The genetic toxicity of the peroxisome proliferator class of rodent hepatocarcinogen

Peroxisome proliferators comprise a structurally diverse class of chemicals. Some of the members of this class show evidence of genetic toxicity (most evidently the in vitro clastogen Wyeth 14,643, WY), while others do not (most evidently methyl clofenapate, MCP). When attempting to understand the mechanism of rodent hepatocarcinogenesis of this class of chemicals the possible role of genetic toxicity should be assessed on a class-wide basis, i.e., if just one peroxisome proliferator is shown to be unequivocally inactive as a genetic toxin, genetic toxicity cannot be implicated in the carcinogenic activity of peroxisome proliferators as a class. In an earlier paper, we established MCP as inactive in a range of in vitro and in vivo genetic toxicity assays. However, the top dose level of MCP that could be tested for induction of chromosome aberrations (clastogenicity) in human lymphocytes and CHO cells was limited by the relative insolubility of the test agent in the assay medium. Methyl clofenapate was not toxic up to a dose that produced precipitate, so cannot be directly compared with WY, which induced aberrations only at toxic dose levels. In the present paper, we have evaluated the clastogenicity of the carcinogenic peroxisome proliferator nafenopin (NAF) at dose levels up to those that are toxic to CHO cells, and found no evidence of chromosome aberration induction. These data isolate further the genetic toxicity of WY from other peroxisome proliferators, and increase confidence in the proposal that genetic toxicity does not play a critical role in the hepatocarcinogenicity of peroxisome proliferators.     

Species differences in ciprofibrate induction of hepatic cytochrome p450 4A1 and peroxisome proliferation

Six species (CD-1 mouse, Fischer 344 rat, Syrian golden hamster, Duncan-Hartley guinea pig, half-lop rabbit and marmoset monkey) were treated orally with ciprofibrate, a potent oxyisobutyrate hypolipidaemic drug for 14 days. A dosedependent liver enlargement was observed in the mouse and rat and at the high dose level in the hamster. A marked dose-dependent increase in the 12-hydroxylation of lauric acid was observed in the treated mouse, hamster, rat, and rabbit, associated with a concomitant elevation in the specific content of cytochrome P-450 4A1 apoprotein, determined by an ELISA technique. Similarly, in these responsive species, an increase in mRNA levels coding for cytochrome P450 4A1 was observed. Lauric acid 12-hydroxylation was unchanged in the guinea pig and marmoset after ciprofibrate pre-treatment, and cytochrome P-450 4A1 was not detected immunochemically in liver microsomes from these latter species. In the untreated mouse, hamster, rat, and rabbit, the 12-hydroxylation of lauric acid was more extensive than the 11-hydroxylation, whereas in the guinea pig and marmoset the activity ratios were reversed, with 11-hydroxylation predominating. Peroxisomal fatty acid β-oxidation was markedly induced in the mouse, hamster, rat, and rabbit on treatment at the higher dose level (39-, 3-, 13- and 5-fold, respectively) and was slightly increased in the marmoset (2-fold), yet was unchanged in the guinea pig following treatment. In the marmoset the increase in peroxisomal β-oxidation was 3- to 4-fold at the high dose level; however, the dose levels used in the marmoset were 20 and 100 mg/kg as opposed to 2 and 20 mg/kg in the other species. The differences in the foregoing hepatic enzyme responses to ciprofibrate between the species examined in our studies indicate a specific pattern of enzyme changes in responsive species. In the responsive species (rat, mouse, hamster, and rabbit), cytochrome P-450 4A1 specific content and related enzyme activity were increased concomitant with elevated peroxisomal β-oxidation. By contrast, the marmoset and guinea pig lack the coordinate hepatic induction of peroxisomal and microsomal parameters and may be categorized as less responsive species. Accordingly, the rat hepatic responses to peroxisome proliferators cannot confidently be used to predict biological responses in primates, with obvious implications for the extrapolation of animal data to man.     

Ultrastructural analysis of the effect of ethane dimethanesulphonate on the testis of the rat,guinea pig,hamster and mouse

Summary The morphological response of the testis of rats, guinea pigs, Syrian hamsters and mice to treatment with the cytotoxin ethane dimethanesulphonate was examined using light and electron microscopy. One to two days after a single administration of ethane dimethanesulphonate to adult rats, guinea pigs, and hamsters, the Leydig cells showed marked ultrastructural alterations suggestive of degeneration and cell death. The former alterations included karyopyknosis, cytoplasmic vesiculation and accumulation of lipid inclusions and large lipofuscin bodies. Fragments of necrotic Leydig cells were often engulfed by the interstitial tissue macrophages. The morphology of the seminiferous epithelium of these three species was unchanged from the morphology observed in vehicle-injected control animals. In contrast, multiple injections of ethane dimethanesulphonate given to mice produced no ultrastructural alterations to Leydig cells yet the seminiferous epithelium exhibited disruption of spermatogenesis. Although the Leydig cells of the mouse appear resistant to ethane dimethanesulphonate, this agent exerts a selective cytotoxic action upon Leydig cells of the rat, guinea pig and hamster thus identifying ethane dimethanesulphonate as a useful chemical for future endocrine and physiological studies of testicular function in three common laboratory species.     

Comparative physiology of rodent pulmonary macrophages: in vitro functional responses

Since toxicological testing of inhaled materials frequently requires utilization of several species, we have investigated pulmonary macrophage (PM) functional responses and compared the rat model with other rodents. Two strains of rats, three strains of mice, and one strain each of hamster and guinea pig were used in this study. The numbers of recovered cells by bronchoalveolar lavage generally correlated with animal body weight. The one exception was the Syrian Golden hamster from which increased numbers of macrophages were recovered. Cellular differential data obtained from lavaged cytocentrifuge preparations demonstrated that PM's account for greater than 97% of recoverable free lung cells for all species except the guinea pig, which contains a resident population of eosinophils. Cell morphology studies indicated that hamster PM exhibited the highest proportion of ruffled PM and demonstrated the highest phagocytic activity, whereas mouse PM phagocytic activity was significantly reduced compared with the other three species. In addition, chemotaxis studies showed that rat PM migrated best to zymosan-activated, complement-dependent chemoattractants, whereas hamster PM demonstrated an enhanced chemotactic response to N-formyl peptides. The results of these studies suggest that the rat may be the most efficient species for clearing inhaled particles, whereas hamsters and guinea pigs may best respond to bacteria.     

Immunofluorescent research on the species specificity of the antigens of hematopoietic cells in rodents

The bone marrow cells of mice, rats, guinea pigs, Syrian and dwarf hamsters exhibit a positive immunofluorescence reaction with antisera against insoluble antigens of the bone marrow cells of mice, Syrian and dwarf hamsters and, hence, contain common "cross-reacting" antigens. The use of different methods of antiserum absorption made it possible to reveal, in addition, antigens of "narrow" specificity in (1) mice, Syrian and dwarf hamsters, (2) Syrian and dwarf hamsters, as well as species specific antigens of the bone marrow cells of mice, Syrian and dwarf hamsters.     

Species differences in cytotoxic and genotoxic effects of phenacetin and paracetamol in primary monolayer cultures of hepatocytes

The cytotoxic and genotoxic effects of phenacetin and paracetamol were examined in monolayer cultures of hepatocytes isolated from the mouse, hamster, rat and guinea pig. No marked increase in unscheduled DNA synthesis (UDS) after exposing hepatocytes from any of the species to phenacetin was observed. At cytotoxic concentrations of paracetamol, an increased UDS in mouse hepatocytes in vitro was observed. Pretreatment of the mice by inducers of drug-metabolizing enzymes, such as 3-methylcholanthrene and Aroclor 1254, lowered the concentration threshold for the toxic responses. With rat hepatocytes only a minor increase in UDS was noted, while with hepatocytes from hamsters and guinea pigs in fact a decrease was seen. The narrow range observed between the cytotoxic and genotoxic effects of paracetamol makes it difficult to predict whether the initial DNA damage could lead to a mutation or whether the cells will die before the mutation is expressed. With respect to the cytotoxic effects, hamster hepatocytes were found to be most susceptible to paracetamol, followed by mouse, while rat and guinea pig were less affected. These data were in accordance with in vivo findings (Davis et al., 1974), indicating the potential value of hepatocyte culture when screening for possible liver toxic substances.     

Induction of microsomal 1-acylglycerophosphocholine acyltransferase by peroxisome proliferators in rat kidney; co-induction with peroxisomal beta-oxidation

Induction of microsomal 1-acyl-glycerophosphocholine (GPC) acyltransferase in rat tissues by four peroxisome proliferators, clofibric acid, tiadenol, DEHP and PFOA, was examined. Among the nine tissues examined, kidney, liver and intestinal mucosa responded to the challenges by the peroxisome proliferators to induce the enzyme. The treatment of rats with various dose of clofibric acid, tiadenol, DEHP or PFOA resulted in an induction of kidney microsomal 1-acyl-GPC acyltransferase in a dose-dependent manner. Despite the structural dissimilarity of peroxisome proliferators, the induction of microsomal 1-acyl-GPC acyltransferase was highly correlated with the induction of peroxisomal beta-oxidation. The activity of microsomal 1-acyl-GPC acyltransferase was not affected by changes in hormonal (adrenalectomy, diabetes, hyperthyroidism and hypothyroidism) and nutritional (starvation, starvation-refeeding, fat-free-diet feeding and high-fat-diet feeding) states. The induction of renal microsomal 1-acyl-GPC acyltransferase was seen in mice subsequent to the administration of clofibric acid and tiadenol and in guinea pigs subsequent to the administration of tiadenol. These results may indicate that kidney microsomal 1-acyl-GPC acyltransferase is a highly specific parameter responsive to the challenges by peroxisome proliferators and may suggest that the possibility that the inductions by peroxisome proliferators of microsomal 1-acyl-GPC acyltransferase and peroxisomal beta-oxidation in kidney are co-regulated.     

Subacute inhalation of cigarette smoke by pregnant and lactating rodents: AHH changes in perinatal tissues

To study the transplacental acquisition of tobacco smoke products and the effects on fetal tissue enzymes, pregnant rats, guinea pigs, and hamsters were exposed to freshly generated cigarette smoke via a nose-only inhalation system on a daily basis through the latter one-third (guinea pigs) or latter half (rats, hamsters) of the gestational period. Following euthanasia on the day of parturition, microsomal aryl hydrocarbon hydroxylase (AHH) activities were determined in the lungs, livers, and kidneys of both dams and fetuses. The possible acquisition of tobacco smoke products via the milk was studied by exposing lactating dams to cigarette smoke daily for either 4 or 14 days (rats), 4 or 7 days (guinea pigs), or 10 days (hamsters), with analysis of tissues from the euthanized pups for AHH. Pups were also exposed directly (nose only) to cigarette smoke. In the treated pregnant and lactating rat, maternal pulmonary, hepatic, and renal AHH was significantly increased but only fetal lung and the liver of 14-day-old pups showed a marked induction of AHH activity. In the pregnant and lactating guinea pig, only the pulmonary and renal AHH activities were increased following exposure, whereas in the fetuses and nursing pups, none of the tissue AHH activities was significantly altered by exposure. In the pregnant and lactating hamster, only the pulmonary AHH was increased following exposure to cigarette smoke, whereas the activity in fetal and pup tissues remained unchanged from the levels observed in control animals.(ABSTRACT TRUNCATED AT 250 WORDS)     

Calcium content of peripheral and central mitochondria in the guinea pig myocardium: electron probe analysis

We quantitated subcellular elemental concentrations in stimulated and resting guinea pig myocardium to determine whether species-specific properties of guinea pigs or the subcellular localization of mitochondria accounted for reports of higher mitochondrial Ca in guinea pigs than in other species. Small papillary muscles or trabeculae isolated from guinea pig ventricles were stimulated to raise cytosolic [Ca(2+)](i) by two methods: (1). tetanizing by rapid pacing preparations in which Ca(2+) uptake by the sarcoplasmic reticulum was inhibited with cyclopiazonic acid or (2). freeze trapping paced muscles near-peak systole. Electron probe X-ray microanalysis showed no significant difference between the (low, approximately 0.4 mmol/kg dry weight) mitochondrial Ca content of stimulated guinea pig hearts, compared to mitochondria of other species, such as rat and hamsters, and the Ca contents of peripheral and central mitochondria were also not significantly different.     

Ammonium Sulfate Fractionation of Sera: Mouse, Hamster, Guinea Pig, Monkey, Chimpanzee, Swine, Chicken, and Cattle

Optimal (NH(4))(2)SO(4) concentrations were sought for serum fractionation in order to obtain the gamma globulin as free as possible from other serum components while maintaining a reasonable recovery. Various ammonium sulfate concentrations were used to fractionate sera from mice, hamsters, guinea pigs, monkeys, chimpanzees, swine, chicken, and cattle. All precipitates and supernatants were analyzed by electrophoresis to study the effects of various treatments on the composition of these materials. Approximately 75% of all the gamma globulins were recovered when each serum was fractionated with its optimal sulfate concentration. These optimals were determined to be as follows: three precipitations in 35% saturated ammonium sulfate (SAS) for hamster, chimpanzee, swine, and chicken serum; one precipitation in 35% SAS followed by two in 40% SAS for mouse and guinea pig serum; one precipitation in 30% SAS and then two in 40% SAS for monkey serum; and one precipitation in 30% SAS followed by two in 35% SAS for cattle serum.     

Effect of phenobarbital and the peroxisome proliferator ciprofibrate on γ-glutamyltranspeptidase activity and leukotriene C4 concentration in cultured rat hepatocytes

Peroxisome proliferators induce hepatocellular carcinomas in rodents by an unknown mechanism. γ-Glutamyltranspeptidase (GGT), a biochemical marker for identifying putative preneoplastic lesions in the liver, is highly expressed in phenobarbital (PB)-promoted altered hepatic foci but not in those promoted by peroxisome proliferators. One of the substrates of GGT is the eicosanoid LTC₄. Because peroxisome proliferators and PB have differing effects on eicosanoid metabolism in vivo, we hypothesized that PB would similarly increase LTC₄ concentrations, whereas the peroxisome proliferator ciprofibrate (CIP) would not. Cultured hepatocytes were treated with the peroxisome proliferator ciprofibrate (CIP: 100 and 400 μM) or PB (PB: 0.5 and 2 mM). Competitive radioimmunoassay (RIA) was used to determine the concentration of LTC₄ in extracts of cultured hepatocytes. CIP decreased the concentration of LTC₄ throughout the culture period, but PB increased the LTC₄ concentration. Both doses of CIP significantly inhibited the induction of GGT activity at 48 and 72 hours, whereas PB enhanced GGT activity. We therefore hypothesized that LTC₄, a substrate of GGT, may induce GGT activity. LTC₄, however, did not enhance GGT activity and inhibited it at very high concentrations. The results of this experiment show that CIP and PB have different effects on GGT activity and LTC₄ concentration. LTC₄, however, does not induce GGT in cultured hepatocytes. © 1996 John Wiley & Sons, Inc.     

Cytopathic effect of heat-labile toxin of Bordetella parapertussis on aortic smooth muscle cells from pigs or guinea pigs

The in vitro effect of the heat-labile toxin (HLT) of Bordetella parapertussis on HeLa, baby hamster kidney (BHK), chinese hamster ovary (CHO), myeloma (Pa-NS-1), human embryonic lung (HEL-R66) cells, erythrocytes, adipocytes and lymphocytes from guinea pigs, mice or rats, or aortic smooth muscle cells from pigs or guinea pigs was examined. Within 8, 6, 4, 2, and 2 hr after the exposure to 1, 3, 10, 30, and 100 MNDs/ml of HLT, respectively, the cultured smooth muscle cells only showed a cytopathic change. When the cells exposed to HLT were washed out within 60 min post-exposure, the change could be induced with an extend period of lag. Histamine, KCl or norepinephrine caused similar change in the cells, but the period of lag was within 30 min. The HLT activity was neutralized by an anti-B. parapertussis- or B. bronchiseptica-HLT guinea pig IgG. HLT had no effects on any other cells tested.     

Inhaled radon-induced genotoxicity in Wistar rat,Syrian hamster,and Chinese hamster deep-lung fibroblasts in vivo

This study was performed (1) to provide a comparison of the genotoxin effects of inhaled radon and radon progeny, referred to as radon in this paper, among three species of rodents: Wistar rats, Syrian hamsters, and Chinese hamsters; (2) to determine if initial chromosome damage was related to the risk of induction of lung cancer; and (3) to evaluate the tissue repair and long-term presence of cytogenetic damage in respiratory tract cells. These species were selected because Syrian hamsters are very resistant to radon induction of lung cancer and Wistar rats are sensitive; no literature is available on the in vivo effects of radon in the Chinese hamster. Exposure-response relationships were established for the rats and Syrian hamsters while the Chinese hamsters received a single exposure of radon. At 4 h (0.2 days), 15 days, and 30 days after the highest WLM exposure to radon, Wistar rats, Chinese hamsters, and Syrian hamsters were killed, and lung fibroblasts were isolated and grown in culture to determine the frequency of induced micronuclei. Animals at each level of exposure showed an increase in the frequency of micronuclei relative to that in controls (P < 0.05). The exposure-response relationship data for rats and Syrian hamsters killed 0.2 days after the end of exposure were fit to linear equations (micronuclei/1000 binucleated cells = 15.5±14.4+0.53±0.06 WLM and 38.3±15.1+0.80±0.08 WLM, respectively). For the single exposure level used (496 WLM) in Chinese hamsters killed at 0.2 days after exposure, the frequency of micronuclei/1000 binucleated cells/WLM was 1.83±0.02. A comparison of the sensitivity for induction of micronuclei/WLM illustrated that Chinese hamsters were three times more sensitive than rats. The Syrian hamsters also showed a significantly elevated response (P < 0.05) relative to rats. These data suggest that initial chromosome damage is not the major factor responsible for the high rate of radon-induced cancer in rats relative to Syrian hamsters. The frequency of micronuclei in radon-exposed rats, Syrian hamsters, and Chinese hamsters significantly decreased (P < 0.05) as a function of time after the exposure. The rate of loss of damaged cells from the lung was greatest in the Chinese hamsters, followed by Wistar rats and Syrian hamsters, respectively. Our experiments demonstrated that the mammalian lung fibroblast/micronucleus method has the potential to (1) detect species differences in the induction of in vivo genotoxic damage in the lungs by inhaled environmentalal agents; (2) evaluate exposure-response relationships for in vivo induction of genetic damage; and (3) determine the persistence in vivo of preclastogenic and premutagenic lesions in cell populations.     

Susceptibility of inbred Syrian golden hamsters (Mesocricetus auratus) to lethal disease by lymphocytic choriomeningitis virus

An acutely lethal LCMV disease model has been established in the Syrian golden hamster (Mesocricetus auratus) in which lethality and disease are dependent upon both the inbred hamster strain and the LCMV strain. Young adult inbred, male and female, hamsters were tested for lethal-disease susceptibility by lymphocytic choriomeningitis virus (LCMV) strains, WE or Armstrong (ARM). With WE inocula, PD4 and MHA inbred hamsters were highly susceptible to a wasting disease. LVG and LHC inbred hamsters were intermediate in susceptibility; some of these animals died of wasting illness, and others exhibited minimal disease and survived. CB and LSH hamsters were highly resistant to any disease by WE. Mean survival times of susceptible hamsters given lethal WE inocula approximated 2.5 weeks and were not dependent on virus dose. By 1.5 weeks after WE inoculation wasting disease signs were notable and consisted of lethargy, progressive body weight loss, and diarrhea. The LCMV strain, ARM, was avirulent for all hamster strains, causing neither death nor disease. Hamsters surviving WE or ARM inoculation appeared healthy, produced LCMV antibody, and acquired resistance to further lethal WE challenge. Despite hamster-lethality differences. WE and ARM appeared comparably immunogenic for all hamster strains, based on host antibody titers. A number of other differences between the LCMV strains were, however, noted which could be relevant to virus virulence and lethality for hamster hosts. These included guinea pig lethality, temperature sensitivity, and plaque morphology.     

The lamina cribrosa in the eyes of rats, hamsters, gerbils and guinea pigs

The scleral lamina cribrosa in the eyes of adult rats, hamsters, gerbils and guinea pigs was examined by ordinary histology and by scanning electron microscopy after soft tissue digestion. The complexity of the lamina, when mounted for scanning electron microscopy, was graded on a scale of 0 to 4.5 by three independent observers under X 60 magnification in a stereo microscope. The observers were unaware of the species and were offered the 44 specimens twice in random order. The average variance attributable to an observer was 22 +/- 3% (SE) of the total variance of the gradings. The rat eyes had the least developed lamina cribrosa, with only 1-2 layers of sparse connective tissue. The mean complexity grading of 12 rat eyes was 1.6 +/- 0.15. The lamina cribrosa of the eyes of gerbils and guinea pigs was much more developed with at least 3 layers of abundant connective tissue, the mean grades of complexity being 3.4 +/- 0.09 and 3.5 +/- 0.15, respectively, in 12 eyes of each species. The lamina cribrosa in the hamster eyes was somewhat more developed than that of the rat, but much less than that of the gerbil and guinea pig. The mean grade of complexity was 2.4 +/- 0.14 in 8 eyes. In 6 pairs of rat eyes there was no correlation in grade of laminar complexity between the two eyes of the same animal. The present study makes the rat eye a candidate for experiments where a possible influence of the lamina cribrosa as such is undesired.     

Formaldehyde-Induced Fluorescence as a Means for Differentiating Epinephrine Cells from Norepinephrine Cells in Adrenal Medulla

The Falck and Hi Harp technique for the cellular localization of catecholamines by formaldehyde-induced fluorescence was applied to rat, mouse, and Syrian hamster adrenal. Some medullary cells revealed an unexpected orange-brown fluorescence. In guinea pig or rabbit adrenals, which store predominantly epinephrine, orange-brown fluorescence was not readily observed. It was found in Syrian hamsters only at the medullary periphery, where norepinephrine-producing cells are known to occur. Orange-brown fluorescence was depleted by administration of reserpine and intensified by nialamide plus DOPA. The same cell clusters which stained specifically for norepinephrine with ferric ferricyanide were found in adjacent sections to exhibit orange-brown fluorescence. Only by reducing the temperature of the formaldehyde reaction could sections of hamster adrenal showing only yellow-green fluorescence be obtained. These data suggest that the orange-brown fluorescence might result from' polymerization, oxidation, or both, of the isoquinoline produced by the norepinephrine-formaldehyde reaction under conditions slightly more vigorous than optimal and in the presence of high concentrations of norepinephrine     

Induction and peroxisomal appearance of gulonolactone oxidase upon clofibrate treatment in mouse liver

Various antihyperlipemic peroxisome proliferators are known to be carcinogenic in rodents but not in human, other primates and guinea pig, which species lost their ability to synthesize ascorbate due to mutations in the gulonolactone oxidase gene. Ascorbate synthesis is accompanied by H2O2 production, consequently its induction can be potentially harmful; therefore, the in vivo effect of the peroxisome proliferator clofibrate was investigated on gulonolactone oxidase expression in mouse liver. Liver weights and peroxisomal protein contents were increased upon clofibrate treatment. Elevated plasma ascorbate concentrations were found in clofibrate-treated mice due to the higher microsomal gulonolactone oxidase activities. Remarkable gulonolactone oxidase activity appeared in the peroxisomal fraction upon the treatment. Increased activity of the enzyme was associated with an elevation of its mRNA level. According to the present results the evolutionary loss of gulonolactone oxidase may contribute to the explanation of the missing carcinogenic effect of peroxisome proliferators in humans.     

Immunohistochemical localization of renin, NO synthase-1, and cyclooxygenase-2 in rodent kidney

The renin-angiotensin system (RAS) and tubuloglomerular feedback (TGF) are central to the maintenance of blood pressure and body fluid composition. Renin, NO synthase-1 (NOS-1), and cyclooxygenase-2 (COX-2) are key regulators of the RAS and TGF. In the present study, to investigate species-specific differences in the RAS and TGF, we immunohistochemically and morphometrically investigated the localization of renin, NOS-1, and COX-2 in the kidneys of various laboratory rodents and comparing males with females (DBA/2Cr mice, F344/N rats, Syrian hamsters, MON/JmsGbs gerbils and Hartley guinea pigs). In all animals, renin-positive immunoreactions were observed in the vascular walls of afferent arterioles. Renin immunoreactions appeared to be more widely distributed in mice. Mice had a greater number of renin-positive arterioles than other species. NOS-1-positive reactions were detected in the macula densa (MD) of all animals. Mice had the greatest number of NOS-1-positive MD cells. In addition to NOS-1-positive reactions, COX-2-positive reactions were observed in the MD of mice, rats, hamsters and gerbils. Interestingly, guinea pigs had no COX-2-positive MD cells. Rats had the greatest number of COX-2-positive MD cells. In nephron segments excluding the MD, the immunohistochemical localization of NOS-1 and COX-2 differed markedly among not only species but also sexes within the same species. In conclusion, we determined that localization of renin, NOS-1, and COX-2 showed large species- and sex-related differences. These data suggest that the regulation mechanisms of the RAS and TGF via renin, NOS-1, and COX-2 differ among rodents.     

In vitro metabolism of delta-11-tetrahydrocannabinol in the mouse, rat, guinea pig, rabbit, hamster, gerbil and cat

1. Liver microsomes were prepared from rats, rabbits, guinea pigs, hamsters, gerbils, a cat and three strains of mice, and were incubated with delta-11-tetrahydrocannabinol (delta-11-THC). The extracted metabolites were separated by chromatography on Sephadex LH-20 and examined by gas chromatography and combined gas chromatography/mass spectrometry. 2. Eleven metabolites were identified; these were formed by aliphatic hydroxylation of all positions of the pentyl chain, allylic hydroxylation at C-10 and C-8 (alpha and beta), and by the epoxide-diol pathway. 3. The ratio of the metabolites varied considerably between the species. Mice and rats favoured hydroxylation at C-8-alpha with very little hydroxylation of the pentyl chain. 4. In the guinea pig, however, hydroxylation of the pentyl chain, particularly at C-4', produced the major metabolites; very little hydroxylation occurred at C-8. 5. Side-chain hydroxylation was also favoured by the gerbil. 6. In the cat and hamster, 8-beta-hydroxylation was by far the major metabolic route, accounting, in the cat, for nearly 70% of the recovered metabolites. 7. The rabbit, on the other hand, favoured the epoxide-diol pathway with over 70% of the recovered metabolites being accounted for by the 9,11-dihydro-diols. 8. The results emphasise the need to make appropriate choices of animal models for metabolic and toxicological studies in humans.     

Preparation and characterization of monoclonal antibodies against a form of hamster liver cytochrome P-450 highly specific to aflatoxin B1

Monoclonal antibodies were prepared against a form of cytochrome P-450 (designated as cytochrome P-450-I) purified from 3-methylcholanthrene-treated hamster livers which is highly specific to aflatoxin B1. The cytochrome P-450-I was detected in ELISA and Western blots in liver microsomes from 3-methylcholanthrene-treated hamsters and also from non-treated and phenobarbital-treated hamsters in smaller amounts. However, none of the liver microsomes from 3-methylcholanthrene-treated rat, rabbit, guinea pig and Suncus murinus contained the cytochrome P-450-I. These results suggest that cytochrome P-450-I is specific to hamster and is induced mainly by 3-methylcholanthrene.