demonstrated a high antioxidant capacity in vitro and decreased the parkinsonian effects, such as behavioral changes, motor alterations, and body weight loss. MP-1 was also able to control the upregulated levels of nitrosative stress and lipid peroxidation. These findings suggest MP-1 as a diterpene with high antioxidant capacity which might be used to development of new approach against Parkinson's disease.

Protein N-pyrrolation, which converts lysine residues to N^ε-pyrrole-l-lysine (pyrK), is a naturally occurring covalent modification. The pyrrolated proteins have a unique property of binding to DNA-staining agents, such as SYBR Green I (SG), and anti-DNA antibodies, suggesting a physiologically relevant modification that gives rise to DNA mimic protein. These properties of pyrrolated protein are suggested to be associated with innate and autoimmune responses. Short-chain aldehydes derived from lipid peroxidation are thought to be involved in the formation of pyrK. We now report that similar lysine N-pyrrolation also occurs during the metal-catalyzed oxidation of proteins with ascorbate. When human serum albumin (HSA) was incubated with Fe²⁺/ascorbate in the presence and absence of docosahexaenoic acid, the protein was converted to SG-binding proteins even without the polyunsaturated fatty acid. The formation of SG-binding proteins by Fe²⁺/ascorbate was accompanied by the formation of pyrK, which was also detected in ascorbate-treated hemoglobin. Moreover, the metal-catalyzed oxidation of ascorbate produced the pyrrolation factors, glycolaldehyde and glyoxal. These results and the observations that sera from autoimmune-prone MRL-lpr mice recognized modified proteins with Fe²⁺/ascorbate and with glycolaldehyde/glyoxal suggest that the autoxidation of ascorbate, as well as lipid peroxidation, can be a source of autoantigenic N-pyrrolated proteins. Our findings revealed a possible function of ascorbate as an endogenous source of pyrrolated proteins and suggested that the pyrK residues generated in proteins may play a role in the innate and autoimmune responses associated with the oxidative metabolism of ascorbate.