Styrene trimer may increase thyroid hormone levels via down-regulation of the aryl hydrocarbon receptor (AhR) target gene UDP-glucuronosyltransferase.

BACKGROUND: Styrene trimers (STs) are polystyrene-container-eluted materials that are sometimes detected in packaged foods. Although the possible endocrine-disrupting effects of STs, such as estrogenic activities, have been reported, their potential thyroid toxicity, such as that caused by the related endocrine disruptor 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), has not been studied in detail. OBJECTIVE: Using wild-type and aryl hydrocarbon receptor (Ahr)-null mice, we investigated whether 2,4,6-triphenyl-1-hexene (ST-1), an isomer of STs, influences thyroxin (T(4)) levels in the same manner as TCDD, which induces UDP-glucuronosyltransferase (UGT) via the AhR, resulting in a decrease in T(4) levels in the plasma of mice. METHODS: Both wild-type and Ahr-null mice (five mice per group) were treated for 4 days by gavage with ST-1 (0, 32, or 64 micromol/kg). RESULTS: High-dose (64 micromol/kg) ST-1 decreased the expression of AhR, cytochrome P450 (CYP) 1A1/2, UGT1A1/A6, and CYP2B10 mRNAs and the enzyme activity for CYP1A and UGT1A only in the wild-type mice. This dose decreased AhR DNA binding, but paradoxically increased AhR translocation to the nucleus. In contrast, a high dose of ST-1 increased T(4) levels in the plasma in wild-type mice but did not influence T(4) levels in AhR-null mice. CONCLUSIONS: Although ST-1 treatment might cause an increase in AhR levels in the nucleus by inhibiting AhR export, this chemical down-regulated AhR mRNA, thus leading to down-regulation of AhR target genes and an increase in plasma T(4) levels.

Quaternary (triphenyl-) phosphonium compounds: Environmental behavior and toxicity.

An analytical method based on high resolution mass spectrometry coupled with liquid chromatography (LC-HRMS) for 25 quaternary phosphonium compounds (QPCs) and derived phosphine oxides (POs) was developed and validated. To investigate the occurrence and fate of QPCs in the aquatic environment, water, suspended solids and sediments from the rivers Rhine and Elbe (upper and middle Elbe as well as tidal Elbe) were analyzed, as well as samples from tributaries bearing significant loads of QPCs. For the first time, the quaternary phosphonium compound tetrabutylphosphonium (Bu4P+) was detected. In the river Elbe concentrations were determined of up to 4700 ng/L (surface water) and 1000 µg/kg (sediment), respectively. Analysis of a time series of suspended solids (2005-2015) showed that QPCs have been present in the Elbe and Rhine catchment for at least one decade, with partly rising tendency. A degradation experiment with Rhine sediment revealed that triphenylphosphonium compounds (R-Ph3P+) and Bu4P+ are persistent in contact with sediment and suspended solids and tend to sorb onto sediment particles. Toxicological studies (reactive oxygen species (ROS) after substance exposure, Ames test, Micronucleus test, determination of cytotoxicity) with selected QPCs confirmed that all of them exhibit cytotoxicity and some even genotoxic potential at elevated concentrations, which emphasizes the need for an emission regulation of these compounds.

Hepatocarcinogenicity of polychlorinated terphenyl (PCT) in ICR mice and its enhancement by hexachlorobenzene (HCB).

Studies were made on the carcinogenic effects of hexachlorobenzene (HCB) and polychlorinated terphenyl (PCT) singly and in combination when given orally to mice for 24 weeks. PCT alone induced liver tumors, nodular hyperplasias and hepatocellular carcinomas. HCB alone had no effect, but it enhanced the induction of liver tumors by PCT.

Organophosphorus-induced neurotoxicity in the absence of neuropathy target esterase inhibition: the effects of triphenyl phosphine in the European ferret.

Abou-Donia et al. (in Toxicologist, Vol. 30, 1996) have reported that repeated oral administration of the organo-phosphorus compound triphenyl phosphine (TPPn) to the domestic chicken results in neuropathological changes in the spinal cord and peripheral nerves, accompanied by ataxia and paralysis. This study also noted that single doses of TPPn resulted in no inhibition of the enzymes neuropathy target esterase (NTE) and acetylcholinesterase (AChE). We undertook the present study to determine the biochemical, neuropathological, and clinical effects of single doses of TPPn in the European ferret, a mammalian species shown to be susceptible to organophosphorus-induced neurotoxicity. Eight 12-week-old ferrets were each injected subcutaneously with either 250 mg TPPn/kg bw or 500 mg TPPn/kg bw, or with the peanut oil/ethyl ether vehicle. Twenty-four h after dosing, the brains of 5 animals from each dose group were examined for NTE and AChE activities. The remaining 3 animals in each group were observed for 6 days for the development of clinical signs, after which their brains were processed for the presence of axonal degeneration using the Fink-Heimer silver impregnation method. Single injections of TPPn had no effect on the activities of whole-brain NTE or AChE 24 h after injection. The animals observed for clinical signs showed increasing trunk and hindlimb ataxia beginning 4 days after injection, culminating in fore-and hindlimb paralysis 6 days after injection. All brains exposed to either dose of TPPn showed widespread axonal degeneration extending from the brainstem and cerebellum into midbrain and forebrain areas. The results of this study support the hypothesis that TPPn-induced neurotoxicity is a separate and distinct form of organophosphorus-induced neurotoxicity not dependent on NTE inhibition, and therefore not a variant of organophosphorus-induced delayed neurotoxicity (OPIDN).

Polychlorinated terphenyls: alterations in liver morphology and induction of cytochrome P-450.

Exposure of male rats to the polychlorinated terphenyl (PCT) mixtures Aroclor 5460 and Aroclor 5432 containing 60% and 32% (w/w) of chlorine, respectively, showed that the PCT mixture with a low degree of chlorination, Aroclor 5432, was a potent inducer of liver microsomal cytochrome P-450, aryl hydrocarbon hydroxylase (AHH) and 7-ethoxyresorufin O-deethylase whereas Aroclor 5460 and the unchlorinated isomers o-, m- and p-terphenyl were weak inducers. Ultrastructurally, proliferation of SER but not RER, frequent occurrence of lysosomes containing partially degraded lipid material as well as an increased number and size of cytoplasmic lipid droplets were observed. These changes were most pronounced in Aroclor 5432 treated rats. Competition experiments with [3H]2,3,7,8-tetrachloro-p-dibenzodioxin (TCDD) for binding to the cytosolic receptor protein indicated the presence in Aroclor 5432, but not in Aroclor 5460, of components with receptor affinity. These components represent only a minor fraction of the mixture as judged by the 1900-fold excess necessary to displace 50% of the specific [3H]TCDD binding. Based on the biochemical results and the ultrastructural findings it is concluded that the PCT mixture Aroclor 5432 is a mixed type inducer of hepatic cytochrome P-450 in the rat. The presence in Aroclor 5432 of compounds capable of inducing AHH in vivo and of binding to the TCDD-receptor might be highly relevant with regard to the potential toxicity in view of the apparent correlation between affinity for the TCDD-receptor, induction of AHH activity and toxic properties for chlorinated aromatic hydrocarbons.

Cytotoxicity evaluation of a new radiopaque resin additive--triphenyl bismuth.

Triphenyl bismuth (Ph3Bi) is a promising new additive for making biomedical resins visible on x-ray images. We evaluated the cytotoxicity of Ph3Bi, both alone and as a component of a denture resin, as an initial step in determining its biocompatibility. These experimental materials were compared with several types of dental materials that are in current clinical use (PMMA denture acrylic resin, two photo-cured sealants, and two glass-ionomer cements). Human embryonic lung fibroblast tissue cultures (WI-38 cells) were exposed to 24-hour aqueous extracts of the materials. Changes in cell growth, cell viability, and the visual appearance of cells were used for the assessment of toxic response. Only a slight degree of cytotoxicity was observed for Ph3Bi, both alone and in combination with self-cured PMMA. All clinical materials showed a higher level of cytotoxicity than did Ph3Bi. The sealants and cements exhibited the most cytotoxicity and PMMA acrylic the least. The cytotoxicity of PMMA was elevated slightly by inclusion of Ph3Bi, probably due to decreased monomer conversion. When stored in water, the already low levels of cytotoxicity of both PMMA and PMMA with added Ph3Bi were reduced even further. From these results, we can predict a high level of safety for Ph3Bi as a radiopaque additive for biomedical resins. Any toxicity associated with Ph3Bi-containing resins can be reduced or avoided by prior extraction. Alternatively, curing conditions can be selected that would drive the polymerization reaction to a higher level of conversion.

Effect of triphenyldioxane on phase I xenobiotic metabolism enzymes in the liver of rats and rabbits.

We studied the effect of triphenyldioxane on phase I xenobiotic metabolism enzymes in the liver of rats and rabbits. Total cytochrome P450 content, protein concentration, and catalytic activity of CYP2B, CYP3A, and CYP2C isoforms were measured. Triphenyldioxane significantly increases specific activity of CYP2B and CYP2C in the liver of rats and rabbits, respectively. Immunoblotting analysis of microsomal enzymes in the liver of animals showed that the increase in specific activity of CYP is related to high content of apoenzymes. We showed for the first time that rats and rabbits are characterized by interspecies differences in the induction of cytochrome P450 isoforms under the influence of triphenyldioxane.

Synthesis and identification of small molecules that potently induce apoptosis in melanoma cells through G1 cell cycle arrest.

Late-stage malignant melanoma is a cancer that is refractory to current chemotherapeutic treatments. The average survival time for patients with such a diagnosis is 6 months. In general, the vast majority of anticancer drugs operate through induction of cell cycle arrest and cell death in either the DNA synthesis (S) or mitosis (M) phase of the cell cycle. Unfortunately, the same mechanisms that melanocytes possess to protect cells from DNA damage often confer resistance to drugs that derive their toxicity from S or M phase arrest. Described herein is the synthesis of a combinatorial library of potential proapoptotic agents and the subsequent identification of a class of small molecules (triphenylmethylamides, TPMAs) that arrest the growth of melanoma cells in the G1 phase of the cell cycle. Several of these TPMAs are quite potent inducers of apoptotic death in melanoma cell lines (IC(50) approximately 0.5 muM), and importantly, some TPMAs are comparatively nontoxic to normal cells isolated from the bone marrow of healthy donors. Furthermore, the TPMAs were found to dramatically reduce the level of active nuclear factor kappa-B (NFkappaB) in the cell; NFkappaB is known to be constitutively active in melanoma, and this activity is critical for the proliferation of melanoma cells and their evasion of apoptosis. Compounds that reduce the level of NFkappaB and arrest cells in the G1 phase of the cell cycle can provide insights into the biology of melanoma and may be effective antimelanoma agents.

Cytotoxic effects of triphenylbismuth on rat thymocytes: comparisons with bismuth chloride and triphenyltin chloride.

The biomedical and industrial uses of organobismuth compounds have become widespread, although there is limited information concerning their cytotoxicity. Therefore, the actions of triphenylbismuth on rat thymocytes were examined using a flow cytometer with ethidium bromide, annexin V-FITC, fluo-3-AM, and 5-chloromethylfluorescein (5CMF) diacetate. Triphenylbismuth at 3-30 microM increased the population of cells stained with ethidium, indicating a decrease in cell viability. Organobismuth at 30 microM increased the population of cells positive to annexin V, suggesting an increase in the population of apoptotic cells. Triphenylbismuth at 3 microM or more decreased cellular glutathione content (5CMF fluorescence intensity) and increased intracellular Ca(2+) concentration ([Ca(2+)](i), fluo-3 fluorescence intensity) in a dose-dependent manner. Because an increase in [Ca(2+)](i) is linked to cell death or cell injury and a decrease in cellular glutathione content increases cell vulnerability to oxidative stress, the triphenylbismuth-induced changes in cellular parameters may be responsible for triphenylbismuth-induced cytotoxicity. Bismuth chloride at 10-30 microM did not significantly affect cell viability. These results suggest that triphenylbismuth at micromolar concentrations exerts cytotoxic action on rat thymocytes, possibly related to a health hazard. Although the cytotoxicity of triphenylbismuth was less than that of triphenyltin, one of the environmental pollutants, it is necessary to direct our attention to the use and disposal of organobismuth compounds.

Acute and subacute inhalation toxicities of phosphine, phenylphosphine and triphenylphosphine.

The four-hour LC50 values for ChR-CD male rats for phosphine, phenylphosphine and triphenylphosphine have been determined to be 0.44 micromoles per liter (muM/liter), 1.56 muM/liter and 47.8 muM/liter respectively. The dose-death curves are parallel. During exposure, all three caused clinical signs indicative of mild respiratory irritation. Triphenylphosphine also caused severe weight loss immediately after exposure, followed by normal rate of weight gain. No histopathologic effects due to exposure were seen in any of the tissues examined after single exposures to any of the three compounds. Phosphine and triphenylphosphine caused mild weight loss during a 10-day exposure period followed by normal rate of weight gain during a 14-day recovery period. The phenylphosphine exposures caused a decreased rate of weight gain during the exposure period which returned to normal during the recovery period. Silver nitrate paper was found to be unsuitable for field analysis of phenylphosphine.

Evaluation of laser dye mutagenicity using the Ames/Salmonella microsome test.

Twenty-five laser dyes and four analogs were tested for mutagenicity in the Ames/Salmonella test. Seven dyes and two analogs gave positive mutagenic responses with bacterial strains TA1538 and TA98. Of two widely used families of laser dyes (coumarins and rhodamines), four coumarin samples, but none of the rhodamine samples, were mutagenic. All mutagenic compounds require enzyme activation for positive response except two terphenyl analogs, which are mutagenic with or without activation. Using high-performance liquid chromatography (HPLC), it was determined that five mutagenic dye samples had multiple components. The dyes themselves may not be the mutagenic agents in all cases (as with Nile Blue) but may contain impurities that are mutagenic. One dye, adicyanomethylene (DCM) (greater than or equal to 95% pure), was mutagenic at doses below 0.5 micrograms/plate on strains TA1538 and TA98. DCM also induced reversions in strains TA96, TA97, TA100, TA102, and TA104, although less efficiently. This study indicates the need for further toxicological testing of these types of compounds. The mutagenic components of these dye mixtures, whether it is the dye or a contaminant, presents a possible hazard to those handling them. Therefore, practices and procedures for the safe handling of specific dyes should be reviewed in light of these findings.

The effects of polybrominated biphenyls and perchlorinated terphenyls on in vitro fertilization in the mouse.

Polybrominated biphenyls (PBBs) and perchlorinated terphenyls (PCTs) are industrial chemicals that are long-lasting environmental contaminants. Although in vivo effects of PBBs on reproduction are documented, no information is available on the effects of these chemicals on sperm-egg interactions or fertilization. The present study was undertaken to determine the toxic potential of PBBs and PCTs on in vitro fertilization (IVF) in the mouse. 2-Bromobiphenyl, 4-bromobiphenyl, o-terphenyl, m-terphenyl, and p-terphenyl were added to the IVF medium at various concentrations. Oocytes collected from superovulated B6D2F1 mice were maintained in a medium containing the chemicals. Capacitated sperm were then added and the dishes cultured in a humidified atmosphere at 37 degrees C in 5% CO2 + 95% air. Oocytes were assessed for fertilization 20-24 h after insemination. polybrominated biphenyls and PCTs reduced the IVF rate at the higher dosages. Furthermore, an increased incidence of abnormal two-cell embryos and degenerative oocytes was observed at the 1 and 10 micrograms/ml concentrations of PBBs and PCTs. These results indicate that PBBs and PCTs adversely effect IVF and increase the incidence of abnormal embryos and oocyte degeneration in the mouse.

Impaired nerve-muscle interaction by percutaneous terphenyl exposure.

Rat tail muscle responses decreased in amplitude and showed polyphasia in intermittent (3 h daily for 4 to 8 wks) percutaneous exposure to a commercial terphenyl mixture. The gross conduction times did not change excluding demyelination as a mechanism. The exposure technique allowed an exact estimation of the absorbed dose (0.23 +/- 0.05 g/12 cm2 skin in 3 h). This could be altered by changing the skin contact time. The described technique may used also in the detection of local toxicity of other lipid-soluble compounds.

Comparative activity of 4,4'-diaminobiphenyl (benzidine) and its terphenyl analogue, 4,4'-diaminoterphenyl, in two in vitro assays for potential carcinogenicity.

The carcinogen 4,4'-diaminobiphenyl (benzidine) has been compared in vitro with its terphenyl analogue 4,4''-diaminoterphenyl using the Salmonella reverse mutation assay and the BHK cell-transformation assay. The responses observed, taken together with a consideration of chemical structures, indicate that the terphenyl compound is a potential carcinogen. These findings may contribute to an understanding of the mechanism of action of benzidine as a carcinogen.

Subchronic inhalation and oral toxicity of hydrogenated terphenyls in rats.

The subchronic toxicity of a commercial blend of partially hydrogenated terphenyl was evaluated in rats by inhalation and oral routes of exposure. Animals were exposed to target concentrations of 0, 10, 100, or 500 mg/m3 for 6 hr/day, 5 days/week or were offered diets daily with concentrations of 0, 50, 200, or 2000 ppm. Each study lasted approximately 14 weeks. The study designs included observations for clinical signs, body weights, ophthalmic exams, hematology and clinical chemistry, major organ weights, and gross and histopathology. No treatment-related effects were noted in the ophthalmic exams. Body weights were slightly depressed in high-dose males from the inhalation study and high-dose females in the dietary study. Liver and liver/body weights were increased in high-dose animals of both sexes and high- and mid-dose males in the dietary and inhalation studies, respectively. In the high-dose females of the dietary study, kidney and kidney/body weights were increased with increased adrenal and adrenal/body weights were also observed. No compound-related gross lesions nor pathological correlates to the organ weight changes were observed in either study. The no-adverse effect levels were considered to be 100 mg/m3 and 200 ppm (15.9 mg/kg) for the inhalation and dietary studies, respectively. These data indicate that a wide margin of safety exists for hydrogenated terphenyl workplace exposure.

Effects of polychlorinated terphenyls and paraffins on rat liver microsomal cytochrome P-450 and in vitro metabolic activities.

The polychlorinated terphenyl Aroclor 5460 and the polychlorinated paraffins Witaclor 171 P and Witaclor 149 increased to different degrees the total microsomal concentration of cytochrome P-450 in the rat liver after intraperitoneal injection of 0.3, 1.0, and 1.0 g . kg-1 body weight, respectively, each day for four days. The multiple forms of cytochrome P-450 were affected differently with an induction of RLvMc P-450(50) and RLvMc P-450(54) by all chemicals, and an additional induction of RLvMc P-450(55) by the polychlorinated terphenyl. The rat liver weights were extensively increased after treatment with the polychlorinated paraffins. Alterations were found in the in vitro metabolism of biphenyl, benzo(a)pyrene and the steroid hormones, 4-androstene-3,17-dione and 5 alpha-androstane-3 alpha, 17 beta-diol, after exposure to all chemicals. Changes in the in vitro formation of benzo(a)pyrene metabolites were found to correlate with changes in the multiple forms of cytochrome P-450. The present study demonstrate that only limited information can be obtained from alterations in the total concentration of cytochrome P-450 and show the importance of studying changes in the multiple forms and the metabolism of different substrates. Our results further indicate that exposure to any of the investigated polychlorinated chemicals may alter the biological effects of other environmental contaminants, drugs and endogenous substances which are metabolized by the cytochrome P-450 enzyme system.

[Tercuronium, a new nondepolarizing myorelaxant with high activity and selective action]. / Terkuronii--novyi nedepoliarizuiushchii miorelaksant s vysokoi aktivnost'iu i izbiratel'nost'iu deistviia.

Tercuronium is p',p"-bis-triethylammonium-p-terphenyl dibenzosulfonate. As a curarelike agent tercuronium is 4--8 times as potent as (+)-tubocurarine. The time of the development and lasting of the blocking effect of tercuronium is approximately the same as that of (+)-tubocurarine. In a blocking dose tercuronium does not exert any effect either on the vegetative ganglia or arterial blood pressure. Partial blocking of the transmission through the vegetative ganglia as well as an insignificant and short-term drop of arterial blood pressure are recorded after intravenous injection of 10 myoparalytic doses of tercuronium. The antagonism of neostigmine against tercuronium was more pronounced than against (+)-tubocurarine, pancuronium and gallamine.

Biochemical toxicology and disposition of Therminol 66 heat transfer fluid after inhalation or after dietary administration to male Sprague-Dawley rats.

The objectives of this study were to determine the disposition of Therminol 66 in rats and to determine the effects of this heat-transfer fluid on liver and kidney microsomal drug-metabolizing enzymes. Therminol 66 was administered to male Sprague-Dawley rats at various doses as either a single oral administration at 0, 100, or 300 mg/kg, or as a single 6-h inhalation exposure at 0 or 350 mg/m3. Animals were killed 48 h after gavage or after termination of inhalation exposure. Additional groups of animals were exposed to Therminol 66 via the diet at 0, 100, 500, or 5000 ppm for 14 d, or via repeated inhalation exposure at 0, 25, 250, or 1200 mg/m3 for 6 h/d for 14 d. These exposure scenarios represent approximately equivalent doses of Therminol 66 by the different routes of administration. No change in body weight was observed after acute oral or inhalation exposure, and little change in body weight was observed in animals administered Therminol 66 via the diet except at the highest dose. There was no change in kidney weight, and liver weights were increased only at the higher doses of Therminol 66. The body weight gain of animals exposed to Therminol 66 via inhalation decreased in a dose-dependent manner over the 2-wk exposure period. Results from the disposition study indicated that Therminol 66 did not appear to accumulate in the tissues examined and did not appear to be extensively absorbed after a single oral dose of 300 mg/kg. The whole-body elimination half-life was approximately 14 h and occurred primarily via the feces. There was no significant induction of hepatic aryl hydrocarbon hydroxylase (AHH) activity after single oral or inhalation exposures to Therminol 66. Ethoxycoumarin O-deethylase (ECOD) was significantly induced only in animals exposed to 350 mg/m3 via inhalation. Repeated dietary and inhalation exposures resulted in AHH and ECOD induction only at the highest doses, and the kidney appeared to be less sensitive than the liver. Animals exposed via inhalation demonstrated a greater hepatic inductive effect than did animals exposed via the diet, which may be due to absorption differences.(ABSTRACT TRUNCATED AT 400 WORDS)