Determination of the place conditioning and locomotor effects of amperozide in rats: a comparison with cocaine.

Amperozide promotes social interactions in rats (Rattus norvegicus) and reduces craving for cocaine and alcohol without producing adverse side effects. Amperozide administration produced a place preference in the dose range from 3.0 to 10.0 mg/kg. Locomotor behavior tended to be suppressed on the days of amperozide pairings, elevated on the days of vehicle pairings, and elevated on test days. Administration of 20.0 mg/kg but neither 2.0 nor 0.2 mg/kg cocaine produced place preference. Rats exhibited a greater place preference for the chamber paired with 2.0 mg/kg and 20.0 mg/kg but not 0.2 mg/kg cocaine compared to the chamber paired with 5.0 mg/kg amperozide. The results indicate that amperozide is appetitive. The most appetitive dose of amperozide (5.0 mg/kg) is less appetitive than the most appetitive dose of cocaine (20.0 mg/kg). More research is needed before conclusions regarding the degree of abuse potential for amperozide can be made.

Activation of the human aldehyde oxidase (hAOX1) promoter by tandem cooperative Sp1/Sp3 binding sites: identification of complex architecture in the hAOX upstream DNA that includes a proximal promoter, distal activation sites, and a silencer element.

Aldehyde oxidase (AOX) is a member of the molybdenum iron-sulfur flavoproteins and is of interest for its role in clinical drug metabolism and as a source of reactive oxygen species (ROS) potentially involved in human pathology. The ROS derived from AOX contribute significantly to alcohol-induced hepatotoxicity. Therefore, expression of AOX could determine both the susceptibility of certain cells and tissues to clinically important pharmacologic agents and the levels of ROS produced under certain pathophysiological conditions. Although some pharmacologic agents regulate AOX enzyme activity, very little is known about the activation or regulation of the human AOX gene (hAOX). In the present study, we sought to identify features in the upstream DNA of hAOX that could confer regulation of the gene, to locate and characterize the basal promoter apparatus activating hAOX, and to identify transcription factors that could mediate activation or regulation. We transfected promoter fusion constructs into epithelial cells from the lung and the mammary gland that express AOX in cell culture. The hAOX gene was found to possess a structurally complex region in the upstream DNA that contained sequences for a proximal promoter, enhancer sites, and silencer elements. In addition, we identified an essential role for the transcription factors Sp1 and Sp3 in the proximal promoter. Unexpectedly, hAOX was activated in lung and mammary epithelial cells by indistinguishable mechanisms. These observations reveal a potentially complex mode of hAOX gene expression in epithelial cells that is dependent on Spl and Sp3 transcription factors.

cDNA cloning, sequencing, and characterization of male and female rat liver aldehyde oxidase (rAOX1). Differences in redox status may distinguish male and female forms of hepatic APX.

Molecular characterization of male and female rat liver aldehyde oxidase is reported. As described for the mouse liver, male and female rat liver expressed kinetically distinct forms of aldehyde oxidase. Our data suggest that the two forms arise as a result of differences in redox state and are most simply explained by expression of a single gene encoding aldehyde oxidase in rats. In support of this argument we have sequenced cDNAs from male and female rat liver. We examined mRNA expression by Northern blot analysis with RNA from males and females, from several tissues, and following androgen induction. Purified rat liver enzyme from males or females revealed a single 150-kDa species consistent with cDNA sequence analysis. Both male and female forms were reactive to the same carboxyl-terminal directed antisera. Km(app) values obtained in crude extracts of male or female rat liver and post-benzamidine-purified aldehyde oxidase differed substantially from each other but could be interconverted by chemical reduction with dithiothreitol or oxidation with 4,4'-dithiodipyridine. Our data indicate that a single gene is most likely expressed in male or female rat liver and that the kinetic differences between male and female rat liver aldehyde oxidases are sensitive to redox manipulation.

Intra-arterial administration of carboplatin and the blood brain barrier permeabilizing agent, RMP-7: a toxicologic evaluation in swine.

RMP-7 is a bradykinin B2 receptor agonist shown to permeabilize the blood-brain barrier, especially that associated with brain tumors, when administered via both intracarotid and intravenous routes. Both routes of administration are currently being tested in human trials in combination with the chemotherapeutic agent carboplatin as therapy for gliomas. As an essential prerequisite to the initial intracarotid clinical trials, the potential neurotoxicity of intra-arterial administration of RMP-7 (at a high or low dose), alone and in combination with carboplatin, was assessed in anesthetized Red Duroc swine. Five treatment groups were evaluated with each pig receiving a series of alternating, intra-arterial infusions of RMP-7 (or saline) followed by carboplatin (or saline), as follows: (1) vehicle control: saline/saline; (2) carboplatin only control: saline/carboplatin (50 mg total); (3) RMP-7 only control: RMP-7 (750 ng/kg)/saline; (4) low dose combination: RMP-7 (75 ng/kg)/carboplatin (50 mg total); and (5) high dose combination: RMP-7 (750 ng/kg)/carboplatin (50 mg total). For each subject, one of the alternating dosing sequences (above) was repeated four times during a single dosing session which lasted approximately 40 minutes. Assessments during the in-life phase of the study in the pre- and post-treatment periods consisted of heart rate, arterial blood pressure (systolic, diastolic, and mean), blood gases, body weight, general clinical observations (including evaluation for neurological deficit) and clinical pathology (including a comprehensive battery of standard blood coagulation, hematological and serum chemistry tests). In addition, during the time of treatment, heart rate and arterial blood pressure were monitored. The animals were terminated two weeks after dosing and the brain and rete mirabile (distal to site of infusion) were evaluated for gross and histopathological abnormalities. The histopathology analysis included a reader-blinded analysis using low and high power light microscopic examination of both H&E and Kluver-Berrera stained sections through several key cortical and subcortical brain regions. Transient decreases in arterial blood pressure (mean of 10-25 mmHg) were observed in both groups receiving the high dose of RMP-7 (i.e., 750 ng/kg). No other side effects attributable to RMP-7 and/or carboplatin were observed, and clinical observations revealed no evidence of neurologic deficits. Post-mortem examination revealed no evidence of CNS or cerebral vascular pathology attributable to carboplatin and RMP-7. This study demonstrates that intracarotid administration of the maximum tolerated dose of RMP-7 (750 ng/kg) alone, or in combination with carboplatin (50 mg) is not accompanied by any serious adverse effect, apparent cerebrovascular abnormality or neuropathologic consequence and offers further evidence for the safety of this novel therapeutic approach for enhancing delivery of chemotherapeutics to brain tumors.

Rarely occurring spontaneous metastasizing testicular tumors in rats. Histopathologic and ultrastructural features.

Testicular tumors are not uncommon in aging rats. However, metastasis from these neoplasms to several distant visceral organs has not been reported. Testicular tumors were observed in a total of 18 (1.34%) Sprague-Dawley rats out of 1340 males examined. Out of these 18 neoplasms, interstitial tumors were observed in 12 rats, of which 11 were benign and one malignant. Spermatic seminomas were observed in two (0.15%) cases (one benign and one malignant). Both rats with malignant testicular neoplasms were sacrified in extremis. Remaining four tumors were classified as mesotheliomas. Detailed gross, histopathologic and electron microscopic features of primary testicular neoplasms with metastases to several distant visceral organs are described and discussed.

Spontaneous renal neoplasms in rats.

Spontaneous renal neoplasms in the rat are uncommon. This paper presents the incidence and histopathological features of various long-term developing renal tumors observed in control rats from 17 carcinogenicity studies (1340 males and 1329 females) in Sprague-Dawley rats and 10 carcinogenicity studies (530 males and 530 females) in Fischer-344 (F-344) rats. Renal cell adenoma (0.08% in Sprague-Dawley and 0.28% in F-344), lipoma/liposarcoma (0.37% in Sprague-Dawley and 0.75% in F-344) and transitional cell carcinoma (0.07% in Sprague-Dawley and 0.09% in F-344) were observed in both Sprague-Dawley (0.49%) and F-344 (1.13%) rats. Pulmonary metastasis was observed from one case of transitional cell carcinoma. Renal cell carcinoma with metastasis to the lung and liver was seen in one F-344 rat. In addition, transitional cell papilloma and nephroblastomas were also observed in Sprague-Dawley rats. Metastases from nephroblastoma were seen in the lungs and renal lymph node in two cases. In both rat strains, the tumor incidence was higher in males than in females.

Spontaneous neoplasms in aged Sprague-Dawley rats.

Incidence of neoplastic lesions in untreated Sprague-Dawley rats (1340 males and 1329 females) used as controls in 17 carcinogenicity studies are tabulated and evaluated. In male rats, the most common neoplasms were benign pheochromocytomas and keratoacanthomas (4.0% in each case) followed by pancreatic islet cell adenomas (3.7%), thyroid parafollicular cell adenomas (3.6%), fibromas and squamous cell papillomas of the skin and hepatocellular adenomas (2.0% in each), malignant lymphoma lymphocytic (1.9%), histiocytic sarcomas (1.4%), and adrenal cortical adenomas (1.2%). In female rats, the most common neoplasms were of mammary gland origin (31.3%: fibroadenoma 19.0%, adenocarcinomas 8.8%, and adenomas 3.5%) followed by thyroid parafollicular cell adenomas (2.9%), uterine endometrial stromal polyps (2.6%), adrenal cortical adenomas (1.9%), malignant lymphoma lymphocytic (1.6%), fibromas in the skin (1.3%), and pancreatic islet cell adenoma (1.1%). Metastases were observed from pheochromocytomas, hepatocellular carcinomas, nephroblastomas, renal pelvis transitional cell carcinoma, interstitial cell tumor and seminoma of the testes, Zymbal's gland adenocarcinomas, and mammary adenocarcinomas.

Clear-cell carcinoma in the trachea of a rat.

Necropsy examination of a control Sprague-Dawley male rat from a carcinogenicity study disclosed the presence of a neoplastic mass in the trachea. Microscopic examination disclosed that the tumor consisted of closely packed cells arranged in lobules and cords separated by thin connective tissue stroma. The cells had distinct cell borders and abundant pale cytoplasm. Electron microscopic examination further disclosed that the cytoplasm contained numerous argyrophilic neurosecretory granules measuring 298 nm in diameter. These granules were positive for Grimelius silver stain. The histopathologic and electron microscopic features of this tumor are consistent with a diagnosis of clear-cell carcinoma.

Incidence and pathology of spontaneous renal pelvis transitional cell carcinomas in rats.

Spontaneous transitional cell carcinoma (TCC) originating from renal pelvis is very rare. Metastasis from this tumor having been reported only once previously. Data from control groups of 17 carcinogenicity studies in Sprague-Dawley and 10 carcinogenicity studies in Fischer 344 (F-344) rats were reviewed and evaluated to determine the incidence and pathology of TCC. Renal pelvis TCC were observed in 1 male and 1 female Sprague-Dawley rat and in 1 male F-344 rat. Metastasis from TCC to the lung was observed in 1 male Sprague-Dawley rat. Detailed histopathological features of this tumor are described and discussed.

One-year inhalation toxicity study of chlorine in rhesus monkeys (Macaca mulatta).

Chlorine (Cl2) gas is a potentially lung-damaging irritant which is used in the chemical, plastics, and paper industries. There are no data published using experimental animals on the chronic inhalation toxicity of chlorine. The purpose of this study was to investigate the chronic effects of Cl2 inhalation in Rhesus monkeys (Macaca mulatta). Rhesus monkeys were exposed to concentrations of 0, 0.1, 0.5, or 2.3 ppm Cl2 for 6 hr per day. 5 days per week for 1 year. Pulmonary physiology (pulmonary diffusing capacity and distribution of ventilation), body weights, urinalysis, electrocardiographs, hematology, and clinical chemistry were evaluated monthly during the study. Blood gas evaluations were performed at 3-month intervals during the study. Histopathologic, ophthalmologic, and neurologic parameters were evaluated after the 1-year exposure period. Monkeys exposed to 2.3 ppm Cl2 exhibited signs of ocular irritation during the daily exposures and a superficial conjunctival irritation was present in the 2.3 ppm group after the 1-year exposure regimen. Treatment-induced lesions revealed by histopathology were confined to the respiratory tract. Lesions associated with the nasal parasite Anatrichosoma spp. were present in the region of squamous epithelium of the nasal vestibule and did not interfere with interpretation of Cl2-induced effects. Treatment-induced histopathologic changes were found in the respiratory epithelium of the nasal passages and trachea and were limited to focal, concentration-related epithelial hyperplasia with loss of cilia and decreased numbers of goblet cells in affected areas. These changes in the nose and trachea were focal and mild in monkeys exposed to 2.3 ppm and were not found in all animals in these exposure groups. Tracheal lesions were confined to the 2.3 ppm group. The lesions observed at 2.3 ppm were not present in all animals. At the lower Cl2 concentrations, similar though less prominent respiratory epithelial lesions were observed. The latter changes were very minimal and were confined to the nasal passages of some treated monkeys and one male control animal. The results of this study indicate that 2.3 ppm chlorine acts as an upper respiratory irritant in monkeys, while 0.5 and 0.1 ppm induce changes of questionable clinical significance. Furthermore, the monkey appears to be less sensitive than the rat to chlorine toxicity.

Acute target organ toxicity of 1-nitronaphthalene in the rat.

1-Nitronaphthalene (1-NN) produced respiratory distress and centrilobular liver necrosis in male Sprague-Dawley rats after a single intraperitoneal injection (100 mg kg-1). Microscopic examination of the lungs of rats killed 24 h after the injection revealed a highly selective non-ciliated bronchiolar (Clara) cell necrosis as the only remarkable lesion. Pretreatment of animals with phenobarbital offered complete protection from the respiratory distress induced by 1-NN, but increased the severity of the hepatotoxicity. Pretreatment with SKF-525A protected against 1-NN-induced liver necrosis, but did not alter the incidence or severity of the respiratory distress. Under similar conditions, this pattern of toxicity was not seen with the structural analogue 2-nitronaphthalene.

Urine and tissue oxalate and hippurate levels in ethylene glycol intoxication in the dog.

Oxalate, hippurate and ethylene glycol were measured in dogs given 5 ml/kg ethylene glycol orally. Eight hours after administration, oxalate levels were 7.2-9.1 ppm in renal tissue and 10-100 ppm in urine. Hippurate was 40-90 ppm in urine. Ethylene glycol was 10-100 ppm in renal tissue and 5,300-27,000 ppm in urine. The above substances were measured in urine and renal tissue from normal dogs, dogs in non-toxic renal failure and clinical cases of ethylene glycol intoxication. Urine was analyzed for ethylene glycol by gas chromatography using direct injection. Ethyl ether extraction provided good recovery of ethylene glycol from tissues and blood. Results were confirmed by gas chromatography/mass spectrometry (GC/MS). Oxalic acid (CA oxalate) was extracted with acidic methanol, methylated and analyzed by GC. Results were confirmed by GC/MS. Hippuric acid (calcium hippurate) was extracted with acidic methanol and chromatographed by thin layer chromatography (TLC). Ultraviolet spectrometry and mass spectroscopy were used to confirm the results. High performance liquid chromatography (HPLC) and TLC with densitometry were used to quantitate hippuric acid.

Haemophilus parasuis infection in swine.

Septicemic disease occurred in 49 of 126 pigs several days after being transported 80 km. All affected pigs died. The main changes in acutely affected pigs were skin discoloration, pulmonary edema, arthritis, meningitis, and renal glomerular thrombosis. In peracute cases, gross findings were minimal. Haemophilus parasuis was isolated from multiple organ sites in most affected pigs. Haemophilus parasuis was isolated from nasal swab specimens from 17 of 20 clinically normal pigs on the farm of origin. Fatal acute septicemia was reproduced in 2 pigs by intravenous or intratracheal exposure to an isolant of H parasuis obtained from 1 of of the 49 fatally affected pigs. Aerosol exposure of 5 pigs resulted in mild pneumonia in 4 pigs and severe pneumonia, pleurisy, pericarditis, and terminal septicemia in 1 pig.