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Lack of oxidative DNA damage or initiation of carcinogenesis in the kidneys of male F344 rats given subchronic exposure to p-dichlorobenzene (pDCB) at a carcinogenic dose.

p-Dichlorobenzene (pDCB) is a male rat kidney carcinogen believed to act through alpha2u-globulin nephropathy. Recent data on metabolism, however, suggest a potential for generating oxidative stress. To examine possible mechanisms of kidney carcinogenesis, pDCB was studied for ability to produce 8-oxodeoxyguanosine (8-oxodG) in kidney nuclear DNA and for initiating activity in a two-stage renal carcinogenesis model. F344 male rats were given pDCB by intragastric instillation, 5 days/week for 13 weeks at 300 mg/kg per day, which is a carcinogenic dose with chronic administration. To assess initiation after exposure, trisodium nitrilotriacetic acid (NTA), a kidney tumor promoter was given in the drinking water at 1,000 ppm for 39 weeks. At the end of the exposure segment, pDCB did not produce an increase of 8-oxodG levels in the kidney nuclear DNA in contrast to potassium bromate (KBrO3). Following NTA promotion, no neoplastic lesions occurred in rats given pDCB, although diethylnitrosamine carcinogenesis was enhanced. Thus, pDCB did not produce oxidative DNA damage in the rat kidney or effect initiation of kidney carcinogenesis. These data suggest that oxidative stress is not involved in pDCB-induced renal carcinogenesis. The alpha2u-globulin-mediated chronic nephropathy probably acts as a promoter, not an initiation of renal carcinogenesis. Accordingly, pDCB is assessed to have no cancer hazard to humans who are not susceptible to the alpha2u-globulin nephropathy.

8-Hydroxy-2'-Deoxyguanosine↗

Determination of trace platinum by supramolecular catalytic kinetic spectrofluorimetry of beta-cyclodextrin-platinum-KBrO3-salicylaldehyde furfuralhydrazone.

A supramolecular catalytic kinetic spectrofluorimetric method was developed for the determination of platinum(IV) and the possible mechanism of catalytic reaction was discussed. The method was based on the fluorescence-enhancing reaction of salicylaldehyde furfuralhydrazone (SAFH) with potassium bromate, which was catalysed by platinum(IV) in a water-ethanol medium. beta-Cyclodextrin (beta-CD) obviously sensitized the determination at pH 5.20 and 25 degrees C. Under optimum conditions, the beta-CD-platinum-KBrO3-SAFH supramolecular kinetic catalytic reaction system had excitation and emission maxima at 372 and 461 nm, respectively. The linear range of this method was 0.60-180 ng ml-1 with a relative standard deviation of 1.2%, and the detection limit was 0.18 ng ml-1. Investigation of the mechanism and the effects of interferences is presented. The proposed method was applied successfully to determine trace platinum(IV) in the chemotherapeutic drug cisplatin and serum from patients with satisfactory results.

Journal Article↗

Photokinetic voltammetric method for the determination of thiocyanate.

Thiocyanate traces have a strong inhibitory effect on the oxidation of Neutral Red by potassium bromate under UV irradiation in diluted phosphoric acid. Neutral Red exhibits a sensitive second derivative oscillopolarographic wave at -0.6 V(vs. SCE) in diluted phosphoric acid and sodium acetate solution. The oscillopolarographic behavior of Neutral Red was selected as indicator component for its photo-activated oxidation. The photochemical reaction rate equation was determined. A detection limit of 0.3 ng mL(-1) (3sigma/k) and a linear calibration curve from 2.0-48.0 ng mL(-1) thiocyanate were obtained. The method was applied to the determination of thiocyanate in urine, saliva and serum with satisfactory results.

Journal Article↗

Direct determination of iron in ferritin using the polarographic catalytic current.

A new method for the determination of iron in ferritin using the polarographic catalytic current has been developed. Iron is released in NaOH and triethanolamine (TEA) solution and subsequently determined based on the polarographic catalytic wave in the presence of bromate with the peak potential of -1.0 V (vs.SCE). The 2.5-derivative peak height of iron is linearly proportional to its concentration in the range of 8.0 x 10(-8)-1.0 x 10(-6) g/ml with a correlation coefficient of 0.9991. The method has been applied to the determination of iron in human fetal liver and horse-spleen ferritin and the recovery is 95-112%.

Journal Article↗

Analysis of oxyhalide disinfection by-products and other anions of interest in drinking water by ion chromatography.

The US Environmental Protection Agency is developing regulations for various drinking water disinfection by-products (DBPs). This effort involves developing analytical methods for the DBPs formed as a result of different disinfection treatments and collecting occurrence data for these species. Ion chromatography is one method being used to analyze drinking water samples for the following inorganic DBPs: chlorite, chlorate and bromate. These anions, however, are difficult to separate from common interfering anions of chloride, carbonate and nitrate. A method is therefore presented by which tetraborate/boric acid is used to separate these anions. Method detection limits of the order of 10 micrograms/l, using conductivity and UV detection were obtained. Stability studies of chlorite showing the effectiveness of ethylenediamine as a preservative and summary data for an occurrence of nitrite, nitrate and the DBP precursor bromide are presented.

Anions↗

Difference between intraperitoneal and oral gavage application in the micronucleus test. The 3rd collaborative study by CSGMT/JEMS.MMS. Collaborative Study Group for the Micronucleus Test/Mammalian Mutagenesis Study Group of the Environmental Mutagen Society of Japan.

In the third collaborative study organized by the Collaborative Study Group for the Micronucleus Test (CSGMT), a task group belonging to the Mammalian Mutagenesis Study subgroup of the Environmental Mutagen Society of Japan (JEMS.MMS), intraperitoneal (i.p.) injection and oral (p.o.) gavage were compared as routes of administration of test chemicals. Two mouse strains, MS/Ae and CD-1, and 17 chemicals with various modes of action were used. The chemicals were 1-beta-D-arabinofuranosylcytosine, 6-mercaptopurine monohydrate, benzo[a]pyrene, 7,12-dimethylbenz[a]anthracene, 2-acetylaminofluorene, phenacetin, cyclophosphamide, ethyl methanesulfonate, N-ethyl-N-nitrosourea, methyl methanesulfonate, mitomycin C, colchicine, vincristine sulfate, potassium bromate, potassium chromate(VI), benzene, and procarbazine hydrochloride. On the basis of the findings of an acute toxicity test and a pilot experiment for dose and sampling time, a full-scale micronucleus test was performed on each chemical. Almost all the chemicals showed a positive response in micronucleus induction by both routes of administration in both mouse strains. Contradictory outcomes were obtained between the i.p. and p.o. routes on potassium chromate in both strains (i.p.: positive, p.o.: negative). In the CD-1 mice, benzene potently induced micronuclei when administered p.o., but gave only a marginal response when administered i.p. Generally, the chemicals induced micronuclei at lower dose levels (mg/kg) when administered i.p. This tendency, however, was decreased or even reversed when the dose was expressed as a percentage of the LD50. Although the i.p. route, an artificial exposure route, is useful to detect the inducibility of micronuclei of a test chemical per se at a small dose, the p.o. route seemed sensitive and valuable enough to evaluate the test chemicals. When the dose levels of chemicals are adjusted on the basis of the LD50, both i.p. injection and p.o. gavage are acceptable as routes of administration in the micronucleus test.

Administration, Oral↗

Mechanism of dioxygen formation catalyzed by vanadium bromoperoxidase from Macrocystis pyrifera and Fucus distichus: steady state kinetic analysis and comparison to the mechanism of V-BrPO from Ascophyllum nodosum.

Vanadium bromoperoxidase (V-BrPO) catalyzes the oxidation of bromide by hydrogen peroxide, which results in the bromation of appropriate organic substrates or the formation of dioxygen, in the absence of an organic substrate and under certain other conditions. The mechanism of the bromide-assisted disproportionation of hydrogen peroxide catalyzed by V-BrPO, which is the reaction that forms dioxygen, has been investigated for V-BrPO isolated from two new marine algal sources, Macrocystis pyrifera and Fucus distichus. The steady state kinetic studies have been performed under conditions of 0.02-40 mM H2O2, 1-500 mM Br- and pH 4.0-8.0. The rate data is consistent with a substrate-inhibited bi bi ping pong mechanism, in which the substrate bromide, is also an inhibitor by a noncompetitive-type mechanism. Bromide inhibits V-BrPO from M. pyrifera most strongly at pH 5.0-5.5 and V-BrPO from F. distichus most strongly at pH 5.5-6.0. The steady state mechanism of the Macrocystis and the Fucus enzymes are compared to the mechanism of the bromide-assisted disproportionation of hydrogen peroxide catalyzed by V-BrPO from Ascophyllum nodosum. In addition, the substrate hydrogen peroxide can also inhibit V-BrPO.

Catalysis↗

Regulation of food-related carcinogens in Japan.

Use of hazardous environmental substances in Japan is regulated, based on their risk assessment, by laws of various ministries, depending upon the chemicals concerned. Food additives and food contaminants are controlled by the Food Sanitation Law of the Ministry of Health and Welfare. When manufacture or import of new food additives is intended, their physicochemical and biological data are evaluated by the Food Sanitation Investigation Council. When new data become available on the carcinogenicity of food additives that have been approved and are in use, these compounds are reevaluated. Between 1974 and 1986, twelve chemicals were proved to be carcinogenic in Japan by task groups for evaluation of the carcinogenicity of food additives, medical drugs and pesticides, which were organized by the Ministry of Health and Welfare. Of these compounds, 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide was banned from use, but use of hydrogen peroxide and potassium bromate was permitted, provided that these compounds were decomposed or removed from the final products. The use of butylated hydroxyanisole within fixed limits in some foodstuffs was also permitted. Decisions on the use of these compounds were based on the prevailing opinion on carcinogens at the time. This paper reports these facts with a consideration of future problems on the regulation of food carcinogens.

Animals↗

Primary mutagenicity screening of food additives currently used in Japan.

Salmonella/microsome tests (Ames tests) and chromosomal aberration tests in vitro using a Chinese hamster fibroblast cell line were carried out on 190 synthetic food additives and 52 food additives derived from natural sources, all of which are currently used in Japan. Fourteen out of 200 tested in the Ames assay showed positive effects and 54 out of 242 were positive in the chromosome test. Three additives (erythorbic acid, chlorine dioxide and beet red) were positive only in the Ames test, although their mutagenic potentials were relatively weak, while 43 additives were positive only in the chromosome test. Eleven additives (calcium hypochlorite, cinnamic aldehyde, L-cysteine monohydrochloride, Food Green No. 3 (Fast Green FCF), hydrogen peroxide, potassium bromate, sodium chlorite, sodium hypochlorite, sodium nitrite, cacao pigment and caramel) were positive in both the Ames test and the chromosome test. The usefulness of such primary screening tests combining two different genetic end-points, gene mutation and chromosomal aberration, and some correlation between mutagenicity and carcinogenicity of food additives are discussed.

Animals↗

Synchronization in two interacting oscillatory systems.

Nonlinear phenomena arising from the interaction of two oscillating systems of chemical reactions are studied experimentally. The system of two connected flow-through continuous stirred tank reactors (cells) with controlled exchange of reaction mixture is used. The Belousov reaction (oxidation of malonic acid by bromate in sulphuric acid with ceric/cerous ions as catalyst) served as model system. The frequency of oscillations was controlled by change of the reaction temperature. Phenomena such as synchronization of oscillations at a common frequency, synchronization at multiples of a common frequency, rhythm splitting and amplitude amplification were observed, depending on the degree of interaction and the differences in the original oscillation frequencies. Mathematical modelling of the above phenomena failed, probably due to insufficient knowledge of a kinetic model.

Cell Physiological Phenomena↗

Determination of vitamin C in effervescent tablets containing other vitamins together with trace elements.

A simple, rapid method is reported for the determination of vitamin C in effervescent tablets containing other vitamins and several trace elements such as Mg(II), Zn(II), Fe(II), Mn(II), Cu(II) and Mo(VI). The procedure was developed on the basis of the bromate ion-iodide ion-ascorbic acid clock reaction (Landolt reaction). Interference effects of air oxygen and metal ions and the role of pH are discussed in detail.

Ascorbic Acid↗

Heterogeneous photocatalysed reaction of three selected pesticide derivatives, propham, propachlor and tebuthiuron in aqueous suspensions of titanium dioxide.

Heterogeneous photocatalysed reaction of three selected pesticide derivatives such as propham (1), propachlor (2) and tebuthiuron (3) has been investigated in aqueous suspensions of titanium dioxide by monitoring the change in substrate concentration employing UV Spectroscopic analysis and depletion in Total Organic Carbon (TOC) content as a function of irradiation time. The degradation kinetics was studied under different conditions such as pH, catalyst concentration, substrate concentration, different types of TiO(2) and in the presence of electron acceptors such as hydrogen peroxide (H(2)O(2)), potassium bromate (KBrO(3)) and ammonium persulphate (NH(4))(2)S(2)O(8) besides molecular oxygen. The degradation rates were found to be strongly influenced by all the above parameters. The photocatalyst Degussa P25 was found to be more efficient as compared with other photocatalysts. The pesticide derivative propham (1) was found to degrade faster as compared to propachlor (2) and tebuthiuron (3). An attempt has also been made to identify the products formed during the photooxidation process through GC/MS analysis technique. All the model pollutants showed the formation of several intermediate products, which were identified on the basis of molecular ion and mass spectrometric fragmentation pattern. A probable mechanism for the formation of the products has been proposed.

Acetanilides↗

Separation and identification by gel filtration and high-performance liquid chromatography with UV or electrochemical detection of the disulphides produced from cysteine and glutathione oxidation.

Methods for quantification of oxidised and reduced forms of glutathione (GSSG and GSH) and cysteine (CSSC and CSH) and the disulphide glutathione-cysteine (GSSC) resulting from the oxidation of the mixture of CSH and GSH are performed by RP-HPLC with coulometric and UV detection after separation of these compounds by size-exclusion fast protein liquid chromatography. The fractionation of the disulphides (GSSG, GSSC and CSSC) was achieved by size exclusion using a Superdex peptide column coupled with an UV detection at 254 nm. The conditions of separation of these compounds by RP-HPLC were optimised using the response surface methodology. Optimal peak resolution and retention times were obtained on a C18 YMC ODS AQ column with 20 mM of ammonium phosphate at pH 2.5 and 2% of acetonitrile in the elution phase. In these experimental conditions, CSH, CSSC, GSH and GSSG were eluted within 20 min. Coulometric detection enabled a sensitivity 100 times higher for the disulphides than the UV detection at 220 nm. These methods were applied to follow the consumption of thiols and the disulphide formation by three oxidising systems, sulphydryl oxidase, glutathione dehydroascorbate oxidoreductase and potassium bromate. This study revealed that the relative proportions of the disulphides formed were similar for the three oxidising systems when the reactions are in their state of equilibrium.

Chromatography, Gel↗

Electrochemically modulated liquid chromatographic separations of inorganic anions.

Inorganic anion retention on a porous graphitic carbon (PGC) stationary phase is investigated by electrochemically modulated liquid chromatography (EMLC). Through various combinations of the potential applied (Eapp) to the PGC packing and/or changes in the composition (sodium salts of tetrafluoroborate, sulfate, and fluoride) and concentration (10, 25, and 50 mM) of an aqueous mobile phase, conditions for the separation of two different inorganic anion mixtures (iodate, bromide, nitrite, and nitrate or iodate, bromate, and chlorate) are developed. Results show that retention was affected by both variables, with the analyte retention factor, k', changing in a few cases by as much as a factor of ca. six. Moreover, plots of In k' are linearly dependent on both Eapp and In [SE], where [SE] is the supporting electrolyte concentration. Based on these findings, insights into the retention mechanism are briefly discussed by drawing on the theory for ion exchange chromatography.

Anions↗

Fast ion chromatography of inorganic anions and cations on a lysine bonded porous silica monolith.

A 0.46 cm x 10.0 cm silica monolith column was modified through the in situ covalent attachment of lysine (2,6-diaminohexanoic acid) groups. Due to the zwitterionic nature of the resultant stationary phase, the modified monolithic column contained both cation and anion exchange capacity. In the case of cation exchange, the capacity was found to be relatively low at between 5 and 6.5 micromoles Me2+ per column. However, as expected, the lysine monolith exhibited a higher anion exchange capacity at 12-13 micromoles A- per column (at pH 3.0), which was found to be dependent upon column pH, due to the dissociation of the weak acid carboxylic acid groups. High-performance separations of transition metal cations and inorganic anions were achieved using the modified monolith, with the effects of eluent concentration, pH and flow rate evaluated. Using elevated flow rates of up to 5 mL/min the separation of nitrite, bromate, bromide, nitrate, iodide and thiocyanate was possible in approximately 100 s with peak efficiencies of between 50 and 100,000 N/m and retention time %RSD of under 0.3%.

Anions↗

Rapid determination of acrylamide contaminant in conventional fried foods by gas chromatography with electron capture detector.

Gas chromatography coupled with electron capture detector (GC-ECD) was successfully developed and applied for the rapid determination of acrylamide in conventional fried foods, such as potato crisps, potato chips, and fried chicken wings. The method included defatting with n-hexane, extraction with aqueous solution of sodium chloride (NaCl), derivatization with potassium bromate (KBrO3) and potassium bromide (KBr), and liquid-liquid extraction with ethyl acetate. The final acrylamide extract was analyzed by GC-ECD for quantification and by GC-MS for confirmation. The chromatographic analysis was performed on the HP-INNOWax capillary column, and good retention and peak response of acrylamide were achieved under the optimal conditions (numbers of theoretical plates N = 83,815). The limit of detection (LOD) was estimated to be 0.1 microg kg(-1) on the basis of ECD technique. Recoveries of acrylamide from conventional samples spiked at levels of 150, 500 and 1000 microg kg(-1) (n = 4 for each level) ranged between 87 and 97% with relative standard deviations (RSD) of less than 4%. Furthermore, the GC-ECD method showed that no clean-up steps of acrylamide derivative would be performed prior to injection and was slightly more sensitive than the MS/MS-based methods. Validation and quantification results demonstrated that this method should be regarded as a new, low-cost, and robust alternative for conventional investigation of acrylamide.

Acrylamide↗

Human cytochrome P450 reductase can act as a source of endogenous oxidative DNA damage and genetic instability.

Studies with repair-deficient mice and other experiments suggest that oxidative DNA modifications are generated in all types of cells even under physiological conditions and that this type of endogenous DNA damage contributes to spontaneous cancer incidence. However, the cellular sources of reactive oxygen species that are relevant for nuclear oxidative DNA damage are largely unknown. Here, we report that expression of human NADPH-cytochrome P450 reductase (hOR) in cultured V79 Chinese hamster cells gives rise to elevated basal levels of oxidative purine modifications after depletion of glutathione. Also, the basal levels of micronuclei are increased in the hOR-expressing cells, and again the effect is enhanced when the antioxidant defense system of the cells is diminished by depletion of glutathione. The oxidative DNA damage is increased when duroquinone, a substrate of hOR, is added, both in the presence and absence of glutathione. In contrast, hOR-expressing cells are similarly sensitive as the parental cells when oxidative DNA damage and micronuclei are induced by a mechanism independent of hOR, i.e., exposure to bromate. The results identify hOR as a potential source of endogenous oxidative DNA damage and subsequent genetic instability in mammalian cells.

Animals↗

Electrochemistry of ITO electrode modified by multilayer ultrathin films based on crown-shaped polyoxomolybdate.

The electrochemical multilayer films of crown-shaped polyoxomolybdate Na21{[Na5(H2O)14] intersection[Mo(V)(20)Mo(VI)(26)O134(OH)10(mu-CH3COO)4]}.CH3COONa.90H2O (Mo46) and polyelectrolytes by layer-by-layer assembly were investigated. The stable multilayer films were assembled by alternate adsorption of negatively charged POM and positively charged polyelectrolytes is from their aqueous dispersions. UV-vis spectroscopy and cyclic voltammetry were used to monitor the regular growth of the multilayer films. The multilayer films-modified ITO electrode was used for the detection of electrocatalytic activity toward the reduction of nitrite, bromate, and hydrogen peroxide. The proposed novel immobilized method exhibited good stability, reproducibility and high sensitivity for the determination of electrocatalytic, which is important for practical application.

Letter↗