Reactive Oxygen Species Species Research Articles

Red blood cell membrane biomimetic nanoprobes for ratiometric imaging of reactive oxygen species level in atherosclerosis

Atherosclerosis (AS) is a major pathological factor to cardiovascular diseases, wherein reactive oxygen species (ROS) serve as the initiating link for the development of AS. However, direct and accurate assessment of ROS levels in AS plaque sites remains challenging. Herein, by embedding a ROS-responsive fluorescent small molecule (HDB with positive response or Cy7 with negative response) and a stable ROS-independent fluorescent dye CH1055 into red blood cell membrane (RBCM)-biomimetic liposomes, two biomimetic nanoprobes (R-Lipo@HDB/CH1055 and R-Lipo@Cy7/CH1055) were developed for indicating the existence of AS plaques and direct in situ ratiometric imaging of ROS at plaque sites. The fused RBCM endows the nanoprobes with attractive immune escape properties and the ability to accumulate at the AS sites. They not only can indicate the presence of plaques by the second near-infrared (NIR-II) fluorescence of CH1055, but also enable the accurate response to ROS levels through the upward or downward ratiometric fluorescence of HDB or Cy7 to CH1055, avoiding the interference of probe accumulation fluctuation or metabolism at the disease site and thus allowing for more accurate imaging compared to traditional monochromatic imaging. Via the specific responses of HDB and Cy7 to different species of ROS, we verified the upregulation of these ROS at the AS site using these two nanoprobes, and demonstrated their potential to assess the therapeutic effect of AS drugs. This study paves a feasible way to construct a ratiometric imaging platform using RBCM biomimetic liposomes to simultaneously encapsulate several functional molecules, achieving the disease diagnosis, pathological analysis and drug effect evaluation.

Construction of Z-scheme NiO/BiOBr heterojunction for facilitating photocatalytic degradation of oxytetracycline and 2-mercaptobenzothiazole

Z-scheme NiO/BiOBr heterojunction was prepared by mechanical ball milling method and applied for the photodegradation removal of oxytetracycline (OTC) and 2-mercaptobenzothiazole (MBT). The Z-scheme electron transport pathway optimizes the separation of photogenerated carriers, thereby significantly increasing the generation capacity of reactive oxygen species (ROS). The photocatalytic degradation experiments demonstrate that the optimized OTC and MBT removal efficiency of NiO/BiOBr (NBOB) composites could reach 72.6% (120 min) and 97.7% (9 min), respectively. Photoluminescence (PL) spectra and PL decay curves indicate that NBOB-2 composites exhibit low photoinduced electron-hole recombination rate and accelerated electron separation efficiency. In addition, ESR spectra confirm that the Z-scheme NiO/BiOBr heterojunction can generate a larger quantity of ROS species. The possible photodegradation pathways and toxicity analysis for the OTC and MTB photodegradation intermediates are elucidated by HPLC-MS measurement and T.E.S.T. application. The manuscript provides a reference for the macro-preparation of bismuth-based photocatalysts with superior performance and high stability.

Electrocatalytic reduction of hydrogen peroxide in solution in the presence of polyacrylic acid and copper ions: Application for gaseous hydrogen peroxide detection

A comprehensive voltammetric investigation into the electrocatalytic electrode mechanism for the reduction of hydrogen peroxide in an aqueous polyacrylic acid solution, in the presence of Cu2+ ions, is presented. The primary objective is to lay the groundwork for the development of a straightforward electrochemical sensor for the detection of gaseous H2O2. Various electrode reaction pathways have been observed depending on the potential applied to the glassy carbon electrode. At potentials approximately −0.400 V vs. Ag/AgCl (3 mol L−1 KCl), an electrochemical reduction of H2O2 occurs. In this context, the Cu2+/Cu+ redox couple, stabilized by polyacrylic acid, acts as a redox mediator, following the EC’ reaction scheme. Additionally, a complex Fenton-like spontaneous redox chemistry takes place between Cu2+/Cu+ and H2O2 within the bulk of the polyacrylic aqueous solution. This results in the generation of a variety of reactive oxygen species (ROS), with the superoxide anion and hydroxyl radical being the most probable intermediates. The electrochemical reduction of ROS species effectively recycles H2O2 near the electrode surface, constituting an electrocatalytic mechanism of the second order. This particular electrocatalytic process is evident as a prominent reduction current tail at potentials more negative than −0.400 V. This, in conjunction with the adsorption and stabilizing influence of polyacrylic acid, facilitates exceptional analytical sensitivity for the detection of gaseous H2O2. The sensor exhibits the capability to detect concentrations of gaseous H2O2 as low as 10−11 mol L−1, accomplished through the utilization of square-wave voltammetry and other advanced voltammetric techniques.

Unravelling the Mechanistic Understanding of Metal Nanoparticle-Induced Reactive Oxygen Species Formation: Insights from a Cu Nanoparticle Study.

Humans can be exposed to engineered and nonintentionally formed metal and metal oxide nanoparticles (Me NPs) in occupational settings, in public transportation areas, or by means of contact with different consumer products. A critical factor in the toxic potency of Me NPs is their ability to induce oxidative stress. It is thus essential to assess the potential reactive oxygen species (ROS) formation properties of Me NPs. A common way to assess the relative extent of ROS formation in vitro is to use fluorescence spectroscopy with the DCFH-DA (2',7'-dichlorofluorescein diacetate) probe, with and without HRP (horseradish peroxidase). However, this method does not provide any information about specific ROS species or reaction mechanisms. This study investigated the possibility of using complementary techniques to obtain more specific information about formed ROS species, both the type and reaction mechanisms. Cu NPs in PBS (phosphate buffered saline) were chosen as a test system to have the simplest (least interference from other components) aqueous solution with a physiologically relevant pH. ROS formation was assessed using fluorescence by means of the DCFH-DA method (information on relative amounts of oxygen radicals without selectivity), the Ghormley's triiodide method using UV-vis spectrophotometry (concentrations of H2O2), and electron paramagnetic resonance with DMPO as the spin-trap agent (information on specific oxygen radicals). This approach elucidates that Cu NPs undergo ROS-generating corrosion reactions, which previously have not been assessed in situ. In the presence of H2O2, and based on the type of oxygen radical formed, it was concluded that released copper participates in Haber-Weiss and/or Fenton reactions rather than in Fenton-like reactions. The new combination of techniques used to determine ROS induced by Me NPs provides a way forward to gain a mechanistic understanding of Me NP-induced ROS formation, which is important for gaining crucial insight into their ability to induce oxidative stress.

Abstract 14665: Cybrid Analysis Delineates Nuclear and Mitochondrial Contributions to Oxidative Stress in Pulmonary Arterial Hypertension

Rationale : Heritable pulmonary arterial hypertension (HPAH) is a serious vasculopathy caused by remodeling of pulmonary arterioles. It is inherited in an autosomal dominant pattern with reduced penetrance, suggesting additional triggers are needed for disease development. Past study shows increased DNA damage and total reactive oxygen species (ROS) in lymphoblastoid cells (LCL) from PAH patients and unaffected relatives, indicating a potential genetic component. Hypothesis: Increased oxidative stress is an independent genetically-determined modifier of PAH susceptibility. Methods: LCL from patients, relatives and unrelated controls were analyzed for total and mitochondrial (mt) ROS and mt function (Seahorse). Untargeted metabolomics were performed by Metabolon. Nuclear vs mt gene contribution was studied using cybrids created by transferring the mitochondria from high (HC) and low ROS LCL (LC) to mt negative 143B Rho0 cells. Results: Metabolomics revealed significant differences between PAH and control LCL in the TCA cycle and glutathione metabolism relating to oxidative stress. HPAH LCL showed significant upregulation of both ROS species compared to controls. LCL from relatives clustered into two groups, one with increased mtROS, the other comparable to controls . Like PAH lung vascular cells, LCL with increased mtROS had reduced spare respiratory capacity (SRC) suggesting dysfunctional mitochondria, but no glycolytic switch. Cybrid analysis revealed that HCs had significantly higher basal respiration (p=0.02), with SRC trending higher compared to LCs, indicating a functional electron transport chain. These cells also had elevated mtROS (p=0.04), suggesting reduced antioxidants. Conclusions: HPAH is a complex disease, and mutation status alone doesn’t determine its susceptibility. Patient LCL recapitulate some of the metabolomic pathways dysregulated in lung vascular cells. Oxidative stress in LCL extends to some unaffected relatives, suggesting this is an independent genetic trait that modifies PAH risk. Our cybrid analysis, novel in PAH highlights the importance of nuclear-mt crosstalk. WGS is underway to identify nuclear variants that could affect the mt, and act as potential modifiers and second hits in PAH pathogenesis.

Magnetically recyclable Cu2+ doped Fe3O4@biochar for in-situ inactivation of Microcystis aeruginosa: Performance and reusability

Harmful cyanobacterial blooms in eutrophic water bodies have frequently occurred worldwide and become a major environmental concern. Therefore, it is imperative to develop a stable and efficient algaecide to solve this issue. In this study, our purpose was to investigate the efficacy and mechanism of a newly developed Cu2+ doped Fe3O4@Biochar magnetic composite (Cu-Mag-BC) in in-situ inactivation of Microcystis aeruginosa (M. aeruginosa). We successfully synthesized the Cu-Mag-BC by coating Cu2+ onto Fe3O4@Biochar. Cu-Mag-BC exhibited superparamagnetic behavior and was uniformly impregnated by Cu2+. Cu-Mag-BC (5 mg/L), rapidly inactivated chlorophyll-a (Chl-a) in M. aeruginosa with low Fe and Cu leaching, during which time the OD264 value and malondialdehyde (MDA) content increased, while the activities of superoxide dismutase (SOD) and catalase (CAT) first increased and then decreased, due to oxidative stress induced by over-generated reactive oxygen species (ROS). Quantitative results showed that ·O2− and ·OH were the main ROS species produced from Cu-Mag-BC. Inactivation efficiency was maintained at approximately 80 % after three consecutive runs and total Chl-a removal efficiency reached 2.84 g/g, indicating good reusability and stability. A possible inactivation mechanism is proposed; amino groups and adipose chain were the primary oxidation sites. Thus, Cu-Mag-BC shows potential as a candidate for simultaneously inactivating harmful cyanobacteria and preventing secondary pollution.

In Vitro Antioxidant and In Vivo Antigenotoxic Features of a Series of 61 Essential Oils and Quantitative Composition-Activity Relationships Modeled through Machine Learning Algorithms.

The antioxidant activity of essential oils (EOs) is an important and frequently studied property, yet it is not sufficiently understood in terms of the contribution of EOs mixtures' constituents and biological properties. In this study, a series of 61 commercial EOs were first evaluated as antioxidants in vitro, following as closely as possible the cellular pathways of reactive oxygen species (ROS) generation. Hence, EOs were assessed for the ability either to chelate metal ions, thus interfering with ROS generation within the respiratory chain, or to neutralize 2,2-diphenyl-1-picrylhydrazyl (DPPH•) and lipid peroxide radicals (LOO•), thereby halting lipid peroxidation, as well as to neutralize 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid cation radicals (ABTS•+) and hydroxyl radicals (OH•), thereby preventing the ROS species from damaging DNA nucleotides. Showing noteworthy potencies to neutralize all of the radicals at the ng/mL level, the active EOs were also characterized as protectors of DNA double strands from damage induced by peroxyl radicals (ROO•), emerging from 2,2'-azobis-2-methyl-propanimidamide (AAPH) as a source, and OH•, indicating some genome protectivity and antigenotoxicity effectiveness in vitro. The chemical compositions of the EOs associated with the obtained activities were then analyzed by means of machine learning (ML) classification algorithms to generate quantitative composition-activity relationships (QCARs) models (models published in the AI4EssOil database available online). The QCARs models enabled us to highlight the key features (EOSs' chemical compounds) for exerting the redox potencies and to define the partial dependencies of the features, viz. percentages in the mixture required to exert a given potency. The ML-based models explained either the positive or negative contribution of the most important chemical components: limonene, linalool, carvacrol, eucalyptol, α-pinene, thymol, caryophyllene, p-cymene, eugenol, and chrysanthone. Finally, the most potent EOs in vitro, Ylang-ylang (Cananga odorata (Lam.)) and Ceylon cinnamon peel (Cinnamomum verum J. Presl), were promptly administered in vivo to evaluate the rescue ability against redox damage caused by CCl4, thereby verifying their antioxidant and antigenotoxic properties either in the liver or in the kidney.

Enhanced Antibacterial Activity at Ag-Cu Nanojunctions: Unveiling the Mechanism with Simple Surfaces of CuNPs-on-Ag Films.

Deposition of CuNPs on silver film gives rise to the formation of active Ag-Cu interfaces leading to dramatic enhancements in antibacterial activity against Escherichia coli. Transmission electron microscopy (TEM) and energy-dispersive X-ray spectroscopy (EDAX) analyses reveal that CuNPs are covered in a thin Cu2O shell, while X-ray photoelectron spectroscopy measurements (XPS) reveal that the Ag film samples contain significant amounts of Ag2O. XPS analyses show that the deposition of CuNPs on Ag films leads to the formation of a photoactive Ag2O-Cu2O heterostructure. Following a Z-scheme mechanism, electrons from the conduction band of Ag2O recombine with photogenerated holes from the valence band of Cu2O. Consequently, electrons at Cu2O's conduction band render Cu reduced and cause reductive activation of surface oxygen species on Cu forming reactive oxygen species (ROS). Interaction between metallic Cu and ROS species leads to the formation of a Cu(OH)2 phase. Both ROS and Cu(OH)2 species have previously been reported to lead to enhanced antibacterial properties. Holes on Ag2O produce a highly oxidized AgO phase, a phase reported to exhibit excellent antibacterial properties. Quantitative analysis of Cu and Ag high-resolution X-ray photoelectron spectroscopy (HR-XPS) spectra directly reveals several-fold increases in these active phases in full agreement with the observed increase in antibacterial activities. This study provides insight and surface design parameters by elucidating the important roles of Ag and Cu's bifunctionality as active antibacterial materials.

Cell cycle and enzymatic activity alterations induced by ROS production in human neuroblastoma cells SH-SY5Y exposed to Fumonisin B1, Ochratoxin A and their combination

The presence of mycotoxins such as Fumonisin B1(FB1) and Ochratoxin A (OTA) in food and feed has become a threat to human and animal health since they can produce several afflictions. Different mechanisms of action by which they exercise their cytotoxic activity have been attributed to them, including the production of reactive oxygen species (ROS). For this reason, a measurement of the production of ROS species, and an evaluation of the intrinsic cell enzymatic antioxidant activity, including glutathione peroxidase (GPx), glutathione transferase (GTS), and catalase (CAT) together with a cytotoxicity and cell cycle assay have been performed in undifferentiated SH-SY5Y cells exposed to FB1, OTA and [FB1 + OTA] after 24 h and 48 h. FB1 and OTA. Monitoring of intracellular ROS production was carried out by the H2-DCFDA probe; while spectrometry analysis of absorbances was used for measuring GPx, GST and CAT activity. Finally, cell proliferation and cell cycle distribution were studied by flow cytometry. When cells were treated with OTA, an increase in GPx and GST activity was observed compared to FB1 and [FB1 + OTA]; conversely, a decrease in CAT activity was observed when cells were exposed to OTA coinciding with the results observed for ROS measurement. Regarding the cell cycle, when cells were exposed to OTA, a decrease in G0/G1 was detected, revealing an arrest of cell division for SH-SY5Y cells at the concentrations studied.

Review of optical reporters of radiation effects in vivo: tools to quantify improvements in radiation delivery technique.

Radiation damage studies are used to optimize radiotherapy treatment techniques. Although biological indicators of damage are the best assays of effect, they are highly variable due to biological heterogeneity. The free radical radiochemistry can be assayed with optical reporters, allowing for high precision titration of techniques. We examine the optical reporters of radiochemistry to highlight those with the best potential for translational use in vivo, as surrogates for biological damage assays, to inform on mechanisms. A survey of the radical chemistry effects from reactive oxygen species (ROS) and oxygen itself was completed to link to DNA or biological damage. Optical reporters of ROS include fluorescent, phosphorescent, and bioluminescent molecules that have a variety of activation pathways, and each was reviewed for its in vivo translation potential. There are molecular reporters of ROS having potential to report within living systems, including derivatives of luminol, 2'7'-dichlorofluorescein diacetate, Amplex Red, and fluorescein. None have unique specificity to singular ROS species. Macromolecular engineered reporters unique to specific ROS are emerging. The ability to directly measure oxygen via reporters, such as Oxyphor and protoporphyrin IX, is an opportunity to quantify the consumption of oxygen during ROS generation, and this translates from in vitro to in vivo use. Emerging techniques, such as ion particle beams, spatial fractionation, and ultra-high dose rate FLASH radiotherapy, provide the motivation for these studies. In vivo optical reporters of radiochemistry are quantitatively useful for comparing radiotherapy techniques, although their use comes at the cost of the unknown connection to the mechanisms of radiobiological damage. Still their lower measurement uncertainty, compared with biological response assay, makes them an invaluable tool. Linkage to DNA damage and biological damage is needed, and measures such as oxygen consumption serve as useful surrogate measures that translate to in vivo use.

Meticulously Designed Carbon Dots as Photo-Triggered RNA-Destroyer for Evoking Pyroptosis.

An ideal photosensitizer for photodynamic therapy should not only possess high reactive oxygen species (ROS) generation efficiency but also maximize utilization of the in situ produced ROS species, where the latter is closely related to its intracellular location. However, rational design of such photosensitizer without tedious conjugation procedures remains a grand challenge. Here, we report the one-pot preparation of carbon dots (CDs)-based photosensitizer from levofloxacin and neutral red featuring both high 1O2 quantum yield (φΔ = 38.85%) and superior RNA selectivity. Moreover, the φΔ value shows a further 40% improvement and reaches 54.33% in response to RNA binding. Owing to these combined attributes, the CDs could exert great damage to the cellular RNA system (termed the RNA-destroyer) under extremely low dosage of light irradiation (15 mW cm-2, 1 min). It induces pyroptotic cell death and causes rapid release of different cytokines that served as molecular markers in photodynamic immunotherapy. This work represents the meticulously designed CDs with high ROS generation and utilization efficiency via good organization of the photosensitive and targeting modularity. Moreover, it is the first CDs-based pyroptosis inducer to the best of our knowledge.

Semiconducting Polymer Nanoporous Thin Films as a Tool to Regulate Intracellular ROS Balance in Endothelial Cells.

The design of soft and nanometer-scale photoelectrodes able to stimulate and promote the intracellular concentration of reactive oxygen species (ROS) is searched for redox medicine applications. In this work, we show semiconducting polymer porous thin films with an enhanced photoelectrochemical generation of ROS in human umbilical vein endothelial cells (HUVECs). To achieve that aim, we synthesized graft copolymers, made of poly(3-hexylthiophene) (P3HT) and degradable poly(lactic acid) (PLA) segments, P3HT-g-PLA. In a second step, the hydrolysis of sacrificial PLA leads to nanometer-scale porous P3HT thin films. The pore sizes in the nm regime (220-1200 nm) were controlled by the copolymer composition and the structural arrangement of the copolymers during the film formation, as determined by atomic force microscopy (AFM) and transmission electron microscopy (TEM). The porous P3HT thin films showed enhanced photofaradaic behavior, generating a higher concentration of ROS in comparison to non-porous P3HT films, as determined by scanning electrochemical microscopy (SECM) measurements. The exogenous ROS production was able to modulate the intracellular ROS concentration in HUVECs at non-toxic levels, thus affecting the physiological functions of cells. Results presented in this work provide an important step forward in the development of new tools for precise, on-demand, and non-invasive modulation of intracellular ROS species and may be potentially extended to many other physiological or pathological cell models.

Surface-bound sulfate radical-dominated degradation of sulfamethoxazole in the CuFeAl-LDH/peroxymonosulfate system: The abundant hydroxyl groups enhancing efficiency mechanism

In this study, a fast and efficient Cu2Fe0.5Al0.5-LDH/PMS water purification system was constructed to degrade sulfamethoxazole (SMX). The optimized Cu2Fe0.5Al0.5-LDH/PMS reaction system degraded 50% SMX for only 0.5 min, and 98.25% SMX for 10 min. The advanced characterization techniques proved that the abundant hydroxyl groups of Cu2Fe0.5Al0.5-LDH can form ≡M−(O)OSO3− complexes with PMS, resulting in reactive oxygen species (ROS) and promoting SMX degradation. Moreover, the electron transfer between the active bimetallic components played a synergistic role in the generation of ROS species. Compared with the electron transfer degradation mechanism, ROS mechanism contributed more to SMX degradation, in which surface-bound SO4− was the dominated active species. The possible degradation pathway and intermediate toxicity of SMX were proposed. This work provided a strategy to fully explore the catalytic degradation mechanism of heterogeneous catalysts in wastewater purification.

Type-I organic photosensitizers with two complementary reactive oxygen species based on donor-acceptor (D-A) molecules

Type-I photosensitizers (PSs) are considered to be a better choice for photodynamic therapy (PDT) because they can efficiently generate reactive oxygen species (ROS) in the hypoxia condition of tumor, showing less oxygen dependence. However, there are relatively few reported highly efficient type-I organic PSs, especially those simultaneously generated two or more species of type-I ROS. Herein, based on electron-acceptor of 9,10-phenanthrenequinone (PQ), a D-A-D structural type-I PS of PPQ constructed from electron-donor of 9-phenyl-carbazole (PCZ) was firstly developed and could simultaneously produce two type-I ROS under white light irradiation. As the generated ROS of hydroxyl radical (•OH) and superoxide anion radical (O2•−) possessed complementary oxidizing ability and radical lifetime, PPQ showed excellent PDT effects for tumor cells under normoxia or hypoxia. The ability of ROS generation and PDT performance of PPQ were better than those of another PQ derivative (TPQ) constructed with common electron-donor triphenylamine (TPA). More importantly, experiments in aqueous solution and vitro both revealed that •OH generated not only from the well-known transformation of O2•− but also direct electron transition from water to triplet excited PS, which demonstrated the oxygen dependence of type-I PSs more clearly. This new type-I PS offers an opportunity to obtain deep insights of the mechanism of type-I PSs, and provides a new strategy for development of type-I PSs.

An efficient reactive oxygen species/reactive nitrogen species generator for dual imaging-guided orthotopic glioblastoma therapy through intrathecal delivery

The treatment of glioblastoma is still an enormous challenge in the clinic, and treatment based on reactive oxygen species (ROS) has exhibited good potential for its therapy. However, the weak toxicity and short lifetime (e.g., 1O2 10−6 s,·OH 10−2 s) as great obstacles of the ROS-based antitumor strategy, as well as the blood-brain barrier (BBB) obstruction of most drugs into the brain, have immensely constrained the therapeutic effect. Herein, we construct a novel theranostic nanoagent (GLIF) based on Gd2(WO4)3:Nd3+ nanoparticles loaded with ploy-L-arginine (PLA), indocyanine green (ICG), as well as lactoferrin (LF) to enable efficient low-dose drug therapy via ROS and reactive nitrogen species (RNS) using intrathecal injection. The PLA and ICG loading not only enhances GLIF’s ability to generate NO, inhibit cell migration and support ROS/RNS to kill tumors, but also produces ONOO- with greater toxicity and longer lifetime (10−2 s) than 1O2. Moreover, intrathecal injection is adopted to bypass the BBB and the modification of LF promotes accumulation in lesions, causing tremendous side effects with long blood circulation. The significant therapeutic effect of GLIF has been proved in vitro and in vivo and confirmed for the first time in glioblastoma. The excellent performances of GLIF as a second near-infrared fluorescence (NIR-II) and magnetic resonance (MR) imaging contrast agent for glioblastoma diagnosis at a lower injection dose also prove the superiority of intrathecal injection over intravenous injection. These findings show that GLIF has great potential for dual-modal imaging and effective therapy of glioblastoma by combining intrathecal injection and the new pathways of ROS/RNS.

Thermally enhanced biodegradation of benzo[a]pyrene and benzene co-contaminated soil: Bioavailability and generation of ROS

In this study, a set of comprehensive experiments were conducted to explore the effects of temperature on the biodegradation, bioavailability, and generation of reactive oxygen species (ROS) by thermally enhanced biodegradation (TEB) under benzene and BaP co-contaminated conditions. The biodegradation rates of benzene increased from 57.4% to 88.7% and 84.9%, and the biodegradation efficiency of BaP was enhanced from 15.8% to 34.6% and 28.6%, when the temperature was raised from the ambient temperature of 15 °C to 45 °C and 30 °C, respectively. In addition, the bioavailability analysis results demonstrated that the water- and butanol-extractable BaP increased with elevated temperatures. High enzymatic activities and PAH-RHDα gene in gram-positive bacteria favored the long-term elevated temperatures (30 and 45 °C) compared to gram-negative bacteria. Moreover, ROS species (O2•− and •OH) generation was detected which were scavenged by the increased superoxide dismutase and catalase activities at elevated temperatures. Soil properties (pH, TOC, moisture, total iron, Fe3+, and Fe2+) were affected by the temperature treatments, revealing that metal-organic-associated reactions occurred during the TEB of benzene–BaP co-contamination. The results concluded that biodegradation of benzene–BaP co-contamination was greatly improved at 45 °C and that microbial activities enhanced the biodegradation under TEB via the increased bioavailability and generation and degradation of ROS.

Abstract 4781: New generation of fluorescent probes for cell-based measurements of caspase activation and mitochondrial superoxide

Abstract Cell health and stress readouts are critical indicators of altered or impaired function in normal and diseased states of cells, and work has been underway to develop improved small molecule sensor dyes compatible with traditional imaging and High Content Analysis (HCA) interrogation of apoptotic and mitochondrial stress pathways. The CellEvent™ Caspase Green dye effectively reports caspase activation, but suffers complications in assay configuration when attempting to multiplex with the Green Fluorescent Protein (GFP), calcein, or other 488 laser line tools in fluorescence microscopy. Here, we describe the testing and functional characterization of a new candidate molecule for measuring apoptosis in living cells. Our sensor is comprised of a fluorogenic reporter dye that is liberated from a DEVD peptide substrate by caspase activation, but operates in the Texas Red, 590nm excitation band, with an emission peak near 610 nm, permitting easy multiplex with GFP or calcein stained neurons in both traditional and HCA microscopy configurations. Similarly, mitochondrial superoxide accompanying cell stress is probed in microscopy with the MitoSOX™ Red Mitochondrial Superoxide Indicator dye, which localizes to mitochondria and reports superoxide generation, ignoring other Reactive Oxygen Species (ROS) and Reactive Nitrogen Species (RNS). This dye has an unusually long Stokes’ shift, requiring specialized microscopy and HCA filters that excite at 405nm, and capture emission at 610nm for specific superoxide detection. This unconventional spectroscopic profile prevents the dye’s use on many imaging platforms and promotes phototoxicity. To this end, our team has produced a dye with the same level of specificity for superoxide that will operate in one of the traditional fluorescence microscopy channels. Our candidate dye, here named MitoSOX™ Green Mitochondrial Superoxide Indicator also localizes to mitochondria of live cells and selectively reports superoxide generation, while ignoring other ROS and RNS species in ex vivo testing. With an Excitation/Emission profile in the GFP/FITC microscopy channel, a series of comparative studies in immortalized and neural cells are shown, highlighting photostability, specificity and signal amplitude from the dye. These reagents are research use only, not for diagnostic purposes Citation Format: Bhaskar S. Mandavilli, Daniel Beacham, Yi Zhen Hu, Jongtae Yang, Aimei Chen. New generation of fluorescent probes for cell-based measurements of caspase activation and mitochondrial superoxide. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4781.

Overview and New Insights into the Metabolic Syndrome: Risk Factors and Emerging Variables in the Development of Type 2 Diabetes and Cerebrocardiovascular Disease.

Metabolic syndrome (MetS) is considered a metabolic disorder that has been steadily increasing globally and seems to parallel the increasing prevalence of obesity. It consists of a cluster of risk factors which traditionally includes obesity and hyperlipidemia, hyperinsulinemia, hypertension, and hyperglycemia. These four core risk factors are associated with insulin resistance (IR) and, importantly, the MetS is known to increase the risk for developing cerebrocardiovascular disease and type 2 diabetes mellitus. The MetS had its early origins in IR and syndrome X. It has undergone numerous name changes, with additional risk factors and variables being added over the years; however, it has remained as the MetS worldwide for the past three decades. This overview continues to add novel insights to the MetS and suggests that leptin resistance with hyperleptinemia, aberrant mitochondrial stress and reactive oxygen species (ROS), impaired folate-mediated one-carbon metabolism with hyperhomocysteinemia, vascular stiffening, microalbuminuria, and visceral adipose tissues extracellular vesicle exosomes be added to the list of associated variables. Notably, the role of a dysfunctional and activated endothelium and deficient nitric oxide bioavailability along with a dysfunctional and attenuated endothelial glycocalyx, vascular inflammation, systemic metainflammation, and the important role of ROS and reactive species interactome are discussed. With new insights and knowledge regarding the MetS comes the possibility of new findings through further research.

Lipid membrane antioxidant/PLA2-cleavage protection by natural biocompounds.

A wide variety of natural bio-compounds (NBCs), ubiquitous in fruits, vegetables and spices have been studied for their therapeutic potential. NBCs have been associated with reduction of the incidence of cancer, cardiovascular, neurological, respiratory, and age-related diseases. Among these compounds, particularly flavonoid and non-flavonoid polyphenols such as quercetin, resveratrol, oleanolic acid and pristimerin have shown antioxidant and anti-inflammatory properties. In recent years however, the growing evidence of NBCs interaction with lipid membranes evoke questions such as how these NBCs interact and protect lipid membranes from specific reactive oxygen species (ROS) and how NBCs can protect from an inflammatory-pathway inducer such as phospholipase A2 (PLA2) enzyme. In this study, we use spectrofluorometry to characterize individual ROS species absorption of several NBCs, as well as systematically study their lipid membrane interaction and antioxidant protection. In addition, using differential scanning calorimetry (DSC), we performed a thermodynamic study to evaluate the PLA2-cleaving kinetics of lipid membranes and the protective effect of NBCs. Our results suggest that signature chemical parameters (e.g., pKa values and polarity) of such molecules provide them unique antioxidant properties and the ability to hinder PLA2-cleavage during lipid membrane interaction.

SOD2 in platelets: with age comes responsibility

SOD2 in platelets: with age comes responsibility