trans-Nonachlor and cis-nonachlor toxicity in Sprague-Dawley rats: comparison with technical chlordane.

cis-Nonachlor and trans-nonachlor are bioaccumulating components of the pesticide chlordane, which can be detected in various environmental biota and in humans. Existing studies have focused on the potential adverse health effects of the parent chlordane mixture. Comparable toxicity data are nonexistent for individual chlordane constituents such as trans-nonachlor, cis-nonachlor, or oxychlordane, which are among the most common chlordane-related environmental contaminants and tissue residues. In this study, rats were administered cis-nonachlor, trans-nonachlor, or technical chlordane by gavage for 28 days at doses of 0.25 to 25 mg/kg body weight. Residue analyses indicated that trans-nonachlor accumulation in adipose was greater than cis-nonachlor when rats were administered each chemical under identical conditions of dose and exposure. For all test chemicals, the major metabolite oxychlordane accumulated in adipose tissue. Adipose tissue residue levels of all test chemicals and the major metabolite were higher in female rats. The liver was a target organ in male and female rats, indicated by increased liver weight and histopathological changes consistent with microsomal enzyme induction. Hepatic changes were most pronounced in rats treated with trans-nonachlor. Elevated kidney weights and depressed organic ion transport were observed in males treated with trans-nonachlor and chlordane. Although in general, changes in target organs and clinical chemistry endpoints were similar for all 3 test chemicals, the approximate toxicity ranking from most to least toxic was trans-nonachlor > technical chlordane > cis-nonachlor.

Glutathione S-transferase-placental form expression and proliferation of hepatocytes in fumonisin B1-treated male and female Sprague-Dawley rats.

Fumonisin B1 (FB1), a mycotoxin produced by a common corn contaminant Fusarium moniliforme and a hepatocarcinogen in rats, has been previously suggested to act as a poor initiator, but a better promoter of gamma-glutamyltranspeptidase (GGT)-positive rat liver preneoplastic lesions. Using glutathione S-transferase-placental form (GSTP) as a more sensitive marker of initiation, we have further evaluated the initiating capacity of various doses of purified FB1 administered (a) intraperitoneally (i.p.) to male Sprague-Dawley (SD) rats for 4 days and (b) orally (PO) to male and female SD rats for 11 days. Compared to their respective controls, significant increases in GSTP-positive hepatocytes were observed in male rats administered FB1 i.p. at 10 mg/kg body weight/day for 4 days, as well as in male and female rats treated with 35 and 75 mg/kg body weight/day FB1 p.o. for 11 days. The percentage section area of liver occupied by GSTP-positive mini-foci comprising of three to 12 cells was increased significantly in male rats given 10 mg/kg FB1 i.p., or in p.o.-treated males and females with 75 mg/kg FB1. Both i.p. and p.o. FB1 treatments resulted in dose-related enhanced hepatocyte proliferation as measured by proliferating cell nuclear antigen (PCNA) labeling with significant increases in the number of PCNA-positive nuclei at the same i.p. and p.o. dose levels where the number of GSTP-positive cells were elevated. In all studies, enhanced PCNA and GSTP expression occurred at FB1 doses which, based on serum biochemical and histopathological data previously reported from our laboratory, were shown to be hepatotoxic. Therefore, our data suggest that in a manner similar to known genotoxic carcinogens, FB1 has the capacity to initiate GSTP-positive hepatocytes with their subsequent development into GSTP mini-foci at exposure levels that induce enhanced hepatocyte proliferation in response to liver toxicity. In SD rats, this occurs as early as within 4 days of i.p. treatment or 11 days of p.o. treatment.

Gavage administration of the fungal toxin fumonisin B1 to female Sprague-Dawley rats.

The fungal toxin fumonisin B1 (FB1) is a contaminant of corn-based foods and feeds produced by members of the genus Fusarium. Fumonisin B1 toxicity was examined using gavage administration of purified toxin to female Sprague-Dawley rats. For 11 consecutive days each rat received a single dose of FB1 at the following concentrations: control (saline), 1, 5, 15, 35, or 75 mg FB1/kg body weight/d. Significantly depressed body weight and food consumption occurred at 35 and 75 mg FB1/kg/d. By the end of the dosing period there were no significant changes in food consumption. Kidneys and bone marrow were most sensitive to FB1 exposure. Changes in renal morphology were observed from 5 to 75 mg FB1/kg/d, accompanied by transient changes in urine osmolality and urine enzyme levels. Increased cellular vacuolation was the primary change associated with bone-marrow toxicity, starting at doses of 5 mg FB1/kg/d. Hepatotoxicity was indicated by reduced liver weight, elevated serum alanine amonitransferase (ALT), and mild histopathological changes occurring at doses of 15 mg FB1/kg/d and higher. Increased cytoplasmic vacuolation of adrenal cortex cells occurred in rats treated with 15 mg FB1/kg/d and higher, indicating that the adrenals are also potential targets of FB1. Elevated serum cholesterol, which is a consistent response to FB1 was observed at 5 mg FB1/kg/d and higher. Based on responses in this study, gavage is an appropriate substitute for longer feeding studies. Compared to previous work with male rats, gender-related difference in FB1 responses lacked consistency but indicated that males may be marginally more sensitive than female Sprague-Dawley rats.

Toxicity of fumonisin B1 to B6C3F1 mice: a 14-day gavage study.

Fumonisin B1 (FB1) is a fungal toxin produced by members of the genus Fusarium. Ingestion of FB1 causes species-specific neurotoxic, nephrotoxic, hepatotoxic and pulmonary effects. The clinical, haematological and pathological responses of adult male and female B6C3F1 mice to FB1 were assessed following 14 daily gavage doses ranging from 1 to 75 mg FB1/kg body weight/day. There were no consistent sex-related changes. Although all responses were modest, the most notable effects of FB1 were on the liver, bone marrow, adrenals and kidneys. In the liver, hepatocellular single cell necrosis, mitosis and anisokaryosis were observed, accompanied by elevated serum ALT. In the kidneys, minor histopathological changes were confined to female mice, while mild decreases in ion transport and increases in blood urea nitrogen were seen only in males. Small changes in glutathione levels were observed in the kidneys and livers of male mice. Adrenal cortical cell vacuolation was observed at 15 mg FB1/kg and higher in females and from 35 mg FB1/kg in males. Serum cholesterol was elevated in both male and female mice, possibly due to FB1-induced changes in lipid metabolism in the liver and adrenals. Although bone marrow cell numbers were unchanged, increases in vacuolated myeloid cells and lymphocytes were observed in female mice. In general, the degree of changes observed indicate that mice are not as sensitive a model of FB1 toxicity as rats.

The effects of fumonisin B1 on several markers of nephrotoxicity in rats.

Rats were injected intraperitoneally with saline or fumonisin B1 (FB1) at doses of 7.5 and 10.0 mg FB1/kg for 4 days. For each day of dosing, 24-hr urine samples were collected and analyzed for creatinine and protein content and the enzymes gamma-glutamyl-transpeptidase, lactate dehydrogenase, and N-acetyl-beta-D-glucosaminidase. Twenty-four hours after the last dose, animals were killed and kidneys removed for ion transport measurement and histopathology. Significant increases in urine volume and decreases in urine osmolality were observed in both FB1 dose groups. Creatinine excretion was decreased only in the 10 mg FB1/kg group on the final day of the study. Urine protein excretion was elevated in both treated groups and found to be due primarily to high-molecular-weight proteins indicative of increased glomerular permeability. Enzymuria, a marker of tubular cell damage, was also observed with increases in the urinary excretion of all three enzymes measured. In renal cortical slices tubular transport of the anion p-aminohippuric acid was reduced by 75-80% and cationic transport of tetraethylammonium was reduced by 40% in the FB1-treated animals. While these results suggest significant alterations in renal function, only minor histopathologic changes were observed in the kidneys of both dose groups. Results of the present study indicate that urine volume, proteinuria, enzymuria, and ion transport are sensitive indicators of early FB1-induced nephrotoxicity.

Renal clearance of domoic acid in the rat.

The renal clearance (Clr) of the seafood toxin domoic acid (DA) was investigated in the rat. Following cannulation of the right femoral artery, the left femoral vein and the bladder of anaesthetized rats, a single bolus injection of either [3H]DA, [14C]p-aminohippuric acid (PAH) or [3H]inulin was administered through the venous cannula. Blood samples were taken from the arterial cannula at 1, 2, 10, 30, 50, 70, 90, 110 and 130 min following injection, and urine samples were collected at 20-min intervals starting from the time of bolus injection. Based on plasma concentration-time profiles, the total clearances (Clt) for DA, PAH and inulin were 9.12, 33.17 and 7.50 ml/min/kg body weight, respectively. The Clr calculated from urinary excretion rates were not significantly different from the Clt. Probenecid significantly reduced the Clr of PAH but did not affect that of DA. When DA was given at doses of 0.5 ng, 0.5 mg and 2.0 mg/kg body weight, the pharmacokinetic parameters Clt, Clr, elimination-rate constant and apparent volume of distribution at steady state were not statistically different between doses. The entire dose of 3H was recovered in the urine by 160 min after dosing, and analysis of urine samples by HPLC confirmed that the radiolabel (3H) was associated predominantly with the parent form of DA. The results of the present study demonstrate that DA is cleared from plasma primarily through the kidneys. DA clearance occurs primarily by renal glomerular filtration since its Clt is comparable with that of inulin, is less than that of PAH and is not affected by probenecid.

Systemic toxicity of the heavy fraction of a coal coprocessing product in male rats following subchronic dermal exposure.

The systemic toxicity of a coal coprocessing product [heavy gas oil II (HGOII)] following subchronic, dermal exposure in male Sprague-Dawley rats was investigated. HGOII was applied to the dorsal skin daily at doses of 8.7, 20.8, 50.0, or 120.0 mg/kg body weight (bw) for 13 weeks. Another group of rats treated with a medium boiling coal liquefaction product (CLP) served as positive controls. Growth suppression and decreased food consumption were noted in the groups exposed to HGOII at 20.8 mg/kg and higher, and to CLP starting at the third week of treatment. Relative liver, kidney, and brain weights in the 20.8 mg/kg HGOII group and up were higher than those of the control. Increased spleen weight was observed in all HGOII-treated groups. CLP treatment also caused increased relative kidney and brain weights. Serum cholesterol was elevated in the HGOII-treated groups starting at 8.7 mg/kg while increased uric acid and lactate dehydrogenase were observed at 20.8 mg/kg and up. Decreased erythrocyte, hemoglobin, and platelet counts were observed at 20.8 mg/kg and higher. All HGOII-treated groups had elevated reticulocytes. These biochemical and hematological changes were not observed in the CLP-treated group. Mild to marked histological changes were observed in the thyroid, thymus, liver, spleen, and bone marrow of HGOII groups. In contrast, morphological changes were relatively mild in CLP-treated animals. Data from the present study demonstrated that the hematological endpoints were sensitive to the liquid fuels and that HGOII was more toxic than CLP.(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of enzymuria as an indicator of amikacin-induced renal damage in guinea pigs.

Guinea pigs were injected subcutaneously for 10 days with amikacin (AK) at a dose of 0, 100, 200 or 400 mg/kg body wt. per day. The total daily dose was administered in either a single injection or divided equally and given as two daily injections. After the 10 days of AK treatment, uptake of the organic cation, tetraethylammonium (TEA) into renal cortical slices was inhibited in a dose-related manner. Changes in renal tubular morphology also increased with higher doses. The urinary excretion of the enzymes, N-acetyl-beta-D-glucosaminidase (NAG), and alkaline phosphatase (ALP) significantly increased during the course of AK treatment, however, due to the large intragroup variability, the daily fluctuations and the absence of any distinct trends in urinary enzyme excretion it was difficult to establish a dose-relationship between AK-induced renal damage and the resultant enzymuria. At doses of 100 and 200 mg/kg body wt., the two-injection regimen resulted in the greater renal accumulation of AK and damage as reflected by a greater inhibition of TEA uptake and greater changes in renal tubular morphology. In contrast, this difference in toxicity could not be detected with enzymuria again due to the large intragroup variability and the absence of discernable excretion patterns of NAG and ALP. Thus, neither NAG nor ALP appear to be suitable quantitative markers of AK-induced nephrotoxicity.

The effect of guanidine on the accumulation of amikacin in guinea pig renal cortical slices.

The role of a recently identified organic ion transport system in the accumulation of the aminoglycoside (AG), amikacin (AK) in the kidney was investigated in the present study. Because this transport system has been characterized as being a carrier for the organic cation, guanidine, the effect of guanidine on the uptake of AK into renal slices from guinea pig was examined. Renal slices incubated in medium containing AK concentrated the drug against a concentration gradient (i.e. slice:medium ratio (S/M) greater than 1.0). This uptake was significantly reduced when an equimolar concentration (1 x 10(-5) M) of another AG, gentamicin was added to the incubation medium. In contrast, AK uptake was relatively insensitive to the presence of the cation, tetraethylammonium (TEA) in the medium. Guanidine was also ineffective at inhibiting AK uptake into slices and reduced AK uptake by only 22% at guanidine concentrations of 1 x 10(-2) M. In comparison, TEA was slightly more sensitive to the presence of guanidine in the incubation media since TEA uptake was reduced by 22% at guanidine concentrations of 1 x 10(-3) M and reduced by approximately 70% at guanidine concentrations of 1 x 10(-2) M. Thus, the results of the present study suggest that the guanidine transport system does not play a role in the renal accumulation of AK since the presence of guanidine in the incubation medium had little effect on the accumulation of AK into renal cortical slices.

The interactions of cis-diamminedichloroplatinum with metallothionein and glutathione in rat liver and kidney.

The involvement of metallothionein (MT) in the nephrotoxicity of cis-diamminedichloroplatinum (c-DDP) was investigated in rats using enzyme excretion and histology as indicators of renal damage. In addition, the effects of renal glutathione (GSH) depletion on the nephrotoxicity of c-DDP was assessed by organic anion transport in renal cortical slices. A dose of 6.0 mg c-DDP/kg body wt, i.p. was administered to rats either as a single injection of 6.0 mg/kg or as six daily injections of 1.0 mg/kg. Concentrations of platinum (Pt) after c-DDP injection in both dosing regimens were approximately 12 micrograms/g in kidney and 2 micrograms/g in liver. However, there were no increases in either hepatic or renal concentrations of MT after both series of c-DDP injections. Fractionation of kidney cytosols from c-DDP injected rats on Sephadex G-75 columns revealed that 60-70% of cytosolic Pt was associated with proteins of high molecular weight and 15-20% of the Pt associated with the low molecular weight ligands. No discernable Pt peak was detected in the elution volume of MT. Pretreatment of rats with ZnSO4 increased both hepatic and renal concentrations of MT, but there was no Pt associated with the MT fraction after a subsequent injection of c-DDP. Small increases in the urinary excretion of the lysosomal enzyme, N-acetyl-beta-D-glucosaminidase and two brush border enzymes, alkaline phosphatase and gamma-glutamyltranspeptidase were observed 2 and 3 days after a single injection of c-DDP (6.0 mg/kg body wt, i.p.). Urinary creatinine excretion decreased by 50% 1 day after c-DDP injection and continued to decrease for the next 2 days. On the third day after c-DDP treatment, a small but significant decrease in body weight was also observed in the c-DDP injected animals. Pretreatment with Zn did not alter the c-DDP-induced enzymuria or renal tubular damage but slightly attenuated both the decrease in creatinine excretion and the loss in body weight. Uptake of the organic anion, p-aminohippuric acid (PAH) was reduced at 12 and 24 h after c-DDP injection. Reduction of tissue GSH concentrations by pretreatment with buthionine sulfoxime (BSO), resulted in only a slight increase in the c-DDP-induced inhibition of PAH uptake at 24 h after c-DDP injection. These results suggest that, in rats, neither MT nor GSH appear to play major roles in the binding or nephrotoxicity of c-DDP.