Inhibition of glutathione S-transferase zeta and tyrosine metabolism by dichloroacetate: a potential unifying mechanism for its altered biotransformation and toxicity.

Dichloroacetate (DCA) inhibits its own metabolism and is converted to glyoxylate by glutathione S-transferase zeta (GSTz). GSTz is identical to maleylacetoacetate isomerase, an enzyme of tyrosine catabolism that converts maleylacetoacetate (MAA) to fumarylacetoacetate and maleylacetone (MA) to fumarylacetone. MAA and MA are alkylating agents. Rats treated with DCA for up to five days had markedly decreased hepatic GSTz activity and increased urinary excretion of MA. When dialyzed cytosol obtained from human liver was incubated with DCA, GSTz activity was unaffected. In contrast, DCA incubation inhibited enzyme activity in dialyzed hepatic cytosol from rats. Incubation of either rat or human hepatic cytosol with MA led to a dose dependent inhibition of GSTz. These data indicate that humans or rodents exposed to DCA may accumulate MA and/or MAA which inhibit(s) GSTz and, consequently, DCA biotransformation. Moreover, DCA-induced inhibition of tyrosine catabolism may account for the toxicity of this xenobiotic in humans and other species.

Toxicity of the styrene metabolite, phenylglyoxylic acid, in rats after three months' oral dosing.

Male Wistar rats were dosed with 0, 1250, 3750 or 5000 mg/l of phenylglyoxylic acid (PGA) (CAS no. 611-73-4) in the drinking water ad libitum for 3 months. During the entire treatment period, there were no gross signs of toxicity related to PGA. No changes in neurobehavior were found after using a functional observational battery or radial arm maze. An increased relative kidney weight was seen in the highest dose-group (Controls: 0.504 +/- 0.031 g/100 g b.wt.; 5000 mg PGA/l: 0.579 +/- 0.033 g/100 g b.wt.; p<0.01). No other organ weights were affected. Histopathology revealed no change in kidney structure. No changes in clinical biochemistry. In the highest dose-group three animals out of ten showed reduction in peripheral nerve myelin sheath thickness. No such changes were seen in the control group. The study revealed no changes in auditory brain stem response but minor changes in electroretinography. The noradrenaline (NA) concentration decreased in pons and thalamus whereas it increased in medulla oblongata and whole brain. The dopamine (DA) concentration increased in cerebellum, hippocampus, pons, and whole brain. The most marked DA increase was seen in hippocampus (Controls: 0.56 +/- 0.10 nmol/g tissue; 5000 mg/l: 1.04 +/- 0.11 nmol/g tissue; p<0.001). The 5-hydroxytryptamine (5-HT) concentration decreased in cerebellum, cerebral cortex, hippocampus, and medulla oblongata, whereas it increased in thalamus. The yield of synaptosomal protein, synaptosomal NA, DA, and 5-HT concentrations, and DA uptake rate were not affected. When dosed males were mated with naive females, there were no differences between groups in the pregnancy rate, number of corpora luteae, implantations, live or dead fetuses, resorptions, preimplantation loss, or postimplantation loss. It is concluded that a part of the effects on kidney, peripheral nerves, and vision, which have previously been reported after exposure to styrene, might be induced by the styrene metabolite, PGA. If PGA has ototoxic effects in rats, the dosing in the present study is not sufficient to induce the necessary ototoxic concentration in blood. Alternatively, the ototoxicity of styrene, like toluene, may be caused the parent compound itself and not by a metabolite like PGA.

Determination of the urinary metabolites of styrene: estimation of the method evaluation function and evaluation of reference values in Danish subjects.

A European study on styrene exposure was initiated in 1989 to evaluate the health effects of environmental and occupational exposure. A part of this study included the development of an analytical method for use in a biological monitoring program. The urinary metabolites of styrene, mandelic acid (MA) and phenylglyoxylic acid (PGA) were quantitated by a direct and convenient high-performance liquid chromatography method. Urine samples were diluted with eluent and analysed by HPLC with a C8 reversed-phase column and a buffer to acetonitrile (9:1) eluent with a counterion added. The detector used was a variable UV detector and the wavelength was lambda = 210 nm. The method was statistically evaluated by a method evaluation demonstrating no systematic error. The uncertainty was 23.8 mumol/l and 11.5 mumol/l for MA and PGA, respectively. The limit of detection (LOD) of MA is 71.4 mumol/l and the LOD of PGA is 34.5 mumol/l, sufficiently low for the measurement of styrene exposure at a low exposure level. The present study indicates that reference values for MA and PGA are low. The fraction of reference values below LOD was 0.80 for MA and 0.66 for PGA; consequently, the reference values were described by a non-parametric one-sided tolerance interval. The 95% one-sided upper tolerance limits calculated for MA and PGA were 31.0 mumol/mmol creatinine and 20.1 mumol/mmol creatinine, respectively, with the coverage 0.95 +/- 0.045 for both metabolites. The method has been used for biological monitoring in several studies of environmentally and occupationally exposed subjects in concentrations up to 200 mumol/mmol creatinine for MA and 150 mumol/mmol creatinine for PGA.

Kinetics of styrene urinary metabolites: a study in a low-level occupational exposure setting in Singapore.

Biological monitoring of styrene exposure commonly involves measurement of styrene metabolites, mainly mandelic acid (MA) and phenylglyoxylic acid (PGA), in the urine of exposed subjects. Previous studies on the kinetics of styrene metabolites in urine were mostly conducted in a controlled environment on subjects exposed to high concentrations of styrene. In this study, we examined subjects exposed to low levels of styrene in a fiber-reinforced plastics (FRP) plant to see whether the excretion kinetics of styrene metabolites are similar under field conditions. Eight healthy Chinese male volunteers were exposed to styrene for 4 h with a mean environmental concentration of 11 ppm. Urine samples were collected continuously for 20 h after termination of the exposure and concentrations of urinary MA and PCA were determined. The results showed that MA was rapidly excreted in urine after the exposure, with a half-life of 2.1 h or 1.9 h when corrected with urine creatinine. The excretion of PGA followed that of MA and the half-life was 8.1 h or 5.1 h after correction with creatinine. The half-lives are considerably shorter compared to those in previous reports, suggesting that environmental factors, exposure conditions, or ethnic differences may affect the excretion kinetics of styrene metabolites. The fast excretion of styrene metabolites is also consistent with the observation that urine MA and PGA levels correlated better with the half-day time-weighted average (TWA) concentration of environmental styrene than with the whole-day TWA concentration. Our findings thus underscore the need for information on excretion kinetics in order to develop an appropriate biological monitoring scheme for specific exposure settings and subjects.

[A thin-layer chromatography method for determining the styrene metabolites mandelic and phenylglyoxylic acid in urine]. / Eine dünnschichtchromatographische Methode zur Bestimmung der Styrenmetaboliten Mandel- und Phenylglyoxylsäure im Urin.

A thin-layer chromatographic method is described for the determination of mandelic and phenyglyoxillic acid on silicagel (Silufol UV 254) after extraction from urine of styrene exposed workers. The quantitative determination was performed after eluting the spots. Phenylglyoxilic acid was measured at 255 nm and mandelic acid by derivative spectroscopically estimation of the .CH(OH).COOH -chromophore at 217 nm or by a three-wavelength mode, respectively. The recovery in urine was 80-104% for phenylglyoxilic acid and 99-105% for mandelic acid.

High-performance liquid chromatographic determination of glyoxylic acid in urine.

A high-performance liquid chromatographic (HPLC) method for the determination of urinary glyoxylic acid is proposed. The system is based on the precolumn derivatization of alpha-keto acids by means of phenylhydrazine, separation of the phenylhydrazone formed by HPLC and spectrophotometric detection at 324 nm. The method is precise and allows the determination of 0.5 mumol/l glyoxylate. The poor stability of glyoxylate under all conventional preservation conditions requires the analysis to be carried out as soon as possible after urine collection. Results of determinations on urine samples from healthy controls and from patients with idiopathic calcium stone disease and type I primary hyperoxaluria are reported.