Ozone produced by corona discharge in the presence of water

Significant levels of ozone have been detected in a reaction flask that was designed for studies using negative air ions generated from corona discharge. While diluting the gas in the vessel at the rate of 1.5 liters/min., more than 1.0 ppm O₃ was measured in the vessel when grounded water was present. Ozone levels were much less (15 ppb) in ambient air near the ion generator. Even in the absence of water, O₃ levels were only 7 ppb in the reaction flask. Without the corona discharge apparatus, levels of ozone in the gas phase (with water present) averaged 6 ppb. These results demonstrate that when negative air ions are generated from corona discharge in the presence of water, significant levels of ozone are produced. Therefore, O₃ could be directly responsible for many of the oxidizing effects that have been reportedly due to negative air ions.     

Photoluminescence characteristics of novel Sm3+ ions-doped La1.4Al22.6O36 phosphor for n-UV w-LED

The combustion procedure was used to synthesize La_1.4Al_22.6O₃₆:Sm³⁺ phosphors. The X-ray diffraction (XRD) patterns and morphological and photoluminescence properties were investigated. The XRD patterns consisted of a hexagonal crystal structure. At 405 nm, the maximum excitation intensity was obtained. Following 405 nm excitation, three different emission peaks at 573, 604, and 651 nm were seen. Concentration quenching occurred at 1.5 mol% Sm³⁺ ions. The Commission Internationale de l'éclairage coordinates for the La_1.4Al_22.6O₃₆ phosphor with Sm³⁺ doping were 604 nm (x = 0.644, y = 0.355) falling in the red region. The findings implied that the prepared phosphor may be used to develop w-light-emitting diodes.     

The stimulatory effect of negative air ions and hydrogen peroxide on the activity of superoxide dismutase

The activity of erythrocyte cytosolic superoxide dismutase from rat, bovine, man and duck was considerably increased when measured after preparation or incubation in media pretreated with negative air ions (mostly superoxide) from electroeffluvial ion generator. 0.5–1.0 μM H₂O₂ was found in incubation medium after treatment with air ions. The stimulatory effect of air ions on superoxide dismutase activity was mimicked by addition of 0.5–6 μM H₂O₂. The primary physicochemical mechanism of beneficial biological action of negative air ions is suggested to be related to the stimulation of superoxide dismutase activity by micromolar concentrations of H₂O₂.     

Evolution of the negative ion composition in the afterglow of a streamer discharge in air

Evolution of the neutral and charged particle compositions in the afterglow of a single streamer channel in atmospheric-pressure air is simulated numerically. It is shown that, in the course of charge-exchange reactions, O₂⁻ ions rapidly transform into ions with a larger electron bound energy (such as O₃⁻ and NO₃⁻); in humid air, they also transform into hydrated ions NO₃⁻(H₂O) _n . As a result, the efficiency of primary electron production in the streamer head due to destruction of residual negative ions generated in the previous pulse decreases substantially.     

Effects of Ambient and Acute Partial Pressures of Ozone on Leaf Net CO(2) Assimilation of Field-Grown Vitis vinifera L

Mature, field-grown Vitis vinifera L. grapevines grown in open-top chambers were exposed to either charcoal-filtered air or ambient ozone partial pressures throughout the growing season. Individual leaves also were exposed to ozone partial pressures of 0.2, 0.4, or 0.6 micropascals per pascal for 5 hours. No visual ozone damage was found on leaves exposed to any of the treatments. Chronic exposure to ambient O₃ partial pressures reduced net CO₂ assimilation rate (A) between 5 and 13% at various times throughout the season when compared to the filtered treatment. Exposure of leaves to 0.2 micropascals per pascal O₃ for 5 hours had no significant effect on A; however, A was reduced 84% for leaves exposed to 0.6 micropascals per pascal O₃ when compared to the controls after 5 hours. Intercellular CO₂ partial pressure (c_i) was lower for leaves exposed to 0.2 micropascals per pascal O₃ when compared to the controls, while c_i of the leaves treated with 0.6 micropascals per pascal of 0₃ increased during the fumigation. The long-term effects of ambient O₃ and short-term exposure to acute levels of O₃ reduced grape leaf photosynthesis due to a reduction in both stomatal and mesophyll conductances.     

Measurement of protein by spectrophotometry at 205 nm

A method is described for the measurement of protein concentration by using the peptide bond absorption at 205 nm. ?₂₀₅ is estimated, allowing for the absorption due to Trp and Tyr residues, by measuring the absorbance at 280 nm as well as at 205 nm. The estimated ?₂₀₅ is compared with the actual ?₂₀₅ for a number of proteins, the mean error being less than 2%. This is about three times better than using an average ?₂₀₅^{1 mg/ml} of 31 and approaches the range of experimental error inherent in any method of protein estimation.     

The effects of air ions on brain levels of serotonin in mice

Mice were maintained in a controlled pollutant-free microenvironment and were exposed for 12, 24, 48 and 72 hr to 3 different concentrations of small positive or negative air ions: 2–4 × 10³ ions/cm³, 3–4 × 10⁴ ions/cm³ or 3.5–5 × 10⁵ ions/cm³. Spectrophotofluorometric assays of brain serotonin levels of air ion-treated mice showed statistically significant differences as early as 12 hours from those of mice kept in untreated pollutant-free air. Essentially no deviation from control values were observed at 24 and 48 hours. After 72 hours of exposure sharp decreases took place in all groups with the single exception of the animals exposed to 3–4 × 10⁴ positive ions/cm³. The hypothesis that alterations in mood and affect associated with certain meteorological conditions, e.g. winds such as the foehn, sirocco, etc. might depend upon air ion-induced changes in brain levels of serotonin was examined in the light of recent advances in neurophysiology and neuropharmacology.     

Brain Monoamine Changes in Rats After Short Periods of Ozone Exposure

We have shown in our laboratory that cat's and rat's sleep disturbances are produced by 24 h of ozone (O₃) exposure, indicating that the central nervous system is affected by this gas. To demonstrate the probable changes in brain neurotransmitters, we evaluated the monoamine contents of the midbrain and striatum of rats exposed to 1 part per million O₃ for 1 or 3 hours periods. The results were compared with rats exposed to fresh air and to those exposed to 3 hours of O₃ followed by 1 or 3 hours of fresh air. We found a significant increase in dopamine (DA) and its metabolites noradrenaline (NA) and 3,4 dihydroxyphenylacetic acid (DOPAC), as well as an increase in the 5-hydroxyindolacetic acid (5-HIAA) contents of the striatum. There were no changes in homovanillic acid (HVA) and serotonin (5-HT) levels during O₃ exposure. Additionally, an increase in DA, NA and 5-HIAA in the midbrain during O₃ exposure was observed. Turnover analysis revealed that DA increased more than its metabolites in both the midbrain and striatum. However, the metabolite of 5-HT, i.e. 5-HIAA, increased more than its precursor, this reaching statistical significance only in the midbrain. These findings demonstrate that O₃ or its reaction products affect the metabolism of major neurotransmitter systems as rapidly as after 1 h of exposition.     

Simultaneous photoreduction and Raman spectroscopy of red blood cells to investigate the effects of organophosphate exposure

Simultaneous photoreduction and Raman spectroscopy with 532 nm laser has been used to study the effects of organophosphate (chlorpyrifos [CPF]) exposure on human red blood cells (RBCs). Since in RBCs, auto‐oxidation causes oxidative stress, which, in turn, is balanced by the cellular detoxicants, any possible negative effect of CPF on this balance should results in an increased level of damaged (permanently oxygenated) hemoglobin. Therefore, when 532 nm laser, at a suitable power, was applied to photoreduce the cells, only common oxygenated form of hemoglobin got photoreduced leaving the permanently oxygenated hemoglobin detectable in the Raman spectra simultaneously excited by the same laser. Using the technique effects of CPF to build up oxidative stress on RBCs could be detected at concentrations as low as 10 ppb from a comparison of relative strengths of different Raman bands. Experiments performed using simultaneously exposing the cells, along with CPF, to H₂O₂ (oxidative agent) and/or 3‐Aminotriazole (inhibitor of anti‐oxidant catalase), suggested role of CPF to suppress the cellular anti‐oxidant mechanism. Since the high level of damaged hemoglobin produced by the action of CPF (at concentrations >100 ppm) is expected to cause membrane damage, atomic force microscopy (AFM) was used to identify such damages.Upper panel: Raman spectra of normal, photoreduced CPF exposed and unexposed RBCs. Lower panel: The weak Fe‐O₂ Raman band for CPF exposed cells shown on the left. The AFM images of unexposed and exposed cells are shown on the right. Scale bar, 2.5 μm.      

Purification,characterization, and coal depolymerizing activity of lignin peroxidase from <Emphasis Type="Italic">Gloeophyllum sepiarium</Emphasis> MTCC-1170

Lignin peroxidase from the liquid culture filtrate of Gloeophyllum sepiarium MTCC-1170 has been purified to homogeneity. The molecular weight of the purified enzyme was 42 kDa as determined by SDS-PAGE. The K _m values were 54 and 76 μM for veratryl alcohol and H₂O₂, respectively. The pH and temperature optima were 2.5 and 25°C, respectively. Depolymerization of coal by the fungal strain has been demonstrated using humic acid as a model of coal. Depolymerization of humic acid by the purified lignin peroxidase has been shown by the decrease in absorbance at 450 nm and increase in absorbance at 360 nm in presence of H₂O₂. Depolymerization of humic acid by the purified enzyme has also been demonstrated by the decrease in the viscosity with time of the reaction solution containing humic acid, H₂O₂, and the purified lignin peroxidase. The influence of NaCl and NaN₃ and inhibitory effects of various metal chelating agents on the lignin peroxidase activity were studied.     

Grassland yield declined by a quarter in 5 years of free-air ozone fumigation

The sensitivity of seminatural grasslands to ozone (O₃) pollution is not well known, in spite of the important function of these common ecosystems for agriculture and nature conservation. A 5‐year field experiment was carried out at a rural, mid‐elevation site at Le Mouret (Switzerland) to investigate the effect of elevated O₃ on yield and species composition of a permanent, extensively managed, species‐rich old pasture. Using a free‐air fumigation system, circular plots of 7 m diameter were exposed to either ambient air (control plots) or to air containing O₃ of approximately 1.5 × ambient concentrations. The resulting accumulated O₃ exposure over the threshold of 40 ppb for one season ranged from 13.3 to 59.5 ppm h in the elevated O₃ plots, and from 1.0 to 20.7 ppm h in the control plots. Subplots in each ring were harvested three times each year, and harvested biomass was separated into functional groups (FGs) (grasses, legumes, forbs). Productivity in both treatments decreased over time, but the yield of O₃‐exposed plots decreased faster than that of the control plots, with the reduction being twice as large by the end of the fifth season. Compared with the ambient air control, loss in annual dry matter yield was 23% after 5 years. The change in annual biomass production because of O₃ stress was accompanied by a change in the fractions of FGs, with the legume fraction showing a strong negative response. Such long‐term effects of O₃ stress could have negative implications for the maintenance of biological diversity in rural landscapes across large areas of Europe. The results from this first long‐term experiment show that a moderately elevated O₃ level reduces the productivity of intact grasslands during a 5‐year exposure under real field conditions.     

The persistent microbicidal effect in water exposed to the corona discharge

This article describes and particularly explains a new phenomenon of persistent microbicidal effect of water previously exposed to the low-temperature plasma, which cannot be attributed to the acidification only. The direct microbicidal action of plasma is well documented, being mediated by number of reactive particles with a short lifetime. However, we observed the microbicidal effect also in exposed water stored for a month, where it must be mediated by stable particles. In water and in phosphate-buffered saline, the formation of NO_x and corresponding acids, H₂O₂ and O₃ was confirmed after exposition to the low-temperature plasma generated in air by DC negative glow corona and positive streamer discharge. The time course of acidification, H₂O₂ and O₃ formation were deremined. Except uncertain traces of HCN, SIFT-MS analysis of exposed liquids reveals no additional reactive compounds. The microbicidal effect persists almost unchanged during 4 weeks of storage, although O₃ completely and H₂O₂ almost disappears. Staphylococcus epidermidis and Escherichia coli were inactivated within 10 min of incubation in exposed liquids, Candida albicans needs at least 1 h. The solutions prepared by artificial mixing of reactive compounds mimic the action of exposed water, but in lesser extent. The acid milieu is the main cause of the microbicidal effect, but the possibility of still unidentified additional compound remains open.     

Absence of harmful effects of protracted negative air ionisation

The absence of harmful effects of protracted negative air ionisation was studied in 5 weather-sensitive women and 5 normal men chosen at random. Negative ions were generated by the Modulion of Amcor-Amron (Herzliya, Israel). The patients were exposed separately during 8 sleeping hours and 8 working hours to the apparatus at 1–2 m distance in a 4 × 4 m room, for 2 months. Thus they were exposed to a daily uptake of 1 × 10⁴ negative ions/cm³ for 16 h/day during 2 months. Urinary 17-KS, 17-OH, adrenaline and noradrenaline excretion was not affected by the negative ionisation. However serotonin, 5-HIAA, histamine and thyroxine excretion — if increased before — diminished by 50% on an average. There were no changes in body weight, blood pressure, pulse, respiratory rate, oral morning temperature, dynamometer grip strength, routine liver function tests, urinary pH, albumen, glucose, ketones, bilirubin, or occult blood, red and white blood count and ECG records. The EEG revealed the typical changes due to negative air ionisation: stabilising of frequency, increased amplitudes, spreading of brainwaves from the perceptive occipital area to the conceptive frontal area and synchronisation of both hemisphere tracings.     

The role of the superoxide anion as a toxic species in the erythrocyte.

The toxic action of the superoxide anion (O₂^?) toward the erythrocyte was investigated with O₂^? generated through the autooxidation of dihydroxyfumaric acid (DHF). A suspension of human red cells exposed to DHF undergoes a rapid breakdown of the cellular hemoglobin to methemoglobin and other green pigments. This hemoglobin breakdown is inhibited by superoxide dismutase (SOD) or catalase (CAT) and is accelerated by lactoperoxidase (LP) added externally to the red cell medium. Associated with the hemoglobin breakdown is a hypotonic hemolysis also inhibited by SOD or CAT and initially accelerated but later inhibited by LP. Conversion of the red cell hemoglobin to carbonmonoxyhemoglobin in an aerated medium results in no hemoglobin breakdown or hypotonic lysis in the presence of DHF, even though O₂^? can be demonstrated in the medium. Although no evidence for membrane sulfhydryl oxidation or lipid peroxidation can be demonstrated in red cells exposed to DHF, the membranes of these cells were found to retain a green pigment. The presence of this green pigment in red cell membranes was inhibited by SOD, CAT, or conversion of the cellular hemoglobin to carbonmonoxyhemoglobin, but was not inhibited by LP. These results have been interpreted as a peroxide-dependent formation of O₂^? by DHF, followed by attack of O₂^? on hemoglobin. The reaction of O₂^? with hemoglobin leads to the formation of a hemoglobin-breakdown product that binds to the red cell membrane, resulting in an increased osmotic fragility of the cell.     

The effects of negative air ions on various physiological functions during work in a hot environment

The effects of negative air-ions on human physical performance has been investigated. Twenty-one healthy males, 20–25 years old (X=23.6±2.6) were exposed to two 180-min rest and exercise sessions two weeks apart. The subjects were randomly assigned into either an experimental group (n=12) or to a control group (n=9). The experimental group performed the first session in neutral air conditions and the second one in air containing 1.36 to 1.90×10⁵ negative air ions and 1.40 to 1.66×10² positive air ions/ml. The control group performed both sessions under neutral air conditions. All sessions were held at T_a=40±1C and 25±5% RH. Each session included one hour of resting under the respective ionization conditions, followed by 3 30-min cycle ergometer work bouts, separated by 7-min rest periods. The mechanical work-load during the bicycle exercise was 1.64±0.6 W/kg BW. The experimental group showed a significant reduction with negative air-ions in heart rate (HR), in rectal temperature, and in the rating of perceived exertion (RPE), all when compared with their own neutral session. The control group showed no significant changes between the first and the second exposure. Although not statistically significant, being exposed to negative air-ions seems also to reduce total sweat rate and minute ventilation (V_E), and to increase O₂ pulse. It is suggested that under the conditions of this study negative air ions canimprove various cardiovascular and thermoregulatory functions as well as subjective feelings during physical effort. It is felt that such positive influences may be augmented by increasing the exposure time to negative ionized air and/or prolonging the stressful conditions.With the technical assistance of M. Sheinovitz.     

Variable growth and reproduction response of the spruce shoot aphid, Cinara pilicornis, to increasing ozone concentrations

Spruce shoot aphid, Cinara pilicornis Hartig (Homoptera: Lachnidae), is an aphid species that has shown enhanced performance on trees exposed to SO₂ or mixtures of air pollutants, whereas results with ozone have been contradictory. Using a 4-week chamber fumigation experiment, we tested how different population dynamic parameters of aphids are affected by ozone (O₃). Mean relative growth rate (MRGR), development time and reproduction rate of C. pilicornis were determined using 7h day-time O₃ concentrations of 0, 40, 80 and 160 ppb. Development period from birth to reproductive stage was fastest at 80 ppb during early shoot elongation, while reproduction and intrinsic rate of population increase was not significantly affected by ozone concentration. There was a significant negative correlation between ozone concentration and the MRGR of first instar nymphs in the third fumigation week. In feeding test performed on seedlings after fumigation, the MRGR was reduced on shoots which had been exposed to 80 ppb O₃. The results suggest that elevated O₃ concentration during early shoot elongation period may stimulate population development of C. pilicornis, but on maturing shoots, high O₃ concentration has a negative effect on aphid performance. This might be due to accelerated ageing of O₃ exposed shoots.     

Conjugation of nattokinase and lumbrukinase with magnetic nanoparticles for the assay of their thrombolytic activities

Two important thrombolytic enzymes, nattokinase (NK) and lumbrukinase (LK), were immobilized onto fine magnetic Fe₃O₄ nanoparticles using 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide (EDC) as the coupling reagent, and their thrombolytic activities were studied. The Fe₃O₄ nanoparticles and NK- and LK-conjugated magnetic nanoparticles were characterized by transmission electron microscopy, Fourier transform infrared spectrophotometry, vibrating sample magnetometry, X-ray diffraction, and UV–vis absorption spectroscopy. Dual kinetic absorbance measurements at 405 and 630 nm were employed to measure their thrombolytic activity. Analysis of protein amount showed that the optimum conditions for NK and LK binding to nanoparticles were respectively at a mass ratio of 2:1:1, 2:1:2 (magnetic nanoparticles:protein:EDC), and pH 6.00. Thrombolytic activity assay showed that the best thrombolytic activity could reach 91.89% for NK–nanoparticle conjugates and 207.74% for LK–nanoparticle conjugates, which are much higher than the pure enzymes (NK, 82.86%; LK, 106.57%).     

Effects of negative air ions,noise, sex and age on maze learning in rats

An equal number of male and female rats of the King-Holtzman hybrid breed, divided into two age groups (group A₁ : 21–30 days old and group A₂ : 90–100 days old), were exposed to two levels of noise: N₀ = 30 db and N₁ = 90 db, (Ref. 0.0002μ bar), and three levels of negative air ions: I_O = no measurable ion concentration, I₁ = 7 × 10⁶ ions/cm³ and I₂ = 7 × 10⁷ ions/cm³. Time and error scores of 240 rats running a modified Lashley left-right maze with an escape-from-water motive served as criteria. A randomized complete blocks design with replications (2×3×2×2×10) was selected for treatment by analysis of variance. The results indicate that (a) the males show significantly lower error score in negatively ionized air; and (b) the females swim significantly faster than males under all investigated conditions with no apparent effect of noise or ions on their performance.

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Zusammenfassung Eine gleiche Anzahl m?nnlicher und weiblicher Ratten (n = 120) des King-Holtzman Stammes, unterteilt in 2 Altersgruppen (21–30 und 90–100 Tage) wurden zwei Ger?uschintensit?ten (30 db und 90 db) und 3 Konzentrationen negativer Luftionen I₀: nicht messbar niedrig, I₁:7×10⁶, I₂:7×10⁷ Ionen/cm³ ausgesetzt. Die Wirkung wurde geprüft an der Schwimmzeit und der Anzahl Fehler der Tiere im Lasley-Links-Rechts-Labyrinth. Das Motiv zum Schwimmen war, aus dem Wasser zu entkommen. Die Auswertung durch Varianzanalyse erfolgte unter Nachbildung randomisierter kompletter Blockentwürfe. Die Ergebnisse zeigen, dass (a) die M?nnchen bei negativ ionisierter Luft signifikant weniger Fehler machten und (b) die Weibchen unter allen Bedingungen schneller schwammen als die M?nnchen, ohne dass das L?rmniveau oder die Luftionisation darauf einen Einfluss hatten.

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Resume Un nombre égal de rats males et femelles (120 de chaque) de la race King-Holtzman futdivisé en deux groupes d'age (21 à 30 jours et 90 à 100 jours). Ils furent exposés à 2 intensités auditives (30 et 90 db) et à 3 concentrations d'ions négatifs: I₀ non mesurables (faible), I₁ : 7 × 10⁶ et I₂: 7 × 10⁷ ions/cm³. Les effets d'un tel traitement furent examinés au moyen de deux critères: le temps pris pour sortir de l'eau et le nombre de fautes commises dans un labyrinthe droitegauche de Laslay. Pour l'analyse des variances, on a adopté une répartition due au hasard avec répétitions (2×3×2×2×10). On peut en conclure que: (a) les males font moins d'erreurs dans l'air ionisé négativement et cela de fa?on significative et (b) les femelles ont nagé plus vite que les males dans toutes les conditions de l'essai. Ni le bruit ni le degré d'ionisation ne semblent avoir d'influence sur leur comportement.

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This study was supported in part by a grant from the Faculty Research Fund of the University of Oklahoma.     

CO-binding pigments and the functional terminal oxidase of the trypanosomatid hemoflagellate Crithidia fasciculata

CO-difference absorbance spectra of both intact cells and of mitochondrial preparations isolated from Crithidia fasciculata were obtained after anaerobiosis was attained either with substrates or with dithionite. Under both sets of conditions, the CO-difference spectrum of cytochrome a₃, with difference absorbance maxima at 430 and 589 nm and minima at 443 and 612 nm, was readily identified in both the intact cells and in the mitochondria. In addition to the difference absorbance bands of cytochrome a₃-CO, three difference absorbance maxima at 417, 538 and 570 nm and a minimum at 556 nm were observed. The magnitude of the maximum at 570 nm relative to the maximum of cytochrome a₃-CO at 589 nm was less for mitochondria rendered anaerobic with substrate than for mitochondria rendered anaerobic with dithionite. This difference was taken to define operationally two groups of mitochondrial CO-binding pigments: Group I is that group observed on anaerobiosis with substrate: Group II is the additional group observed on anaerobiosis with dithionite. The Group I CO-binding pigments were virtually absent from submitochondrial particles derived by sonication, but the Group II pigments remained.Photochemical action spectra were obtained with isolated mitochondria and intact cells to ascertain if cytochrome o was present. These action spectra, obtained in CO plus O₂ atmospheres, had maxima only at 432, 550 and 588 nm, attributable to the photodissociation of cytochrome a₃-CO. Even after suppression of cytochrome a₃ activity to 10% of the normal value, no contribution of cytochrome o activity to the photochemical action spectrum was observed. Cytochrome a₃ is therefore the only functional terminal oxidase present in the mitochondria of Crithidia fasciculata.     

Quantitative generation of singlet (1Δg) oxygen from acidified aqueous peroxynitrite produced by the reaction of nitric oxide and superoxide anion

Simple acidification of aqueous alkaline peroxynitrite quantitatively generates singlet (¹Δ_g) molecular oxygen, detected and quantitated spectroscopically (1270 nm). This observation provides a chemical basis for physiological cytotoxicity of ONOO^? generated in the diffusion - controlled reaction of cellular NO^? and O. The experiments consist of (i) chemical generation of ONOO^? from NO^? gas and KO₂ powder in alkaline aqueous solution; (ii) absorption spectral identification of ONOO^? in the near-UV with maximum at 302 nm; (iii) spectroscopic identification of ¹O₂ by its emission band at 1200–1340 nm with maximum at 1275 nm; and (iv) quantitation of ¹O₂ generated in ONOO^?/H⁺ reaction by comparison of the chemiluminescence intensity at 1270 nm with that from H₂O₂/OCl^? reaction that generates ¹O₂ with unit efficiency at alkaline pH. ¹O₂ was generated with unit efficiency with respect to ONOO^? concentration by the ONOO^?/H⁺ reaction.