Image- and Fluorescence-Based Test Shows Oxidant-Dependent Damages in Red Blood Cells and Enables Screening of Potential Protective Molecules

An increase of oxygen saturation within blood bags and metabolic dysregulation occur during storage of red blood cells (RBCs). It leads to the gradual exhaustion of RBC antioxidant protective system and, consequently, to a deleterious state of oxidative stress that plays a major role in the apparition of the so-called storage lesions. The present study describes the use of a test (called TSOX) based on fluorescence and label-free morphology readouts to simply and quickly evaluate the oxidant and antioxidant properties of various compounds in controlled conditions. Here, TSOX was applied to RBCs treated with four antioxidants (ascorbic acid, uric acid, trolox and resveratrol) and three oxidants (AAPH, diamide and H₂O₂) at different concentrations. Two complementary readouts were chosen: first, where ROS generation was quantified using DCFH-DA fluorescent probe, and second, based on digital holographic microscopy that measures morphology alterations. All oxidants produced an increase of fluorescence, whereas H₂O₂ did not visibly impact the RBC morphology. Significant protection was observed in three out of four of the added molecules. Of note, resveratrol induced diamond-shape “Tirocytes”. The assay design was selected to be flexible, as well as compatible with high-throughput screening. In future experiments, the TSOX will serve to screen chemical libraries and probe molecules that could be added to the additive solution for RBCs storage.     

Asymmetric Epoxidation of Allylic Alcohols Catalyzed by (≡SiO) x Ta(OEt)3-x (Dialkyl Tartrate Diolate): Influence of the Reaction Conditions

Silica-supported chiral tantalum alkoxides are active catalysts for the asymmetric epoxidation of propenol and trans hex-2-en-1ol. The influence of different parameters on their catalytic performance was followed: the impregnation duration by a tartrate (step in their preparation); the nature of the solvent (CH₂Cl₂, pentane, toluene) and of the oxidant (TBHP, CHP, H₂O₂); poisoning effects by water or t-butanol; the reaction temperature and the substrate concentration.     

Surface and structural characterization of multi-walled carbon nanotubes following different oxidative treatments

Six commonly used wet chemical oxidants (HNO₃, KMnO₄, H₂SO₄/HNO₃, (NH₄)₂S₂O₈, H₂O₂, and O₃) were evaluated in terms of their effects on the surface chemistry and structure of MWCNTs using a combination of analytical techniques. X-ray photoelectron spectroscopy (XPS) and energy dispersive spectroscopy (EDX) were used to characterize the extent of surface oxidation, while chemical derivatization techniques used in conjunction with XPS allowed the concentration of carboxyl, carbonyl, and hydroxyl groups at the surface to be quantified for each MWCNT sample. Our results indicate that the distribution of oxygen-containing functional groups was insensitive to the reaction conditions (e.g., w/w% of oxidant), but was sensitive to the identity of the oxidant. MWCNTs treated with (NH₄)₂S₂O₈, H₂O₂, and O₃ yielded higher concentrations of carbonyl and hydroxyl functional groups, while more aggressive oxidants (e.g., HNO₃, KMnO₄) formed higher fractional concentrations of carboxyl groups. IR spectroscopy was unable to identify oxygen-containing functional groups present on MWCNTs, while Raman spectra highlighted the frequently ambiguous nature of this technique for measuring CNT structural integrity. TEM was able to provide detailed structural information on oxidized MWCNT, including the extent of sidewall damage for different oxidative treatments.     

Infrared study of acetone and nitrogen oxides on Cu-ZSM-5

The oxidation of bulky thioethers has been carried out on Ti-Beta and Ti-MCM-41 catalysts, using H₂O₂ andt-butyl hydroperoxide (TBHP) as oxidants. The intrinsic activity of Ti-Beta was higher than that of Ti-MCM-41 for the oxidation of methyl phenyl sulfide which can penetrate in the pores of Beta, while in the case of the larger isopentyl phenyl sulfide, which diffuses more slowly in Ti-Beta zeolite, Ti-MCM-41 gives a larger activity. Ti-Beta is able to perform better for the more demanding oxidation of sulfoxides to sulfones giving, therefore, higher selectivities to sulfones than Ti-MCM-41. Similar results were obtained when using either H₂O₂ or TBHP as oxidants. However, the sterical effects were enhanced when TBHP was used as oxidant.     

Role of oxidant during phosphoric acid activation of lignocellulosic material

Lignin-based and wood-based activated carbons were prepared by phosphoric acid activation conducted using the oxidant ammonium persulfate ((NH₄)₂S₂O₈) as the activation additive. Results showed that at a (NH₄)₂S₂O₈ dosage of less than 10%, the oxidant increased the activated carbon yield and promoted pore development. However, at a (NH₄)₂S₂O₈ dosage of higher than 10%, an over-oxidation condition was created so that the activated carbon yield was reduced and pore development was negatively affected. It is suggested that oxidants play two roles during phosphoric activation, promotion of phosphate ester formation on the matrix and over-oxidation of carbon.     

Complementary uses of chlorine dioxide and ozone for drinking water treatment

Application of both chlorine dioxide (ClO₂) and ozone (O₃) at a single drinking water treatment plant is rare in the United States but not in other countries. European utilities that use both oxidants commonly add ozone at one or more points prior to filtration and then apply ClO₂ after filtration. In this mode, the oxidant demand is considerably reduced prior to ClO₂ addition, thus reducing the ClO₂ requirement for maintaining a concentration of 0.1 mg/L to 0.2 mg/L in the distribution system. The combined use of ClO₂ and O₃ has merit in many situations, but the way in which the oxidants are sequenced (e.g. pre‐ozonation followed by ClO₂ treatment or vice versa) is critical in terms of finished water quality. The objectives of this paper are: (1) to review the most common water‐treatment goals associated with ClO₂ and O₃, and (2) to discuss the potential consequences and benefits of applying both oxidants at various points in the treatment train.     

Reaction Kinetics of C3H6 Oxidation for Various Reaction Pathways Over Diesel Oxidation Catalysts

Reaction kinetics studies of C₃H₆ oxidation over Pt/Al₂O₃ and Pt/SiO₂ catalysts were characterized using temperature-programmed oxidation with different oxidants: O₂, NO₂ and surface nitrates. Activation energies and conversion performance were compared in order to determine which hydrocarbon oxidation reaction pathway(s) is relevant in diesel exhaust gas aftertreatment applications. NO _x adsorption did not occur on the SiO₂ surface so the reaction between C₃H₆ and NO₂ could be isolated, i.e. no nitrate effect would complicate the analysis and their significance could be decoupled. These results were then compared with Pt/Al₂O₃ where surface nitrates did form upon exposure to NO _x . The onset of C₃H₆ oxidation was observed at a lower temperature with O₂ than with NO₂, but the activation energy was lower with NO₂. This apparent discrepancy is related to the different oxidant concentrations used and the different adsorption pathways. The results indicate that hydrocarbons must be activated first for oxidation to begin, for either NO₂ or O₂. In analyzing the reaction between C₃H₆ and nitrates, the reaction did not occur until NO _x started to desorb from the catalyst at higher temperatures, i.e. when nitrates became unstable and decomposed, thus providing a readily available oxidant source. However, when O₂ was added to the nitrate/C₃H₆ system, the reaction began at even lower temperature than with just C₃H₆ and O₂. Nitrate consumption was also observed once oxidation began. The presence of the combination of nitrates and O₂ resulted in a lower C₃H₆ oxidation activation energy.     

Synthesis, characterization and optimum reaction conditions of oligo-2-amino-3-hydroxypyridine and its Schiff base oligomer

The oxidative polycondensation reaction conditions of 2-amino-3-hydroxypyridine (AHP) and 2-[benzilydeneimino] pyridine-3-ol (BIP) were studied by oxidants such as with air O₂, NaOCl and H₂O₂. Oligo-2-amino-3-hydroxypyridine (OAHP) was synthesized from the oxidative polycondensation of AHP with air O₂, NaOCl and H₂O₂ in an aqueous acidic and alkaline medium at 30-90 °C. BIP was synthesized from condensation of 2-amino-3-hydroxypyridine with benzaldehyde. Oligo-2-[benzilydeneimino] pyridine-3-ol (OBIP) was synthesized from the oxidative polycondensation of BIP with air O₂, NaOCl and H₂O₂ in an aqueous alkaline medium at 40-90 °C. About 95% BIP was converted to OBIP. The number average molecular weight, (M_n) weight average molecular weight (M_w) and polydispersity index (PDI) values of OAHP and OBIP (for air O₂ oxidant) were found to be 1433, 1912 g mol⁻¹, 1.33 and 2637, 5106 g mol⁻¹ and 1.94, respectively. At the optimum reaction conditions, the yield of OAHP was found to be 86.0% (for air O₂ oxidant), 43.0% (for H₂O₂ oxidant) and 85.0% (for NaOCl oxidant). At the optimum reaction conditions, the yield of OBIP was found to be 91.0% (for air O₂ oxidant), 92.0% (for H₂O₂ oxidant) and 95.0% (for NaOCl oxidant). The OHAP and OBIP were characterized by FT-IR, UV-Vis, ¹H and ¹³C-NMR elemental analysis. TG and DTA analyses were shown to be unstable of OAHP and OBIP against thermo-oxidative decomposition. According to TG analyses, the weight loss of OAHP and OBIP was found to be 97.35 and 96.60% at 520 and 685 °C, respectively.     

Fusion with Promiscuous Gα16 Subunit Reveals Signaling Bias at Muscarinic Receptors

A complex evaluation of agonist bias at G-protein coupled receptors at the level of G-protein classes and isoforms including non-preferential ones is essential for advanced agonist screening and drug development. Molecular crosstalk in downstream signaling and a lack of sufficiently sensitive and selective methods to study direct coupling with G-protein of interest complicates this analysis. We performed binding and functional analysis of 11 structurally different agonists on prepared fusion proteins of individual subtypes of muscarinic receptors and non-canonical promiscuous α-subunit of G₁₆ protein to study agonist bias. We have demonstrated that fusion of muscarinic receptors with Gα₁₆ limits access of other competitive Gα subunits to the receptor, and thus enables us to study activation of Gα₁₆ mediated pathway more specifically. Our data demonstrated agonist-specific activation of G₁₆ pathway among individual subtypes of muscarinic receptors and revealed signaling bias of oxotremorine towards Gα₁₆ pathway at the M₂ receptor and at the same time impaired Gα₁₆ signaling of iperoxo at M₅ receptors. Our data have shown that fusion proteins of muscarinic receptors with α-subunit of G-proteins can serve as a suitable tool for studying agonist bias, especially at non-preferential pathways.     

Synthesis,characterization, and thermal degradation of oligo‐2‐(morpholinoiminomethyl)phenol and its Pb(II) complex compound

The oxidative polycondensation reaction conditions of 2‐(morpholinoiminomethyl)phenol were studied with H₂O₂, air O₂, and sodium hypochloride (NaOCl) oxidants in an aqueous alkaline medium between 40 and 90°C. The structure of oligo‐2‐(morpholinoiminomethyl)phenol was characterized with ¹H‐ and ¹³C‐NMR, Fourier transform infrared, ultraviolet–visible, size exclusion chromatography, and elemental analysis techniques. Under the optimum reaction conditions, the yield of oligo‐2‐(morpholinoiminomethyl)phenol was 28% for the H₂O₂ oxidant, 12% for the air O₂ oxidant, and 58% for the NaOCl oxidant. According to the size exclusion chromatography analysis, the number‐average molecular weight, weight‐average molecular weight, and polydispersity index of oligo‐2‐(morpholinoiminomethyl)phenol were 2420 g/mol, 2740 g/mol, and 1.187 with H₂O₂, 1425 g/mol, 2060 g/mol, and 1.446 with air O₂, and 1309 g/mol, 1401 g/mol, and 1.070 with NaOCl, respectively. Thermogravimetry/dynamic thermal analysis showed that the oligo‐2‐(morpholinoiminomethyl)phenol–lead complex compound was more stable than 2‐(morpholinoiminomethyl)phenol and oligo‐2‐(morpholinoiminomethyl)phenol against thermal degradation. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 102:3795–3804, 2006     

Environment Friendly Protocol for Enantioselective Epoxidation of Non-Functionalized Alkenes Catalyzed by Recyclable Homochiral Dimeric Mn(III) Salen Complexes with Hydrogen Peroxide and UHP Adduct as Oxidants

Environment friendly oxidants e.g. H₂O₂ and anhydrous urea hydrogen peroxide adduct (UHP) epoxidize non-functionalized alkenes (epoxide yield 40–99%) with high enantioselectivity (up to 94% ee) in 4–15 h using two recyclable homochiral dimeric Mn(III) salen complexes in presence of PyN-oxide with the catalysts reusability several times when UHP was used as an oxidant.     

Hydrogen production by oxidative methanol reforming with various oxidants over Cu-based catalysts

In order to improve the start-up property of a small hydrogen producer with a micro methanol reformer, oxidative methanol reforming (OMR) with various oxidants over copper-based catalysts was examined. The addition of Fe to a Cu/ZnO/Al₂O₃ catalyst resulted in higher catalyst durability, with a slight improvement in catalytic activity, for OMR with air. However, the addition of larger amounts of Fe inhibited further improvement of catalytic performance, possibly due to the formation of less active CuFe₂O₄ spinel in the catalyst. The production of hydrogen by OMR with hydrogen peroxide as an alternative oxidant, which has the potential to provide concentrated hydrogen without nitrogen dilution, was also considered. It was found that hydrogen peroxide is an effective oxidant for OMR over copper-based catalysts due to its ability to suppress CO formation and its improving effect on methanol conversion.     

Oligomerization and Nitration of the Grass Pollen Allergen Phl p 5 by Ozone,Nitrogen Dioxide,and Peroxynitrite: Reaction Products,Kinetics, and Health Effects

The allergenic and inflammatory potential of proteins can be enhanced by chemical modification upon exposure to atmospheric or physiological oxidants. The molecular mechanisms and kinetics of such modifications, however, have not yet been fully resolved. We investigated the oligomerization and nitration of the grass pollen allergen Phl p 5 by ozone (O₃), nitrogen dioxide (NO₂), and peroxynitrite (ONOO^–). Within several hours of exposure to atmospherically relevant concentration levels of O₃ and NO₂, up to 50% of Phl p 5 were converted into protein oligomers, likely by formation of dityrosine cross-links. Assuming that tyrosine residues are the preferential site of nitration, up to 10% of the 12 tyrosine residues per protein monomer were nitrated. For the reaction with peroxynitrite, the largest oligomer mass fractions (up to 50%) were found for equimolar concentrations of peroxynitrite over tyrosine residues. With excess peroxynitrite, the nitration degrees increased up to 40% whereas the oligomer mass fractions decreased to 20%. Our results suggest that protein oligomerization and nitration are competing processes, which is consistent with a two-step mechanism involving a reactive oxygen intermediate (ROI), as observed for other proteins. The modified proteins can promote pro-inflammatory cellular signaling that may contribute to chronic inflammation and allergies in response to air pollution.     

Comparative Proteomic Analysis of Puccinellia tenuiflora Leaves under Na2CO3 Stress

Soil salt-alkalinization is a widespread environmental stress that limits crop growth and agricultural productivity. The influence of soil alkalization caused by Na₂CO₃ on plants is more severe than that of soil salinization. Plants have evolved some unique mechanisms to cope with alkali stress; however, the plant alkaline-responsive signaling and molecular pathways are still unknown. In the present study, Na₂CO₃ responsive characteristics in leaves from 50-day-old seedlings of halophyte Puccinellia tenuiflora were investigated using physiological and proteomic approaches. Comparative proteomics revealed 43 differentially expressed proteins in P. tenuiflora leaves in response to Na₂CO₃ treatment for seven days. These proteins were mainly involved in photosynthesis, stress and defense, carbohydrate/energy metabolism, protein metabolism, signaling, membrane and transport. By integrating the changes of photosynthesis, ion contents, and stress-related enzyme activities, some unique Na₂CO₃ responsive mechanisms have been discovered in P. tenuiflora. This study provides new molecular information toward improving the alkali tolerance of cereals.     

Proteomic Analysis Identifies an NADPH Oxidase 1 (Nox1)-Mediated Role for Actin-Related Protein 2/3 Complex Subunit 2 (ARPC2) in Promoting Smooth Muscle Cell Migration

A variety of vascular pathologies, including hypertension, restenosis and atherosclerosis, are characterized by vascular smooth muscle cell (VSMC) hypertrophy and migration. NADPH oxidase 1 (Nox1) plays a pivotal role in these phenotypes via distinct downstream signaling. However, the mediators differentiating these distinct phenotypes and their precise role in vascular disease are still not clear. The present study was designed to identify novel targets of VSMC Nox1 signaling using 2D Differential In-Gel Electrophoresis and Mass Spectrometry (2D-DIGE/MS). VSMC treatment with scrambled (Scrmb) or Nox1 siRNA and incubation with the oxidant hydrogen peroxide (H₂O₂; 50 μM, 3 h) followed by 2D-DIGE/MS on cell lysates identified 10 target proteins. Among these proteins, actin-related protein 2/3 complex subunit 2 (ARPC2) with no previous link to Nox isozymes, H₂O₂, or other reactive oxygen species (ROS), was identified and postulated to play an intermediary role in VSMC migration. Western blot confirmed that Nox1 mediates H₂O₂-induced ARPC2 expression in VSMC. Treatment with a p38 MAPK inhibitor (SB203580) resulted in reduced ARPC2 expression in H₂O₂-treated VSMC. Additionally, wound-healing “scratch” assay confirmed that H₂O₂ stimulates VSMC migration via Nox1. Importantly, gene silencing of ARPC2 suppressed H₂O₂-stimulated VSMC migration. These results demonstrate for the first time that Nox1-mediated VSMC migration involves ARPC2 as a downstream signaling target.     

Mathematical Modeling and Genetic Algorithm Optimization of Reactive Absorption of H2S

A rate‐based mathematical model was developed for the reactive absorption of H₂S in NaOH, with NaOCl or H₂O₂ as the chemical oxidant solutions in a packed column. A modified mass transfer coefficient in the gas phase was obtained by genetic algorithm and implemented in the model to correct the assumption of instantaneous reactions. The effects of different operating variables including the inlet H₂S concentration, inlet gas mass flux, initial NaOH, concentrations of the chemical oxidants in the scrubbing solutions, and liquid‐to‐gas ratio on the H₂S removal efficiency were studied. A genetic algorithm was employed to optimize the operating variables in order to obtain maximum removal efficiency of H₂S. The model results were in good agreement with the experimental data.     

Mechanism of GAPDH Redox Signaling by H2O2 Activation of a Two−Cysteine Switch

Oxidation of glyceraldehyde−3−phosphate dehydrogenase (GAPDH) by reactive oxygen species such as H₂O₂ activate pleiotropic signaling pathways is associated with pathophysiological cell fate decisions. Oxidized GAPDH binds chaperone proteins with translocation of the complex to the nucleus and mitochondria initiating autophagy and cellular apoptosis. In this study, we establish the mechanism by which H₂O₂−oxidized GAPDH subunits undergo a subunit conformational rearrangement. H₂O₂ oxidizes both the catalytic cysteine and a vicinal cysteine (four residues downstream) to their respective sulfenic acids. A ‘two−cysteine switch’ is activated, whereby the sulfenic acids irreversibly condense to an intrachain thiosulfinic ester resulting in a major metastable subunit conformational rearrangement. All four subunits of the homotetramer are uniformly and independently oxidized by H₂O₂, and the oxidized homotetramer is stabilized at low temperatures. Over time, subunits unfold forming disulfide−linked aggregates with the catalytic cysteine oxidized to a sulfinic acid, resulting from thiosulfinic ester hydrolysis via the highly reactive thiosulfonic ester intermediate. Molecular Dynamic Simulations provide additional mechanistic insights linking GAPDH subunit oxidation with generating a putative signaling conformer. The low−temperature stability of the H₂O₂−oxidized subunit conformer provides an operable framework to study mechanisms associated with gain−of−function activities of oxidized GAPDH to identify novel targets for the treatment of neurodegenerative diseases.     

Performance of a photo‐impinging streams reactor for the phenol degradation process

BACKGROUND: Photocatalysis is one of the advanced oxidation processes that has gained in importance over recent years owing to its ability to decompose a wide range of organic and inorganic pollutants at ambient temperature and pressure. However, there are two essential issues regarding photocatalytic processes, i.e. limitations on photon transfer and on mass transfer. In the present study, a novel photo‐impinging streams reactor, which can minimize such limitations, has been utilized in the photocatalytic degradation of phenol. The design and operating parameters such as type of nozzle, flow rate, catalyst loading, pH, initial phenol concentration and light intensity were found to have the expected impact on the efficiency of the process. The effects of two different co‐oxidants, H₂O₂ and Na₂S₂O₈ on the photocatalysis were also examined. RESULTS: Results indicated that 100 mg L^?1 of phenol in a 750 cm³ solution was completely degraded within 2.5 h reaction time in the presence of TiO₂ without a co‐oxidant present; and within 1 h in the presence of a co‐oxidant. CONCLUSION: A comparison between the current data and those available in the literature revealed higher efficiency and increased performance of the present reactor relative to conventional apparatus. Copyright © 2010 Society of Chemical Industry     

Complexity of NAC Action as an Antidiabetic Agent: Opposing Effects of Oxidative and Reductive Stress on Insulin Secretion and Insulin Signaling

Dysregulated redox balance is involved in the pathogenesis of type 2 diabetes. While the benefit of antioxidants in neutralizing oxidative stress is well characterized, the potential harm of antioxidant-induced reductive stress is unclear. The aim of this study was to investigate the dose-dependent effects of the antioxidant N-acetylcysteine (NAC) on various tissues involved in the regulation of blood glucose and the mechanisms underlying its functions. H₂O₂ was used as an oxidizing agent in order to compare the outcomes of oxidative and reductive stress on cellular function. Cellular death in pancreatic islets and diminished insulin secretion were facilitated by H₂O₂-induced oxidative stress but not by NAC. On the other hand, myotubes and adipocytes were negatively affected by NAC-induced reductive stress, as demonstrated by the impaired transmission of insulin signaling and glucose transport, as opposed to H₂O₂-stimulatory action. This was accompanied by redox balance alteration and thiol modifications of proteins. The NAC-induced deterioration of insulin signaling was also observed in healthy mice, while both insulin secretion and insulin signaling were improved in diabetic mice. This study establishes the tissue-specific effects of NAC and the importance of the delicate maintenance of redox balance, emphasizing the challenge of implementing antioxidant therapy in the clinic.