Redox Changes Research Articles

Multifaceted Proteome Analysis at Solubility, Redox, and Expression Dimensions for Target Identification.

Multifaceted interrogation of the proteome deepens the system-wide understanding of biological systems; however, mapping the redox changes in the proteome has so far been significantly more challenging than expression and solubility/stability analyses. Here, the first high-throughput redox proteomics approach integrated with expression analysis (REX) is devised and combined with the Proteome Integral Solubility Alteration (PISA) assay. The whole PISA-REX experiment with up to four biological replicates can be multiplexed into a single tandem mass tag TMTpro set. For benchmarking this compact tool, HCT116 cells treated with auranofin are analyzed, showing great improvement compared with previous studies. PISA-REX is then applied to study proteome remodeling upon stimulation of human monocytes by interferon α (IFN-α). Applying this tool to study the proteome changes in plasmacytoid dendritic cells (pDCs) isolated from wild-type versus Ncf1-mutant mice treated with interferon α, shows that NCF1 deficiency enhances the STAT1 pathway and modulates the expression, solubility, and redox state of interferon-induced proteins. Providing comprehensive multifaceted information on the proteome, the compact PISA-REX has the potential to become an industry standard in proteomics and to open new windows into the biology of health and disease.

In vivo redox imaging of plasma-induced skin-inflammation in mice

Cold atmospheric plasma (CAP) generates reactive oxygen species (ROS) which induce biological effects on living cells. CAP has potential applications in medicine, but its highly reactive nature can lead to adverse skin complications. A noninvasive technique to examine redox changes in skin is needed for monitoring the treatment process. This study was conducted to develop a skin-inflammation model triggered by CAP-derived ROS and to monitor its progression noninvasively by in vivo dynamic nuclear polarization-MRI (DNP-MRI). The model was successfully developed by exposing the skin to both direct and remote modes of CAP. In vivo DNP-MRI imaging revealed faster reduction rates of TEMPOL in plasma-irradiated skin-inflammation areas, particularly in the remote mode plasma-irradiated skin. MRI revealed high-intensity areas in both the superficial and deep layers of the plasma-irradiated skin. The study highlights the potential importance of DNP-MRI in imaging skin-inflammation models and could improve the use of CAP in medical treatments.

Abstract Tu138: Tandem Mass Tagging (TMT)-based Identification of Proteome Signatures for Ischemia-Reperfusion Injury in Swine

Background: The composition of proteins in blood plasma mirrors the physical changes occurring in the course of disease pathogenesis. Currently available clinical markers are insufficient to predict the actual damage and the redox changes in response to ischemia. Hence, we identified novel redox markers for ischemia/reperfusion (I/R) in swine using TMT-proteomics. Methodology: We employed TMT-10plex isobaric labeling-based quantitative proteomics to track the plasma proteome in Sus Scorfa pigs subjected to surgical I/R at different time points. Plasma samples were collected before ischemia, at 1-hr after ischemia (baseline), and 4-hr, 24-hrs, and 7 days following reperfusion. Results: An average of 4853 spectra and a total of 457 proteins were identified, ensuring a 1% false discovery rate for peptides and proteins with at least two distinct peptides. Pre-ischemia plasma samples served as the control group in all the analyses. TMT analysis revealed temporal changes in the plasma proteome with a prominent shift at 4 hours after I/R insult. To investigate the significantly changed proteins (>1.5 fold), a temporal analysis was carried out. Notably, endopin-1, a serpin protein family member, and fibronectin B were detectable in the plasma following 4 hours of ischemia-reperfusion, indicating the release of these proteins in the circulation during the ischemia-reperfusion. A positive correlation was observed between the redox proteins (i.e., catalase and glutathione peroxidase) with the serpin proteins in the plasma. Biochemical analysis of circulating glutathione, a small-molecular antioxidant, revealed a reductive redox after 60 minutes of Ischemia (hypoxic condition) but this was shifted to an oxidative milieu at 4hrs following reperfusion (hyperoxia) in the adult pigs subjected to I/R insult. Interestingly, we noticed a significant increase in GSH levels and a decrease in catalase levels at 24 hours following I/R, suggesting a compensatory biochemical response in association with repair after I/R insult/injury in the heart. These results correlated with the increased circulating LDH levels indicating the rate of tissue damage following I/R. Conclusion: The present study reveals the presence of a previously unexplored Serpin protein member, endopin-1, during I/R. Understanding the kinetics of endopin-1 expression along with the shift in redox state can help unravel its role in the progression and resolution phases of I/R injury.

Redox changes in the Iapetus Ocean during the Late Ordovician extinction crises

The cause of the Late Ordovician mass extinction (LOME) is widely debated, with glaciation, volcanism and oceanic redox fluctuations being proposed as possible drivers. Here, we apply a multi-proxy approach to deep-water Iapetus Ocean samples from Dob's Linn, Scotland, to determine oceanic redox conditions and changes in chemical weathering intensity. We document major redox fluctuations between anoxic ferruginous and oxic conditions during the first, end-Katian extinction pulse, whereas the end-Hirnantian extinction phase witnessed more persistent anoxia. These two episodes were separated by oxic conditions and a major, short-lived decline in chemical weathering during the Hirnantian glaciation, suggesting that although global cooling may have placed stress on certain biota, it was unlikely to be the cause of the extinction crisis. Late Hirnantian anoxia persisted into the Silurian, with widespread euxinia resulting in global drawdown of redox-sensitive trace metals. Recent studies have identified a mid-Katian biotic crisis and recovery prior to the LOME, although the precise stratigraphic position is not yet defined. The mid-Katian record at Dob's Linn shows a major redox change, with dysoxic to anoxic ferruginous deep ocean waters giving way to well-oxygenated conditions at this time. However, links between the mid-Katian biotic crisis and these redox changes remain unclear.

The impact of chemical pollution across major life transitions: a meta-analysis on oxidative stress in amphibians.

Among human actions threatening biodiversity, the release of anthropogenic chemical pollutants which have become ubiquitous in the environment, is a major concern. Chemical pollution can induce damage to macromolecules by causing the overproduction of reactive oxygen species, affecting the redox balance of animals. In species undergoing metamorphosis (i.e. the vast majority of the extant animal species), antioxidant responses to chemical pollution may differ between pre- and post-metamorphic stages. Here, we meta-analysed (N = 104 studies, k = 2283 estimates) the impact of chemical pollution on redox balance across the three major amphibian life stages (embryo, tadpole, adult). Before metamorphosis, embryos did not experience any redox change while tadpoles activate their antioxidant pathways and do not show increased oxidative damage from pollutants. Tadpoles may have evolved stronger defences against pollutants to reach post-metamorphic life stages. In contrast, post-metamorphic individuals show only weak antioxidant responses and marked oxidative damage in lipids. The type of pollutant (i.e. organic versus inorganic) has contrasting effects across amphibian life stages. Our findings show a divergent evolution of the redox balance in response to pollutants across life transitions of metamorphosing amphibians, most probably a consequence of differences in the ecological and developmental processes of each life stage.

Dolostone-barite-phosphorite sequence in the basal Doushantuo Formation: Origin and implications for post-Marinoan ocean chemistry and phosphogenesis

Barite (BaSO4) represents a major component of many post-Marinoan cap dolostones worldwide, recording important information on climate upheavals during the terminal Neoproterozoic. However, its origins and relations with other lithofacies remain to be explored. Here we report new occurrences of barite in the basal Ediacaran Doushantuo Formation at a major phosphate mine in Weng'an, South China, and present results of detailed petrographic and geochemical investigations on the barites and related phases. At the outcrop scale, barite is mainly present as a 1–2 cm-thick layer at the contact of the cap dolostones and overlying phosphorites. Detailed petrographic observations reveal a lithofacies transition from dolostone through a mixture of barite, dolomite and apatite to barite and finally to granular phosphorite. The barites can be categorized into three generations in stratigraphic order, which are present as loose, randomly-oriented blades (B1), dense aggregates (B2), and radially aligned crystal fans (B3), respectively. B1 is dispersed in a matrix of apatite and dolomite, which has an average δ13Ccarb of −0.61‰. This, along with the common presence of organic matter (OM) in this barite, point to a possible source of Ba from organic degradation. B3 forms a centimeter-scale layer of barite fans that laterally extends over the entire mining pit, whereas B2 is locally distributed between B1 and B3, and occurs as relatively unoriented aggregates. δ34S values of B3 show a narrow range between +25.9‰ and +30.9‰, which is similar to that of contemporaneous seawater and indicate a seawater source of sulfate in barite. Based on the present investigation and a compilation of published data, we propose that B1 is diagenetic barite with a main source of Ba from degradation of organic matter and possibly also from dissolution of previous pelagic barite, whereas B2 and B3 represent seafloor precipitates that resulted from interactions between deep, anoxic Ba-rich waters and oxic, sulfate-rich surface waters during the post-glacial transgression. This episode of barite precipitation was immediately followed by massive accumulations of phosphorites in Weng'an and many areas of South China. The consecutive deposition of cap dolostone, barite and phosphorite signifies the transition from an alkalinity-rich, stagnant anoxic ocean to a dynamic marine regime featuring oxygenated surface waters and upwelling currents. These pH and redox changes likely facilitated the cycling and enrichment of phosphate in the shallow waters that was necessitated by the unprecedented accumulation of Ediacaran phosphorites in South China.

Enrichment pattern of tungsten in sediments under methane seepage environments: Applicability as a proxy for tracing and reconstructing (paleo-)methane seepage

The release of hydrogen sulfide by sulfate-driven anaerobic oxidation of methane (SD-AOM) during methane seepage activities has a strong impact on the evolution of marine redox conditions and the cycles of redox-sensitive elements (e.g. Mo and U). Consequently, the sedimentary enrichment or depletion of these elements is often used to trace and reconstruct (paleo-)methane seepage activity. In recent years, tungsten (W) has shown promise as an indicator of marine redox changes, but its applicability to trace methane seepage activity is unknown. This study identified significant differences in the W content of sediments in methane seepage environments compared to suboxic environments not influenced by methane seepage and oxic environments in the South China Sea. The sediments subjected to methane seepage were characterized by low W contents, similar to those of euxinic Black Sea sapropels. It is believed that the sulfidic environment resulting from SD-AOM is the primary cause of W depletion in sediments subjected to methane seepage. By examining W's interaction with its twin element, Mo, we present WMo enrichment patterns in various environments. In methane seepage environment (sulfidic environments), sediments had high MoEF (> 10) and low WEF (< 2) with a high Mo/W molar ratio (> 10); in suboxic environments not influenced by methane seepage, sediments exhibited a WMo enrichment pattern similar to the UCC (the MoEF and WEF values are predominantly below 1 and 2, respectively, with a low Mo/W molar ratio, <1.5); in oxic environments (enrichment of FeMn (oxy)hydroxides), sediments exhibited high MoEF (2 < MoEF < 10) and high WEF (> 2), with a medium Mo/W molar ratio (1.5–10). This further indicates that the Mo/W molar ratio has great potential in tracing and reconstructing (paleo-)methane seepage activities.

First lacustrine application of the diatom‐bound nitrogen isotope paleo‐proxy reveals coupling of denitrification and N2 fixation in a hyper‐eutrophic lake

AbstractPast changes in the input/output, and internal cycling, of bioavailable nitrogen (N) in marine and lacustrine environments can be reconstructed by analyzing the N isotopic composition (δ15N) of organic matter in the sedimentary record. To verify, and eliminate, potential biases of bulk sedimentary δ15N (δ15Nbulk) signatures by diagenetic alteration and external N inputs, we applied, for the first time, the diatom‐bound N isotope (δ15Ndb) paleo‐proxy to lake sediments. By comparing δ15Nbulk and δ15Ndb in a sedimentary record from eutrophic Lake Lugano (Switzerland), we demonstrate that changing redox conditions influence the degree of N‐isotopic alteration of the bulk sediment, emphasizing the need for caution when interpreting δ15Nbulk in paleolimnological studies. Furthermore, in combining δ15Ndb measurements with X‐ray fluorescence scanning and state‐of‐the‐art molecular biomarker analyses, we reconstruct nutrient cycling and paleoenvironmental conditions in the lake over the past ~ 125 yr. Coeval with the period of severe eutrophication in Lake Lugano in the 1960s, our proxy data indicate that export production, δ15Ndb, and the concentration of heterocyst glycolipids (a biomarker for N2‐fixing cyanobacteria) increased simultaneously. Together, these data suggest that the rise in δ15Ndb is likely the result of enhanced water‐column denitrification in response to increased phytoplankton productivity. We hypothesize that greater export production during eutrophication led to anoxic conditions in the hypolimnion as a result of enhanced organic matter remineralization, raising water‐column denitrification. Enhanced N loss and remobilization of phosphorous (P) from the sediments under anoxic conditions lowered the N : P ratio in the lake, fostering cyanobacterial N2 fixation in surface waters.

A Redox-Shifted Fibroblast Subpopulation Emerges in the Fibrotic Lung.

Idiopathic pulmonary fibrosis (IPF) is an aggressive and thus far incurable disease, characterized by aberrant fibroblast-mediated extracellular matrix deposition. Our understanding of the disease etiology is incomplete; however, there is consensus that a reduction-oxidation (redox) imbalance plays a role. In this study we use the autofluorescent properties of two redox molecules, NAD(P)H and FAD, to quantify changes in their relative abundance in living lung tissue of mice with experimental lung fibrosis, and in freshly isolated cells from mouse lungs and humans with IPF. Our results identify cell population-specific intracellular redox changes in the lungs in experimental and human fibrosis. We focus particularly on redox changes within collagen producing cells, where we identified a bimodal distribution of NAD(P)H concentrations, establishing NAD(P)Hhigh and NAD(P)Hlow sub-populations. NAD(P)Hhigh fibroblasts exhibited elevated pro-fibrotic gene expression and decreased collagenolytic protease activity relative to NAD(P)Hlow fibroblasts. The NAD(P)Hhigh population was present in healthy lungs but expanded with time after bleomycin injury suggesting a potential role in fibrosis progression. We identified a similar increased abundance of NAD(P)Hhigh cells in freshly dissociated lungs of subjects with IPF relative to controls, and similar reductions in collagenolytic activity in this cell population. These data highlight the complexity of redox state changes in experimental and human pulmonary fibrosis and the need for selective approaches to restore redox imbalances in the fibrotic lung.

Fluid-mediated Cu and Zn isotope fractionation in subduction zones and implications for arc volcanism: Constraints from high pressure veins within eclogites in the Dabie Orogen

High-pressure (HP) veins in eclogites are the products of fluid-rock interaction and provide insight into the composition and evolution of fluids in subduction zones. We here present the Cu and Zn isotope data for different types of HP veins and their host eclogites from Ganghe and Hualiangting in the Dabie Orogen to reveal the behavior of Cu and Zn isotopes during fluid-rock interaction and fluid evolution. The HP veins include omphacite-epidote (Omp-Ep), epidote-quartz (Ep-Qtz), and kyanite-epidote-quartz (Ky-Ep-Qtz) veins. The Omp-Ep veins first crystallized from eclogite-derived, solute-rich fluids with the Ep-Qtz and Ky-Ep-Qtz veins successively crystallizing from the residual fluids after the Omp-Ep vein formation. The early Omp-Ep veins have variably lower δ65Cu (−1.32 to −1.12‰ at Ganghe, −0.70 to −0.66‰ at Hualiangting) but higher δ66Zn (0.36 to 0.38‰ at Ganghe, 0.40 to 0.41‰ at Hualiangting) relative to the host eclogites (δ65Cu: −0.71 vs. 1.84‰, δ66Zn: 0.22 vs. 0.33‰), indicating CuZn isotope fractionation during slab dehydration in subduction zones with the lighter Cu and heavier Zn isotopes preferentially entering the vein-forming fluids from the eclogites. Systematic increase of δ65Cu from the Omp-Ep (−0.70 to −0.66‰) through Ep-Qtz (0.01‰) to Ky-Ep-Qtz veins (0.46 to 0.95‰) can be attributed to isotope fractionation induced by redox changes during the evolution of metamorphic fluids, as revealed by the negative correlations of δ65Cu with redox-sensitive ratios of Fe3+/ΣFe and (Eu/Eu*)N in those veins. Additionally, the higher δ66Zn of the Omp-Ep veins relative to the Ky-Ep-Qtz veins can be explained by equilibrium isotope fractionation between crystallized minerals and evolved metamorphic fluids. Our results thus demonstrate that CuZn isotope fractionation occurred during the evolution of slab-derived metamorphic fluids in subduction zones. Binary mixing calculations show that fluids derived from dehydration of mafic rocks in subducted slabs, represented by the multistage HP veins in the present study, cannot account for the heavier Cu and lighter Zn isotope compositions in most arc magmas than in mid-ocean ridge basalts. This offset can be resolved by addition of forearc serpentinite-derived fluids enriched in heavy Cu and light Zn isotopes into the mantle source of arc magmas.

Probing ligands to reaction centers to limit the photocycle in photosynthetic bacterium Rubrivivax gelatinosus

Light-induced electron flow between reaction center and cytochrome bc1 complexes is mediated by quinones and electron donors in purple photosynthetic bacteria. Upon high-intensity excitation, the contribution of the cytochrome bc1 complex is limited kinetically and the electron supply should be provided by the pool of reduced electron donors. The kinetic limitation of electron shuttle between reaction center and cytochrome bc1 complex and its consequences on the photocycle were studied by tracking the redox changes of the primary electron donor (BChl dimer) via absorption change and the opening of the closed reaction center via relaxation of the bacteriochlorophyll fluorescence in intact cells of wild type and pufC mutant strains of Rubrivivax gelatinosus. The results were simulated by a minimum model of reversible binding of different ligands (internal and external electron donors and inhibitors) to donor and acceptor sides of the reaction center. The calculated binding and kinetic parameters revealed that control of the rate of the photocycle is primarily due to 1) the light intensity, 2) the size and redox state of the donor pool, and 3) the unbinding rates of the oxidized donor and inhibitor from the reaction center. The similar kinetics of strains WT and pufC lacking the tetraheme cytochrome subunit attached to the reaction center raise the issue of the physiological importance of this subunit discussed from different points of view. SignificanceA crucial factor for the efficacy of electron donors in photosynthetic photocycle is not just the substantial size of the pool and large binding affinity (small dissociation constant KD = koff/kon) to the RC, but also the mean residence time (koff)−1 in the binding pocket. This is an important parameter that regulates the time of re-activation of the RC during multiple turnovers. The determination of koff has proven challenging and was performed by simulation of widespread experimental data on the kinetics of P+ and relaxation of fluorescence. This work is a step towards better understanding the complex pathways of electron transfer in proteins and simulation-based design of more effective electron transfer components in natural and artificial systems.

Shallow-water redox changes and nitrogen cycles in the early Cambrian Nanhua Basin, South China

Widespread oceanic anoxia has been frequently invoked for metazoan extinctions during the Cambrian. However, this hypothesis remains to be debated. In this study, we measured total organic carbon (TOC), total bulk nitrogen (TNbulk), iron speciation, major and trace elements, and isotopic compositions of sedimentary bulk nitrogen (δ15Nbulk) for the Shiyantou Formation in the Meishucun section. Our results suggest ferruginous conditions and high productivity in the lower member (LM: 0–16.0 m), dominantly suboxic conditions and low productivity in the middle member (MM: 16.0–23.7 m), and oxic conditions and low productivity in the upper member (UM: 23.7–28.0 m). Low δ15Nbulk values (average 1.0 ‰) in the LM suggest quantitative assimilation of NH4+ and strong nitrogen fixation as a consequence of high primary productivity and widespread water-column reducing conditions in the Nanhua Basin. δ15Nbulk values close to 0 ‰ in the MM suggest intense nitrogen fixation owing to quantitative denitrification and anammox in an oligotrophic environment. Low δ15Nbulk values (average 1.2 ‰) in the UM are indicative of marine nitrogen limitations in an oligotrophic environment. These findings suggest a similar range of low δ15Nbulk values might result from different environments. In comparison with marine redox conditions and δ15Nbulk data from the other coeval sections in the Nanhua Basin, we propose that widespread anoxic conditions might cause the extinction of small shelly faunas. More importantly, the oligotrophic environment and anoxic conditions might delay the recovery of small shelly faunas in the shallow and deep waters of the Nanhua Basin, respectively.

Memristors Based on Ni(II)‐tetraaza[14]annulene Complexes: Toward an Unconventional Resistive Switching Mechanism

AbstractIn this work, a family of Ni‐based dibenzotetraaza[14]annulene (dtaa) complexes are investigated for their application in memristors (memory resistors). A series of four Ni(II) complexes with different peripheral substituents of the dtaa ligand are successfully synthesized. Based on these compounds, two‐terminal thin‐film devices are fabricated in planar architecture. Four metals with different work functions are tested: Mg, Cu, Ni, and Au. It is demonstrated that ITO|[Ni(Me4dtaa)]|Cu devices show hysteretic behavior and offer stable, robust, and reproducible switching between high‐ and low‐resistive states. An in‐depth spectroscopic characterization of the Ni complex is performed, using radiation from infrared, through visible and ultraviolet, to tender X‐rays. Operando X‐ray fluorescence spectroscopy is used to monitor redox and structural changes upon the polarization of the studied memristor with the external electric field. Density functional theory calculations are used to better understand the electronic structure of the studied material, as well as structural rearrangement after electron injection that may be responsible for the modulation of electric conductivity. Finding a unique case of filamentary‐type resistive switching involving redox reactions of stationary molecules within a molecular solid is postulated. Yet, the formation of these filaments is not related to any significant configurational changes at the atomic scale.

Simultaneous removal of ammonia nitrogen and manganese by anoxic/oxic (A/O) process from electrolytic manganese residue leachate

Ammonia nitrogen and manganese contained in the leachate of electrolytic manganese residue are causing serious damage to the ecological environment. In this study, anaerobic/oxic (A/O) process as a biological method to achieve simultaneous removal of nitrogen and manganese produced during the actual production of manganese. Results indicated that after 137 d of operation, the system achieved 96.73 ± 1.25%, 98.42 ± 0.60%, 96.97 ± 0.19%, and 100% removal of Mn(II), NH4+-N, TN, and chemical oxygen demand (COD), respectively. Extracellular polymeric substances (EPS) secreted by microorganisms was converted to carbon source to promote denitrification. Scanning electron microscopy (SEM) showed that Mn(II) formed manganese crystals in the process of complete reflection with sludge, acting as electron donor to achieve simultaneous removal of nitrogen and manganese, and removal of Mn(II) was found by X-ray photoelectron spectroscopy (XPS) through redox and valence changes. 16S rRNA indicated that Proteobacteria, Deinococcus-Thermus, Bacteroidete, and Firmicutes were involved in the process of removal nitrogen and manganese. The presence of Limnobacter allowed a more stable accumulation of NO2−-N in reactor, thus achieving shortcut nitrification-denitrification, and Flavihumibacter promoted the secretion of EPS. Collectively, the study reveals that the interaction of autotrophic denitrifying bacteria (Chryseolinea), heterotrophic denitrifying bacteria (Fusibacter, Thauera), and manganese-oxidizing bacteria (Pseudaminobacter, Bacillus, Pedomicrobium, Aeromonasr) achieved simultaneously removal of nitrogen and manganese from the reactor shortcut nitrification-denitrification. This study provides a new insight for the treatment of electrolytic manganese residue leachate produced in the process of manganese production.

In planta imaging of pyridine nucleotides using second-generation fluorescent protein biosensors.

Redox changes of pyridine nucleotides in cellular compartments are highly dynamic and their equilibria are under the influence of various reducing and oxidizing reactions. To obtain spatiotemporal data on pyridine nucleotides in living plant cells, typical biochemical approaches require cell destruction. To date, genetically encoded fluorescent biosensors are considered to be the best option to bridge the existing technology gap, as they provide a fast, accurate, and real-time readout. However, the existing pyridine nucleotides genetically encoded fluorescent biosensors are either sensitive to pH change or slow in dissociation rate. Herein, we employed the biosensors which generate readouts that are pH stable for in planta measurement of NADH/NAD+ ratio and NADPH level. We generated transgenic Arabidopsis lines that express these biosensors in plastid stroma and cytosol of whole plants and pollen tubes under the control of CaMV 35S and LAT52 promoters, respectively. These transgenic biosensor lines allow us to monitor real-time dynamic changes in NADH/NAD+ ratio and NADPH level in the plastids and cytosol of various plant tissues, including pollen tubes, root hairs, and mesophyll cells, using a variety of fluorescent instruments. We anticipate that these valuable transgenic lines may allow improvements in plant redox biology studies.

Detection of fucosylated extracellular vesicles miR-4732-5p related to diagnosis of early lung adenocarcinoma by the electrochemical biosensor

Our preliminary investigation has identified the potential of serum fucosylated extracellular vesicles (EVs) miR-4732-5p in the early diagnosis of lung adenocarcinoma (LUAD) by a fucose-captured strategy utilizing lentil lectin (LCA)-magnetic beads and subsequent screening of high throughput sequencing and validation of real-time quantitative polymerase chain reaction (RT-qPCR). Considering the relatively complicated procedure, expensive equipment, and stringent laboratory condition, we have constructed an electrochemical biosensor assay for the detection of miR-4732-5p. miR-4732-5p is extremely low in serum, down to the fM level, so it needs to be detected by highly sensitive electrochemical methods based on the Mg2+-dependent DNAzyme splitting nucleic acid lock (NAL) cycle and hybridization chain reaction (HCR) signal amplification. In this study, signal amplification is achieved through the dual amplification reactions using NAL cycle in combination with HCR. In addition, hybridized DNA strands bind to a large number of methylene blue (MB) molecules to enhance signaling. Based on the above strategy, we further enhance our signal amplification strategies to improve detection sensitivity and accuracy. The implementation of this assay proceeded as follows: initially, miR-4732-5p was combined with NAL, and then Mg2+-dependent DNAzyme splitted NAL to release auxiliary DNA (S1) strands, which were subsequently captured by the immobilized capture probe DNA (C1) strands on the electrode surface. Following this, abundant quantities of DNA1 (H1) and DNA2 (H2) tandems were generated by HCR, and S1 strands then hybridized with the H1 and H2 tandems through base complementary pairing. Finally, MB was bonded to the H1 and H2 tandems through π–π stacking interaction, leading to the generation of a signal current upon the detection of a potential capable of inducing a redox change of MB by the electrode. Furthermore, we evaluated the performance of our developed electrochemical biosensor assay. The results demonstrated that our assay is a reliable approach, characterized by its high sensitivity (with a detection limit of 2.6 × 10−17 M), excellent specificity, good accuracy, reproducibility, and stability. Additionally, it is cost-effective, requires simple operation, and is portable, making it suitable for the detection of serum fucosylated extracellular vesicles miR-4732-5p. Ultimately, this development has the potential to enhance the diagnostic efficiency for patients with early-stage LUAD.

Hydrogen on Colloidal Gold Nanoparticles.

Colloidal gold nanoparticles (AuNPs) have myriad scientific and technological applications, but their fundamental redox chemistry is underexplored. Reported here are titration studies of oxidation and reduction reactions of aqueous AuNP colloids, which show that the AuNPs bind substantial hydrogen (electrons + protons) under mild conditions. The 5 nm AuNPs are reduced to a similar extent with reductants from borohydrides to H2 and are reoxidized back essentially to their original state by oxidants, including O2. The reactions were monitored via surface plasmon resonance (SPR) optical absorption, which was shown to be much more sensitive to surface H than to changes in solution conditions. Reductions with H2 occurred without pH changes, demonstrating that hydrogenation forms surface H rather than releasing H+. Computational studies suggested that an SPR blueshift was expected for H atom addition, while just electron addition likely would have caused a redshift. Titrations consistently showed a maximum redox change of the 5 nm NPs, independent of the reagent, corresponding to 9% of the total gold or ∼30% hydrogen surface coverage (∼370 H per AuNP). Larger AuNPs showed smaller maximum fractional surface coverages. We conclude that H binds to the edge, corner, and defect sites of the AuNPs, which explains the stoichiometric limitation and the size effect. The finding of substantial and stable hydrogen on the AuNP surface under mild reducing conditions has potential implications for various applications of AuNPs in reducing environments, from catalysis to biomedicine. This finding contrasts with the behavior of bulk gold and with the typical electron-focused perspective in this field.

Oxidative Stress in Breast Cancer: A Biochemical Map of Reactive Oxygen Species Production.

This review systematizes information about the metabolic features of breast cancer directly related to oxidative stress. It has been shown those redox changes occur at all levels and affect many regulatory systems in the human body. The features of the biochemical processes occurring in breast cancer are described, ranging from nonspecific, at first glance, and strictly biochemical to hormone-induced reactions, genetic and epigenetic regulation, which allows for a broader and deeper understanding of the principles of oncogenesis, as well as maintaining the viability of cancer cells in the mammary gland. Specific pathways of the activation of oxidative stress have been studied as a response to the overproduction of stress hormones and estrogens, and specific ways to reduce its negative impact have been described. The diversity of participants that trigger redox reactions from different sides is considered more fully: glycolytic activity in breast cancer, and the nature of consumption of amino acids and metals. The role of metals in oxidative stress is discussed in detail. They can act as both co-factors and direct participants in oxidative stress, since they are either a trigger mechanism for lipid peroxidation or capable of activating signaling pathways that affect tumorigenesis. Special attention has been paid to the genetic and epigenetic regulation of breast tumors. A complex cascade of mechanisms of epigenetic regulation is explained, which made it possible to reconsider the existing opinion about the triggers and pathways for launching the oncological process, the survival of cancer cells and their ability to localize.

The paleoredox context of early eukaryotic evolution: insights from the Tonian Mackenzie Mountains Supergroup, Canada.

Tonian (ca. 1000-720 Ma) marine environments are hypothesised to have experienced major redox changes coinciding with the evolution and diversification of multicellular eukaryotes. In particular, the earliest Tonian stratigraphic record features the colonisation of benthic habitats by multicellular macroscopic algae, which would have been powerful ecosystem engineers that contributed to the oxygenation of the oceans and the reorganisation of biogeochemical cycles. However, the paleoredox context of this expansion of macroalgal habitats in Tonian nearshore marine environments remains uncertain due to limited well-preserved fossils and stratigraphy. As such, the interdependent relationship between early complex life and ocean redox state is unclear. An assemblage of macrofossils including the chlorophyte macroalga Archaeochaeta guncho was recently discovered in the lower Mackenzie Mountains Supergroup in Yukon (Canada), which archives marine sedimentation from ca. 950-775 Ma, permitting investigation into environmental evolution coincident with eukaryotic ecosystem evolution and expansion. Here we present multi-proxy geochemical data from the lower Mackenzie Mountains Supergroup to constrain the paleoredox environment within which these large benthic macroalgae thrived. Two transects show evidence for basin-wide anoxic (ferruginous) oceanic conditions (i.e., high FeHR/FeT, low Fepy/FeHR), with muted redox-sensitive trace metal enrichments and possible seasonal variability. However, the weathering of sulfide minerals in the studied samples may obscure geochemical signatures of euxinic conditions. These results suggest that macroalgae colonized shallow environments in an ocean that remained dominantly anoxic with limited evidence for oxygenation until ca. 850 Ma. Collectively, these geochemical results provide novel insights into the environmental conditions surrounding the evolution and expansion of benthic macroalgae and the eventual dominance of oxygenated oceanic conditions required for the later emergence of animals.

Fundamentals of redox regulation in biology.

Oxidation-reduction (redox) reactions are central to the existence of life. Reactive species of oxygen, nitrogen and sulfur mediate redox control of a wide range of essential cellular processes. Yet, excessive levels of oxidants are associated with ageing and many diseases, including cardiological and neurodegenerative diseases, and cancer. Hence, maintaining the fine-tuned steady-state balance of reactive species production and removal is essential. Here, we discuss new insights into the dynamic maintenance of redox homeostasis (that is, redox homeodynamics) and the principles underlying biological redox organization, termed the 'redox code'. We survey how redox changes result in stress responses by hormesis mechanisms, and how the lifelong cumulative exposure to environmental agents, termed the 'exposome', is communicated to cells through redox signals. Better understanding of the molecular and cellular basis of redox biology will guide novel redox medicine approaches aimed at preventing and treating diseases associated with disturbed redox regulation.