Reactive oxygen species responsible for beta-glucan degradation

The presence of iron(II) in beta-glucan in solution causes the formation of hydroxyl radical, which further oxidises the polysaccharide. This degradation can be enhanced by the presence of a reducing agent, such as ascorbic acid. In this study we investigated the effect the iron(II) concentration on the hydroxyl radical-mediated degradation of beta-glucan and identified the intermediate species involved in the formation of hydroxyl radicals. An increase in the iron(II) concentration did not have a significant effect on the degradation in the presence of a reducing agent (ascorbic acid), while in the mere presence of iron(II) it accelerates the degradation. The addition of catalase and superoxide dismutase (SOD) prevented the hydroxyl radical driven-degradation of beta-glucan induced by iron(II) or ascorbic acid/iron(II), demonstrating the involvement of both superoxide and hydrogen peroxide in the hydroxyl radical formation. SOD, which catalyses the dismutation of superoxide into hydrogen peroxide, should have stimulated the formation of radicals, since these radicals are generated from the reaction between hydrogen peroxide and iron(II). In the present study, we hypothesise the mechanism of the inhibition of beta-glucan degradation by superoxide dismutase.     

Visible light-induced degradation of carbon tetrachloride on dye-sensitized TiO2

This study investigated an application of TiO2 photocatalyst sensitized with tris(4,4'-dicarboxy-2,2'-bipyridyl)ruthenium-(II) complex to CCl4 degradation under visible light irradiation. By injecting electrons from the photoexcited sensitizer to the conduction band, the sensitized TiO2 degraded CCl4 under the irradiation of lambda > 420 nm. The quantum yield of CCl4 dechlorination was about 10(-3). The dechlorination rate of CCl4 was reduced in the presence of dissolved O2 due to its competition for conduction band electrons. The photolysis rate was dependent on pH due to the strong pH dependence of the sensitizer adsorption on TiO2 surface with a maximum degradation rate achieved at pH approximately 3. A two-site Langmurian model successfully described the adsorption of the sensitizer on TiO2 particles. The monolayer coverage was achieved at the added sensitizer concentration of 10 microM at [TiO2] = 0.5 g/L. However, the photolysis rate of CCl4 showed a maximum at a sensitizer surface coverage of 0.3 monolayer. Since the photoinduced electron injection gradually depleted active sensitizer molecules on TiO2, sacrificial electron donors to regenerate the sensitizer were sought. 2-Propanol as an electron donor was efficient in the present RuIIL3/TiO2/CCl4 system, which showed no sign of deceleration in the dechlorination rate up to 6 h of irradiation.     

Chemistry and reactions of reactive oxygen species in foods

Reactive oxygen species (ROS) are formed enzymatically, chemically, photochemically, and by irradiation of food. They are also formed by the decomposition and the inter-reactions of ROS. Hydroxy radical is the most reactive ROS, followed by singlet oxygen. Reactions of ROS with food components produce undesirable volatile compounds and carcinogens, destroy essential nutrients, and change the functionalities of proteins, lipids, and carbohydrates. Lipid oxidation by ROS produces low molecular volatile aldehydes, alcohols, and hydrocarbons. ROS causes crosslink or cleavage of proteins and produces low molecular carbonyls from carbohydrates. Vitamins are easily oxidized by ROS, especially singlet oxygen. The singlet oxygen reaction rate was the highest in ss-carotene, followed by tocopherol, riboflavin, vitamin D, and ascorbic acid.     

Fullerol-sensitized production of reactive oxygen species in aqueous solution

The relative production rate of reactive oxygen in aqueous solution sensitized by fullerol (a polyhydroxylated, water-soluble form of the fullerene C60) was measured and compared to known reactive oxygen sensitizers using an oxygen consumption method. The solutions were irradiated by polychromatic visible and ultraviolet light. Reactive oxygen species were generated under both visible and ultraviolet light sources. The greatest rates of oxygen consumption were observed at acidic pH. We show for the first time evidence of both singlet oxygen and superoxide production by fullerol under both UV and polychromatic light sources.     

The role of reactive oxygen species in the electrochemical inactivation of microorganisms

Electrochemical disinfection has emerged as one of the most promising alternatives to the conventional disinfection of water in many applications. Although the mechanism of electrochemical disinfection has been largely attributed to the action of electro-generated active chlorine, the role of other oxidants, such as the reactive oxygen species (ROS) *OH, O3, H2O2, and *O2- remains unclear. In this study, we examined the role of ROS in the electrochemical disinfection using a boron-doped diamond (BDD) electrode in a chloride-free phosphate buffer medium, in order to avoid any confusion caused by the generation of chlorine. To determine which species of ROS plays the major role in the inactivation, the effects of several operating factors, such as the presence of *OH scavenger, pH, temperature, and the initial population of microorganisms, were systematically investigated. This study clearly showed that the *OH is the major lethal species responsible for the E. coli inactivation in the chloride-free electrochemical disinfection process, and that the E. coli inactivation was highly promoted at a lower temperature, which was ascribed to the enhanced generation of O3.     

油茶总皂甙抑菌活性及清除活性氧能力初步研究

用溶剂法提取油茶饼中油茶总皂甙,经柱层析制备标准品标定有效成分含量为82.5%。以油茶皂甙为材料对枯草杆菌、大肠杆菌、金黄色葡萄球菌进行抑菌试验,结果表明均有抑制作用;其中对枯草杆菌和金黄色葡萄球菌抑制作用较强,20%浓度对三种细菌均具有致死作用,抑菌圈直径大小与油茶总皂甙浓度呈正相关;且初步测定油茶总皂甙清除活性氧能力强于维生素C。     

加热蒜氨酸形成活性硫产物的鉴定

为研究加热蒜氨酸后所形成的活性硫(RSS)化合物,在363.15 K温度下加热纯蒜氨酸溶液后,用液相色谱-质谱联用法(LC-MS)、串联质谱法(MS/MS)和液相色谱-高分辨质谱法(UPLC-HRMS)联用鉴定所形成的RSS产物。结果表明,蒜氨酸受热分解、转化可产生烯丙基丙氨酸基二硫烷、三硫烷和四硫烷、双丙氨酸基二硫烷、三硫烷和四硫烷等RSS化合物;其中烯丙基丙氨酸基四硫烷应是新发现的有机含硫化合物。提出了蒜氨酸加热形成RSS和生成S-烯丙基-L-半胱氨酸的新途径。结果表明,厌氧、加热蒜氨酸时,可形成多种RSS化合物;RSS化合物有助于揭示黑蒜、蒜泥、腌制蒜等大蒜加工品的生物活性与保健作用。     

Generation of reactive oxygen species mediated by humic-like substances in atmospheric aerosols

Particulate matter (PM)-mediated reactive oxygen species (ROS) generation has been implicated in health effects posed by PM. Humic-like substances (HULIS) are an unresolved mixture of water-extracted organic compounds from atmospheric aerosol particles or isolated from fog/cloudwater samples. In this study, we use a cell-free dithiothreitol (DTT) assay to measure ROS production mediated by HULIS. The HULIS samples are isolated from aerosols collected at a rural location and a suburban location in the Pearl River Delta, China. In our experiments, ROS activities by residue metal ions in the HULIS fraction are suppressed by including a strong chelating agent in the DTT assay. Under conditions of DTT consumption not exceeding 90%, the HULIS-catalyzed oxidation of DTT follows the zero-order kinetics with respect to DTT concentration, and the rate of DTT oxidation is proportional to the dose of HULIS. The ROS activity of the aerosol HULIS, on a per unit mass basis is 2% of the ROS activity by a reference quinone compound, 1,4-naphthoquinone and exceeds that of two aquatic fulvic acids. The HULIS fraction in the ambient samples tested exhibits comparable ROS activities to the organic solvent extractable fraction, which would contain compounds such as quinones, a known organic compound class capable of catalyzing generation of ROS in cells. HULIS was found to be the major redox active constituent of the water-extractable organic fraction in PM. It is plausible that HULIS contains reversible redox sites, thereby serving as electron carriers to catalyze the formation of ROS. Our work suggests that HULIS could be an active PM component in generating ROS and further work is warranted to characterize its redox properties.     

壳聚糖处理对马铃薯块茎组织活性氧代谢的影响

以大西洋马铃薯(cv.Atlantic)块茎为试材,研究了壳聚糖处理及干腐病菌Fusarium sulphureum挑战接种对马铃薯块茎组织活性氧(ROS)代谢的影响.结果表明:壳聚糖处理能显著提高马铃薯切片过氧化氢(H2O2)、超氧阴离子(O-2)和抗坏血酸(AsA)含量及过氧化氢酶(CAT)和谷胱甘肽还原酶(GR)活性,而降低超氧化物歧化酶(SOD)和抗坏血酸过氧化物酶(APX)活性,F.sulphureum挑战接种可进一步提高壳聚糖处理及对照马铃薯切片的H2O2和O2-的含量及CAT、APX和GR的活性.可见壳聚糖和病原物可通过诱导氧进发而启动马铃薯组织的抗病反应.     

Effect of cis-urocanic acid on bovine neutrophil generation of reactive oxygen species

Neutrophils play a fundamental role in the host innate immune response during mastitis and other bacterial-mediated diseases of cattle. One of the critical mechanisms by which neutrophils contribute to host innate immune defenses is through their ability to phagocytose and kill bacteria. The ability of neutrophils to kill bacteria is mediated through the generation of reactive oxygen species (ROS). However, the extracellular release of ROS can be deleterious to the host because ROS induce tissue injury. Thus, in diseases such as mastitis that are accompanied by the influx of neutrophils, the generation of large quantities of ROS may result in significant injury to the mammary epithelium. cis-Urocanic acid (cis-UCA), which is formed from the UV photoisomerization of the trans isoform found naturally in human and animal skin, is an immunosuppressive molecule with anti-inflammatory properties. Little is known about the effect of cis-UCA on neutrophils, although one report demonstrated that it inhibits human neutrophil respiratory burst activity. However, the nature of this inhibition remains unknown. Because of the potential therapeutic use that a molecule such as cis-UCA may have in blocking excessive respiratory burst activity that may be deleterious to the host, the ability of cis-UCA to inhibit bovine neutrophil production of ROS was studied. Further, because neutrophil generation of ROS is necessary for optimal neutrophil bactericidal activity, a response which is critical for the host innate immune defense against infection, the effects of cis-UCA on bovine neutrophil phagocytosis and bacterial killing were assayed. cis-Urocanic acid dose-dependently inhibited the respiratory burst activity of bovine neutrophils as measured by luminol chemiluminescence. Subsequently, the effect of cis-UCA on the production of specific oxygen radicals was investigated using more selective assays. Using 2 distinct assays, we established that cis-UCA inhibited the generation of extracellular superoxide. In contrast, cis-UCA had no effect on the generation of intracellular levels of superoxide or other ROS. At concentrations that inhibited generation of extracellular superoxide, bovine neutrophil phagocytosis and bacterial activity remained intact. Together, these data suggest that cis-UCA inhibits the tissue-damaging generation of extracellular ROS while preserving neutrophil bactericidal activity.     

Effects of iron purity and groundwater characteristics on rates and products in the degradation of carbon tetrachloride by iron metal

Carbon tetrachloride (CT) batch degradation experiments by four commercial irons at neutral pH indicated that iron metal (Fe0) purity affected both rates and products of CT transformation in anaerobic systems. Surface-area-normalized rate constants and elemental composition analysis of the untreated metals indicate that the highest-purity, least-oxidized Fe0 was the most reactive on a surface-area-normalized basis in transforming CT. There was also a trend of increasing yield of the hydrogenolysis product chloroform (CF) with increasing Fe0 purity. Impurities such as graphite in the lower purity irons could favor the alternate CT reaction pathway, dichloroelimination, which leads to completely dechlorinated products. High pH values slowed the rates of CT disappearance by Peerless Fe0 and led to a pattern of decreasing CF yields as the pH increased from 7 to 12.9. The Fe/O atomic ratio vs depth for Peerless Fe0 filings equilibrated at pH 7 and 9.3, obtained by depth profiling analysis with X-ray photoelectron spectroscopy, indicated differences in the average oxide layer composition as a function of pH, which may explain the pH dependence of rate constants and product yields. Groundwater constituents such as HS-, HCO3-, and Mn2+ had a slight effect on the rates of CT degradation by a high-purity Fe0 at pH 7, but did not strongly influence product distribution, except for the HS amended Fe0 where less CF was produced, possibly due to the formation of carbon disulfide (CS2).     

Source apportionment of in vitro reactive oxygen species bioassay activity from atmospheric particulate matter

Recent atmospheric particulate matter health studies have suggested that the redox activity is an important factor in particulate matter toxicology, and that reactive oxygen species (ROS) activity may be an important characteristic of particulate matter that is associated with adverse health effects. In this study, associations between atmospheric particulate matter sources and in vitro ROS activities are investigated. Ambient concentrations of fine particle water-soluble elements and total organic and elemental carbon were measured daily in Denver for the 2003 calendar year. The data were used in a multivariate factor analysis source apportionment model, positive matrix factorization (PMF), to determine the contributions of nine sources or factors: a mobile source factor, a water soluble carbon factor, a sulfate factor, a soil dust source, an iron source, two point sources characterized by water soluble toxic metals, a pyrotechnique factor, and a platinum group metal factor. Aqueous leachates, including water soluble and colloidal components, as well as insoluble particles that pass through a 0.2 microm pore size filter, of 45 randomly selected PM samples, were assayed to quantify ROS activity using an in vitro rat alveolar macrophage assay. Results show that PM-stimulated in vitro ROS production was significantly positively correlated with the contributions from three sources: the iron source, the soil dust source and the water soluble carbon factor. The iron source accounted for the greatest fraction of the measured variability in redox activity, followed by the soil dust and the water-soluble carbon factor. Seventy-seven percent of the in vitro ROS activity was explained by a linear combination of these three source contributions.     

Neferine induces reactive oxygen species mediated intrinsic pathway of apoptosis in HepG2 cells

Evidence has accumulated concerning the medicinal application of Nelumbo nucifera in the treatment of various diseases. Neferine, an alkaloid from N. nucifera was found to exert cytotoxicity on liver cancer cells HepG2 in a dose-dependent manner. We evaluated its anticancer potential by studying its effect on mitochondrial membrane potential, intracellular calcium levels [Ca²⁺]_i, cell membrane integrity, apoptotic body formation and DNA fragmentation in cultured HepG2 cells. The reactive oxygen species level has been increased upon neferine treatment with concomitant decrease in reduced glutathione. Our data further indicate reduction of ΔψM and increased [Ca²⁺]_i during apoptosis induction by neferine with increased expression of apoptotic proteins such as Bax, Bad, cleaved forms of caspase 3, caspase 9 and PARP, with the downregulation of anti-apoptotic protein Bcl2 in HepG2 cells. Moreover, the expressions of tumour suppressor proteins p53 and PTEN were upregulated along with the downregulation of P-Akt. In addition, expression levels of TNF-α, p38 and ERK1/2 MAP kinases were increased upon neferine treatment. These results imply that mitochondrial-mediated ROS generation induced by neferine leads to caspase-dependent apoptosis in HepG2 cells.     

Photochemical production of reactive oxygen species by C60 in the aqueous phase during UV irradiation

The objective of this study was to investigate photochemical production of singlet oxygen (1O2) and superoxide radical anion (02*-) by C60 in water. It was demonstrated that photoexcited C60 in the aqueous phase efficiently mediated transfer of absorbed energy to oxygen and produced singlet oxygen when associated with surfactant (Triton X100 and Brij 78) or polymer (polyvinylpyrrolidone), which is consistent with previously observed behavior in organic solvents. However, when C60 was present as colloidal aggregate suspension, prepared through solvent exchange or sonication, this intrinsic character was lost. Similarly, C60 associated with surfactant mediated electron transfer from electron donor (triethylamine) to oxygen producing superoxide radical, while C60 aggregates and C60 associated with polymer did not. These results suggestthat the ability of C60 to mediate energy and electron transfer may be affected by the degree of C60 aggregation in the aqueous phase as well as characteristics of associated stabilizing molecules. Dependence of photochemical reactivity of C60 on its dispersion status in the aqueous phase is critical in assessing environmental impact and cytotoxicity of this material, as C60 associated with model natural organic matter was found to exist in aggregate form and did not produce reactive oxygen species under UV irradiation.     

Evaluation of reactive oxygen species generating AirOcare system for reducing airborne microbial populations in a meat processing plant

The microbial contamination of meat and meat products is of continuing concern to the meat industry and regulatory agencies. Air has been established as a source of microbial contamination in slaughter and processing facilities. The objective of this research was to determine the efficacy of reactive oxygen species (ROS) generating AirOcare equipment in reducing airborne bacteria in a meat processing environment. Bacterial strains found in ground beef were used to artificially contaminate the air using a 6-jet Collison nebulizer. Airborne bacterial populations in the meat processing room were monitored every 24 h at multiple locations using a Staplex 6 stage air sampler. Total aerobic, Gram-negative, and lactic acid bacterial populations were determined by sampling on R₂A agar, MacConkey agar and Lactobacilli MRS agar, respectively. Approximately 3 log reductions of lactic acid bacteria and Gram-negative bacteria were observed after 24 h of treatment (p < 0.05) compared to ~1.5 log reduction in the control treatment. Further exposure with ROS significantly reduced lactic acid bacteria and Gram-negative bacteria in the air at 48 and 96 h sampling intervals. These findings reveal that reactive oxygen species treatment using AirOcare unit significantly reduces airborne contamination in a meat processing environment. Mention of trade names or commercial products does not imply recommendation or endorsement to the exclusion of other products by the U.S. Department of Agriculture.