dinitrobenzenes and Body-Weight

Dinitrotoluene (DNT) is a major commodity chemical; over six hundred million pounds of DNT were used in the chemical industry in 1980. Interest in the toxicology of this important chemical was greatly increased when separate oncogenicity assays yielded the conflicting results that DNT was either not hepatocarcinogenic or produced a 100% incidence of hepatocellular carcinomas in male rats in one year. Research revealed pronounced differences in the activity of the DNT isomers and provided the reason for the dissimilar results of the various carcinogenicity studies. Cell culture genetic toxicology assays failed to predict the potent carcinogenic activity of any isomer of DNT. Only when the complex pattern of metabolic activation of DNT began to unfold and genotoxic activity was assessed in the appropriate target organ in the intact treated animal was the potent genotoxic activity of DNT revealed, and the reasons for the negative in vitro results understood. The DNTs have been extensively tested for reproductive effects in animals and humans, and the metabolism and disposition of each of the six possible isomers have been studied. This work has provided valuable information in establishing the risk of these compounds to humans.

Topics: Animals; Body Weight; Carcinogens; Cells, Cultured; Dinitrobenzenes; DNA Repair; Humans; Liver; Male; Mutagenicity Tests; Mutagens; Neoplasms, Experimental; Nitrobenzenes; Reproduction; Spermatozoa

As part of a collaborative study by the Mammalian Mutagenicity Study Group of the Environmental Mutagen Society of Japan, we examined micronucleus induction in hepatocytes following oral administration of 2,6-dinitrotoluene (2,6-DNT) at 30, 40, and 50mg/kg/day for 14 days or at 20, 30, and 40mg/kg/day for 28 days to young adult male rats. This compound is known to be a rat liver carcinogen. The formation of micronucleated hepatocytes was confirmed to be dose-dependent with statistically significant increases observed in both treatments. In contrast, no statistically significant changes in the percentage of micronucleated immature erythrocytes were observed in any dose group in the bone marrow micronucleus assay. These results indicated that the repeated-dose liver micronucleus assay has the potential to detect genotoxic hepatocarcinogens and can be integrated into general toxicological studies.

Topics: Administration, Oral; Age Factors; Animals; Body Weight; Bone Marrow; Carcinogens; Chromosome Aberrations; Cooperative Behavior; Dinitrobenzenes; Dose-Response Relationship, Drug; Drug Administration Schedule; Hepatocytes; Humans; Japan; Liver; Male; Micronucleus Tests; Organ Specificity; Rats; Rats, Sprague-Dawley; Reticulocytes; Societies, Pharmaceutical

The liver micronucleus assay using young adult rats has the potential to detect liver carcinogens by repeated dosing, and could be expected to be integrated into repeated-dose toxicity studies using a hepatocyte isolation method without the traditional in situ collagenase perfusion. In this study, to assess the performance of the repeated-dose liver micronucleus assay, 2,4-dinitrotoluene (DNT), which is a rodent liver carcinogen, was administered orally to male rats at doses of 50, 100 and 200 mg/kg/day once daily for 14 or 28 consecutive days, and the frequencies of micronucleated hepatocytes (MNHEPs) and micronucleated immature erythrocytes (MNIMEs) were examined. Significant increases in the MNHEPs were observed at 50 mg/kg/day or more in the 14-day treatment, and 50 and 100 mg/kg/day in the 28-day treatment. These increases were dependent on both the dose and the number of administrations, which indicates the possibility that the MNHEPs accumulate as a result of repeated dosing. In contrast, no increase in the MNIMEs was observed. In conclusion, the repeated-dose liver micronucleus assay using young adult rats is sufficiently sensitive to detect the genotoxicity of 2,4-DNT at a low dose.

Topics: Administration, Oral; Animals; Body Weight; Bone Marrow; Carcinogens; Chromosome Aberrations; Cooperative Behavior; Dinitrobenzenes; Dose-Response Relationship, Drug; Drug Administration Schedule; Hepatocytes; Humans; Japan; Liver; Male; Micronucleus Tests; Organ Specificity; Rats; Rats, Sprague-Dawley; Reticulocytes; Societies, Pharmaceutical

2,4-dinitrotoluene (2,4-DNT) is a nitroaromatic used in industrial dyes and explosives manufacturing processes that is found as a contaminant in the environment. Previous studies have implicated antagonism of PPARα signaling as a principal process affected by 2,4-DNT. Here, we test the hypothesis that 2,4-DNT-induced perturbations in PPARα signaling and resultant downstream deficits in energy metabolism, especially from lipids, cause organism-level impacts on exercise endurance. PPAR nuclear activation bioassays demonstrated inhibition of PPARα signaling by 2,4-DNT whereas PPARγ signaling increased. PPARα (-/-) and wild-type (WT) female mice were exposed for 14 days to vehicle or 2,4-DNT (134 mg/kg/day) and performed a forced swim to exhaustion 1 day after the last dose. 2,4-DNT significantly decreased body weights and swim times in WTs, but effects were significantly mitigated in PPARα (-/-) mice. 2,4-DNT decreased transcript expression for genes downstream in the PPARα signaling pathway, principally genes involved in fatty acid transport. Results indicate that PPARγ signaling increased resulting in enhanced cycling of lipid and carbohydrate substrates into glycolytic/gluconeogenic pathways favoring energy production versus storage in 2,4-DNT-exposed WT and PPARα (-/-) mice. PPARα (-/-) mice appear to have compensated for the loss of PPARα by shifting energy metabolism to PPARα-independent pathways resulting in lower sensitivity to 2,4-DNT when compared with WT mice. Our results validate 2,4-DNT-induced perturbation of PPARα signaling as the molecular initiating event for impaired energy metabolism, weight loss, and decreased exercise performance.

Topics: Animals; Body Weight; Dinitrobenzenes; Dose-Response Relationship, Drug; Energy Metabolism; Environmental Pollutants; Genomics; Glycogen; Liver; Mice, Inbred C57BL; Mice, Knockout; Physical Endurance; PPAR alpha; PPAR gamma; Signal Transduction; Swimming

Chronic aqueous exposures were conducted using bullfrog (Rana catesbeiana) tadpoles (8 d old) exposed to TNT (0-4 mg/L), 2,4-DNT (0-4 mg/L), and 2,6-DNT (0-8 mg/L) for 90 d. Survival of tadpoles examined using Cox proportional hazard models was reduced at all concentrations tested. Percent of abnormal swimming and other morphological abnormalities after sublethal exposure to TNT, 2,4-DNT, and 2,6-DNT at 2 mg/L were also evaluated. The effects of TNT, 2,4-DNT, and 2,6-DNT on wet body mass, snout vent length (SVL), and developmental stage of surviving tadpoles were examined. Only 2,4-DNT did not have a significant effect on body mass or SVL, but all three compounds tested had significant effects on survival. Long-term continuous exposure to these compounds at concentrations of 0.25 mg/L could lead to significant changes in growth and survival of larval amphibians.

Topics: Animals; Behavior, Animal; Body Weight; Dinitrobenzenes; Explosive Agents; Female; Larva; Male; Rana catesbeiana; Swimming; Trinitrotoluene; Water Pollutants, Chemical

2,4,6-Trinitrotoluene (TNT) is one of the most prevalent high explosives in the environment. 2,4-Dinitrotoluene (2,4-DNT) and 2,6-dinitrotoluene (2,6-DNT) are the most common isoforms of dinitrotoluene. The goal of this study was to determine the acute toxic effects of TNT, 2,4-DNT, and 2,6-DNT in adult male bullfrogs. The LD(50) for TNT was 1,060 mg/kg BW while the LD(50 )for 2,4-DNT and 2,6-DNT was 1,098 mg/kg BW. All three compounds elicited similar symptoms of toxicity including changes of skin color, body weight, development of seizures, liver and kidney necrosis, and lung cyanosis. Relative organ weights did not show significant change.

Topics: Animals; Body Weight; Cyanosis; Dinitrobenzenes; Dose-Response Relationship, Drug; Explosive Agents; Kidney; Lethal Dose 50; Liver; Longevity; Lung; Male; Necrosis; Rana catesbeiana; Seizures; Skin Pigmentation; Trinitrotoluene

Dinitrotoluenes are used as propellants and in explosives by the military and as such have been found at relatively high concentrations in the soil. To determine whether concentrations of 2,4-dinitrotoluene (2,4-DNT) in soil are toxic to amphibians, 100 red-backed salamanders (Plethodon cinereus) were exposed to either 1500, 800, 200, 75 or 0mg 2,4-DNT/kg soil for 28 days and evaluated for indicators of toxicity. Concentrations of 2,4-DNT were less than targets and varied with time. Most salamanders exposed to concentrations exceeding 1050 mg/kg died or were moribund within the first week. Salamanders exposed to soil concentrations exceeding 345 mg/kg lost >6% of their body mass though no mortality occurred. Overt effects included a reduction in feed consumption and an increase in bucco-pharyngeal oscillations in salamanders. These results suggest that only high soil concentrations of 2,4-DNT have the potential to cause overtly toxic effects in terrestrial salamanders.

Topics: Animals; Blood Cell Count; Body Weight; Dinitrobenzenes; Environmental Exposure; Female; Liver; Male; Soil; Soil Pollutants; Urodela

Explosives and their breakdown products are commonly found in soils at U.S. military installations. Many areas where these compounds are found represent useful habitat for ground-foraging birds. Because these birds likely are exposed to such compounds, we evaluated the oral toxicity of 2,6-dinitrotoluene (DNT) in a representative ground-foraging species of management concern. Adult male and female northern bobwhite (Colinus virginianus) were exposed to either 0, 5, 10, 40, or 60 mg/kg/d via gavage for 60 d (subchronic) following determination of the median acute lethal dose (320 mg/kg). Circulating levels of red blood cells and hemoglobin were significantly decreased in a dose-dependent manner; however, levels remained within normal ranges. Plasma concentrations of total protein, albumin, globulin, aspartate aminotransferase, and potassium, sodium, and chlorine ions were significantly decreased, and circulating levels of uric acid were significantly increased. Decreased body weight, enlarged gallbladders, edematous gastrointestinal tracts, pale kidneys, pale and fibrous livers, and loose stools were consistent observations. The effects found in the clinical chemistries taken together with histopathological abnormalities observed in sections of hepatic and renal tissue suggest that the liver and kidneys are major targets for 2,6-DNT. Oral exposures to 2,6-DNT appear to affect northern bobwhite in a manner similar to that of the other main DNT isomer, 2,4-DNT, but in more subtle ways, adversely affecting the gastrointestinal system and leading to diarrhea and, ultimately, emaciation. The lowest-observed-adverse-effect level is 40 mg/ kg/d based on hematological measures, and the no-observed-adverse-effect level is 10 mg/kg/d based on the absence of results indicative of adverse effects.

Topics: Animals; Body Weight; Colinus; Dinitrobenzenes; Dose-Response Relationship, Drug; Eggs; Organ Size

Shrews are abundant in most areas of toxic chemical contamination and bioaccumulate pollutants at much higher rates than sympatric rodent species. As a part of studies to provide information concerning the toxicity of 1,3-dinitrobenzene (DNB) in least shrews (Cryptotis parva), groups of 10 females and 10 males received DNB at 0 (control), 0.7, 2.9, 11.6, and 46.3 microl/L (equivalent mean daily dosage of 0, 0.26, 1.06, 4.26, and 17.0 mg/kg body wt in each sex) in their diet for 14 d. Leukocytosis present at the 0.26 mg/kg body weight/d dosage established the lowest-observed-adverse effect level (LOAEL). Adrenal enlargement was noted at the 1.06 mg/kg body weight/d level. Splenic enlargement and reductions in hematocrit and hemoglobin values occurred at the 4.26 mg/kg body weight/d treatment. Enlargements in the liver and heart and reductions in brown fat weight, granulocyte numbers, and alanine aminotransferase levels were present at high dose levels. Histopathologic examinations showed Kupffer's cell hemosiderosis and suggested testicular damage at the two highest tested doses but failed to confirm brain lesions. Least shrews do not follow standard scaling estimates for lifespan or metabolic rates. The LOAEL calculated from the standard terrestrial screening benchmark equation was higher than our findings, suggesting that these estimates must be viewed with caution.

Topics: Animals; Body Weight; Dinitrobenzenes; Dose-Response Relationship, Drug; Environmental Pollutants; Female; Liver; Male; Reference Values; Shrews; Splenomegaly; Testis

Seasonal changes in day length enhance or suppress immune function in individuals of several mammalian species. Siberian hamsters (Phodopus sungorus) are long-day breeders that adjust reproductive physiology and behavior, body mass, and immune function following exposure to short photoperiods. Photoperiods of intermediate-duration, encountered in nature by juvenile hamsters born in early-spring and by those born in mid-summer, trigger gonadal development in the former cohort and inhibit the onset of puberty in the latter. Divergent reproductive responses to the same intermediate photoperiod depend on a photoperiod history, communicated during gestation. These experiments assessed whether photoperiod history during gestation likewise impacts immunological responses to intermediate photoperiods. Male hamsters were gestated in long photoperiods and remained in long photoperiods postnatally, or were transferred to an intermediate-duration or a short-duration photoperiod; other males were gestated in short days and transferred to an intermediate-duration photoperiod at birth. Long days stimulated, and short days inhibited, somatic and reproductive development; intermediate day lengths either accelerated or inhibited somatic and reproductive development, depending on whether hamsters were gestated in short days or long days, respectively. By contrast, photoperiod during gestation did not affect most immune endpoints. The data suggest that photoperiodic mechanisms that enhance and suppress several aspects of immunity in young-adult hamsters are not responsive to prenatally communicated photoperiod history information.

Topics: Age Factors; Analysis of Variance; Animals; Animals, Newborn; Behavior, Animal; Body Weight; Cricetinae; Dinitrobenzenes; Environment; Enzyme-Linked Immunosorbent Assay; Female; Hair; Hemocyanins; Immune System; Immunization; Immunoglobulins; Leukocyte Count; Leukocytes; Lymphocyte Activation; Male; Organ Size; Phodopus; Photoperiod; Pregnancy; Seasons; Testis; Time Factors

As part of a collaborative work, male rats were administered 1,3-dinitrobenzene (1,3-DNB) daily at 0, 25 and 50 mg/kg/day from the age of 6 weeks for 4 weeks (4-week exp.), or at 25, 50 and 75 mg/kg/day from the age of 8 weeks for 2 weeks (2-week exp.). After the end of each administration period, all survivors were sacrificed, and their testes and epididymides were removed, weighed and examined histopathologically. The following results were obtained. In the 4-week exp.: At 50 mg/kg/day, the weights of testes and epididymides showed decrease with macroscopic atrophy. The testicular spermatogenic epithelium showed decrease in the number of sperm-spermatocytes, degeneration/necrosis, giant cell formation and vacuolation, reduction in sperm counts also being evident in the ducts of the epididymides. In the 2-week exp.: At 50 and 75 mg/kg/day, the weights of testes and/or epididymides showed decrease with macroscopic atrophy. Several histopathological changes in the testes and epididymides were essentially the same changes as in the group given 50 mg/kg/day in the 4-week exp., with a clear relation. These results indicate that a 2-week administration period is sufficient to detect testicular and epididymal histopathological changes induced by 1,3-dinitrobenzene in male rats.

Topics: Administration, Oral; Animals; Atrophy; Body Weight; Dinitrobenzenes; Dose-Response Relationship, Drug; Epididymis; Male; Organ Size; Rats; Rats, Sprague-Dawley; Seminiferous Epithelium; Specific Pathogen-Free Organisms; Sperm Count; Testis; Time Factors; Toxicity Tests

2,4,5-Trichlorophenoxyacetic acid (2,4,5-T) and 2,6-dinitrotoluene (2,6-DNT) are hazardous chemicals that have potential harmful effects. 2,6-DNT is recognized as a hepatotoxicant while 2,4,5-T, a component of Agent Orange, is also suspect. 2,6-DNT requires both oxidative and reductive metabolism to elicit genotoxic effects. To determine what effect 2,4,5-T had on 2,6-DNT metabolism, intestinal enzymes, microbial populations, and urine mutagenicity were examined during 2,4,5-T treatment. Weanling Fischer 344 male rats were treated daily with 54.4 mg/kg 2,4,5-T by gavage for 4 weeks. One, two, and four weeks after the initial 2,4,5-T dose, rats were administered (po) 2,6-DNT (75 mg/kg) and urine was collected for 24 hr in metabolism cages. Azo reductase, nitroreductase, beta-glucuronidase, dechlorinase, and dehydrochlorinase activities were examined concurrently. Treatment of rats for 1 week reduced the transformation of 2,6-DNT to mutagenic urinary metabolites. This was accompanied by a decrease in the fecal anaerobic microorganisms. The elimination of Lactobacillus fermentum from the small intestine and cecum of treated animals accompanied a significant increase in oxygen-tolerant lactobacilli and other unidentified aerobic microorganisms. However, there were no significant alterations in the intestinal enzyme activities examined. By 2 weeks of 2,4,5-T treatment, microbiota and urine genotoxicity returned to the levels observed in control animals. This trend continued for the duration of the experiment. After 2 weeks, while cecal nitroreductase and azo reductase activities increased, small intestinal beta-glucuronidase activity decreased. By 4 weeks, treated and untreated animal intestinal enzyme activities were indistinguishable.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: 2,4,5-Trichlorophenoxyacetic Acid; Animals; Biotransformation; Body Weight; Dinitrobenzenes; Drug Interactions; Feces; Genes; Intestinal Mucosa; Intestines; Liver; Male; Mutagenicity Tests; Rats; Rats, Inbred F344

Musk ketone, musk xylene, musk tibetene and moskene, synthetic musks used in fragrances, were applied dermally to rats in daily doses of 240 (musk ketone and musk xylene only), 75, 24 or 7.5 mg/kg body weight for 90 days. The chemically related musk ambrette, a known neurotoxin in rats, was used as a positive control. While musk ambrette was clearly neurotoxic and caused testicular atrophy, as had been previously reported, the other compounds tested caused neither effect. The only effects of application of these materials were some organ weight changes at the higher doses, but these were not associated with histopathological changes in any of the tissues. The no-effect levels were: musk ketone, 75 mg/kg for males and females; musk xylene, 75 mg/kg for males and 24 mg/kg for females; moskene, 24 mg/kg for males and 75 mg/kg (highest dose administered) for females; and musk tibetene, 75 mg/kg (highest dose) for males and females.

Topics: Administration, Topical; Animals; Body Weight; Dinitrobenzenes; Female; Indans; Kidney; Liver; Male; Molecular Structure; Nervous System; Organ Size; Perfume; Rats; Rats, Inbred Strains; Skin Absorption; Xylenes

These studies evaluated the reproductive response and the possible influence of testicular maturation on the reproductive parameters, in male rats treated with 1,3-dinitrobenzene (m-DNB). Young adult male rats (75 or 105 days of age) were given a single oral dose of 0, 8, 16, 24, 32, or 48 mg/kg of m-DNB and killed at 14 days post-treatment. Mortality and neurotoxicity were observed at 48 mg/kg, but only in the older animals. Epididymis weight, testicular sperm head counts, cauda sperm reserves, and sperm morphology were affected at 16 and 24 mg/kg and higher in the older and younger animals, respectively. Testis weight and sperm motility were affected at 24 mg/kg and higher in both age groups. Histologic changes included maturation depletion of mid and late spermatids at 16 mg/kg and higher, atrophy of a few to many seminiferous tubules at 24 mg/kg and higher, and immature germ cells in the epididymis. The movement and/or mixing of luminal elements in the epididymis appeared to be influenced by severe testicular effects. In separate groups given only the 48 mg/kg dosage, fertilizing ability was lost by 5-6 weeks post-treatment and several animals failed to recover in 5 months. In the breeder males, minimal to extensive degrees of seminiferous tubule atrophy and sloughed germ cells in the epididymis were still present after 175 days. The studies indicated that the lowest dosage to produce reproductive changes was 16 mg/kg with a no-effect level of 8 mg/kg. A few animals suffered protracted or permanent reproductive damage. Since the older animals were more susceptible to both the general and the reproductive toxicity of m-DNB, the less severe reproductive changes in the younger animals cannot be attributed solely to maturational differences in the testis.

Topics: Aging; Animals; Body Weight; Breeding; Dinitrobenzenes; Dose-Response Relationship, Drug; Epididymis; Male; Nitrobenzenes; Rats; Rats, Inbred Strains; Reproduction; Sperm Count; Testis

Exposure of prepubertal, pubertal, and adult mice to 0, 8, 16, 32, 40, or 48 mg 1,3-dinitrobenzene (m-DNB)/kg body weight and measuring responses 1-25 d posttreatment (dpt) demonstrated significant effects on testicular function only at 48 mg/kg dosage. m-DNB had no effect on body or testis weights with the exception of reduced adult mouse testis weights at 22 dpt with 48 mg/kg (p less than .05). None of the exposures resulted in detectable levels of germinal epithelial cells in the ductus epididymis. Exposure of prepubertal and pubertal mice to m-DNB caused only minimal nonsignificant changes in the relative percent of testicular cell types present up to 25 dpt. The adult mice testicular cell type ratios, in particular the round and elongating spermatid populations, changed significantly at doses of 48 mg/kg. Also, a reduction in the percent tetraploid cells occurred at d 1, suggesting these cells may be a primary target of m-DNB action. Caput and caudal sperm from mice exposed to m-DNB prior to puberty did not demonstrate an increased susceptibility to DNA denaturation when analyzed by the sperm chromatin structure assay. However, in pubertal mice, m-DNB exposure further exaggerated the abnormal chromatin structure that normally characterizes sperm during the onset of sperm production. In adult mice, 48 mg/kg resulted in increased susceptibility to DNA denaturation of caput sperm chromatin at 11 dpt (p less than .05) and in caudal sperm at 22 dpt (p less than .01). The abnormal chromatin structure of cauda sperm from adult mice was highly correlated with sperm head morphology abnormalities (ABN; 0.82 to 0.95, p less than .01, 11 and 22 dpt, respectively), but showed lower correlations with dose (0.60 to 0.79, p less than .01, 11 and 22 dpt, respectively). For pubertal mice, a positive relationship was also observed between the variation of sperm chromatin structure abnormalities and ABN. The effect of m-DNB on testicular function in prepubertal and pubertal mice appear to be less pronounced than in adult mice. Furthermore, following exposure to the same dosage, the effect of m-DNB is less severe in adult mice than that observed for adult rats as reported in the companion paper.

Topics: Animals; Body Weight; Chromatin; Dinitrobenzenes; Diploidy; Haploidy; Male; Mice; Mice, Inbred Strains; Mutation; Nitrobenzenes; Organ Size; Sexual Maturation; Spermatogenesis; Testis

Exposure of 100-d old rats to 1,3-dinitrobenzene (m-DNB) at dosages up to 48 mg/kg resulted in disruption of spermatogenesis as measured by flow cytometry (FCM) of acridine orange-stained sperm and testis cells. One day (d 1) after a single exposure to 48 mg/kg m-DNB, FCM measurements of caput epididymal fluid cells demonstrated the presence of testicular germinal epithelial cells apparently sloughed off into the epididymis. Also, at d 1 after the same exposure, a decrease in pachytene spermatocytes was observed. By d 16 after exposure to 32 or 48 mg/kg, testicular damage was evidenced by an alteration of cell type ratios in FCM-analyzed populations of testicular cells. Extensive recovery of cell type ratios occurred by d 32. At d 16, dosages of 32 and 48 mg/kg caused alterations of sperm chromatin structure as determined by the flow cytometric sperm chromatin structure assay (SCSA); 48 mg/kg caused alterations at both d 16 and d 32. Exposure to m-DNB caused a dose response increase in percent sperm head morphology abnormalities (%ABN) assessed in cauda epididymal and vas sperm. A slightly higher correlation existed between dose and SCSA alpha t values (d 16, .78; p less than .01) than between dose and %ABN (d 16, .70; p less than .01). Also, a higher correlation existed between standard deviation of alpha t (SD alpha t) values and %ABN (.97; p less than .01) than between dose and %ABN (.70; p less than .01). This study demonstrated rapid and unique FCM procedures originally derived for reproductive toxicology studies in mice to be equally useful for studies in rats.

Topics: Animals; Body Weight; Chromatin; Dinitrobenzenes; Diploidy; Epididymis; Flow Cytometry; Haploidy; Male; Nitrobenzenes; Organ Size; Rats; Rats, Inbred Strains; Ribonucleases; Sonication; Spermatids; Spermatogenesis; Testis; Vas Deferens

The present study was undertaken to evaluate the effects of the chemosterilant 2,4-dinitrotoluene (DNT) on the rat testis. Adult male rats were fed control, or 0.1%, or 0.2% DNT for 3 weeks. An ultrastructural study of the testes was performed, serum was assayed for testosterone and gonadotropins, and sperm reserve count was determined. A marked change in Sertoli cell morphology was found after 3 weeks of 0.2% DNT exposure. Varying sized vesicles associated with swollen mitochondria and distended endoplasmic reticulum were visible in cells from DNT-treated animals. Circulating levels of follicle stimulating hormone and luteinizing hormone were increased in 0.2% DNT-treated animals. Reduced weights of the epididymides and decreased epididymal sperm reserves were observed in DNT-treated animals. These results indicate that DNT is capable of inducing testicular injury, of directly or indirectly disturbing pituitary function, and of exerting a toxic effect at the late stages of spermatogenesis. These findings suggest that a locus of DNT action is the Sertoli cell, resulting in both inhibition of spermatogenesis and changes in testicular-pituitary endocrine activity.

Topics: Animals; Body Weight; Dinitrobenzenes; Follicle Stimulating Hormone; Male; Nitrobenzenes; Organ Size; Rats; Rats, Inbred Strains; Sperm Count; Spermatogenesis; Testis

The hepatocarcinogenicity of 2,4-dinitrotoluene [(2,4-DNT) CAS: 121-14-2], 2,6-DNT (CAS: 606-20-2), and a representative technical-grade DNT (TDNT) containing 76% 2,4-DNT and 18% 2,6-DNT was studied in male F344 rats. Rats were fed diets containing 2,4-DNT, 2,6-DNT, or TDNT at concentrations that resulted in doses of 27 mg/kg/day for 2,4-DNT, 7 or 14 mg/kg/day for 2,6-DNT, and 35 mg/kg/day for TDNT. The carcinogenic effects were evaluated after 1 year of treatment. Administration of 2,6-DNT produced hepatocellular carcinomas in 100 and 85% of animals receiving 14 and 7 mg/kg, respectively. In contrast to the 2,6-DNT results, feeding of 2,4-DNT for 1 year caused no hepatic tumors. Treatment with both isomers (TDNT) resulted in a 47% incidence of hepatocellular tumors. The majority of tumors had a trabecular pattern, and pulmonary metastases were present in the 14- and 7-mg/kg 2,6-DNT-fed groups. These results have demonstrated that 2,6-DNT is a potent and complete hepatocarcinogen and that 2,4-DNT, under these conditions, is nonhepatocarcinogenic. In addition, these data indicate that 2,6-DNT accounts for the majority of the carcinogenic activity of TDNT.

Topics: Animals; Body Weight; Carcinogens; Dinitrobenzenes; Epoxide Hydrolases; Isomerism; Liver; Liver Neoplasms, Experimental; Male; Microsomes, Liver; Nitrobenzenes; Organ Size; Phenobarbital; Rats; Rats, Inbred F344

Pectin-induced changes in microflora have been shown to elevate the covalent binding of 2,6-dinitrotoluene (2,6-DNT)-related materials to total rat hepatic macromolecules. Therefore, the effect of diets varying in pectin content on the induction of foci and hepatic tumors induced by 2,6-DNT was studied in male F344 rats. 2,6-DNT (3.0-3.5 and 0.6-0.7 mg/kg/day) was incorporated into NIH-07 (NIH), an open formula cereal-based diet high in pectin content, AIN-76A (AIN), a purified pectin-free diet, or AIN-76A supplemented with 5% pectin (AP). Hepatic foci were scored after histochemical staining for gamma-glutamyl transpeptidase (GGT), canalicular adenosine triphosphatase or glucose-6-phosphatase following administration of test diets for 3, 6 and 12 months. The number of foci per cm3 of liver increased in a dose- and time-department manner following incorporation of 2,6-DNT into test diets with NIH greater than AP greater than AIN. In the NIH diet, 2,6-DNT did not alter the phenotypic distribution of foci. Animals fed control or 2,6-DNT-containing AIN and AP diets had few or no GGT foci throughout the study. Hepatocellular carcinomas and neoplastic nodules were observed only in rats fed NIH containing 2,6-DNT. The concentrations of 2,6-DNT-related material covalently bound to hepatic macromolecules after a single oral dose of radiolabeled 2,6-DNT given after 12 months on the diets increased in control rats and in rats receiving low dose 2,6-DNT in the diet with AIN less than AP less than NIH. These studies show that the carcinogenicity of 2,6-DNT differs depending on whether rats are fed an NIH or AIN (+/- pectin) diet. The results suggest that diet-induced alterations in the covalent binding of 2,6-DNT are not the sole factor in determining the carcinogenic response to 2,6-DNT. Furthermore, unidentified contaminants in cereal-based diets may influence foci and tumor production in rat liver during carcinogen treatment.

Topics: Adenosine Triphosphate; Animals; Body Weight; Dietary Fiber; Dinitrobenzenes; gamma-Glutamyltransferase; Glucose-6-Phosphatase; Liver; Liver Neoplasms, Experimental; Male; Nitrobenzenes; Organ Size; Pectins; Rats; Rats, Inbred F344

The teratogenic potential of the fungicide dinocap was evaluated in CD-1 mice. Pregnant mice were dosed by intubation with dinocap in corn oil on gestation days 7-16. Doses used were 0, 5, 10, 20, 40, 80, and 120 mg/kg/day, based on day 6 weight. Dams were killed on day 18, at which time fetuses were counted, weighed, and preserved for necropsy or skeletal examination. The highest dose killed 80% of the dams dosed, while 29% of the dams in the 80 mg/kg group died during dosing. There was no dose-related maternal mortality at lower doses. Net maternal weight gain was affected only at 80 mg/kg/day. There were no live fetuses at 120 mg/kg/day. The number of live fetuses per litter was decreased and resorptions increased at 80 mg/kg. Dose-related decreases in gravid uterus weight and fetal weight were significant at all doses of dinocap. Cleft palate was found in fetuses at 5(1/234; 0.4%), 20(46/195; 23.6%), 40(140/185; 75.5%), and 80(63/85; 74.1%) mg/kg/day. There was a dose-related increase in supernumerary ribs and a low frequency of exencephaly and umbilical hernia at high doses. This study shows that dinocap is teratogenic in the CD-1 mouse at doses well below those causing maternal toxicity.

Topics: Animals; Body Weight; Dinitrobenzenes; Dose-Response Relationship, Drug; Female; Fetal Growth Retardation; Fungicides, Industrial; Mice; Nitrobenzenes; Pregnancy; Teratogens

Adult male Sprague Dawley rats were gavaged with 2,4-dinitrotoluene (2,4-DNT) dissolved in corn oil at 0, 60, 180, or 240 mg/kg/day for five days. A single oral dose (0.5 mg/kg) of triethylenemelamine was used as a positive control. Induction of dominant lethal events was scored on the basis of early fetal deaths. At the two lower doses, no consistent changes were observed in the numbers of pre-implantation losses, implantation sites, or living or non-living fetuses. The highest dose of 2,4-DNT tested resulted in a marked decrease in the numbers of sperm-positive females (determined by microscopic examination of vaginal smears for sperm) and pregnant females. These two effects diminished in the latter weeks of mating. The low number of pregnant females at the highest dose made meaningful statistical evaluations difficult. The results indicate that 2,4-DNT does not cause dominant lethal mutations but does adversely affect reproductive performance.

Topics: Animals; Body Weight; Corpus Luteum; Dinitrobenzenes; Embryo Implantation; Female; Fertility; Fetal Resorption; Fetus; Genes, Dominant; Genes, Lethal; Male; Mutagenicity Tests; Mutagens; Nitrobenzenes; Pregnancy; Rats; Rats, Inbred Strains; Sexual Behavior, Animal

Topics: Animals; Behavior, Animal; Body Weight; Cells, Cultured; Dinitrobenzenes; Drinking; Eating; Female; Male; Nitrobenzenes; Organ Size; Rats; Sleep; Testis

2, 4-Dinitrotoluene (2, 4-DNT) was incorporated at the level of 0.5% in a standard commercial diet and fed ad lib. to male rate for 6 months. Significant changes were noted in the body weight, organ weight, behavior, mortality, and biochemical analysis of blood and serum of rats ingesting 2, 4-DNT. Furthermore, it was found that the ingestion of 2, 4-DNT affected on the activities of drug metabolizing enzymes in liver microsomes.

Topics: Animals; Behavior, Animal; Body Weight; Diet; Dinitrobenzenes; Enzymes; Male; Methemoglobin; Nitrobenzenes; Organ Size; Rats; Time Factors

Topics: Aging; Animals; B-Lymphocytes; Body Weight; Dinitrobenzenes; Dose-Response Relationship, Immunologic; Female; gamma-Globulins; Humans; Immune Tolerance; Male; Mice; Organ Size; Spleen; T-Lymphocytes