Toxicology Letters

Published by Elsevier
Online ISSN: 0378-4274
Publications
Article
Risk has been defined by the World Health Organization International Program on Chemical Safety as "the probability of an adverse effect in an organism, system or (sub) population caused under specified circumstances by exposure to an agent". This article addresses some of the key components of this definition. Determining risk is dependent on expert judgement of the overall body of evidence relating to adverse effects or hazards. Such evidence can come from a range of experimental and observational studies and reports. Ideally determining risk to humans would be based upon evidence from humans rather than from experimental animals but this is generally not feasible. Even when epidemiological studies are available there can be difficulties in ascribing a disease to a causal chemical agent, and there are inevitable limitations in assessing exposure. Whilst probability refers to a mathematical or statistical quantification of the proportion of a population at risk of adverse effects, it is more common to compare point estimates of exposure with health-based guidance values.
 
Article
Trichloroethylene (TCE) is a common chemical pollutant that exists in air, soil, and drinking water. TCE exposure is known to cause severe hepatotoxicity; however, the mechanisms underlying TCE hepatotoxicity remain poorly understood. In a previous proteomics study, we found that TCE exposure up-regulated the expression of the inhibitor 2 of protein phosphatase 2A (I2PP2A), a potent and specific endogenous inhibitor of protein phosphatase (PP) 2A, in human hepatic L-02 cells. Here, we employed lentivirus-mediated RNA interference (RNAi) to knock down I2PP2A expression in L-02 cells and explored the potential role of I2PP2A in TCE-induced cytotoxicity. We found that TCE treatment of L-02 cells causes decreased cell viability, increased apoptosis and elevated I2PP2A mRNA and protein levels. TCE-treated L-02 cells were also found to have significantly reduced PP2A activity. Lentivirus-mediated I2PP2A knockdown partially prevented the decrease in viability and increased apoptosis induced by TCE treatment. Knockdown of I2PP2A in TCE-treated L-02 cells also suppressed the inhibition of PP2A activity and prevented caspase-3 activation. These data for the first time demonstrate that the up-regulation of I2PP2A could mediate, at least in part, TCE-induced liver cell toxicity through the inhibition of PP2A activity and caspase-3-mediated pathway, and suggest that I2PP2A may play a crucial role in mediating TCE hepatotoxicity.
 
Article
DNA polymerase eta (Polη), the product of the xeroderma pigmentosum variant gene, is required for translesion DNA synthesis, and plays a pivotal role in preventing genome instability after DNA damage induced by genotoxic agents. Studies have previously suggested a link between Polη and susceptibility to hydroquinone (HQ)-induced toxicity. To further address the role of Polη in the response of L-02 cells to HQ, we employed RNA interference to silence Polη expression in L-02 cells and examined the susceptibility of these Polη-deficient cells to the toxic effects of HQ. In this study, cell survival rate was determined using the MTT assay, DNA damage was determined by the Comet assay, apoptosis and cell cycle distribution were determined using flow cytometry, the mRNA expression levels of Polη were determined by real-time PCR, and the protein expression levels of Polη and γ-H2AX were determined by Western blot, γ-H2AX foci were visualized by confocal laser scanning fluorescence microscopy after cells were exposed to HQ at various concentrations for 24h in vitro. The results showed that stable Polη-knockdown cells were successfully constructed and more than 80% inhibition of Polη expression was confirmed. The results also showed that down-regulation of Polη led to a decrease in cell proliferation and an enhanced susceptibility to HQ-induced cytotoxicity. Polη-deficient cells were 2-fold more sensitive to HQ when compared with nonspecific siRNA control cells. Moreover, Polη-silenced L-02 cells treated with HQ displayed an increased level of DNA double-strand breaks as measured by olive tail moment, and an elevated DNA damage response as indicated by the induction of γ-H2AX. In addition, knockdown of Polη resulted in more enhanced apoptosis and more pronounced S phase arrest following HQ treatment. Together, these results show that Polη plays an important role in the response of L-02 cells to HQ-induced DNA damage.
 
Article
Hepatocellular carcinoma (HCC) is a major cause of morbidity and mortality in the world. The aim of the present study is to determine the antitumor effect of PF-04691502, a potent inhibitor of PI3K and mTOR kinases, on the apoptosis and angiogenesis of the hepatoma cancer cells. Our results indicate that treatment of cancer cells with PF-04691502 reduces cell viability and inhibits cell growth in a dose-dependent manner. PF-04691502 triggers apoptosis via a mitochondrial pathway, accompanied by activation of caspase-3, caspase-9, and Poly(ADP-Ribose) Polymerase (PARP). Pre-treatment of hepatoma cells with the caspase-3 inhibitor (z-DEVD-fmk) blocks the PF-04691502-induced death of these cells. In addition, growth factors-induced tube formation and the migration of HUVECs are markedly inhibited by PF-04691502 treatment. The mechanisms of anti-angiogenesis of PF-04691502 are associated with inhibiting the expression of VEGF and HIF-1α. Based on the overall results, we suggest that PF-04691502 reduces hepatocellular carcinoma cell viability, induces cell apoptosis, and inhibits cell growth and tumor angiogenesis, implicating its potential therapeutic value in the treatment of HCC.
 
Article
Oleanolic acid is a triterpenoid compound that has been shown to protect against liver injury produced by some hepatotoxicants. This study was designed to characterize the protective effects of oleanolic acid on carbon tetrachloride-induced hepatotoxicity, and the role of metallothionein in the protection. Oleanolic acid pretreatment (100-400 micromol/kg, s.c.) protected Sprague-Dawley rats and mice from carbon tetrachloride-induced liver injury in a dose- and time-dependent manner, as evidenced by serum alanine aminotransferase and sorbitol dehydrogenase activities, as well as by histopathology. The protection against carbon tetrachloride hepatotoxicity was not evident until animals were pretreated with oleanolic acid 12 h, and lasted for 72 h after a single injection. This suggests that the protection might be due to induction of some adaptive mechanisms. Metallothionein (MT), an acute-phase protein proposed to decrease carbon tetrachloride-induced liver injury, was dramatically induced following oleanolic acid treatment. To examine whether oleanolic acid protection is mediated through MT, MT-I and II knock-out (MT-null) mice were utilized. Oleanolic acid pretreatment increased MT levels in control mice (20-fold), but not in MT-null mice, however, it protected equally against carbon tetrachloride-induced hepatotoxicity in both control and MT-null mice. These data indicate that oleanolic acid is effective in protecting rats and mice from the hepatotoxicity produced by carbon tetrachloride, and the protection is not mediated through induction of MT.
 
Article
1,1-Dichloroethylene and 1,2-dichlorobenzene administered to mice produced liver and/or kidney damage which was quantified in this study by a histochemical method. The in vitro effect of sera obtained from these mice on antibody forming cell (AFC) response and natural killer (NK) cell activity was investigated in parallel with the assessment of sera tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) levels. 1,1-Dichloroethylene (100, 150 and 200 mg/kg) provoked liver and kidney damage. Peak kidney damage occurred 16 h after the dose was administered and at 24 h in the case of the liver. During the peak level of liver damage, a serum-borne immunosuppressive effect was also at its highest level. With respect to sera cytokine levels, an increase of TNF-alpha and IL-6 was detected earlier, i.e. 6 h after toxic administration, followed by a decrease that tended toward a baseline level. There was a relationship between the tissue damage induced by 1,1-dichloroethylene and the immunosuppressive effect of mice sera on AFC response and NK cell activity. 1,2-Dichlorobenzene (300, 500 and 600 mg/kg) provoked only liver damage. Peak liver damage severity was observed 48 h after toxic administration, whereas the highest serum-borne immunosuppressive effect was observed almost immediately, i.e. 6 h after administration. As regards sera cytokine levels, only TNF-alpha could be detected 6 h after administering 500 and 600 mg/kg doses of 1,2 dichlorobenzene. There was a relationship between the liver damage induced by 1,2-dichlorobenzene and the immunosuppressive effect of mice sera on the AFC response. In view of the above results, this study suggests that the immunosuppressive effect in sera of mice treated with 1,1-dichloroethylene and 1,2-dichlorobenzene may result from tissue damage, and that the increased levels of TNF-alpha and IL-6 in sera may contribute to this effect. Further studies are needed to clarify the factor(s) responsible, including transforming growth factor-beta1 (TGF-beta1) causing immunosuppression.
 
Article
Male Swiss OF1 mice were administered orally with a single dose (200 mg/kg) of 1,1-dichloroethylene (DCE). Examination of cryostat kidney sections stained for alkaline phosphatase (APP) revealed damage to about 50% of the proximal tubules at 8 h following DCE administration. Pretreatment with the anionic transport inhibitor probenecid by i.p., (0.75 mmol/kg, 30 min prior to and 10 min and 5 h following DCE administration) and with the gamma-glutamyltranspeptidase (GGT) inactivator acivicin by gavage and i.p. (50 mg/kg, 1 h and 30 min prior to DCE administration) failed to prevent DCE-induced renal toxicity. Pretreatment with the beta-lyase inactivator amino-oxyacetic acid (AOAA) by gavage (100 mg/kg, 30 min prior to and 10 min and 5 h following DCE administration), and with the renal cysteine conjugate S-oxidase inhibitor methimazole by i.p. (40 mg/kg, 30 min prior to DCE administration) reduced the number of damaged tubules by approximately 50 and 60%, respectively in mice treated with DCE. The results suggest that the DCE undergoes biotransformation by NADPH-cytochrome P450 to several reactive species which conjugate with glutathione (GSH). After arriving in the kidneys, the resulting conjugates reach the renal cells by a mechanism which depends on neither GGT, nor on an anionic transport system which is sensitive to probenecid. Once in the cells, the presumed GSH conjugates and/or their derivatives undergo secondary modification by beta-lyase and cysteine conjugate S-oxidase to reactive metabolite(s).
 
Article
To investigate the genotoxic effect of l,1-dichloro-1-fluoroethane (HCFC-141b), which was currently widely used as a cleaning solvent in the electronic parts industry and suggested as a potential reproductive effector, in vivo micronucleus tests were performed. Groups of 10 male and 10 female Sprague-Dawley rats were exposed, by inhalation (6h/day, 5 days/week) to the vapors of HCFC-141b for 13 weeks using whole body exposure chambers at the concentrations of 0 (control), 1500, 3000, and 6000 ppm. The micronuclei frequencies among the polychromatic erythrocytes (PCEs) and the percentage of polychromatic erythrocytes among the total number of erythrocytes were counted in the bone marrow of rats, and body weights, organ weights, histopathology, clinical chemistry and hematologic changes were also observed. Statistically significant and dose-dependant increases were found in the micronuclei frequencies in the male rats (P<0.01), yet not in the females. The decreases in the percentage of polychromatic erythrocytes among the total number of erythrocytes were also statistically significant (P<0.05) in both sexes of the high concentration groups. However, no exposure-related effects of toxicological significance were noted with respect to organ weights, clinical chemistry and histopathology. Apart from it, only slightly decreased mean corpuscular hemoglobin concentration (MCHC) was noted in the females of 6000 ppm group (P<0.05). These results suggest that HCFC 141b can induce the genetic effects, micronuclei in the rat bone marrows, especially in males, at earlier stages before the other general clinical and histopathologic changes occur if with more prolonged exposure.
 
Article
The effects of 1,1-dimethylhydrazine (UDMH) on several early events associated with lymphocyte activation were examined. The concentration of intracellular calcium ([Ca2+]i) and membrane potential of murine lymphocytes were found to be altered upon exposure to UDMH; [Ca2+]i was increased in murine thymocytes, while splenocytes exhibited membrane hyperpolarization. In addition, interleukin-2 receptor expression induced by in-vitro concanavalin A stimulation of murine splenocytes at 24 and 48 h in the presence of UDMH was not affected. UDMH may interfere with the ability of these two distinct lymphocyte populations to regulate normal ionic fluctuations, thus contributing to altered immune responsiveness.
 
Article
A renal toxicity of 4 h inhalation exposure to 1,1-dichloroethylene (vinylidene chloride; VDC) was studied in male Sprague-Dawley rats. Kidney wt./body wt. ratios, serum urea nitrogen and creatinine levels were significantly increased 24 h after exposure to 250 ppm or more of VDC. Histopathologic examination by light microscopy of hematoxylin and eosin (H&E)-stained sections revealed severe tubular necrosis with calcium deposits at the higher exposure concentrations. Specific staining for calcium oxalate was negative, indicating that biotransformation of VDC to oxalate is probably not responsible for its nephrotoxicity. Pretreatment with polychlorinated biphenyl (PCB) induced the level of renal cytochrome P-450. Phenobarbital (PBT) pretreatment did not alter the renal P-450 level, but both PCB and PBT pretreatments antagonized VDC nephrotoxicity. These pretreatments have also been reported to antagonize VDC-induced hepatotoxicity. In summary, inhalation of VDC is nephrotoxic in the rat; the mechanism of nephrotoxicity does not involve calcium oxalate formation, and the magnitude of nephrotoxicity does not correlate directly with the total amount of renal cytochrome P-450.
 
Article
The metabolic activation and covalent binding of 3-methylsulphonyl-2,2-bis (4-chlorophenyl)-1,1-dichloroethene (MeSO2-(14C)DDE)[correction of (14C)DDD)] were studied in vitro in the human adrenal gland. The mitochondrial bioactivation was twice as high as the microsomal, thus the study was focused on the mitochondria. The irreversible binding was time and protein dependent with apparent Km and Vmax values of 1.4 microM and 275 pmol/mg protein/h. As a comparison the activation of 2-(2-chlorophenyl)-2(4-chlorophenyl)-1,1-dichloroethane (o,p'-DDD), a well-known adrenolytic compound in humans, was studied. The irreversible binding of both these DDT metabolites was inhibited by metyrapone, indicating the involvement of cytochrome P-450. Addition of reduced glutathione (GSH) to the incubation medium decreased the irreversible binding of MeSO2-DDE significantly whereas the binding of o,p'-DDD was only slightly affected. The above findings suggest that MeSO2-DDE may be toxic to the human adrenal gland, although further study is necessary to assess this potential.
 
Article
Eosinophilia-myalgia syndrome (EMS) is thought to be caused by the intake of contaminated L-tryptophan products. 1,1'-Ethyliden bis[L-tryptophan] (EBT) is one of the contaminants in the L-tryptophan products and is thought to be a possible cause of EMS. In the present study we report that anti-EBT antibodies are generated by immunizing rabbits with the low molecular weight substance, EBT (molecular weight 434). These antibodies may be available tools for elucidating the mechanisms responsible for the outbreak and/or development of EMS.
 
Body composition of the volunteers in the study (average and range).
Comparison of experimentally observed (dots) and PBPK simulated (lines) time courses of difluoroethane in blood (, , ) and exhaled air (♦, , —) in volunteers exposed to 200 (, ♦) and 1000 (, ) ppm difluoroethane for 2 h during light physical exercise (50 W). Data are shown for the 10 subjects who completed both exposure conditions.  
Human blood:air and tissue:blood partition coefficients used in the PBPK model.
Cumulative excretion of fluorides in urine from 12 volunteers exposed for 2 h to 0, 200 and 1000 ppm of difluoroethane during light physical exercise (50 W). Geometric means and 5–95% confidence intervals were calculated assuming lognormal distribution. Times are given as mid-times.  
Article
The aim of this study was to determine the toxicokinetics of inhaled 1,1-difluoroethane (HFC-152a) in humans. Healthy volunteers were exposed to 0, 200 or 1000 ppm 1,1-difluoroethane for 2h at light exercise in an exposure chamber. Capillary blood, urine and exhaled air were sampled up to 22 h post-exposure and analyzed for 1,1-difluoroethane. Fluoride and other potential metabolites were analyzed in urine. Symptoms of irritation and central nervous system effects were rated and inflammatory markers were analyzed in blood. Within a few minutes of exposure to 200 and 1000 ppm, 1,1-difluoroethane increased rapidly in blood and reached average levels of 7.4 and 34.3 μM, respectively. The post-exposure decreases in blood were fast and parallel to those in exhaled air. The observed time courses in blood and breath agreed well with those obtained with the PBPK model. The PBPK simulations indicate a net uptake during exposure to 1000 ppm of 6.6 mmol (6.7%) which corresponds to the amount exhaled post-exposure. About 20 μmol excess fluoride (0.013% of inhaled 1,1-difluoroethane on a molar basis) was excreted in urine after exposure to 1000 ppm, compared to control. No fluorine-containing metabolites were detected in urine. Symptom ratings and changes in inflammatory markers revealed no exposure-related effects.
 
Article
Exposure to methoxychlor, an agricultural pesticide, has been associated with reduced testicular androgen secretion. However, methoxychlor is converted to 2,2-bis-(p-hydroxyphenyl)-1,1, 1-trichloroethane (HPTE) in the liver, which then acts as its biologically active metabolite. Both methoxychlor and HPTE have been credited with estrogenic properties and have a weak anti-androgenic activity. However, the exact mechanisms of steroidogenic enzyme inhibition remain to be clarified. In the present study, human and rat testis microsomes were employed to investigate the inhibitory activities of methoxychlor and HPTE on 17α-hydroxylase/17,20-lyase (CYP17A1). The CYP17A1 enzyme is critical for androgen biosynthesis and catalyzes conversion of progesterone into androstenedione. The results demonstrated that HPTE directly inhibited human and rat CYP17A1 activities, while methoxychlor had no effects on this enzyme activity even at a concentration of 100μM. The IC50 values of HPTE were 1.13±0.10 (human) and 6.87±0.13μM (rat), respectively. When HPTE was incubated with rat immature Leydig cells, it also inhibited CYP17A1 activity with an IC50 value of 6.29±0.1μM. Results of enzyme inhibition were supported by the observation that HPTE inhibited luteinizing hormone-stimulated 5α-androstane-3α, 17β-diol and testosterone secretion by immature Leydig cells with IC50 values of 6.61±0.03 and 3.78±0.003μM, respectively. The mode of action of HPTE on CYP17A1 activity was determined to be uncompetitive with the substrate progesterone. In conclusion, HPTE, the metabolite of MXC, directly inhibited human and rat testis CYP17A1 activities.
 
Article
The aim of this study was to determine the toxicokinetics and some effects of 1,1,1-trifluoroethane (HFC-143a) in humans. Nine male volunteers were experimentally exposed to 500ppm HFC-143a for 2h during light physical exercise (50W) in an exposure chamber. Blood, urine and exhaled air were sampled before, during and up to 19h after exposure and analysed for HFC-143a by gas chromatography. These data were described by a physiologically based toxicokinetic (PBTK) model. The electrocardiograms of the volunteers were monitored during exposure. Before, during and after exposure the volunteers rated symptoms related to irritation and CNS-symptoms on a visual analogue scale. Inflammatory markers (C-reactive protein, serum amyloid A protein, D-dimer, fibrinogen) and uric acid were analysed in plasma collected before and 21h after exposure. The exposures were performed after informed consent and ethical approval. The plasma concentration of HFC-143a increased promptly at start of exposure, and decreased in the same manner post-exposure. A stable level of 4.8+/-2.0 microM (mean+/-S.D.) was reached within 30min of exposure. The HFC-143a concentration in plasma and exhaled air decreased fast and in parallel when exposure was stopped. The urinary excretion of HFC-143a after exposure was 0.0007% of the inhaled amount. The half-time in urine, calculated from pooled data, was 53min. The experimental and simulated time courses in blood and exhaled air were in agreement. The simulated relative uptake during the exposure was 1.6+/-0.3%. The fibrinogen level in plasma had increased by 11% 1 day post-exposure. No statistically significant increase was seen for the other inflammatory markers or for uric acid. No effects of exposure were seen either in the electrocardiographic monitorings or as symptom ratings on the visual analogue scale.
 
Article
The quantitative relationship between the time-weighted average exposure by inhalation to 1,1,1-trichloroethane and excretion of metabolites in urine at the end of a work shift was investigated in 48 male printing workers exposed to the solvent at levels below 65 ppm. Male control subjects, 10 in all, were also investigated. Statistical analysis showed that there is a linear relationship between the two exposure indicators, suggesting that occupational exposure to 1,1,1-trichloroethane can be bio-monitored by means of urinalysis for either total trichloro-compounds or trichloroacetic acid. The correlation coefficient was higher for total trichloro-compounds (r = 0.86-0.89 depending on correction for urine density) than trichloroacetic acid (r = 0.50-0.71), which suggests that the former is a better indicator of exposure to 1,1,1-trichloroethane than the latter. The calculations based on the present study suggest that, at the end of the shift, only less than 2% of 1,1,1-trichloroethane absorbed will be excreted as metabolites in urine.
 
Article
Methoxychlor (MXC) is primarily used as a pesticide and widely present in the environment. The objective of the present study is to investigate the direct effects of MXC and its metabolite 2-bis(p-hydroxyphenyl)-1,1,1-trichloroethane (HPTE) on two isoforms of 11β-hydroxysteroid dehydrogenase (11β-HSD1 and 11β-HSD2) in vitro. Human liver microsome, rat testis microsome and adult Leydig cells were used for the measurement of 11β-HSD1 activity. Human placental and rat kidney microsomes were used for 11β-HSD2 activity. The IC(50) values on human 11β-HSD1 by MXC and HPTE were 1.91±0.07 and 8.88±0.08μM, respectively. HPTE inhibited rat 11β-HSD1 with IC(50) of 9.15±0.05μM, while MXC did not inhibit the enzyme. MXC and HPTE were competitive inhibitors of 11β-HSD1. HPTE also inhibited human and rat 11β-HSD2 with IC(50) values of 55.57±0.08 and 12.96±0.11μM, respectively, while MXC did not inhibit 11β-HSD2. In summary, our results showed that MXC and its metabolite HPTE inhibited both isoforms of 11β-HSD in a species- and chemical structure-dependent manner.
 
Article
1,1,1,2-Tetrafluoroethane (HFC-134a), which lacks ozone-depleting potential, has been selected as a replacement refrigerant for dichlorodifluoromethane (CFC-12) in air-conditioning and chiller applications, and as a propellant for pharmaceutical aerosols. A variety of paradigms using rats and rabbits have shown that HFC-134a has very little toxic potential. To strengthen the prediction of human hazard associated with HFC-134a exposure, we evaluated the rate of metabolism of this halocarbon by human hepatic microsomes relative to similar tissue preparations derived from rats and rabbits. Human microsomes defluorinated HFC-134a in a cytochrome-P-450-catalyzed reaction, common also to rat and rabbit. In absolute terms, the maximal rate of HFC-134a metabolism by human microsomes was very low, showed little interindividual variation among the samples evaluated (1.3 +/- 0.3 nmol F-/mg protein/15 min, mean +/- SD, n = 10), and did not exceed that in rat or rabbit liver microsomes. These findings support the argument that for characterization of HFC-134a toxicity, especially that which may be mediated by products of halocarbon metabolism, laboratory animals are an adequate surrogate for humans.
 
The physiologically based toxicokinetic (PBTK) model used to describe the kinetics of HFC-134a. From Nihlén and Johanson (1999). 
PBTK model parameters
Sensitivity analyses of the PBTK model of HFC-134a in (A) arterial blood and (B) exhaled air. Normalized sensitivity coefficients were calculated using the exposure condition of the actual experiment, i.e. 2 h exposure at a workload of 50 W. The bars delimit the range of each sensitivity coefficient as it changes over time. BW denotes the body weight, P the partition coefficient (liver:blood, vessel-rich group:blood, fat:blood, muscle:blood and blood:air), CL i the intrinsic metabolic clearance and VPR the ventilation-perfusion ratio. 
Average concentration (geometric mean) of HFC-134a in urine from 10 male volunteers
Article
The aim of this study was to determine the uptake and disposition of inhaled 1,1,1,2-tetrafluoroethane (HFC-134a) in humans. Ten male volunteers were exposed to 500 ppm HFC-134a (2 h, 50 W exercise). The HFC-134a levels were monitored in blood, exhaled air and urine up to 19 h post-exposure. The concentration in blood increased rapidly, reaching a plateau of 9.4+/-1.9 microM (mean+/-S.D.) within 30 min, followed by a fast post-exposure decrease. HFC-134a in expired air decreased rapidly as well and in parallel with that in blood. The post-exposure urinary excretion was 0.002% of the inhaled amount, and the half-time was 58 min (pooled data). A physiologically based toxicokinetic (PBTK) model was developed for further analysis. Experimental and simulated time courses in blood and exhaled air agreed well in all 10 subjects. Further, the late decay in blood was consistent with a wash-out of HFC-134a from fat tissues, with a half-time of 114+/-21 min. The simulated relative uptake during exposure was 3.7+/-0.5%. No remarkable findings were observed in the electrocardiographic recordings. Fibrinogen in plasma increased 1 day after exposure, whereas no effects on C-reactive protein, serum amyloid A protein, D-dimer or uric acid were seen. Further studies are needed to investigate the possible inflammatory response.
 
Article
Acetone potentiation of 1,1,2-trichloroethane (TCEA) hepatotoxicity was found to be strongly dose-dependent for both acetone and TCEA. An oral dose of 0.5 ml of acetone/kg was the most effective potentiating pretreatment when administered 16 h prior to TCEA. Surprisingly, higher doses of acetone did not potentiate the hepatotoxicity of TCEA, but may have decreased the severity of this toxicity. Acetone potentiation of hepatotoxicity was most pronounced at or near the hepatotoxic threshold for TCEA. While acetone treatment alone did not affect hepatic glutathione (GSH) stores the most consistent effect produced by acetone pretreatment was a significant potentiation of TCEA-induced depletion of hepatic GSH. These findings suggest that critical doses of acetone may enhance the subsequent hepatotoxicity of TCEA via mechanisms that affect the interaction of TCEA and hepatic GSH stores.
 
Article
Male Wistar rats were exposed to 200, 1000 or 2000 ppm of 1,1,2-trichloro-1,2,2-trifluoroethane vapour for 2 weeks, 5 day/week, 6 h daily and showed a dose-dependent accumulation of the compound in perirenal fat and brain. In the first week increased NADPH-diaphorase activity was observed and there was decreased cerebral glutathione at the highest dose. During the second week these effects disappeared while RNA tended to increase, and glutathione peroxidase activity to decrease at the highest dose. After a withdrawal period of 7 days, no fluorohydrocarbon was detected and the neurochemical effects had disappeared except that brain RNA at the highest exposure was below the control range.
 
Article
Synthetic musks are present in fine fragrances, cosmetics, soaps and laundry detergents. One of the most important synthetic musks is 7-acetyl-1,1,3,4,4,6-hexamethyl-1,2,3,4-tetrahydro-naphthaline+ ++ (AHTN; annual production: about 1500 metric tons). An increasing number of studies show that AHTN accumulates in surface water and fish and can be detected in human adipose tissue, as well in human milk. In the present report it is shown that a single high dose of AHTN leads to acute hepatic damage in rats, characterized by single cell necrosis, inflammation, swelling of liver parenchymal cells, and the presence of cytoplasmic condensations in the hepatocytes, while at the ultrastructural level disorganization of the rough endoplasmic reticulum and mitochondria as well as focal cytolysis is evident. Furthermore, evidence is presented that AHTN is not genotoxic, does not induce peroxisome proliferation, and does not lead to the induction of drug-metabolizing enzymes as phenobarbital and 3-methylcholanthrene do.
 
Article
1,10-phenanthroline (phen), flufenamic acid, and indomethacin are inhibitors of aldo-keto reductases 1C1 (AKR1C1), but only phen decreased the benzo[a]pyrene (BaP)-induced cytochrome P450 1a1 (Cyp1a1) protein level. Therefore the decrease in the BaP-induced Cyp1a1 protein level was not due to inhibition of Akr1c1, but to phen itself. Phen decreased the BaP-induced Cyp1a1 promoter activity and protein expression, and in contrast, it increased Cyp1a1 mRNA, resulting from an increase in mRNA stability. Phen is also known as a transition metal ion-chelator. Along with the phen study, we also found that Zn(2+), Fe(2+) and Cu(2+) increased Cyp1a1 mRNA and protein stability. Our results show that phen stabilized the mRNA of Cyp1a1, although it decreased cell viability. In addition, Zn(2+) and Fe(2+) highly neutralized phen's suppression of Cyp1a1 protein expression, but they only slightly neutralized phen's promotion of mRNA stability and suppression of cell viability, and had no effect on phen's suppression of promoter activity. Phen's effect on Cyp1a1 expression was reversible, which indicates that phen is non-covalently linked to its target. This report elucidates a new role for phen of stabilizing Cyp1a1 mRNA, and provides information for further studies on mRNA stabilization.
 
Article
Female SWR mice were treated with 1,2-dimethylhydrazine (DMH: 6.8 mg/kg i.p. injection) once weekly for up to 10 weeks, a dosing regime that produced tumours principally within the distal colon (Jackson et al., 1999. Carcinogenesis 20, 509-513). O(6)-Methylguanine (O(6)-MeG) levels, measured using a simple [3H]-based O(6)-alkylguanine-DNA alkyltransferase (ATase) inactivation assay, ranged from 0.6 to 16.7 fmol/microg DNA with: (i) highest levels in the distal colon; and (ii) higher levels after 68 mg/kg total DMH than 6.8 mg/kg DMH. Basal ATase activity varied between 0.97 and 1.22 fmol/microg DNA within the colon but was not associated with adduct levels or tumour induction. After 6.8 mg/kg DMH, the half life of O(6)-MeG in colonic tissue was 36-42 h whereas after 68 mg/kg DMH, t1/2 was approximately 25, 57 and 96 h in the proximal, mid and distal colon, respectively. Tumour induction was thus associated with the levels and persistence of O(6)-MeG in the distal colon.
 
Article
The possible co-carcinogenic effect of ingestion of industrial carbon black was examined in female Sprague-Dawley rats and female CF1 mice treated with 1,2-dimethylhydrazine (DMH) to induce adenocarcinomas of the colon. Carbon black was added to the diet at 2.05 g/kg feed and fed for 52 weeks. No differences (P greater than 0.05) in tumor incidences were seen in rats or mice. Simultaneous 2-year feeding experiments with carbon black alone showed no increase in the development of spontaneous tumors. These findings are consistent with the belief that carbon blacks are ineffective as carcinogens because of their adsorptive properties.
 
Article
Four consecutive intraperitoneal (i.p.) injections with 40 mg/kg of 1,2-dibromo-3-chloropropane (DBCP) reduced the in vitro accumulation of p-aminohippurate (PAH) and tetraethylammonium (TEA) by slices of renal cortex and increased blood urea nitrogen (BUN) concentration in both male and female rats, but elevated serum glutamic pyruvic transaminase (GPT) and glutamic oxaloacetic transaminase (GOT) activities in females only. Four consecutive treatments with 1,2-dibromoethane (EDB) reduced the accumulation of PAH in male rats, but failed to alter TEA accumulation, BUN concentration or GPT and GOT activities in rats of either sex. Single i.p. injections of EDB or DBCP (40 mg/kg, approximately one-half of the acute, i.p. LD50 values) were without effect on serum GPT and GOT activities, BUN concentration or the accumulations of PAH and TEA in male rats when measured 24, 48 or 96 h after treatment, except that PAH accumulation was reduced at 96 h. These results indicate that BUN and the accumulations in vitro of PAH and TEA by renal cortical slices are appropriate endpoints for studying DBCP nephrotoxicity. Measurements of serum GOT and GPT activities detected DBCP hepatotoxicity in female rats only. The nephrotoxicity of EDB was indicated by measurement of TEA accumulation only.
 
Article
The mutagenic properties of 1,2-dibromoethane (DBE) were studied in the Ames Salmonella typhimurium assay using the strains TA 1535 and TA 100. Kidney S9 fraction alone did not modify the direct mutagenic activity of DBE; but an addition of kidney S9 to liver S9 fraction yielded a higher mutagenic activity of DBE than with liver S9 fraction alone. Moreover, the addition of glutathione (GSH) to kidney S9 increased the mutagenic activity of DBE. Methimazole, a competitive inhibitor of the flavin-containing monooxygenase, reduced mutagenic activity suggesting that this enzyme may contribute to renal damage from DBE. No mutagens could be detected in the urine of rats treated with DBE.
 
Article
The environmental carcinogen 5-methylchrysene (5MC) can be activated to mutagenic metabolites by several isozymes of cytochrome P-450 (CYP). The resulting reactive diol-epoxides can be detoxified via conjugation by glutathione S-transferases (GST). We investigated whether expression of human glutathione S-transferase P1 (hGSTP1) would differentially protect cells against the cytotoxicity or mutagenicity of 5MC or its 1,2-dihydrodiol intermediate (5MC-1,2-diol) in V79MZ cells with activation via stably transfected human CYP1B1 (hCYP1B1) as compared to activation by human CYP1A1 (hCYP1A1). The parent compound 5MC was only 2-fold more cytotoxic in the CYP-expressing cell lines than in the V79MZ parental cell line, while 5MC-1,2-dihydrodiol was more than 30-fold more cytotoxic in CYP-transfected cells compared to V79MZ cells. Cells co-expressing either hCYP1B1 or hCYP1A1 together with hGSTP1 were 2-fold less sensitive to 5MC or 5MC-1,2-diol cytotoxicity than their CYP-only parent lines. The 5MC was highly mutagenic with similar potency in both hCYP-transfected cell lines, while 5MC-1,2-diol was 2-fold more mutagenic in hCYP1B1-transfected cells as compared to hCYP1A1 cells. Coexpression of hGSTP1 with either hCYP reduced 5MC or 5MC-1,2-diol mutagenicity by 1.4-4.5-fold compared to the corresponding hCYP-only expressing cell lines. The greater protection against mutagenicity of 5MC is in contrast to our previous studies in which we found greater protection by hGSTP1 against cytotoxicity than mutagenicity of benzo[a]pyrene in cells co-expressing hCYP1A1. Protection against mutagenicity by hGSTP1 was greater with activation of either compound by hCYP1B1 than with hCYP1A1 activation. These studies show that the relative efficacy of protection by hGSTP1 against mutagenicity of 5MC or 5MC-1,2-diol is in part determined by the specific CYP pathway that catalyzes activation to the toxic or mutagenic metabolites.
 
Article
Liver damage resulting from 4 h exposure to bromobenzene (BB) (146-957 ppm) and 1,2-dichlorobenzene (DCB) (245-739 ppm) as model toxicants was evaluated in rats. The modifications considered were the increases in serum glutamate dehydrogenase (GLDH) and sorbitol dehydrogenase (SDH) activities and the decreases in centrolobular liver-cell glucose-6-phosphatase (G6-Pase) staining intensity. A linear inverse relationship was established between the logarithmic values of blood enzyme activities and liver G6-Pase staining intensity. In addition, the levels of exposure to each test chemical were found to be linearly related to liver G6-Pase staining intensity and to the logarithmic values of blood enzyme activities.
 
Article
The present study was performed to investigate comparatively the toxic effects of inhalation exposure of 1-bromopropane, 2-bromopropane, and 1,2-dichloropropane on reproductive physiology, particularly on the estrous cycle and spontaneous ovulation in female F344 rats. The rats received inhalation exposure to different halogenated propanes, and were exposed daily for 8 h throughout almost 3 weeks to 0,50,200 and 1000 ppm of 1-bromopropane or 2-bromopropane, or to 0,50,100 and 200 ppm of 1,2-dichloropropane. Throughout the exposure period of 1-bromopropane or 2-bromopropane, the ratio of the number of estrous cycle of 6 days or longer to the number of all cycles in both 1000 ppm groups were about two-fold the ratio in each control group, however, no significant difference was found between the ratios of exposed and control groups. The ratios of such long estrous cycles in groups exposed to 100 or 200 ppm of 1,2-dichloropropane were six- to seven-fold higher than that of the control group. These ratios in exposed rats differed significantly from those of controls. The number of ovulated ova in rats exposed to 1,2-dichloropropane decreased in a dose-dependent manner, and the number of ovulated ova in the 200 ppm group was significantly different from that of control rats. Such significant changes in ovulation were not observed in rats exposed to 1-bromopropane or 2-bromopropane. The absolute and relative weights of the ovaries and uterus in rats exposed to three halogenated propanes were not significantly different from those in each control. Therefore, the present study clarified that: (1) 1,2-dichloropropane prolonged the length of the estrous cycle and inhibited spontaneous ovulation in F344 rats; and (2) the potency of 1,2-dichloropropane to disturb the female reproductive physiology appeared to be greater compared with that of 1-bromopropane and 2-bromopropane under the present conditions of inhalation exposure.
 
Article
Human polymorphonuclear leukocytes (PMNL) were exposed to 3-phenylamino-1,2-propanediol (PAP) or to its mono- or dioleylesters. The dioleylester of PAP (140 microM) slightly increased the production of reactive oxygen metabolites (ROM) from 8 +/- 2 mV to 18 +/- 3 mV, whereas PAP and its mono-oleylester were without any effect on ROM production in PMNL. None of the compounds were able to modulate formyl-Methionyl-Leucyl-Phenylalanine-induced production of ROM. Moreover, PAP did not have any effect on the production of ROM induced by phorbol myristate acetate (PMA). On the other hand, both mono- and dioleylesters of PAP dose-dependently inhibited PMA-induced production of ROM. The dioleylester of PAP, at a concentration of 14 microM, inhibited PMA-induced production of ROM by 79%, whereas a 2.3 mM concentration of mono-oleylester of PAP was required for 65% inhibition of PMA-induced production of ROM. Moreover, both esters of PAP shifted the peak production of ROM by PMA to a later time-point. Our results suggest that mono- and dioleylesters of PAP modulate the responses of PMNL upon stimulation with PMA, whereas PAP does not have any effect on PMA-induced ROM production in PMNL.
 
Article
Two measurable indices of toxicity that can be correlated with exposure to propylene glycol dinitrate (PGDN) were evaluated along with its metabolism. Propylene glycol dinitrate was administered by rapid i.v. injection to male Fischer-344 rats. These rats demonstrated a dose-response of blood pressure (BP) to doses of PGDN ranging from 0.1 to 30 mg/kg; the maximum fall in systolic BP occurred within 1 min of dosing. The i.v. administration of PGDN to separate groups of animals resulted in an increase in cerebral blood flow that was correlated with the dose, but a clear dose-response was not obtained.
 
Article
The ability of acetone and 3 other ketone vapours to influence the hepatotoxicity of inhaled 1,2-dichlorobenzene (DCB) was examined in rats and mice. Methylethylketone, methylisobutylketone or cyclohexanone increased liver cytochrome P-450 content and glutathione-S-transferase (GST) activity, but did not affect serum glutamate dehydrogenase (GLDH) activity in rats. Pre-exposure to these ketones enhanced DCB-induced increase in serum GLDH activity (8-63-fold), while the increases in cytochrome P-450 content (33-86%) and GST activity (42-64%) were identical to those resulting from exposure to ketones alone. Each of the 3 levels of exposure to acetone elicited cytochrome P-450 and GST responses comparable with those caused by the other ketones. In spite of that, acetone pre-exposure potentiated (4785 ppm), reduced (10670 ppm) or suppressed (14790 ppm) DCB-induced liver toxicity. In mice, the 3 ketones mentioned above interacted with DCB on centrolobular liver glucose-6-phosphatase (G-6-Pase) while acetone pre-exposure elicited an interactive G-6-Pase response in the mediolobular area alone, suggesting topographic change.
 
Article
Male rats treated with a single dose of 1,2-dibromo-3-chloropropane (DBCP) were tested for their ability to carry out the synthesis of liver proteins. In animals treated for 12 h, we found no changes in the uptake of [14C]orotic acid into liver RNA or the uptake of [3H]leucine into liver or serum protein. Uptake of [3H]leucine into the soluble fraction of the enlarged liver increased in proportion to liver size, while the uptake of [14C]orotic acid was unchanged. Examination of the ultrastructure of liver cells from rats treated for 12, 24, or 48 h revealed that the structure of the rough and smooth endoplasmic reticulum (RER; SER) were modified. An absence of ordered stacks of the RER and the presence of tangled nets of SER were noted.
 
Article
Oltipraz (OPZ) is a known inducer of glutathione S-transferases and a mechanism-based inhibitor of cytochrome P450 1A2. Given the detoxification characteristics of this compound, the transcriptional effects of OPZ, along with the related naturally occurring compounds 3H-1,2-dithiole-3-thione (D3T) and sulforaphane (SF), were examined by gene expression profiling in murine BV-2 microglial cells, a neuronal macrophage cell type that mediates inflammatory responses in the brain. We show that the three compounds generate largely overlapping transcriptional changes in genes that are associated with detoxification and antioxidant responses. In addition, induction of an antioxidant/detoxification response in the microglial cells by OPZ, D3T, or SF was also able to protect cells from H2O2 -induced toxicity and to attenuate the production of reactive oxygen species in response to lipopolysaccharide treatment of cells. These results show that OPZ, D3T, and SF activate overlapping changes in gene expression and that they can regulate detoxification/antioxidant responses in multiple cells types, including cell types known to have a role in the production of oxidative stress.
 
Article
The effect of a single intragastric injection of 1,2-dibromoethane was investigated in kidneys of male Wistar rats. DNA synthesis as measured by the incorporation of tritiated thymidine was found to be approximately 5 times greater than that of controls 20-30 h after treatment. DNA synthesis was followed by a striking increase in the mitotic activity with a maximum at 30 h. The labeling and mitotic activities, after an initial increase, fell rapidly 48 h after treatment even though they were still higher than those of control animals. 1,2-Dibromoethane-induced cell proliferation is not a regenerative response because at the dose used in this study, no tubular necrosis was observed by histologic examination.
 
Article
The present study was carried out to investigate the species differences in the nephrotoxic response to S-(1,2-dichlorovinyl)glutathione (DCVG) using rats, hamsters and guinea-pigs. DCVG was given intraperitoneally in physiological saline to groups of 5 animals at doses 0, 165 and 330 mumol/kg. Urine was collected for 24 h and the animals were then sacrificed. Significantly increased levels of urinary glucose, N-acetyl-beta-D-glucosaminidase, gamma-glutamyl transpeptidase, proteins and blood urea nitrogen were observed in rats at both dose levels of DCVG. An increase, but not of similar magnitude, of these biochemical parameters was noted in hamsters only at the higher dose of DCVG. Guinea-pigs showed significant increases in these biochemical parameters at the lower dose, but not at the higher dose. Light-microscopic studies showed increasing proximal tubular necrosis (PTN) in rats with increasing dose of DCVG, but PTN involving straight tubules only was observed at the higher dose in hamsters. PTN was again observed in guinea-pigs at the lower dose, but not at the higher dose of DCVG.
 
Article
Male C57BL/Icrf alpha t mice receiving long courses of weekly subcutaneous injections of 8.25 or 12.5 mg/kg, 1,2-dimethylhydrazine (DMH) developed perianal carcinomas as well as tumours at other sites. Tumours of the perianal gland or its associated hair root sheath and overlying skin included sebaceous gland adenocarcinomas, often with squamous metaplasia, analogous to those in the rat ear, and dysplasia, polyp formation and squamous cell carcinomas (SCC). Of 152 mice treated, 88 remained after 30 injections, and 10 out of a total of 14 tumours developed after 30 treatments. Perianal SCC without definite perianal gland association developed in 2 other animals.
 
DEB concentrations in blood of mice (A, a) and rats (B, b) after exposures (6h) to constant concentrations of BD. Twelve mice, subdivided in 2 groups of 6 animals, were exposed together to each concentration. Four rats were exposed together to BD concentrations of 2.4ppm, 21ppm, and above and two times 4 rats were exposed together to concentrations of 1.1ppm, 5.6ppm, and 11ppm. Whole exposure ranges are represented by the letters A and B, extracts by a and b. Symbols give measured data: (▵) (±)-DEB; (×) meso-DEB; (A, a) two values per exposure concentration (each from pooled blood of 6 mice), identical values at 3 exposure concentrations; (B, b) means±SE (deviation bars, except when smaller than the symbol size) from 4 rats or from 8 rats. Limits of detection were 0.001μmol/l (mouse blood) and 0.0003μmol/l (rat blood). Lines are one-phase exponential association functions (y=y∞[1−e−kx]) that were fitted to the data. The variable x stays for the BD exposure concentration (ppm). Maximum calculated (±)-DEB concentrations in blood (y∞) are 1.92μmol/l in mice (95% confidence intervals 1.62–2.22) and 0.101μmol/l in rats (95% confidence intervals 0.091–0.11). The values of k for (±)-DEB were 0.00284/ppm BD in mice (95% confidence intervals 0.0017–0.0040) and 0.0122/ppm BD in rats (95% confidence intervals 0.0085–0.016). Maximum meso-DEB concentrations in blood (y∞) were 0.0977μmol/l in mice (95% confidence intervals 0.029–0.17) and 0.00478μmol/l in rats (95% confidence intervals 0.0042–0.0054). The values of k for meso-DEB were 0.00175/ppm BD in mice (95% confidence intervals 0.000–0.0043) and 0.0146/ppm BD in rats (95% confidence intervals 0.0087–0.020). The squares of the correlation coefficients (R2) for (±)-DEB were 0.981 in mice and 0.975 in rats. For meso-DEB, they were 0.881 in mice and 0.949 in rats.
Product ion spectrum of the bis(N,N-diethyldithiocarbamoyl) ester of DEB (m/z=385.2). Presumed products are shown. The instrumental parameters are given in the text.
Article
The important industrial chemical 1,3-butadiene (BD; CAS Registry Number: 106-99-0) is a potent carcinogen in B6C3F1 mice and a weak one in Sprague-Dawley rats. This difference is mainly attributed to the species-specific burden by the metabolically formed 1,2:3,4-diepoxybutane (DEB). However, only limited data exist on the DEB blood burden of rodents at BD concentrations below 100 ppm. Considering this, DEB concentrations were determined in the blood of mice and rats immediately after 6h exposures to various constant concentrations of BD of between about 1 and 1200 ppm. Immediately after its collection, blood was injected into a vial that contained perdeuterated DEB (DEB-D(6)) as internal standard. Plasma samples were prepared and treated with sodium diethyldithiocarbamate that derivatized metabolically produced DEB and DEB-D(6) to their bis(dithiocarbamoyl) esters, which were then analyzed by high performance liquid chromatography coupled with an electrospray ionization tandem mass spectrometer. DEB concentrations in blood versus BD exposure concentrations in air could be described by one-phase exponential association functions. Herewith calculated (±)-DEB concentrations in blood increased in mice from 5.4 nmol/l at 1 ppm BD to 1860 nmol/l at 1250 ppm BD and in rats from 1.2 nmol/l at 1 ppm BD to 92 nmol/l at 200 ppm BD, at which exposure concentration 91% of the calculated DEB plateau concentration in rat blood was reached. This information on the species-specific blood burden by the highly mutagenic DEB helps to explain why the carcinogenic potency of BD in rats is low compared to that in mice.
 
Article
7-Hydroxy-5-methyl-3-phenyl-6,7,8,9-tetrahydropyrido[3',2':4,5]imidazo[1,2-a]pyrimidin-5-ium chloride (PhIP-M1) is a newly identified intestinal microbial metabolite from the food carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). Although the mutagenic potential of the endogenous N-hydroxy PhIP derivate has been reported, the risks associated with PhIP-M1 have not yet been explored. In this work, the cytotoxic and genotoxic effects originating from PhIP-M1 were assessed in the epithelial intestinal Caco-2 cell line. PhIP-M1 significantly decreased in a time- and dose-dependent manner mitochondrial dehydrogenase activity and protein synthesis, with IC50 values of, respectively, 180+/-39.4 and 173+/-20.3 microM after 24h, and 33.8+/-3.5 and 37.3+/-10.9 microM after 72 h. Apoptosis within the concentration ranges of cytotoxicity was confirmed by morphological examination, DAPI nuclear staining and annexin V staining. PhIP-M1 provoked cell cycle arrest, characterized by a significant increase in the number of nucleoids in the G2/M phase. A dose-dependent increase in DNA damage, as quantified by the alkaline comet assay, was observed after 3h in the 50-200 microM range. Because these PhIP-M1-induced genomic and cellular events may contribute to the carcinogenicity of PhIP, the potency of the colon microbiota to bioactivate PhIP must be included in future risk assessments.
 
Article
In this study the gonadotoxic effects of 2-hexanone administered as a pretreatment and/or 1,2-dibromo-3-chloropropane administered as a challenge on the activity of several key steroidogenic enzymes in the rat testis were examined. Despite the absence of an effect when either treatment was administered individually, the pretreatment/challenge combination inhibited the steroidogenic capacity of the rat testis. The specific activity of testicular 17 alpha-hydroxylase was significantly reduced (P less than 0.05), with respect to the control, following the pretreatment/challenge combination. There were no significant differences between the control and the individual treatments. The inhibition of 17 alpha-hydroxylase activity occurred in the absence of significant differences in testis weight and testicular protein content. This inhibition of testicular steroidogenic enzyme activity was specific as the activity of another testicular enzyme, namely C17,20-lyase, was not affected by any treatment. The decline in rat testicular 17 alpha-hydroxylase activity 18 h after the 1,2-dibromo-3-chloropropane challenge precedes the reported alterations in the seminiferous epithelium following this same treatment. The results of the present study indicate that the steroidogenic cell of the testis, i.e. the Leydig cell, is a potential site for the primary toxic effects of these agents in the rat testis.
 
Article
The purpose of this experiment was to examine the development of colon tumors after the injection of a single dose of 1,2-dimethylhydrazine (DMH) in rats. Male Fischer-344 rats were injected with 0, 10, 30, 100 or 200 mg/kg of DMH dihydrochloride. No tumors were seen after 3 or 5 months at any dose, but were seen after 7 or 9 months. At 9 months, tumors were induced in a dose-dependent manner, with 64.7% of rats receiving the 200 mg/kg dose developing tumors. This study shows that a high incidence of colon tumors can be induced by a single dose of DMH after a 9-month latency period.
 
Article
1-Keto-1,2,3,4-tetrahydrophenanthrene (THP-1) was reported by Cook et al. in 1933 as the first synthetic estrogen. Estrogenic activity was assessed by the induction of vaginal cornification in ovariectomised rats. The corresponding 4-isomer (THP-4) was shown to be inactive. Both chemicals have been re-synthesised and assessed for hormonal activity. Each chemical bound weakly and to the same extent to isolated estrogen receptors, but only at high concentrations. However, they each lacked estrogenic or anti-estrogenic activity when evaluated in vitro using a yeast hER assay, and both failed to induce vaginal cornification or uterotrophic effects in ovariectomised rats. THP-1, and to a lesser extent THP-4, were shown to possess weak androgenic and anti-androgenic activity in vitro when evaluated using an hAR yeast assay. Estrogenic activity for bisphenol A (BPA) was subsequently demonstrated by [Dodds and Lawson, Synthetic, oestrogenic agents without the phenanthrene nucleus, Nature 137, (1936)] using the same ovariectomised rat protocol, and this activity has been confirmed and supplemented by positive uterotrophic effects for BPA in the same bioassays. The present results illustrate the complexity of deriving conclusions regarding the hormonal activities of chemicals. First, some activities observed in isolated hormonal receptor binding assays may not be expressed in functional hormonal assays. This indicates the need for functional hormonal assays in any screening programme. Second, that activities observed for a chemical in one hormonal assay may not be reflected in related hormonal assays. This indicates the need to define assay protocols with some precision when incorporating them into screening batteries. Finally, that some literature reports of hormonal activity for chemicals may not be capable of independent confirmation under apparently identical conditions of test. This illustrates the need to use lists of hormonally active chemicals with care.
 
Article
The short-term (24 h) effects of 10 mg/kg of the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) on cat catecholamine levels of heart, adrenal gland, retina, and caudate nucleus were studied. Significant differences in catecholamines, including adrenomedullar adrenaline, heart noradrenaline, and retinal dopamine, were observed. No differences were found in caudate nucleus dopamine. In this organ, the levels of dopamine metabolites, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) were significantly lower after MPTP treatment. Biochemical changes caused by MPTP with respect to its effects on the peripheral catecholaminergic organs are discussed.
 
Article
The effect of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), a neurotoxin which selectively destroys the nigrostriatal dopaminergic neurons and produces Parkinson's disease-like syndrome, on striatal dopamine receptors was determined in a mouse strain known to be very sensitive to the neurotoxic effect of MPTP. Daily intraperitoneal administration of MPTP (30 mg/kg) for 7 days to male C57BL/6 mice reduced the concentration of striatal dopamine by 90%. This decrease in dopamine concentration was not associated with changes either in the receptor density (Bmax) or the apparent dissociation constant (Kd) of [3H]spiroperidol to bind to striatal dopamine receptors. It is concluded that in spite of large decrease in striatal dopamine concentration by MPTP the dopamine receptors labeled with [3H]spiroperidol remain intact.
 
Article
A sensitive assay for human plasma BzAO, involving the conversion of 14C-benzylamine to 14C-benzaldehyde, was developed. MPTP and several of its analogues were found to be competitive inhibitors of the enzyme. Ki values for the MPTP analogues in the presence of human plasma BzAO were determined. The analogues had a different rank order of inhibition of human plasma BzAO compared with the rank order of inhibition of bovine plasma BzAO found previously. MPTP and 1-methyl-4-(2-methylphenyl)-1,2,3,6-tetrahydropyridine (2'-CH3-MPTP), which are potent nigrostriatal toxins, were weak inhibitors of human plasma BzAO.
 
Article
The parkinsonism-inducing neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) has been shown to possess marked species as well as strain differences in toxicity on central catecholaminergic systems. In the present study the effects on the peripheral sympathetic nervous system following treatment with MPTP, as well as its metabolite 1-methyl-4-phenylpyridine (MPP+) and the catecholamine neurotoxin 6-hydroxydopamine (6-OHDA) were studied in mice of the NMRI and C57 BL/6 strains, two strains that possess marked difference in MPTP toxicity on central catecholaminergic neurons. No strain differences in the depletions of noradrenaline (NA) in iris and heart auricula and of NA and dopamine (DA) in superior cervical ganglion or in the reduction of the in vitro [3H]NA uptake in iris or heart auricula were found following MPTP treatment (2 X 40 mg/kg s.c., 2 and 7 days). Treatment with the NA uptake blocker desipramine (DMI) did not affect the MPTP-induced NA depletion in either strain. Following treatment with MPP+ (30 mg/kg i.v., 7 days) no differences in the two strains were seen on the reduction of NA levels in iris and heart auricula or decrease in [3H]NA uptake. In addition, no differences were found on NA levels in iris and heart auricula after 6-OHDA treatment (15 mg/kg i.v., 7 days). The data indicate that in the NMRI and C57 BL/6 mice peripheral NA neurons do not possess any notable strain difference in the vulnerability to MPTP or in the mechanism of action of MPTP.
 
Top-cited authors
Chunying Chen
  • Chinese Academy of Sciences
Thomas Heberer
  • Technische Universität Berlin
Tian-Cheng Wang
  • Peking University Third Hospital
Yuliang Zhao
  • Chinese Academy of Sciences
Xiao Chen
  • Wuhan University