Reduced gluconeogenesis in 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-treated rats

The effect of a usually lethal dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD; 125 g/kg) was studied on the conversion of¹⁴C-alanine into¹⁴C-glucose in male Sprague-Dawley rats by established procedures (determination of plasma alanine and blood glucose by enzymatic assays and isolation of¹⁴C-alanine and¹⁴C-glucose from whole blood by column chromatography). TCDD-treated rats converted significantly (p < 0.05) less¹⁴C-alanine into¹⁴C-glucose than did their pair-fed or ad libitum-fed counterparts, indicating reduced gluconeogenesis as a result of TCDD treatment. This finding suggests that reduced gluconeogenesis in TCDD-treated rats contributed to the progressively developing, severe hypoglycemia observed in these animals. Corticosterone, a key hormone in gluconeogenesis, provides partial protection from TCDD-induced toxicity in hypophysectomized rats. Therefore, the conversion of¹⁴C-alanine into¹⁴C-glucose was also determined in hypophysectomized rats dosed with TCDD (125 g/kg) and given corticosterone (25 g/ ml in drinking water). These rats also converted significantly (p <0.05) less¹⁴C-alanine into¹⁴C-glucose than did their pair-fed counterparts. However, in contrast to non-hypophysectomized TCDD-treated rats, these rats maintained marginal normoglycemia even at 64 days after dosing with TCDD, which suggests that the partial protective effect of corticosterone in hypophysectomized, TCDD-treated rats is unrelated to its effect on gluconeogenesis. The protection provided by corticosterone supplementation in TCDD toxicity is more likely due to reduced peripheral utilization of glucose enabling the animals to maintain marginal normoglycemia.Presented in part at the 27th Annual Meeting of the Society of Toxicology, Dallas, TX, 1988     

Corticosterone decreases toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in hypophysectomized rats

Male Sprague-Dawley rats were hypophysectomized by an established surgical technique. Hypophysectomy aggravated the toxicity (mortality and mean time to death) of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD; 125 micrograms/kg ip) when compared to sham-operated rats (100% mortality with 9 +/- 1 d mean time to death vs. 90% mortality with 32 +/- 6 d mean time to death, respectively). However, administration of corticosterone (25 micrograms/ml in drinking water) to hypophysectomized rats resulted in an attenuation of the toxicity (40-60% mortality with 40-90 d mean time to death) to a range of TCDD doses (125, 250, 500 micrograms/kg) much higher than the LD50 (about 60 micrograms/kg TCDD) in nonhypophysectomized rats (about 30 d mean time to death). Furthermore, thyroid hormone supplementation in hypophysectomized rats dosed with 125 micrograms/kg TCDD restored the toxicity of TCDD to approximately "normal." Based on these data it is concluded that one or more as yet unknown key factors that are important in the modulation of the toxicity of TCDD reside in the pituitary.     

Mode of metabolism is altered in 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-treated rats

Male Sprague-Dawley rats were fed either a high-fat (HF) or a high-carbohydrate (HC) diet and subsequently injected with either 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) (125 micrograms/kg) or vehicle (pair-fed controls). In all TCDD-treated animals, a reduction in caloric intake was evident as early as 1 day after dosage. Respiratory quotients (RQ) were determined at 5-day intervals. Their pattern for the HC-fed but not for the HF-fed TCDD-treated rats was different from that of the corresponding pair-fed controls. After an initial parallel decrease the RQ values remained low for TCDD-treated rats whereas they increased again for pair-fed controls. Serum total thyroxine (T4) was significantly lower in TCDD-treated animals and this reduction was not influenced by the composition of the diet. Serum triiodothyronine (T3) was neither altered by diet nor by TCDD. Thymic atrophy was as severe in pair-fed as in TCDD-treated rats fed the HC diet but not in rats fed the HF diet. Our results suggest that TCDD-treated rats are in a different mode of metabolism from pair-fed rats and that this difference is related to gluconeogenesis.     

Learning in monkeys exposed perinatally to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)

TCDD is an extremely toxic chemical pollutant which bioaccumulates in maternal adipose tissue, and is transferred to the developing organism during gestation and lactation. Long-term cognitive deficits have been reported following perinatal exposure to polychlorinated biphenyls, which are structurally and toxicologically similar to TCDD. In the current study, monkeys exposed to TCDD perinatally were later tested in two cognitive paradigms, discrimination-reversal learning (RL) and delayed spatial alternation (DSA). RL detected effects; whereas DSA, as analyzed, did not. RL consisted of a series of simple spatial reversals, followed by spatial reversals with color and shape as irrelevant cues, then by color reversals and finally by shape reversals. TCDD-exposed monkeys exhibited retarded learning of the shape reversals. The deficit was most pronounced on the first reversal following overtraining. There were no group differences on the spatial or color reversals. However, the number of trials the TCDD-exposed monkeys individually took to learn the spatial reversals was positively correlated with TCDD concentration in body fat. Conversely, the number of trials they took to learn the color reversals was negatively correlated with TCDD in body fat.     

Composition of diet modifies toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in cold-adapted rats

The effect of a high carbohydrate, high fat or high protein diet was studied on the acute toxicity of TCDD (125 g/kg) in cold-adapted (4±1° C) rats. Within 10 days after dosing, TCDD-treated rats fed a high carbohydrate or a high protein diet reduced their caloric intake by 25% whereas those fed a high fat diet consumed only 15% fewer kcal/MBS (metabolic body size). TCDD-treated rats fed a high protein diet lost body weight at the same rate as their pair-fed controls, whereas body weight loss in high fat-fed rats was significantly higher than in their pairfed controls. In contrast, TCDD-treated rats fed a high carbohydrate diet effectively maintained their body weight in the 4 days immediately after TCDD dosage, whereas their pair-fed controls lost weight. Mortality in TCDD-treated animals was 100% irrespective of the diet; all pairfed control rats (except one fed a high protein diet) were terminated on days corresponding to the spontaneous death of their TCDD-treated pairs. Mean time to 50% mortality and mean time to death were significantly longer in TCDD-treated rats fed a high carbohydrate diet in comparison with the other two TCDD-treated groups (p<0.05), although caloric intake was comparable. Serum triiodothyronine (T₃) was reduced in TCDD-treated animals fed a high fat or a high carbohydrate diet but not in those fed a high protein diet; serum thyroxine (T₄) was reduced in all the treated groups, irrespective of diet. The relative weight of liver was significantly higher in all TCDD-treated animals; the weight of pancreas was higher only in high carbohydrate-fed TCDD-treated rats and the weight of interscapular brown adipose tissue was lower only in high fat-fed TCDD-treated rats in comparison to pair-fed controls. It is concluded that TCDD-treated cold-adapted rats utilize carbohydrate and fat differently than pair-fed controls and that these changes probably contribute to the development of a wasting syndrome.This work was supported in part by a grant from the Bayerisches Staatsministerium für Landesentwicklung und Umweltfragen, München (Fed. Republic of Germany)     

Characterization of the enhanced responsiveness to postingestive satiety signals in 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-treated Han/Wistar rats.

Previous studies have shown that under free-feeding conditions, TCDD-treated Han/Wistar (H/W) rats consume less sucrose solution but ingest more saccharin solution than their controls thus implying hyperresponsiveness to postingestive satiety signals. In this study, nutrient preloads were employed to further elucidate this phenomenon. Male H/W rats were given a single high but usually non-lethal intraperitoneal dose (1000 micrograms/kg) of TCDD. Feed intake was stimulated by 24 hr feed deprivation at various time points after TCDD exposure. When TCDD-dosed rats were allowed to drink either a 20% sucrose or a 0.25% saccharin solution and then given access to feed, those that had had sucrose ate only about 50% of the amount consumed by the saccharin group. Although the preloads were similar in control rats, no such difference in subsequent feeding occurred. The sucrose solution also produced a longer-lasting suppression of feed intake in TCDD-treated compared with control rats when infused directly into the stomach. By contrast, TCDD-treated H/W rats failed to exhibit an augmented satiety response to parenterally applied glucose independent of testing time. Oral corn oil reduced feed intake in both control and TCDD-exposed rats, but the inhibition was slightly larger in TCDD-treated animals. TCDD did not markedly affect the responsiveness of H/W rats to the suppression of feeding by CCK-8 or bombesin. It is concluded that gastrointestinal factors appear critical to the exaggerated response of TCDD-treated H/W rats to nutrient energy.     

Acute myelotoxic responses in mice exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)

Blood cells are derived from a multitiered system of progenitor stem cells that lose their capacity for proliferation and self-renewal as they continue along pathways of differentiation. Since these hematopoietic events can be readily monitored in vivo and in vitro in the mouse, we have utilized this system to examine altered cellular differentiation associated with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) toxicity. Progenitor cells were suppressed following acute exposure of mice to TCDD at doses as low as 1.0 micrograms/kg body wt. In vitro studies demonstrated that myelotoxicity occurs by a direct inhibition of proliferating stem cells. Genetic studies indicated that the myelotoxic responses to TCDD, both in vivo and in vitro, segregate with the Ah locus. In addition, the in vitro myelotoxicity of various polyhalogenated aromatic hydrocarbon congeners correlated with their previously reported ability to induce hepatic microsomal enzyme activity and to bind to an intracellular receptor for TCDD. TCDD was also found to bind specifically to bone marrow cells from Ah-responsive, but not nonresponsive mice, indicating that bone marrow cells possess a specific receptor for TCDD. These data indicate that the myelotoxic response to TCDD is regulated by the Ah receptor present in the target tissue and demonstrates the utility of this system for examining the cellular and molecular events associated with the toxicity of polyhalogenated aromatic hydrocarbons, the prototype for which is TCDD.     

2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced accumulation of biliverdin and hepatic peliosis in rats.

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a widespread, persistent, and highly toxic environmental pollutant. The most TCDD-sensitive and the most TCDD-resistant rat strains (Long-Evans [Turku/AB] and Han/Wistar [Kuopio], respectively) were crossbred to separate the alleles of two genes (Ahrand an unidentified gene "B") mediating resistance against TCDD toxicity. During crossbreeding, a new type of toxicity in livers of both sexes was detected, characterized macroscopically by intense dark green to black color and swelling that appeared most frequently after a large dose (300 micro g/kg or more as a single intragastric dose) and a follow-up period of more than three weeks. Therefore, studies were undertaken to identify the causative pigment chemically and to examine the hepatotoxicity histologically. The pigment fractions were separated by thin layer chromatography and then analyzed by HPLC and electrospray mass spectrometry. The pigment was found to consist of biliverdin and several biliverdin-related compounds. In liver histopathology carried out on male rats, progressive sinusoidal distension and hepatic peliosis with membrane-bound cysts were seen. The clinical manifestations of pigment accumulation were recorded most often in intermediately resistant rat lines such as line B (homozygous for the gene B), but never occurred in rats expressing only the Han/Wistar (Kuopio)-type Ah receptor with an altered transactivation domain structure.     

Effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on IgA serum and bile levels in rats

Serum IgA is actively transported from blood to bile against a concentration gradient in the liver by the binding of dimeric IgA to secretory component, endocytosis and transport to the bile canaliculus by vesicles. As 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been shown to elicit hepatotoxicity, the effects of TCDD on rat serum and bile IgA levels were investigated. Rats were orally administered 50 micrograms TCDD/kg body weight in 95% corn oil: 5% acetone. At days 5, 10, 15, 20 and 30 after treatment, rats were anesthetized and a cannula inserted into the bile duct for collection of bile. In addition, blood was drawn, and, after euthanasia, the liver and thymus weights were recorded. Enzyme-linked immunosorbent assay (ELISA) techniques were employed to determine IgA in serum and bile and IgG levels in serum. Rocket immunoelectrophoresis was carried out to support ELISA results. It was found that serum IgA increased with time while serum IgG remained unchanged. In addition, while serum IgA levels were increasing, there was a concomitant decrease in biliary IgA. Thymus and liver weight changes were also observed. The data indicate that TCDD affects hepatic clearance of serum dimeric IgA and suggests that liver damage may be reflected by increased serum levels of IgA.     

Hypothalamic site of action of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)

Administration of TCDD produced a significant decrease in the serum concentration of prolactin (PRL) detected in rats after 4 hr compared to pair-fed vehicle controls and noninjected controls. This effect of TCDD was reversed by pimozide, a dopamine receptor antagonist. These data suggest that TCDD decreased the release of PRL from the adenohypophysis either by a direct effect on the gland or by altering the dopamine concentration in the median eminence (ME). Concentrations of TCDD from 5 to 500 ng/ml had no direct effect on the ability of the adenohypophysis to secrete PRL in vitro. However, the dopamine concentration increased to 3.24 +/- 0.07 ng per ME in TCDD-treated rats compared to 2.81 +/- 0.08 ng in vehicle controls. This is a dramatic alteration in the dopamine concentration, since the dopamine is being measured in the portal circulation which exhibits a rapid turnover. The rate constant of dopamine depletion after alpha-methyl-p-tyrosine and the turnover rate were also significantly elevated in the ME of TCDD-treated rats. These data provide the first biochemical evidence for a hypothalamic site of action of TCDD. Since dopamine is inhibitory to PRL release from the adenohypophysis, increased ME steady-state concentrations and turnover of this catecholamine may be responsible for the decreased concentration of serum PRL detected within 4 hr of TCDD injection. Thus, one of the early modes and sites of action of TCDD is to elevate the dopaminergic activity of the tuberoinfundibular nucleus. A hypothalamic site of action for TCDD may result in a number of the endocrinological effects known to be produced by exposure to TCDD.     

Regulation of polyamine biosynthesis by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)

We have examined the effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on tissue polyamine concentrations in CD1 mice. Two days after a low dose treatment with TCDD, polyamine content of the liver and thymus of treated mice showed a 49-82% decrease, but that of spleen was not affected. Based on this finding, we examined the role of alterations in polyamine levels in the toxicity of TCDD. We administered the polyamine biosynthetic inhibitor difluoromethylornithine (DFMO) to animals treated with a toxic dose of TCDD. DFMO dramatically increased the toxicity of TCDD as measured by mortality, ascites and changes in organ weights. In addition, administration of the polyamine putrescine was able to reduce the toxicity of TCDD. These results suggest that a decrease in polyamine concentrations in critical organs may play an important role in the toxic effects of TCDD.     

Percutaneous absorption of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) from soil

Eight dermal absorption experiments (two in vivo; six in vitro) and one intravenous experiment were conducted using 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) either neat (high dose at approximately 250 microg/cm(2) and low dose at 10 ng/cm(2)) or sorbed on a low organic soil (LOS) or high organic soil (HOS) at 1 ppm (10 ng TCDD/10 mg soil/cm(2)). After 96 h the percent of applied dose absorbed (PADA) for the neat low dose was 78% in vivo (rat) and 76% in vitro (rat). PADA for the equivalent TCDD dose sorbed on LOS were 16.3% (rat in vivo), 7.7% (rat in vitro) and 2.4% (human in vitro). The PADA for TCDD sorbed on HOS (1 ppm) was 1.0% (rat in vitro). Generally, rat skin was observed to be three to four times more permeable to TCDD than human skin. At steady state, the dermal flux of TCDD in neat form, sorbed on LOS at 1 ppm, and sorbed on HOS at 1 ppm (all in vitro, rat) was 120, 0.007, and 0.0007 ng/cm(2)/h, respectively (ratio = 1.7 x 10(5):10:1). Making adjustments to account for differences between in vitro and in vivo results and adjusting for application to monolayer loads, the 24-h TCDD absorption for human skin is estimated as 1.9% from LOS (1 ppm) and 0.24% from HOS (1 ppm).     

Thyroid hormones modulate the toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)

These experiments examine the role of thyroxine (T4) and triiodothyronine (T3) on the toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The first experiment is continuation of a study reported previously (Rozman et al., 1984). In this experiment, 60 male Sprague-Dawley rats were divided into 6 equal groups. Four groups of rats were thyroidectomized by 3 mCi Na131 l/kg rat. Five weeks later 2 of the thyroidectomized and 1 of the nonthyroidectomized groups of rats received ip 100 micrograms TCDD/kg body weight in corn oil/acetone, whereas 3 corresponding groups of rats served as vehicle controls. Two days after dosing and every 7 d thereafter, 1 thyroidectomized control group and 1 thyroidectomized TCDD-dosed group were given ip 105 micrograms T4/kg body weight. Mortality and body weight were monitored. The course of TCDD toxicity was similar in nonthyroidectomized and thyroidectomized T4-treated rats but was different in thyroidectomized animals without T4 replacement therapy. At d 90 after TCDD dosage, mortality was still lower and the mean time to death was increased (p less than 0.01) in this group of rats compared to nonthyroidectomized or thyroidectomized T4-treated rats. However, administration of T4 starting at d 91 after dosing with TCDD resulted within 2 wk in the same final mortality in thyroidectomized rats as in nonthyroidectomized or thyroidectomized T4-treated animals, indicating that thyroid hormones modulate the time course of the wasting syndrome but do not affect the ultimate mortality figure. Body weight loss was much slower in thyroidectomized (approximately 1 g/d) than in nonthyroidectomized or thyroidectomized T4-treated rats (approximately 8 g/d). In the second experiment the three vehicle control groups of the first experiment were used. Nonthyroidectomized vehicle controls and thyroidectomized T4-treated controls were maintained as before, whereas thyroidectomized controls received T3 at 5 micrograms/kg daily. One month later each rat was dosed with TCDD at 100 micrograms/kg in corn oil/acetone. Toxicity of TCDD was similar in nonthyroidectomized, thyroidectomized T4-treated, and thyroidectomized T3-treated rats as judged by mortality, body weight, and food intake, indicating no difference between T3 and T4 in the modulation of TCDD toxicity.     

Radial arm maze performance in rats following gestational and lactational exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)

Recently, we reported that in utero and lactational exposure to 2,3, 7,8-tetrachlorodibenzo-p-dioxin (TCDD) resulted in a task-specific reduction of errors on the radial arm maze (RAM), without similar improvements on other spatial learning tasks including the Morris water maze. The effect was more pronounced in males than in females. This study further investigated the effects of in utero and lactational exposure to TCDD on RAM performance by testing male and female TCDD-exposed rats on either an eight-arm RAM with all arms baited or a 12-arm RAM with 8 of the 12 arms baited. If the rats have improved spatial learning or memory on the RAM, then they should be improved on both RAM tasks; whereas, if they are using adjacent arm selection or some other response strategy to solve the task, they should not show enhanced performance on the 12-arm RAM where not all the arms are rewarded. Time-mated Sprague-Dawley dams were gavaged with corn oil vehicle or one of two doses of TCDD in vehicle (0.1 or 0.2 microg/kg body weight) on gestational days 10 to 16. Litters were culled to eight on day 2 and weaned on day 21. Beginning on day 80, one male and female from each litter was tested on the eight-arm RAM with all arms baited. As in our previous studies, the 0.1-microg/kg TCDD-exposed male rats showed a significant decrease in the number of errors. However, the 0.2-microg/kg males did not differ from the controls. Neither group of TCDD-exposed females differed from the controls. None of the TCDD-exposed rats differed from the controls in adjacent arm selection behavior. An additional male and female from each litter were tested on the 12-arm RAM with only 8 of the 12 arms baited. In this task, neither TCDD group differed from the controls. These results suggest that the reduction of errors on the eight-arm RAM may be due to increased response patterning or use of intramaze cues rather than to improved spatial learning or memory. Also, the reduction in errors was only present at the lower dose of TCDD suggesting that the improvement in performance is only present at very low, nonovertly toxic doses of TCDD.     

Biochemical and functional effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on the heart of female rats

Biochemical, functional and morphologic effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on the hearts of female rats were examined. Six days after the treatment of rats with TCDD, the blood pressures and resting heart rates were significantly less than in control animals. Treated animals were also less responsive to the effects of the beta-1 agonist, (-)isoproterenol. No histopathologic changes were observed in the heart although extensive centrilobular necrosis occurred in the liver after TCDD administration. Serum levels of thyroxine were 66% less than in control animals. Marked lipid peroxidation was produced in the liver with small but significant increases occurring in the heart. TCDD administration had no effect on catalase activity in the heart, but produced a 20% decrease in superoxide dismutase activity relative to control animals. The effects of TCDD on cardiac function do not appear to be due to a direct action of the xenobiotic on the heart but possibly to a down-regulation of beta-receptors in the heart as a result of the hypothyroid state.     

Learning and memory in rats gestationally and lactationally exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD).

Recently we reported that in utero and lactational exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) or coplanar polychlorinated biphenyls (PCBs) resulted in a reduction of errors on a radial arm maze (RAM) working memory task. The effect was more pronounced in males than in females. In this study, we further investigated the effects of in utero and lactational exposure to TCDD on learning and memory by testing male and female TCDD-exposed rats on three different spatial learning and memory tasks: the RAM, the Morris water maze (MWM), and spatial discrimination-reversal learning (RL), as well as on a nonspatial learning task, visual RL. Time-mated Sprague-Dawley rats were gavaged with either TCDD (0.1 microg/kg/day) or corn oil vehicle on gestation days 10-16. Litters were culled to eight on day 2 and weaned on day 21. Beginning on day 80, one male and one female from each litter were tested on the same RAM working memory task used in the previous study. Again, the TCDD-exposed male rats displayed a pronounced decrease in errors relative to control males. Following the RAM testing, the same animals were tested on the MWM, but no differences between the exposed and control rats were observed. Another male and female from each litter were tested on spatial RL on a T-maze. There were no differences between the exposed and control rats on this task. Following spatial RL, the same rats were tested on visual RL on the same maze. The exposed animals did not differ from controls on original learning, but took more trials to reach criterion on the first and second reversals. These results demonstrate a reliable, but task-specific, facilitation of spatial learning and memory in male rats exposed to TCDD during gestation and lactation. In contrast, both male and female TCDD-exposed rats showed a deficit in learning on the visual RL task. This pattern is consistent with that seen in earlier monkey studies. Perinatally TCDD-exposed monkeys were facilitated on certain spatial tasks, but impaired on visual RL tasks.     

A metabolite of riboflavin binds to the 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) receptor

A cytosolic receptor to which TCDD binds with high affinity has been reported in a number of rodent tissues, but the endogenous/natural ligand is not known. However, lumichrome, a metabolite of riboflavin, competed with 3H-TCDD for binding to the cytosolic receptor. Neither riboflavin nor riboflavin-5-phosphate competed for binding to the TCDD receptor in rat liver cytosol. Lumichrome is the first known endogenous chemical in the rat to exhibit affinity for the TCDD receptor.     

Effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on thermogenesis in brown adipose tissue of rats

Male Sprague-Dawley rats were treated with a usually lethal dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD; 125 micrograms/kg i.p. in corn oil) or with vehicle alone. Two, 4, and 8 days after treatment the temperature of interscapular brown adipose tissue (IBAT) was monitored during venous infusion of norepinephrine (480 ng/min) for 60 min. The temperature response was about 1.0-1.5 degrees C within 1 h in vehicle-treated, pair-fed and ad libitum-fed controls. In TCDD-treated animals, the response of IBAT decreased with time after TCDD dosage, amounting to only 0.3 +/- 0.1 degree C at 8 days after dosing (differences significant with respect to both controls, P less than 0.05). GDP binding to IBAT mitochondria (a measure of thermogenic capacity) was unchanged in all groups, indicating that the reduced thermogenic response was probably not caused by an impairment of the mitochondrial uncoupling process by TCDD.     

Quantitative cancer risk assessments for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)

State-of-the-art quantitative risk assessment techniques, including consideration of new time-to-response data, have been applied to chronic animal bioassay data on the dietary intake of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The non-linear shapes of the dose-response relationships for the hepatocellular carcinogenic responses have been estimated, and a review of the quantitative impacts of several of the choices involved in the quantitative risk assessment considers, particularly, the definition of the carcinogenic responses of concern, the experimental data set, the pathology evaluation, a biologically effective dose scale versus the administered dose, methods of making the fitted model responsive to the data at the lower experimental doses, consistency in dose-response shapes for different data sets, fitted model values versus bounds, the utilization of time-to-response information incorporating the lateness of the carcinogenic responses, and the method of characterizing the maximum acceptable dose. The estimated virtually safe dose for an increase of 0.000001 (one in a million) in the probability of hepatocellular neoplastic nodule and/or carcinoma in a female rat is approximately 0.1 ng/kg body weight/day in the diet. The estimated mean free dose, corresponding to a reduction in the expected amount of time without hepatocellular neoplastic nodule and/or carcinoma proportional to 1 wk in 70 yr, is in the range of 1-5 ng/kg body weight/day in the diet of a female rat. No species-to-species extrapolations nor human exposure assessments have been made. However, these estimated risks correspond to dietary intakes that are at least 150 times greater than the 0.0006365 ng/kg body weight/day intake described by the Centers for Disease Control as a reasonable level to begin consideration of action to limit human exposure.