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H2O2
Kinds of H2O2 Terms modified by H2O2 Selected AbstractsHydrogen Peroxide and Wound Healing: A Theoretical and Practical Review for Hair Transplant SurgeonsDERMATOLOGIC SURGERY, Issue 6 2008SARA WASSERBAUER MD BACKGROUND In most hair restoration practices, hydrogen peroxide has been routinely used to remove blood during and after hair transplant surgery. In other specialties, hydrogen peroxide is also used in these ways: wound cleaning, prevention of infection, hemostasis, and removal of debris. Despite its widespread use, there are still concerns and controversy about the potential toxic effect of hydrogen peroxide. OBJECTIVE The objective was to review all available literature including in vivo and in vitro effects of hydrogen peroxide, as well as general wound healing research. MATERIAL AND METHODS Literature up to and including the past three decades was investigated. RESULTS Two pilot studies were found, and there are not enough data examining the real impact of using hydrogen peroxide in hair transplant surgery. In other specialties, H2O2 appears to have positive effects, such as stimulation of vascular endothelial growth factor, induction of fibroblast proliferation, and collagen, or negative effects, such as cytotoxicity, inhibition of keratinocyte migration, disruption of scarless fetal wound repair, and apoptosis. CONCLUSIONS There are not enough data in hair restoration surgery about the use of hydrogen peroxide, and it is unknown and unclear what the optimum dilution should be. Positive and negative effects were found in other specialties. Further studies are recommended. [source] The developing embryonic cardiac outflow tract is highly sensitive to oxidant stressDEVELOPMENTAL DYNAMICS, Issue 12 2007Steven A. Fisher Abstract This study tested the hypothesis that the remodeling of the cardiac outflow tract (OFT) may represent a developmental window of vulnerability to reactive oxygen species (ROS). Chick embryos were exposed in ovo or ex ovo to increasing concentrations of the stable oxidant hydrogen peroxide (H2O2). As assessed by trypan blue staining, H2O2 induced cell injury in the stage 25,30 OFT at concentrations as low as 1 nM. Higher concentrations were required to induce cell injury in the ventricular and atrial myocardium. Using DCFDA as an indicator of oxidant stress, H2O2 also induced a greater fluorescent signal in the OFT myocardium. H2O2 at these low concentrations also induced Caspase activity, indicative of activation of the pathway of PCD. Interestingly, the induction of Caspase-3 activity was predominately in the OFT cushion mesenchymal cells. Thus, the developing OFT is particularly sensitive to ROS-mediated injury, suggesting that ROS could play a role in the development of congenital defects of the cardiac OFT. Developmental Dynamics 236:3496,3502, 2007. © 2007 Wiley-Liss, Inc. [source] Protective effect of CPUX1, a progesterone, on hydrogen peroxide-induced oxidative damage in PC12 cells,DRUG DEVELOPMENT RESEARCH, Issue 8 2008Bian-sheng Ji Abstract The protective effect of CPUX1, a novel progesterone analog, on hydrogen peroxide (H2O2)-induced oxidative damage was investigated in rat pheochromocytoma (PC12) cells. Following the exposure of PC12 cells to H2O2, there was a reduction in cell survival and activities of superoxide dismutase (SOD) and mitochondrial membrane potential (MMP) accompanied by increased levels of lactate dehydrogenase (LDH) release, malondialdehyde (MDA) production, and intracellular reactive oxygen species (ROS) and intracellular [Ca2+]i levels. Preincubation of cells with CPUX1 prior to H2O2 exposure attenuated all these changes mentioned and had a protective effect against H2O2 -induced toxicity in PC12 cells, indicating that the compound may have potential therapeutic benefit for CNS disorders influenced by oxidative damage. Drug Dev Res 69: 2008 ©2008 Wiley-Liss, Inc. [source] Prussian Blue-Modified Titanate Nanotubes: A Novel Nanostructured Catalyst for Electrochemical Reduction of Hydrogen PeroxideELECTROANALYSIS, Issue 19 2010Damir Ivekovi Abstract Prussian blue (PB) modified titanate nanotubes (PB-TiNT) have been synthesized by the reaction of Fe2+ -modified TiNT with hexacyanoferrate(III) ions. The rate constant for heterogeneous catalytic reaction between PB-TiNT and H2O2 was found to be k=2×104,dm3,mol,1,s,1, which is an order of magnitude higher than the values of k reported for conventionally prepared, electrochemically deposited PB films. On the PB-TiNT modified electrode with subnanomolar surface concentration of PB (,(PB)=2.8×10,11,mol/cm2), a stable, reproducible and linear response towards H2O2 was obtained in the concentration range 0.02,4,mM, with the sensitivity of 0.10,AM,1,cm,2 at ,150,mV. [source] Fabrication of a Sensitive Cholesterol Biosensor Based on Cobalt-oxide Nanostructures Electrodeposited onto Glassy Carbon ElectrodeELECTROANALYSIS, Issue 24 2009Abdollah Salimi Abstract Electrodeposited cobalt oxide (CoOx) nanomaterials are not only used for immobilization of cholesterol oxidase (ChOx) but also as electron transfer mediator for oxidation of H2O2 generated in the enzymatic reaction. Voltammetry and flow injection analysis (FIA) were used for determination of cholesterol. FIA determination of cholesterol with biosensors yielded a calibration curve with the following characteristics: linear range up to 50,,M, sensitivity of 43.5,nA ,M,1 cm,2 and detection limit of 4.2,,M. The apparent Michaelis-Menten constant and the response time of the biosensor are 0.49,mM and 15,s, respectively. This biosensor also exhibits good stability, reproducibility and long life time. [source] Bioelectrochemical Characterization of Horseradish and Soybean PeroxidasesELECTROANALYSIS, Issue 21 2009Marco Frasconi Abstract Heme peroxidase are ubiquitous enzymes catalyzing the oxidation of a broad range of substrates by hydrogen peroxide. In this paper the bioelectrochemical characterization of horseradish peroxidase (HRP) and soybean peroxidase (SBP), belonging to class III of the plant peroxidase superfamily, was studied. The homogeneous reactions between peroxidases and some common redox mediators in the presence of hydrogen peroxide have been carried out by cyclic voltammetry. The electrochemical characterization of the reactions involving enzyme, substrate and mediators concentrations allowed us to calculate the kinetic parameters for the substrate,enzyme reaction (KMS) and for the redox mediator,enzyme reaction (KMM). A full characterization of the direct electron transfer kinetic parameters between the electrode and enzyme active site was also performed by opportunely modeling data obtained from cyclic voltammetry and square wave voltammetry experiments. The experimental data obtained with immobilized peroxidases show enhanced direct electron transfer and excellent electrocatalytical performance for H2O2. Despite the structural similarities and common catalytic cycle, HRP and SBP exhibit differences in their substrate affinity and catalytic efficiency. Basing on our results, it can be concluded that peroxidase from soybean represents an interesting alternative to the classical and largely employed one obtained from horseradish as biorecognition element of electrochemical mediated biosensors. [source] Reagentless Biosensor for Hydrogen Peroxide Based on the Immobilization of Hemoglobin in Platinum Nanoparticles Enhanced Poly(chloromethyl thiirane) Cross-linked Chitosan Hybrid FilmELECTROANALYSIS, Issue 12 2009Shanshan Jia Abstract An unmediated hydrogen peroxide (H2O2) biosensor was prepared by co-immobilizing hemoglobin (Hb) with platinum nanoparticles enhanced poly(chloromethyl thiirane) cross-linked chitosan (CCCS-PNs) hybrid film. CCCS could provide a biocompatible microenvironment for Hb and PNs could accelerate the electron transfer between Hb and the electrode. Spectroscopic analysis indicated that the immobilized Hb could maintain its native structure in the CCCS-PNs hybrid film. Entrapped Hb exhibited direct electrochemistry for its heme Fe(III)/Fe(II) redox couples at ,0.396,V in the CCCS-PNs hybrid film, as well as peroxidase-like activity to the reduction of hydrogen peroxide without the aid of an electron mediator. [source] An Organic Sol-Gel Film as Modifier to Construct BiosensorELECTROANALYSIS, Issue 2 2009Jian-Feng Wu Abstract A new amperometric biosensor for hydrogen peroxide (H2O2) was developed by adsorbing hemoglobin (Hb) on an organic sol-gel film. The organic sol-gel was prepared using resorcinol and formaldehyde as monomers. This sol-gel film shows a biocompatible microenvironment for retaining the native activity of the adsorbed Hb. The direct electron transfer between Hb and electrode is achieved. Hb adsorbed on the film shows an enzyme-like catalytic activity for the reduction of H2O2. The reduction peak currents are proportional linearly to the concentration of hydrogen peroxide in the range of 6×10,8 to 3.6×10,6,M, with a detection limit of 2.4×10,8,M (S/N=3). This research enlarges the applications of organic sol-gel materials in biosensor field. [source] Microflow Vessel Improving Reproducibility and Sensitivity of Electrochemical MeasurementsELECTROANALYSIS, Issue 23 2008Jan Krejci Abstract A new microflow system was designed and developed for electrochemical measurements. The electrochemical electrodes prepared using thick film technology were used in this arrangement. Results of various measurements such as simple amperometric measurement on the example of H2O2 detection, measurement with glucose oxidase (GOx) biosensor, soluble enzyme activity measurement etc. carried out using this system are reported. It was observed that the sensitivity and reproducibility of the electrochemical measurements is improved significantly. The new device performance was proved on H2O2 detection, activity of GOx measurement and heavy metals detection (measured concentration range: H2O2 10,9 to 10,1,M, glucose 10,6 to 10,2,M, activity of GOx 10,1 to 102,IU, heavy metals (Cu, Pb) 10,4 to 10,3,M). The microflow insert greatly reduces the overall size of the electrolyte vessel and measurements with sample quantity as low as 2,mL can be accomplished. [source] Direct Electrochemistry and Electrocatalysis of Hemoglobin in Lipid Film Incorporated with Room-Temperature Ionic LiquidELECTROANALYSIS, Issue 20 2008Gaiping Li Abstract A facile phospholipid/room-temperature ionic liquid (RTIL) composite material based on dimyristoylphosphatidylcholine (DMPC) and 1-butyl-3-methylimidazolium hexafluorophosphate ([bmim]PF6) was exploited as a new matrix for immobilizing protein. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were adopted to characterize this composite film. Hemoglobin (Hb) was chosen as a model protein to investigate the composite system. UV-vis absorbance spectra showed that Hb still maintained its heme crevice integrity in this composite film. By virtue of the Hb/DMPC/[bmim]PF6 composite film-modified glassy carbon electrode (GCE), a pair of well-defined redox peaks of Hb was obtained through the direct electron transfer between protein and underlying GCE. Moreover, the reduction of O2 and H2O2 at the Hb/DMPC/[bmim]PF6 composite film-modified GCE was dramatically enhanced. [source] Direct Electron Transfer and Electrocatalysis of Hemoglobin on Chitosan-TiO2 Nanorods-Glass Carbon ElectrodeELECTROANALYSIS, Issue 20 2008Xiaoling Xiao Abstract The direct electron transfer between hemoglobin (Hb) and the glassy carbon electrode (GC) can be readily achieved via a high biocompatible composite system based on biopolymer chitosan (CHT) and TiO2 nanorods (TiO2 -NRs). TiO2 -NRs greatly promote the electron transfer between Hb and GC, which contribute to the higher redox peaks. UV-vis spectra result indicated the Hb entrapped in the composite film well keep its native structure. The immobilized Hb remains its bioelectrocatalytical activity to the reduction of H2O2 with a lower detection limit. A novel, sensitive, reproducible and stable electrochemical biosensing platform of H2O2 based on Hb-TiO2 -CHT electrode is explored. [source] Multilayer Assembly of Hemoglobin and Colloidal Gold Nanoparticles on Multiwall Carbon Nanotubes/Chitosan Composite for Detecting Hydrogen PeroxideELECTROANALYSIS, Issue 19 2008Shihong Chen Abstract Chitosan (CS) was chosen for dispersing multi-wall carbon nanotubes (MWNTs) to form a stable CS-MWNTs composite, which was first coated on the surface of a glassy carbon electrode to provide a containing amino groups interface for assembling colloidal gold nanoparticles (GNPs), followed by the adsorption of hemoglobin (Hb). Repeating the assembly step of GNPs and Hb resulted in {Hb/GNPs}n multilayers. The assembly of GNPs onto CS-MWNTs composites was confirmed by transmission electron microscopy. The consecutive growth of {Hb/GNPs}n multilayers was confirmed by cyclic voltammetry and UV-vis absorption spectroscopy. The resulting system brings a new platform for electrochemical devices by using the synergistic action of the electrocatalytic activity of GNPs and MWNTs. The resulting biosensor displays an excellent electrocatalytic activity and rapid response for hydrogen peroxide. The linear range for the determination of H2O2 was from 5.0×10,7 to 2.0×10,3 M with a detection limit of 2.1×10,7 M at 3, and a Michaelis,Menten constant KMapp value of 0.19,mM. [source] Dendritic Silver/Silicon Dioxide Nanocomposite Modified Electrodes for Electrochemical Sensing of Hydrogen PeroxideELECTROANALYSIS, Issue 17 2008Peixi Yuan Abstract A novel biosensor for hydrogen peroxide was prepared by immobilizing horseradish peroxidase (HPR) on newly synthesized dendritic silver/silicon dioxide nanocomposites, which were coated on a glassy carbon electrode. The modified electrode was characterized with XPS, SEM, and electrochemical methods. This biosensor showed a very fast amperometric response to hydrogen peroxide with a linear range from 0.7 to 120,,M, a limit of detection of 0.05,,M and a sensitivity of 1.02,mA mM,1 cm,2. The Michaelis-Menten constant of the immobilized HRP was estimated to be 0.21,mM, indicating a high affinity of the HRP to H2O2 without loss of enzymatic activity. The preparation of the proposed biosensor was convenient, and it showed high sensitivity and good stability. [source] Integrating an Enzyme-Entrapped Conducting Polymer Electrode and a Prereactor in a Microfluidic System for Sensing GlucoseELECTROANALYSIS, Issue 6 2008Po-Chin Nien Abstract In this study, the flow injection analysis was applied to the enzyme-entrapped electrode on a chip for sensing glucose. The on-chip microelectrode was fabricated by the standard photolithography in clean-room environment and the microfluidic channel height of 100,,m on the chip was formed by poly(dimethylsiloxane). The conducting polymer, poly(3,4-ethylenedioxythiophene), PEDOT, was electropolymerized to entrap the coexisting glucose oxidase (GOD) by cyclic voltammetry (CV). The amount of enzyme entrapped in the matrix measured spectroscopically was about 0.101,U/cm2. At a flow rate of 10,ml/hr, the working electrode (Pt/PEDOT/GOD, WE1) was set at 0.7,V (vs. Ag/AgCl) and sensing of H2O2 was carried out by injecting samples with various concentrations of glucose (Glu). A linear relationship between the sensing current and the glucose concentration, ranging from 1 to 20,mM, was obtained with a sensitivity of 8,nA mm,2 mM,1. The response time and the recovery time were about 30 and 230,s, respectively. For a single-potential test, the oxidation currents of 0.08,mM ascorbic acid (AA) and a blend of 0.08,mM AA and 10,mM Glu reached 31.3% and 145.5%, respectively, when compared with the oxidation current of 10,mM Glu alone. However, when a pre-reactor (WE2) was set at the same potential (0.7,V) before the main enzyme integrated electrode (WE1), the oxidation current for the above mixed solution reached 99.6% of the original one. [source] Fabrication of Active Horseradish Peroxidase Micropatterns with a High Resolution by Scanning Electrochemical MicroscopyELECTROANALYSIS, Issue 16 2007Xuemei Li Abstract We used a new reactive species OH, to fabricate active horseradish peroxidase (HRP) micropatterns with a high resolution by scanning electrochemical microscopy (SECM) coupled with a carbon fiber disk electrode as the SECM tip. In this method, except for active HRP micropatterns predesigned other regions on a HRP-immobilized substrate were deactivated by OH, generated at the tip held at ,1.7,V in 1.0,mol/L KCl containing 2.0×10,3 mol/L benzoquinone (BQ) (pH,8.0). The feedback mode of SECM with a tip potential of ,0.2,V was used to characterize the active HRP micropatterns in 1.0,mol/L KCl containing 2.0×10,3 mol/L BQ and 2.0×10,3 mol/L H2O2. [source] Biosensor Based on Self-Assembling Glucose Oxidase and Dendrimer-Encapsulated Pt Nanoparticles on Carbon Nanotubes for Glucose DetectionELECTROANALYSIS, Issue 6 2007Lihuan Xu Abstract A novel amperometric glucose biosensor based on layer-by-layer (LbL) electrostatic adsorption of glucose oxidase (GOx) and dendrimer-encapsulated Pt nanoparticles (Pt-DENs) on multiwalled carbon nanotubes (CNTs) was described. Anionic GOx was immobilized on the negatively charged CNTs surface by alternatively assembling a cationic Pt-DENs layer and an anionic GOx layer. Transmission electron microscopy images and ,-potentials proved the formation of layer-by-layer nanostructures on carboxyl-functionalized CNTs. LbL technique provided a favorable microenvironment to keep the bioactivity of GOx and prevent enzyme molecule leakage. The excellent electrocatalytic activity of CNTs and Pt-DENs toward H2O2 and special three-dimensional structure of the enzyme electrode resulted in good characteristics such as a low detection limit of 2.5,,M, a wide linear range of 5,,M,0.65,mM, a short response time (within 5,s), and high sensitivity (30.64,,A mM,1,cm,2) and stability (80% remains after 30 days). [source] Suitability of Stripping Chronopotentiometry for Heavy Metal Speciation Using Hydrogen Peroxide as Oxidant: Application to the Cd(II)-EDTA-PMA SystemELECTROANALYSIS, Issue 24 2005Núria Serrano Abstract The possibilities of stripping chronopotentiometry (SCP) for heavy metal speciation have been tested in the modality of chemical oxidation using the model systems Cd(II)-polyacrylic acid (PMA), Cd(II)-EDTA and Cd(II)-PMA-EDTA. The use of 0.03% H2O2 as a chemical oxidant provides reliable results from transition times, but peak potentials are dramatically affected by the presence of this reagent. The study suggests that chemical-oxidation SCP can be a technique complementary to other stripping modalities in the study of inert and macromolecular labile metal complexes. [source] Electrochemical, Chemical and Enzymatic Oxidations of PhenothiazinesELECTROANALYSIS, Issue 17 2005B. Blankert Abstract The oxidation of several phenothiazine drugs (phenothiazine, promethazine hydrochloride, promazine hydrochloride, trimeprazine hydrochloride and ethopropazine hydrochloride) has been carried out in aqueous acidic media by electrochemical, chemical and enzymatic methods. The chemical oxidation was performed in acetic acid with hydrogen peroxide or in formate buffers using persulfate. The enzymatic oxidation was performed in acetate or ammonium formate buffer by the enzyme horseradish peroxidase in the presence of H2O2. Molecules with, in the lateral chain, two carbon atoms (2C) separating the ring nitrogen and the terminal nitrogen, showed two parallel oxidation pathways, that is (i) formation of the corresponding sulfoxide and (ii) cleavage of the lateral chain with liberation of phenothiazine (PHZ) oxidized products (PHZ sulfoxide and PHZ quinone imine). Molecules with three carbon atoms (3C) separating the two nitrogens were oxidized to the corresponding sulfoxide. The chemical oxidation of all the studied molecules by hydrogen peroxide resulted in the corresponding sulfoxide with no break of the lateral chain. Oxidation by persulfate yielded, for the 3C derivatives, only the corresponding sulfoxide, but it produced cleavage of the lateral chain for the 2C derivatives. The origin of the distinct oxidation pattern between 2C and 3C molecules might be related to steric effects due to the lateral chain. The data are of interest in drug metabolism studies, especially for the early search. In the case of 2C phenothiazines, the results predict the possibility of an in vivo cleavage of the lateral chain with liberation of phenothiazine oxidized products which are known to produce several adverse side effects. [source] Electroconductive Hydrogels: Electrical and Electrochemical Properties of Polypyrrole-Poly(HEMA) CompositesELECTROANALYSIS, Issue 7 2005Sean Brahim Abstract Composites of inherently conductive polypyrrole (PPy) within highly hydrophilic poly(2-hydroxyethyl methacrylate)-based hydrogels (p(HEMA)) have been fabricated and their electrochemical properties investigated. The electrochemical characteristics observed by cyclic voltammetry suggest less facile reduction of PPy within the composite hydrogel compared to electropolymerized PPy, as shown by the shift in the reduction peak potential from ,472,mV for electropolymerized polypyrrole to ,636,mV for the electroconductive composite gel. The network impedance magnitude for the electroconductive hydrogel remains quite low, ca. 100,,, even upon approach to DC, over all frequencies and at all offset potentials suggesting retained electronic (bipolaronic) conductivity within the composite. In contrast, sustained application of +0.7 V (vs. Ag/AgCl, 3,M Cl,) for typically 100,min. (conditioning) to reduce the background amperometric current to <1.0,,A, resulted in complete loss of electroactivity. Nyquist plots suggest that sustained application of such a modest potential to the composite hydrogel results in impedance characteristics that resembles p(HEMA) without evidence of the conducting polymer component. PPy composite gels supported a larger ferrocene monocarboxylate diffusivity (Dappt=7.97×10,5,cm2,s,1) compared to electropolymerized PPy (Dappt=5.56×10,5,cm2,s,1), however a marked reduction in diffusivity (Dappt=1.01×10,5,cm2,s,1) was observed with the conditioned hydrogel composite. Cyclic voltammograms in buffer containing H2O2 showed an absence of redox peaks for electrodes coated with PPy-containing membranes, suggesting possible chemical oxidation of polypyrrole by the oxidant [source] Composite Multienzyme Amperometric Biosensors for an Improved Detection of Phenolic CompoundsELECTROANALYSIS, Issue 22 2003B. Serra Abstract A biosensor design, in which glucose oxidase and peroxidase are coimmobilized by simple physical inclusion into the bulk of graphite-Teflon pellets, is reported for the detection of phenolic compounds. This design allows the "in situ" generation of the H2O2 needed for the enzyme reaction with the phenolic compounds, which avoids several problems detected in the performance of single peroxidase biosensors as a consequence of the presence of a high H2O2 concentration. So, a much lower surface fouling was found at the GOD-HRP biosensor in comparison with a graphite-Teflon-HRP electrode, suggesting that the controlled generation of H2O2 makes more difficult the formation of polymers from the enzyme reaction products. The construction of trienzyme biosensors, in which GOD, HRP and tyrosinase were coimmobilized into the graphite-Teflon matrix is also reported, and their performance was compared with that of GOD-HRP bienzyme electrodes. The practical applicability of the composite multienzyme amperometric biosensors was evaluated by the estimation of the phenolic compounds content in waste waters from a refinery, and the results were compared with those obtained by using a colorimetric official method based on the reaction with 4-aminoantipyrine. [source] Biosensors Based on Aligned Carbon Nanotubes Coated with Inherently Conducting PolymersELECTROANALYSIS, Issue 13 2003Mei Gao Abstract The use of multiwalled aligned carbon nanotubes provides a novel electrode platform for inherently conducting polymer based biosensors. The example used here to highlight the usefulness of such a platform is the polypyrrole based glucose oxidase system for detection of glucose. The use of these three dimensional electrodes offers advantages in that large accessible enzyme loadings can be obtained within an ultrathin layer. It has also been found that the detection of H2O2 at these new electrode structures containing iron loaded nanotube tips can be achieved at low anodic potentials. The result is a sensitive and selective glucose sensor. [source] Poly(methylmethacrylate) and Topas capillary electrophoresis microchip performance with electrochemical detectionELECTROPHORESIS, Issue 16 2005Mario Castaño-Álvarez Abstract A capillary electrophoresis (CE) microchip made of a new and promising polymeric material: Topas (thermoplastic olefin polymer of amorphous structure), a cyclic olefin copolymer with high chemical resistance, has been tested for the first time with analytical purposes, employing an electrochemical detection. A simple end-channel platinum amperometric detector has been designed, checked, and optimized in a poly-(methylmethacrylate) (PMMA) CE microchip. The end-channel design is based on a platinum wire manually aligned at the exit of the separation channel. This is a simple and durable detection in which the working electrode is not pretreated. H2O2 was employed as model analyte to study the performance of the PMMA microchip and the detector. Factors influencing migration and detection processes were examined and optimized. Separation of H2O2 and L -ascorbic acid (AsA) was developed in order to evaluate the efficiency of microchips using different buffer systems. This detection has been checked for the first time with a microchip made of Topas, obtaining a good linear relationship for mixtures of H2O2 and AsA in different buffers. [source] Age-dependent variations of cell response to oxidative stress: Proteomic approach to protein expression and phosphorylationELECTROPHORESIS, Issue 14 2005Yuri Miura Dr. Abstract We investigated the protein profiles of variously aged rat astrocytes in response to oxidative stress. After H2O2 -exposure of cells at 100,µM for 30,min, the relative intensity of ten protein spots changed on two-dimensional (2-D) gels compared with control gels after silver staining. Matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) analysis after in-gel digestion revealed that six of these spots corresponded to three kinds of proteins, each of which was composed of a protein and its modified form with a different isoelectric point (pI). These three proteins were identified as peroxiredoxins (PRDXs) II and III, and calpactin I light chain (p11). H2O2 -exposure increased the intensity of the spot with lower pI and simultaneously decreased that of the spot with higher pI for both PRDXs II and III. In addition, the expression of annexin VII, S -adenosyl- L -homocysteine hydrolase, elongation factor II fragment (EF-II), and adenosine deaminase was increased by H2O2 -exposure in astrocytes from variously aged rats. Using the Pro-Q® Diamond staining, heat shock protein 60,kDa (Hsp 60) and ,-tubulin were observed to be phosphorylated upon H2O2 -exposure. While phosphorylation of ,-tubulin was correlated positively with age, the changes in abundance of ten protein spots as described above were independent of age. These results suggest that aging does not suppress the responses aimed at limiting injury and promoting repair brought about by severe oxidative stress, and might affect cell dynamics including the formation of microtubules. [source] A systematic investigation into the recovery of radioactively labeled proteins from sodium dodecyl sulfate-polyacrylamide gelsELECTROPHORESIS, Issue 1 2004Shaobo Zhou Abstract We report the results of a systematic investigation designed to optimize a method for quantifying radioactivity in proteins in sodium dodecyl sulfate-polyacrylamide gels. The method involves dissolving appropriately sized pieces of gel in hydrogen peroxide and heating to 70°C overnight followed by liquid scintillation counting. H2O2 had no effect on the count rates of [14C]bovine serum albumin (BSA) when counted in a conventional liquid scintillation system, and the count rates remained stable for several days. Temperatures below 70°C resulted in incomplete extraction of radioactivity from gels containing [14C]BSA, but there was also a significant reduction in count rates in samples incubated at 80°C. At 70°C recovery was not affected by the amount of sample loaded onto the gel or by the staining procedure (Coomassie Brilliant Blue or SYPRO Ruby). Recoveries were in the range of 89,94%, and the coefficient of variation for five replicate samples was 5,10%. This method offers a reliable way of measuring the amount of radioactivity in proteins that have been separated by electrophoresis. It may be useful, for example, in quantitative metabolic labeling experiments when it is necessary to know precisely how much tracer has been incorporated into a particular protein. [source] Application of Exchangeable Biochemical Reactors with Oxidase-Catalase-Co-immobilizates and Immobilized Microorganisms in a Microfluidic Chip-CalorimeterENGINEERING IN LIFE SCIENCES (ELECTRONIC), Issue 5 2008M. Leifheit Abstract Several methods for the quantitative detection of different compounds, e.g., L -amino acids, sugars or alcohols in liquid media were developed by application of an automatic measuring unit including a fluid chip-calorimeter FCC-21. For this purpose, enzymes were immobilized covalently on the inner and outer surface of CPG (controlled porous glass)-spherules with an outer diameter of 100,,m and filled into a micro flow-through reaction chamber (VR = 20,,L). The design of the measuring cell allows for easy insertion into the calorimeter device of a stored series of comfortably pre-fabricated measuring cells. These cells can be filled with different enzyme immobilizates. Different oxidases were used and co-immobilized with catalase for the improvement of the detection sensitivity. A signal amplification could be achieved up to a factor of 3.5 with this configuration. ,- D -glucose, ethanol and L -lysine could be detected in a range of 0.25,1.75,mM using glucose oxidase, alcohol oxidase and lysine oxidase. The group of oxidases in combination with the enzymatic catalysis of the intermediate H2O2 allows the quantitative detection of a large number of analytes. A good measurement and storage stability could be achieved for several weeks by this immobilization method. In addition to enzyme-based detection reactions, it was shown that living microorganisms can be immobilized in the reaction chamber. Thus, the system can be used as a whole-cell biosensor. The quantitative detection of phenol in the range of 10,100,,M could be performed using the actinomycete Rhodococcus sp. immobilized on glass beads by means of embedding into polymers. [source] Potential Applications of Oxidoreductases for the Re-oxidation of Leuco Vat or Sulfur Dyes in Textile DyeingENGINEERING IN LIFE SCIENCES (ELECTRONIC), Issue 3 2008F. Xu Abstract Conventional textile dyeing by vat and sulfur dyes includes reduction and re-oxidation steps (with chemical reductants and oxidants), so that the insoluble dyes can be solubilized in the dyeing solution, adsorbed by the fabric, and fixed onto the dyed fabric. The treatments often involve hazardous chemicals, expensive catalysts, or conditions that are suboptimally effective, energy-intensive, caustic, or polluting. Improving these steps with enzyme technology could be of significant interest in terms of better dyeing, handling of hazardous chemicals, disposal of waste, or production economy. The idea of an enzymatic re-oxidation step for vat and sulfur dyeings was tested under simplified laboratory conditions. Selected vat and sulfur dyes, including Vat Blue,43, Vat Orange,7, Vat Green,3, Vat Orange,2, Vat Red,13, Vat Yellow,2, and Sulfur Black,1, were first chemically reduced. The reduced (leuco) dyes were then re-oxidized by aerated buffer solutions or H2O2, in the presence or absence of an oxidoreductase, selected from seven laccases from Myceliophthora thermophila, Scytalidium thermophilum, Coprinus cinereus, Trametes villosa, Rhizoctonia solani, Pycnoporus cinnabarinus, Botrytis cinerea, a bilirubin oxidase from Myrothecium verrucaria, and a heme peroxidase from Coprinus cineresu. It was shown that the enzymes were able to catalyze and accelerate the re-oxidation of the reduced dyes, even when they were adsorbed on cotton fabric, by dissolved air (O2) or H2O2. Small redox-active mediators could facilitate the enzymatic re-oxidation. For Sulfur Black,1, a higher conversion of the leuco dye was achieved with laccase-catalyzed re-oxidation. The further development of this potential enzyme application is discussed. [source] Arsenate and dimethylarsinic acid in drinking water did not affect DNA damage repair in urinary bladder transitional cells or micronuclei in bone marrow,ENVIRONMENTAL AND MOLECULAR MUTAGENESIS, Issue 9 2009Amy Wang Abstract Arsenic is a human skin, lung, and urinary bladder carcinogen, and may act as a cocarcinogen in the skin and urinary bladder. Possible modes of action of arsenic carcinogenesis/cocarcinogenesis include oxidative stress induction and inhibition of DNA damage repair. We investigated the effects of arsenic in drinking water on DNA damage repair in urinary bladder transitional cells and on micronucleus formation in bone marrow. F344 rats were given 100 ppm arsenate [As(V)] or dimethylarsinic acid [DMA(V)] in drinking water for 1 week. The in vivo repair of cyclophosphamide (CP)-induced DNA damage resulting from a single oral gavage of CP, and the in vitro repair of hydrogen peroxide (H2O2)- or formaldehyde-induced DNA damage, resulting from adding H2O2 or formaldehyde into cell medium, were measured by the Comet assay. DMA(V) effects were not observed on either CP-induced DNA damage induction or on DNA repair. Neither DMA(V) nor As(V) increased the H2O2 - or formaldehyde-induced DNA damage, and neither inhibited the repair of H2O2 -induced DNA damage. Neither DMA(V) nor As(V) increased the micronucleus frequency, nor did they elevate micronucleus frequency resulting from CP treatment above the level observed by the treatment with CP alone. These results suggest that arsenic carcinogenesis/cocarcinogenesis in the urinary bladder may not be via DNA damage repair inhibition. To our knowledge this is the first report of arsenic effects on DNA damage repair in the urinary bladder. Environ. Mol. Mutagen. 2009. Published 2009 by Wiley-Liss, Inc. [source] Antioxidant and antimutagenic effects of the crude foliar extract and the alkaloid brachycerine of Psychotria brachycerasENVIRONMENTAL AND MOLECULAR MUTAGENESIS, Issue 9 2007Naíla Cannes do Nascimento Abstract The monoterpene indole alkaloid brachycerine from Psychotria brachyceras has been shown to be induced by UV and to have in vitro antioxidant activity, indicating a possible protective role against the secondary effects of this radiation. In this work, we have studied the antioxidant properties of brachycerine and a crude foliar extract from P. brachyceras by using Saccharomyces cerevisiae strains proficient and deficient in antioxidant defenses. The mutagenic and antimutagenic potential of these substances were assayed in S.cerevisiae N123 strain in the presence and absence of H2O2. In addition, we tested the antioxidant capacity of brachycerine and a crude foliar extract from P. brachyceras on hydroxyl radicals (OH,) using the hypoxanthine/xanthine oxidase assay. The results show that brachycerine and the crude foliar extract of P. brachyceras have antioxidant and antimutagenic effects in yeast and probably this action is mainly due to the scavenging of OH, radicals. Environ. Mol. Mutagen., 2007. © 2007 Wiley-Liss, Inc. [source] DNA damage and repair measurements from cryopreserved lymphocytes without cell culture,A reproducible assay for intervention studiesENVIRONMENTAL AND MOLECULAR MUTAGENESIS, Issue 7 2006Jyh-Lurn Chang Abstract Single-cell gel electrophoresis (the Comet assay) can be used to measure DNA damage and DNA repair capacity (DRC). However, to test DRC of cryopreserved lymphocytes, published methods include steps for cell culturing and phytohemagglutinin stimulation, which may limit use of this assay in intervention studies. We developed a modified Comet assay protocol that allows us to measure DRC from cryopreserved lymphocytes without these in vitro manipulations. Assay reproducibility was evaluated by performing the assay six times on different dates using six aliquots from one blood draw of one individual. The interindividual variation was assessed by performing the assay using one aliquot from six individuals. When ,-irradiation was used as the mutagen, intra-assay coefficients of variation (CVs.) for baseline DNA damage, damage after ,-irradiation exposure, and DRC,measured as tail moment,were 8, 31, and 10%, respectively. Interindividual CVs. were higher. When H2O2 was used as the mutagen, intra-assay CVs. for damage measurements were lower for a protocol modification that included damage and repair at 37°C (CVs. ranging from 8 to 35%) than for the more standard 4°C protocol. Analyzing moment arm,the average distance of DNA migration within the tail,yielded similar results. DNA repair was successfully detected in each experiment. Comparing freshly isolated lymphocytes to cryopreserved lymphocytes from the same individuals' blood draw indicated that DRC was highly correlated when determined using moment arm values. This modified protocol extends the use of the Comet assay to measuring DRC in intervention studies (e.g., dietary interventions) in that it assesses cellular response after cryopreservation without cell culture or other extensive manipulation. Environ. Mol. Mutagen., 2006. © 2006 Wiley-Liss, Inc. [source] Micronuclei and chromatid buds are the result of related genotoxic eventsENVIRONMENTAL AND MOLECULAR MUTAGENESIS, Issue 1 2001Luis Serrano-García Abstract Chromatin buds (CHB), broken eggs, or budding cell nuclei are structures similar to micronuclei (MN) in shape, structure, and size, which are linked to the main nuclei of cells by a thread or stalks of chromatin. They have been observed in numerous cell types and there are reports of their existence relating them with MN or with genotoxic events. However, there is no systematic study reporting their frequency and no experiment has been done to ascertain whether they are really induced by genotoxins. Furthermore, they have been discarded as genotoxic events with the argument that they are not formed in dividing cells. Studies are presented here that indicate that CHB can be considered as genotoxic events and that their origin is comparable to that of MN. Bromodeoxyuridine (BrdU) was used to label proliferating lymphocytes, which were later identified by means of an immunohistochemical method, using the H2O2,DAB stain. The results show that CHB are consistently formed where MN are seen. CHB were induced by the clastogen mitomycin C (MMC) as well as by the aneuploidogen colcemid, with frequencies similar to MN in both cases, and to multinucleated cells in the case of colcemid. CHB occur in lymphocytes of smokers with frequencies similar to those of MN, and we found that the infection with Taenia solium metacestodes induced a comparable increase of both MN and CHB frequency in lymphocytes from pigs. Environ. Mol. Mutagen. 38:38,45, 2001 © 2001 Wiley-Liss, Inc. [source] | |||||||||||||||||||||||||||||||||||||||||||