Behavioral toxicity of chronic postweaning lead exposure in the rat.

Effects in the rat of chronic postweaning exposure to 50, 300, or 1000 ppm lead acetate in drinking water were assessed on responding maintained by a fixed-interval 30-s food reinforcement schedule. On this schedule, food reinforcement followed the first lever press response occurring at least 30 s after the preceding food delivery. Exposure to 50 and 300 ppm lead acetate increased response rates and intersubject variability, while latency, or time to initiation of responding in the interval, decreased. Exposure to 1000 ppm initially decreased rates and increased latency values. Following termination of exposure to 50 ppm, response rates and latency values gradually returned to control levels. Behavioral effects produced by exposure to 50 and 300 ppm lead were similar in magnitude but varied in time to onset and decline, suggesting time-dependent, rather than concentration-dependent effects of lead.     

Acute kidney toxicity of sodium fluoride in the rat

Single i.p injection of 0, 0.05, 0.10, 0.25, 0.50 and 1.00 mmol NaF/kg to male rats induced a dose-related increase of urine gamma-glutamyl transferase (GGT) elimination on the first day for all doses higher or equal to 0.25 mmol NaF/kg. Kidney damage began between 2--4 h following the injection and lasted only 12 h for the 0.50 mmol NaF/kg dose. There was an increase of diuresis and of phosphaturia even at lower doses.     

Urinary antigens as markers of papillary toxicity. I Identification and characterization of rat kidney papillary antigens with monoclonal antibodies

Monoclonal antibodies were prepared in an attempt to develop diagnostic tools for the identification of toxic damage to the rat renal papilla. One IgG and five IgM monoclonal antibodies, reacting with antigens localized in the papilla were obtained. Three of the IgM class and the IgG class monoclonal antibodies were found to be specific for antigens localized in collecting ducts, two of them staining papillary collecting ducts more intensely than cortical collecting ducts. The IgG class antibody, termed Pap X 5C10, recognizes an antigen located at high density on the luminal side of papillary collecting duct epithelial cells and at lower density in cortical collecting duct cells. One of the IgM class monoclonal antibodies reacts with an antigen localized in epithelial cells of ascending and descending loops of Henle and of connecting tubules. Another of the IgM class monoclonal antibodies reacts with an antigen localized in the interstices of the inner medulla. All these monoclonal antibodies react with their antigens in native frozen as well as in Bouin-fixed and paraffin-embedded tissue slices. Molecular properties of the Pap X 5C10 antigen have been investigated by gel permeation chromatography, SDS-PAGE, Western blotting, and isoelectric focusing. The results indicate that the antigen in both its tissue-derived and urinary form is of large (150–200 kDa) molecular size and can be separated into two molecular species with isoelectric points of pH 7.2 and 7.3 respectively. In the urine the antigens recognized by the monoclonal antibodies form large complexes with Tamm-Horsfall protein. The antigen-containing complexes can be extracted from urine by adsorption to diatomaceous earth and elution with SDS-containing buffer. Using sandwich ELISA-type assays it is possible to determine the concentration of the antigens. In preliminary experiments we were able to show that at least three of the antigens are detected in the urine following toxic insults to the kidney. The monoclonal antibodies prepared and the tests developed thus may provide direct diagnostic access to the renal papilla and allow, for the first time, early detection of papillary damage. Received: 14 May 1996/Accepted: 19 July 1996     

Influence of diet and strain of rat on kidney damage observed in toxicity studies.

The laboratory animal is susceptible to biological variation due to a variety of causes. Thus genetic variation and nutrition have become recognized to influence the reaction of the animal to experimental procedures. Severe kidney injury was observed at our institute in preliminary studies with 1% butylated hydroxytoluene (BHT) incorporated in a semisynthetic diet fed to Wistar rats for one month. These lesions together with nephrocalcinosis, a common finding in rats fed a semisynthetic diet might suggest that the animals in our experiment were predisposed to kidney damage. In order to show the influence of the rat strain and the type of diet a study was performed with both Wistar and Sprague-Dawley rats fed a semisynthetic or commercial diet mixed with 1% BHT. It was observed that the semisynthetic diet had a marked effect on the severity of the kidney lesions and that the lesions were independent of the rat strain used. In addition, an apparent sex variation was found.     

Pharmacological characterization and anatomical localization of prejunctional beta-adrenoceptors in the rat kidney.

1. The subtype and anatomical localization of beta-adrenoceptors mediating facilitation of stimulus-induced overflow of noradrenaline ('prejunctional beta-adrenoceptors') are not conclusively known to date. The present study was undertaken to characterize these receptors by use of pharmacological methods as well as to define their localization (prejunctional or postjunctional) with radio-ligand binding and autoradiography techniques combined with surgical denervation of the sympathetic innervation to the rat kidney. 2. Exposure of the kidney to (-)-isoprenaline, the nonselective beta-adrenoceptor agonist, resulted in a dose-dependent facilitation of stimulus-induced neurotransmitter overflow. This response was inhibited by propranolol, the beta 1- and beta 2-adrenoceptor antagonist, with a pA2 of 9.20 suggesting that the prejunctional beta-adrenoceptors are not of the beta 3-subtype. 3. The rank order of potency and potency ratios of beta-adrenoceptor agonists at renal prejunctional beta-adrenoceptors (EC50 for agonist/EC50 for (-)-isoprenaline) were: (-)-isoprenaline (1) > procaterol (2) > salbutamol (3) > adrenaline (10) > (+)-isoprenaline (25). However, dobutamine, the beta 1-adrenoceptor agonist, failed to enhance stimulus-induced overflow of noradrenaline. These results are indicative of the presence of beta 2-adrenoceptors as prejunctional beta-adrenoceptors. 4. Facilitation elicited by (-)-isoprenaline and procaterol, the selective beta 2-adrenoceptor agonist, was inhibited by ICI 118,551, the selective beta 2-adrenoceptor antagonist, with pKb values of 9.20 and 9.35, respectively at renal prejunctional beta-adrenoceptors. Similarly, the pKb values of metoprolol, the selective beta 1-adrenoceptor antagonist, at renal prejunctional beta-adrenoceptors were determined to be 6.25 and 6.18 against (-)-isoprenaline and procaterol, respectively. These results suggest the presence of a homogeneous population of beta 2-adrenoceptors as prejunctional beta-adrenoceptors. 5. Radio-ligand binding analysis of renal beta-adrenoceptors revealed the prevalence of the beta 1-subtype as compared to the beta 2-subtype (63% vs 37%). However, surgical denervation of the rat kidney, resulting in more than 90% reduction in renal noradrenaline content, selectively reduced the beta 2-adrenoceptor population by 80%, implying the presence of beta 2-adrenoceptors on renal sympathetic nerve terminals. 6. Autoradiographic analysis demonstrated the presence of beta 1-adrenoceptors on cortical structures such as glomeruli and tubules. beta-Adrenoceptors were found to be present on tubules (minor population), collecting tubules in outer medulla and the adventitia and adventitial-medial border of intraparenchymal branches of the renal artery.(ABSTRACT TRUNCATED AT 400 WORDS)     

The chronic toxicity of equine cadmium metallothionein in the rat

The extensive renal tubular necrosis that results in male rats after the intravenous injection of a single, low dose of equine kidney cadmium (Cd), zinc(Zn)-metallothionein (MT) (0.2 mg MT-bound-Cd/kg body wt.) is followed within 72 h by active regeneration. With repeated administration of the same dose at 3-or 4-day intervals, the lesion resolves although, at least initially, the kidney content of Cd increases progressively. At any time during treatment, about 40% of the accumulated Cd is bound as the endogenous (Cd, Cu)MT. The rate of increase in the renal Cd content is dependent on the ratio of CdZn in the injected metalloprotein, and is appreciably less when the constant dose of protein-bound Cd is given as a (2.4 Cd1 Zn)MT, than as a (3.0 Cd1 Zn)MT. On repeated administration of the latter preparation, however, the concentration of Cd in the kidney does not attain a critical concentration, above which persistant tubular damage occurs, but reaches a maximum of about 150–160 g Cd/g wet wt. (after 16 doses) and then declines. After 19 doses of the (2.4 Cd1 Zn)MT under the same conditions, the renal Cd concentration is submaximal and is less (92 g Cd/g wet wt.) than that after either 16 or 27 doses of the (3.0 Cd1 Zn)MT. In animals that are dosed with either of the heterologous MT preparations, the first dose, although not innocuous, seems to protect the kidneys against further damage by subsequent doses. Repeated doses, however, lead to vascular changes, e.g. lymphoid infiltration, periarteriole oedema and dilation of the arcuate veins, and to dilation of the glomerular spaces.     

Lithium and the developing rat kidney in transplacental target organ toxicity

Pregnant female albino rats were treated orally with lithium carbonate dissolved in distilled water. Treatment at 100 mg/kg day 16 until day 20 of gestation caused marked maternal toxicity including polyuria. For the progeny increased rates of prenatal and postnatal mortality were noted. In part of the progeny sacrificed near term by Caesarean section, the visceral examination of the fetuses revealed an enlargement of the renal pelves in association with rudimentary or missing papillae. The renal anomalies are interpreted as being consistent with a developmental retardation due to specific lithium activity. After birth, i.e. after termination of maternal treatment, slight to moderate structural changes of the kidney were apparently compensated. 60 mg/kg through days 16-20 of pregnancy caused moderate maternal toxicity including polyuria. The offspring showed a postnatal development near the normal range, however, and no renal anomalies were recorded. In the view of the nephrotrophic property of lithium as known for the adult rat, the results indicate that possible transplacental effects on the fetal kidney as a target organ should be also considered.     

Chronic low-level lead toxicity in the rat: III. An integrated assessment of long-term toxicity with special reference to the kidney

Male and female rats whose mothers had been exposed to Pb before and during pregnancy and lactation at exposure levels of 0, 0.5, 5, 25, 50, and 250 ppm Pb as Pb-acetate in drinking water were continued on the respective regimens for 6 or 9 months. Body weights of males and females were not significantly different from controls at 6 months of age; however, female body weights were significantly decreased at 250 ppm at 9 months of age. In males at 9 months of age, spleen weights were significantly increased at 250 ppm Pb and kidney weights were increased at 0.5 ppm Pb and above; in females the liver, pituitary, and heart weights were affected at 250 ppm Pb. No significant Pb effects were found in sperm counts or sperm morphology, hematology profiles, or serum chemistries. Blood, brain, femur, and kidney Pb levels as well as urinary ALA excretion were all significantly dose related. Histopathological lesions were noted in the spleen (250 ppm) and in the kidney as evidenced by cytomegaly/karyomegaly (beginning at 5 ppm in males; 25 ppm in females), nuclear inclusion body formation and increased numbers of iron-positive granules within renal proximal tubule cells. These effects were more marked after 9 months exposure. Ultrastructural studies revealed mitochondrial swelling and the presence of increased numbers of lysosomes within renal proximal tubule cells. Energy-dispersive X-ray microanalysis, performed on adjacent sections, showed the highest intracellular Pb concentrations in nuclear inclusion bodies within renal proximal tubule cells. Inhibition of renal mitochondrial respiration for both succinate and NAD-linked substrates was found in 50- and 250-ppm Pb exposure groups at 9 months but not at 6 months. Mitochondrial δ-aminolevulinic acid synthetase and ferrochelatase, but not δ-aminolevulinic dehydratase, were also found to be inhibited at these Pb levels at 9 months. The lowest exposure level resulting in a detectable effect of Pb (cytomegaly/karyomegaly in renal proximal tubule cells) was 5 ppm associated with a median blood Pb concentration of 11 μg/dl.     

A characterization of the acute cardiopulmonary toxicity of fenfluramine in the rat

Fenfluramine (FN) is a potent serotonin-releasing drug used primarily as an anorectic agent. The symptomatology of its acute lethality has been well documented in animal models such as the rat. A very prominent feature of this lethality profile is hypoxia, as demonstrated by the onset of severe cyanosis just prior to death. It is not clear in the literature whether this hypoxia is the result of a direct pulmonary effect or is secondary to cardiac injury. To further characterize this aspect of FN's toxicity, respiratory and electrocardiographic measurements were taken in anaesthetized rats subjected to high doses of FN (129.6 mg/kg, i.p.). Death occurred in these animals within 15 min of drug administration, apparently as the result of abrupt respiratory cessation, followed by cardiac ischaemia. No significant gross or histopathological lesions were evident in these animals. In other trials, prior treatment with diethylcarbamazine (DEC) was found to potentiate the lethality of FN, while cyproheptadine (CHP) pretreatment attenuated FN's toxic effects. Necropsies, conducted 24 h after FN administration, revealed widespread alveolar and pulmonary interstitial haemorrhage in the CHP-pretreated animals. The data suggest that high doses of FN directly result in pulmonary hypertension, which secondarily induces ischaemic cardiac injury.     

Effect of chronic ethanol administration on bromobenzene liver toxicity in the rat

Female Sprague-Dawley rats were pair-fed a nutritionally adequate liquid diet containing either ethanol or isocaloric carbohydrate for 3 weeks. In vitro studies showed that chronic ethanol pretreatment preferentially increased the liver microsomal biotransformation of bromobenzene to p-bromophenol (via the toxic 3,4-epoxide) rather than to o-bromophenol (via the nontoxic 2,3-epoxide) and could thus potentiate bromobenzene hepatotoxicity. Bromobenzene administration (500 mg/kg body weight, ip), after an overnight fast, was associated in ethanol-pretreated rats with greater and accelerated liver glutathione depletion and subsequent decrease in liver cytochrome P-450 content than in controls. As assessed histologically and by determination of the rise in activities of serum enzyme markers of liver necrosis, chronic ethanol pretreatment, however, mainly resulted in earlier onset and resolution of bromobenzene-induced liver necrosis, with only a mild increase in the maximal severity of liver lesions. These results suggest that the twofold increase in liver microsomal bromobenzene 3,4-epoxidation by ethanol, being much less than that seen after phenobarbital pretreatment in our animal model and in that of others, is apparently not sufficient to markedly affect the severity of bromobenzene-induced liver toxicity.     

Induction of drug-metabolizing enzymes and toxicity of trans-stilbene oxide in rat liver and kidney

The effect of trans-stilbene oxide (TSO) on organ function and morphology and on drug-metabolizing enzymes was determined in male Sprague-Dawley rats. TSO (300 or 600 mg/kg) was administered i.p., once daily for 5 consecutive days. At a dose of 3400 mg/kg, TSO did no alter body weight, but increased liver weight. The higher dose (600 mg/kg) markedly decreased body weight. TSO treatment (300 mg/kg) induced several drug-metabolizing enzymes. Epoxide hydrolase activity was enhanced in the liver, kidney and lung. In contrast, arylhydrocarbon hydroxylase activity was not significantly altered. Glutathione S-transferase activity, with 1-chloro-2,4-dinitrobenzene as substrate, and uridine diphosphoglucuronyl transferase activity, with p-nitrophenol as substrate, were also increased in the liver and kidney after TSO treatment. It appears that TSO induces hepatic and renal enzyme activities in a similar manner. Treatment with the higher dose of TSO depressed accumulation of p-amino-hippurate by renal cortical slices and increased blood urea nitrogen concentration. Histological examination of kidney sections after treatment with TSO revealed no abnormality. The lower dose led to negligible alteration in liver and the higher dose resulted in mild to moderate hepatic cellular.     

Curcumin inhibits adenosine deaminase and arginase activities in cadmium-induced renal toxicity in rat kidney

In this study, the effect of enzymes involved in degradation of renal adenosine and l-arginine was investigated in rats exposed to cadmium (Cd) and treated with curcumin, the principal active phytochemical in turmeric rhizome. Animals were divided into six groups (n = 6): saline/vehicle, saline/curcumin 12.5 mg/kg, saline/curcumin 25 mg/kg, Cd/vehicle, Cd/curcumin 12.5 mg/kg, and Cd/curcumin 25 mg/kg. The results of this study revealed that the activities of renal adenosine deaminase and arginase were significantly increased in Cd-treated rats when compared with the control (p < 0.05). However, co-treatment with curcumin inhibits the activities of these enzymes compared with Cd-treated rats. Furthermore, Cd intoxication increased the levels of some renal biomarkers (serum urea, creatinine, and electrolytes) and malondialdehyde level with a concomitant decrease in functional sulfhydryl group and nitric oxide (NO). However, co-treatment with curcumin at 12.5 mg/kg and 25 mg/kg, respectively, increases the nonenzymatic antioxidant status and NO in the kidney, with a concomitant decrease in the levels of malondialdehyde and renal biomarkers. Therefore, our results reinforce the importance of adenosine deaminase and arginase activities in Cd poisoning conditions and suggest some possible mechanisms of action by which curcumin prevent Cd-induced renal toxicity in rats.     

Short-term toxicity of photomirex in the rat.

When photomirex (8-monohydromirex) was fed to rats at dietary levels of 0, 0.5, 5.0, 50, or 500 ppm, all 10 animals receiving the highest dose levels died within 7 days, and two of 10 animals in the 50-ppm group died after 24 days. All other animals (10 per group) survived until the experiment was terminated at 28 days. Animals receiving 50 ppm showed decreased body weight gain and food consumption. Liver hypertrophy and increased microsomal enzyme activity (aniline hydroxylase) were observed in groups of animals receiving 5.0 ppm and greater. Serum sorbitol dehydrogenase activity was increased in animals receiving 5.0 ppm and greater. Photomirex accumulated in all tissues examined, with fat and liver having the highest residues. Histological alterations were observed in the liver at 0.5 ppm and consisted of mild midzonal cytoplasmic enlargement. At a level of 5 ppm, the midzonal cytoplasmic enlargement was more pronounced and was accompanied by mild anisokaryosis and nuclear hyperchromicity. At the highest dose, these alterations were accompanied by perivenous fatty infiltration. Lesions of the thyroid were observed in the group of animals receiving 50 ppm (6.61 mg/kg/day) and consisted of a generalized reduction in follicle size and colloid density. Animals which died showed pronounced thyroid follicular atrophy accompanied by severe colloid depletion and epithelial exfoliation. Animals receiving 50 ppm also had lesions in the testes consisting of a mild to marked reduction in spermatogonia and complete cessation of spermatogenesis.     

Subacute toxicity assessment of annatto in rat.

Increased human use of annatto (Bixa orellana L), a red yellow food colorant, demands generation of toxicity data. The toxic effects of annatto powder (bixin 27%) have been assessed following administration of a subacute regimen (4 weeks, 20 doses) in Wistar male and female rats. A full study with three dose levels was considered unnecessary since no sign of toxicity had been noted in a preliminary experiment with 1000 mg/kg body weight/day as was recommended by the OECD guideline. In this study, annatto administered by gavage at a dose level of 2000 mg/kg/day decreased male body weight gain, but had no effect on either food intake or food conversion efficiency. Haematological and plasma biochemical examination as well necropsy performed at the end of administration (29th day) and observation (43rd day) periods revealed no alterations related with annatto administration. Kidney apoptosis occurred in 20% treated female rats in restricted areas without proliferation or tubular segments modification. The precise nature of apoptosis was not investigated in the present study. These findings suggest that annatto was no toxic to the rat.     

Cellular toxicity of hydrazine in primary rat hepatocytes.

Hydrazine (HzN) is an aircraft fuel and propellant used by the U.S. Air Force. The current study was undertaken to evaluate the acute toxicity of HzN in primary rat hepatocytes in vitro with reference to oxidative stress. The effects of short-term exposure (4 h) of hepatocytes to HzN were investigated with reference to viability, mitochondrial function, and biomarkers of oxidative stress. The viability data showed an increase in lactate dehydrogenase leakage and a decrease in mitochondrial activity with increasing concentration of HzN. The results of studies of oxidative stress biomarkers showed a depletion of reduced glutathione (GSH) and an increase in oxidized GSH, increased reactive oxygen species generation, lipid peroxidation, and reduced catalase activity. Furthermore, depletion of GSH and catalase activity in hepatocytes by buthionine sulfoximine and 3-amino triazole, respectively, prior to exposure to HzN, increased its toxicity. The results suggest that acute HzN-induced cytotoxicity in rat hepatocytes is likely to be mediated through oxidative stress.     

Ninety-day toxicity of photomirex in the male rat.

Photomirex (8-monohydromirex) is a demonstrated environmental contaminant and was observed in previous short-term studies to produce lesions in the liver, thyroid and testes of male rats. The present study was undertaken to confirm those observations and to determine the effects after a longer period of exposure. Male rats were fed photomirex for 13 weeks at levels of 0.20, 1.0, 5.0, 25 and 125 ppm in the diet. Deaths were observed in animals receiving the highest dose. Decreased body weight gain and food intake were also observed in that group. Liver weights were increased at 5.0 ppm photomirex and higher. Photomirex caused changes in several biochemical parameters including serum sorbitol dehydrogenase and hepatic aniline hydroxylase activities. Dose-related histological abnormalities were observed in the thyroid and liver starting at the lowest dose level. These results confirm earlier findings and show that photomirex is a potent hepato- and thyrotoxin.     

Pharmacokinetics and toxicity of idarubicin in the rat.

This study was designed to examine the pharmacokinetics and toxicity of idarubicin (IDA) in rats. In two groups of rats IDA was infused either into the V. iugularis interna or into the A. carotis communis, respectively. The venous plasma concentration of IDA and its primary metabolite idarubicinol (IDOL) were measured up to 48 hours by high-performance liquid chromatography (HPLC) with fluorescence detection. The weights of the rats and the levels of haemoglobin, leukocytes, and thrombocytes were recorded. The plasma concentration-time data were analysed, assuming a biexponential disposition curve, both by the traditional (two-stage) method and by population pharmacokinetic modelling. The basic pharmacokinetic parameters clearance (CL = 27.0 ml min(-1)), mean disposition residence time (MDRT = 519.2 min), and volume of distribution at steady state (Vss = 12.51) were estimated for IDA. The mean residence time (MRT) of the generated IDOL was 2982.5 min. No significant differences between pre- and postpulmonal injection were found in the pharmacokinetics and pharmacodynamics of IDA. The mean survival time of 13.3 days is attributed to a severe myelosuppression.