Species difference in the urinary excretion of isatinecic acid from the pyrrolizidine alkaloid retrorsine

1. The urinary excretion of the metabolites, isatinecic acid and pyrrolic metabolites from the pyrrolizicline alkaloid retrorsine were lower in the resistant species, guinea-pigs, than the susceptible species, mice, hamsters and rats.2. The urinary N-oxides levels, however, were higher in guinea-pigs relative to mice, hamsters and rats.3. These results conform to the postulate that a common metabolic pathway exists between the formation of isatinecic acid and pyrrolic metabolites.4. The resistance of guinea-pigs to PA poisoning is attributed to the high metabolism of PAs to N-oxides combined with a corresponding low conversion to pyrrolic metabolites.     

Metabolite identification of missile fuel JP-10 by gas chromatography mass spectrometry

The identification of the metabolites of the Air Force missile fuel JP-10 was accomplished. 5-Hydroxy-JP-10 was identified as the urinary metabolite of rats, mice and hamsters exposed to JP-10. 5-Keto-JP-10 was a major metabolite found in kidney extracts of male rats exposed to JP-10 but not found in kidney extracts from female rats or from other species. Sex-related differences in the formation of 5-keto-JP-10 in rats were studied.     

Neuroleptanalgesic drug combinations in the anaesthetic management of small laboratory animals.

Trials of different drug combinations for use in rabbits, guinea-pigs, rats, hamsters and mice are described in detail and experience over a 5-year period evaluated. Combinations of fentanyl citrate, fluanisone and diazepam provided exceptionally good anaesthesia in each species and were considered superior to other injectable agents.     

Species differences in circulatory responses to nasopharyngeal perfusion with smoke

Cardiovascular changes were observed in five species of laboratory mammals during upper airway perfusion with cigarette smoke. Apnoea occurred, both before and after vagotomy, in all species. Blood pressure always went up in rabbits, rats and hamsters but was not much affected in cats and guinea-pigs. Before vagotomy heart rate fell in rabbits and rats, rose in hamsters but did not change in cats and guinea-pigs. Vagotomy affected heart rate changes only in rabbits and rats. Results demonstrate markedly different autonomic responses in different species.     

Identification of S-1,2,2-trichlorovinyl-N-acetylcysteine as a urinary metabolite of tetrachloroethylene: bioactivation through glutathione conjugation as a possible explanation of its nephrocarcinogenicity

The elimination and metabolism of [14-C]-tetrachloroethylene (Tetra) was studied in female rats and mice after the oral administration of 800 mg/kg [14-C]-Tetra. Elimination of unchanged Tetra was the main pathway of elimination in both species and amounted to 91.2% of the dose in rats and 85.1% in mice. [14-C]-Carbon dioxide (CO2) was found to be a trace metabolite of [14-C]-Tetra. Only a small part of the applied dose was transformed to urinary (rats = 2.3%, mice = 7.1%) and fecal (rats = 2.0%, mice = 0.5%) metabolites. The urinary metabolites were separated and quantified by high performance liquid chromatography (HPLC) and identified by gas liquid chromatography/mass spectrometry (GC/MS). The following metabolites could be identified: oxalic acid (8.0% of urinary radioactivity in rats, 2.9% in mice), dichloroacetic acid (5.1%, 4.4%), trichloroacetic acid (54.0%, 57.8%), N-trichloroacetyl-aminoethanol (5.4%, 5.7%), trichloroethanol, free and conjugated (8.7%, 8.0%), S-1,2,2-trichlorovinyl-N-acetylcysteine (N-acetyl TCVC) (1.6%, 0.5%), and another conjugate of trichloroacetic acid (1.8%, 1.3%). The structures of the identified metabolites indicate two different pathways operative in Tetra biotransformation: cytochrome P-450-mediated epoxidation forming reactive metabolites in the liver and conjugation of Tetra with glutathione (GSH) catalyzed by glutathione transferase(s). The formation of reactive intermediates by renal processing of the glutathione conjugates may provide a molecular mechanism for the nephrotoxicity and nephrocarcinogenicity of Tetra in male rats.     

Arginine deficiency and orotic aciduria in mammals.

Dietary arginine deficiency in rats causes significant increases in urinary excretion of urea, citric acid and orotic acid independently of feed intake. Urea excretion during arginine deficiency depends upon the diet, sex, age, and species. Thus urea excretion has limitations as an indicator of arginine availability. Although elevated urinary citric acid during arginine deficiency is more consistently observed, it may be influenced by citric acid in natural dietary ingredients. Orotic acid excretion, however, appears to be a reliable indicator of available dietary arginine based upon studies in rats, mice, hamsters, guinea pigs, rabbits, and dogs.     

Pyrrolic and N-oxide metabolites formed from pyrrolizidine alkaloids by hepatic microsomes in vitro: Relevance to in vivo hepatotoxicity

An analytical method of improved sensitivity has enabled measurements to be made of N-oxide as well as pyrrolic metabolites formed from a range of unsaturated pyrrolizidine alkaloids in hepatic microsome preparations. Using microsomes from livers of phenobarbitone-pretreated male Fischer rats, all 13 alkaloids tested were metabolised to both N-oxides and pyrroles. The most lipophilic alkaloids gave enhanced rates of metabolism. No consistent relationship existed between rates of N-oxide and of pyrrole formation. The two pathways appeared to be independent. The ratio of N-oxide to pyrrolic metabolites varied, depending on the type of ester: it was highest for ‘open’ diester alkaloids, lowest for 12 membered macrocyclic diesters and for monoesters. Steric hindrance by the acid moiety could account for these differences, by affecting the balance between microsomal oxidation of the amino alcohol moiety at the nitrogen and C8 positions respectively and could explain the high pyrrole yields given by some macrocyclic diesters. The levels of pyrrolic metabolites bound to liver tissues and responsible for hepatotoxicity in rats given pyrrolizidine alkaloids, did not necessarily reflect the rates of formation of such metabolites measured in vitro. In the animal additional factors could influence the formation and tissue binding of pyrrolic metabolites, including the detoxication of alkaloids by hydrolysis and the chemical reactivity and stability of the toxic metabolites. A comparison of heliotridine esters with retronecine esters showed that the 7-hydroxyl or -ester configuration had a relatively small influence on the balance between formation of pyrrolic metabolites and detoxication by N-oxidation. The results did not support any hypothesis that heliotridine esters should generally be more hepatotoxic than analogous retronecine esters. The structure of the acid moiety was likely to have at least as much influence on toxicity as the base configuration.     

Comparative morphology of the stomach of some laboratory mammals

Histomorphology of the stomach of mouse, rat, hamster, guineapig, gerbil, and rabbit was studied. Although a common structural basis existed in the stomach between these species, the occurrence and distribution of various cells in gastric glands differed considerably between them. In mice, rats, hamsters and gerbils, the lower one-third of the glandular lamina propria was seemingly occupied by a varying proportion of parietal and chief cells. In rabbits, the predominantly occurring chief cells were distributed in the lower three-quarters of the glands intermingling with parietal cells, but in guinea-pigs the chief cells were not discernible. In hamsters, there was, however, a gradual increase of chief cells from the junction between nonglandular-glandular stomach toward the pyloric region. In all these species, parietal cells were the predominant cell type in the upper half to upper one-third of the gastric glands, often extending up to the neck of the glands interspersing between mucus neck cells and occasionally between chief cells.     

Effects of two different loading doses of L-tryptophan on the urinary excretion of tryptophan metabolites in rats, mice and guinea pigs. Correlation with the enzyme activities

The effect of two different loading doses of L-tryptophan (0.5 and 1.0 g/Kg b.w.) on excretion of tryptophan metabolites and the relation to the enzyme activities were studied in rats, mice and guinea pigs. In rats there is no ratio between the dosage used and the levels of the metabolites excreted. Doubling the amount of tryptophan administered, a 5-fold increase in the elimination of the metabolites along the kynurenine pathway is obtained. The 1.0 g/Kg load provides a more complete pattern of the metabolites than with the 0.5 g/Kg b.w. load. Kynurenic acid, kynurenine and xanthurenic acid are the chief metabolites excreted. In mice, the urinary excretion of the metabolites is very low with both loads. In guinea pigs, xanthurenic acid is excreted in the highest amount and kynurenic acid and kynurenine also constitute the large fractions with both loadings. The load of 0.5 g/Kg b.w. is preferable to that of 1.0 g/Kg b.w. for not causing B6-deficiency. Liver tryptophan pyrrolase exists in two forms in rats, while in mice and in guinea pigs it is present only as holoenzyme. This enzyme is more active in rats than in the other two species of animals. Kynureninase activity is lower in guinea pigs, but it apparently correlated to the low levels of excretion of the metabolites following this step. Kynurenine aminotransferase is very active in rats and in mice, while it is apparently depressed in guinea pigs, in contrast with the high excretion of xanthurenic and kynurenic acids, that puts in evidence a B6-deficiency. The excretion of tryptophan metabolites and enzyme activities are better correlated in rats.     

Interspecies differences in the effects of HCG on testicular function among rodents

Adult mice, rats and hamsters were injected with 0 or 0.3 IU hCG/g BW, 24 h before sacrifice. Basal LH receptor concentration was highest in rats and lowest in hamsters (rats greater than mice greater than hamsters). Injection of hCG caused LH receptor down-regulation in rats and mice, and up-regulation in hamsters. Basal plasma progesterone was highest in hamsters and lowest in rats (hamsters greater than mice greater than rats), however, hCG increased plasma progesterone levels in mice and rats, but not in hamsters. Mice had much higher plasma and testicular testosterone levels than other species, but hCG did not induce a relatively more dramatic increase in any species. When testes fragments were incubated with 0 or 12.5 mIU hCG/ml for 4 h, hCG increased media progesterone levels in rats and control mice, but not in hamsters and hCG-injected mice. Also, hCG elevated media testosterone levels in control but not in hCG-injected animals. Furthermore, addition of hCG in vitro partially prevented the elevation of media testosterone induced by in vivo hCG. The present results indicate that the mechanisms for the transduction of the gonadotropic signal by the Leydig cells are species-defined.     

Interspecies differences in testicular LH receptors and in vitro testosterone production among rodents

Testes from rats, mice and hamsters were incubated for 4 h with 0, 3.125 or 12.5 mIU hCG/ml. The LH receptor concentration in incubated testes of rats and mice was higher than that observed in hamsters. Testosterone levels in incubation media were significantly different among species (mice greater than rats greater than hamsters). During the incubation, hCG caused an increase in testosterone levels in all three species, but produced no significant changes in LH receptor concentration. Furthermore, a correlation between LH receptor concentration and testosterone only in hamsters is observed. The efficiency of the LH receptor-steroidogenesis interaction was estimated from the ratio of testosterone levels to receptor concentration under basal conditions and was found to differ among species (mice greater than hamster greater than rats). The levels of PGE and PGF in incubation media were higher in mice than in rats or hamsters, and hCG did not alter prostaglandin levels in any of the species. The present results indicate that acute in vitro hCG stimulation of testosterone synthesis does not involve appreciable changes in testicular LH receptor levels.     

Reduction of tertiary amine N-oxides by liver preparations: function of aldehyde oxidase as a major N-oxide reductase

Reduction of tertiary amine N-oxides to the corresponding amines by liver preparations was investigated with imipramine N-oxide and cyclobenzaprine N-oxide under anaerobic conditions. Rabbit liver cytosol in the presence of an electron donor of aldehyde oxidase exhibited a significant N-oxide reductase activity which is comparable to the activity of the liver microsomes supplemented with NADPH. Rabbit liver aldehyde oxidase also exhibited the N-oxide reductase activity in the presence of its electron donor, indicating that the activity observed in the liver cytosol is due to this cytosolic enzyme. Furthermore, the tertiary amine N-oxide reductase activity of liver cytosols from rats, mice, hamsters and hogs was demonstrated by comparison with that of liver microsomes from these mammalian species.     

Metabolism and toxicity of synthetic analogues of macrocyclic diester pyrrolizidine alkaloids

Seven macrocyclic diesters analogous to hepatotoxic pyrrolizidine alkaloids have been tested in male weanling Wistar rats. The compounds were the succinate (VII), 2,3-dimethylsuccinate (VIII), phthalate (IX), glutarate (X), 2,4-dimethylglutarate (XI), 3,3-dimethylglutarate (XII) and 3,3-pentamethyleneglutarate (XIII) of the synthetic amino dialcohol, synthanecine A. Single doses of these compounds were given i.p. to rats, and liver levels of pyrrolic metabolites were measured 2 h later. For these experiments both normal rats and rats pretreated with the esterase inhibitor tri-orthocresylphosphate (TOCP) were used. In normal rats, low levels of pyrrolic metabolites were formed from compounds VII, IX, X and XI, but these levels were greatly enhanced in rats with inhibited esterase activity. Much higher pyrrole levels were formed from compounds VIII, XII and XIII in normal rats, and esterase inhibition had relatively little effect on their metabolic conversion to pyrroles. This indicated that the last mentioned compounds were relatively resistant to enzymic hydrolysis, whereas VII, IX, X and XI were easily hydrolysed in normal rats, providing an alternative metabolic path which limited their conversion to pyrrolic metabolites. Comparison of results obtained using the 2,4-dimethylglutarate (XI), the 3,3-dimethylglutarate (XII) and the 3,3,-pentamethyleneglutarate (XIII) showed that 3,3-disubstitution but not 2,4-disubstitution in the glutaric acid moiety conferred high resistance to esterase attack. Toxicity tests using four of the compounds confirmed that acute hepatotoxicity was dose related, and associated with the formation of pyrrolic metabolites in the liver. The 3,3-dimethylglutarate (XII) was highly toxic both in normal and in TOCP treated rats, doses of 25-30 mg/kg causing moderate to severe centrilobular necrosis of the liver. In contrast the toxicity of the unsubstituted succinate (VII), glutarate (X) and 2,4-dimethylglutarate (XI) was very low in normal rats but high in rats with inhibited esterase activity. Thus, the glutarate (X) was non-toxic at 200 mg/kg in normal rats, but in TOCP treated rats, in which pyrrolic metabolite formation was enhanced by a factor of 17.5, a 50 mg/kg dose of this compound was severely hepatotoxic. Kidney damage, which was generally limited to the presence of isolated necrotic cells, sometimes accompanied the liver damage caused by these compounds, but acute toxic effects were not observed in any other tissues.(ABSTRACT TRUNCATED AT 400 WORDS)     

Kallikrein-like activity in salivary glands using a new tripeptide substrate, including preliminary secretory studies and observations on mast cells

Summary The tripeptide substrated-val-leu-arg-4-methoxy-2-naphthylamine gave a precise localization of reaction product in cryostat sections of aldehyde-fixed salivary glands from a number of species, with Fast Blue B as the capture reagent. In submandibular glands, there was strong staining of the granules in granular tubules of rats and hamsters and somewhat less in mice. Submandibular striated ducts showed variable periluminal staining in a finer granular form; it was abundant in guinea-pigs, strong in cats but somewhat less pronounced in dogs. Parotid glands contained less reactivity with none detectable in hamsters and guinea-pigs. In the rabbit, neither gland showed any reaction. Mast cells were densely stained in glands from cats and dogs; they were less reactive in rats and unstained in the other species.The closely related 7-amino-4-trifluoromethylcoumarin derivative of the tripeptide has been found highly satisfactory for assessing activity in submandibular saliva from cats. Preliminary functional studies indicate that an extensive rapid secretion of enzyme occurs into saliva on sympathetic stimulation, with a corresponding depletion of reactive material from the striated ducts in tissue sections. Far less mobilization of enzyme occurs into saliva on parasympathetic stimulation with no obvious change in the histochemical reaction of striated ducts. The possible significance of these findings in cats is discussed.Extensive qualitative and quantitative studies are required to evaluate enzyme and substrate specificities in each species. Nevertheless, derivatives ofd-val-leu-arg offer great promise for the functional testing of kallikrein-like reactivity both by histochemical means on cells and biochemically in their secretions.     

Alterations by peroxisome proliferators of acyl composition of hepatic phosphatidylcholine in rats, mice and guinea-pigs. Role of stearoyl-CoA desaturase.

Rats, mice and guinea-pigs were administered p-chlorophenoxyisobutyric acid (clofibric acid) or 2,2'-(decamethylenedithio)diethanol (tiadenol). The treatments of rats and mice with either clofibric acid or tiadenol increased markedly the activities of stearoyl-CoA desaturase, palmitoyl-CoA chain elongation, 1-acylglycerophosphate (1-acyl-GP) acyltransferase and 1-acylglycerophosphocholine (1-acyl-GPC) acyltransferase, but not 2-acylglycerophosphocholine (2-acyl-GPC) acyltransferase in liver microsomes. The treatment of guinea-pigs with clofibric acid did not cause any change in the activities of these enzymes. The treatment of guinea-pigs with tiadenol caused a slight, but significant, increase in the activities of 1-acyl-GP acyltransferase and 1-acyl-GPC acyltransferase. The treatment of rats and mice with either clofibric acid or tiadenol increased markedly the proportion of 18:1 and decreased greatly the proportion of 18:0 in liver microsomal phosphatidylcholine. However, there is a considerable difference in the effects of the two peroxisome proliferators on the composition of polyunsaturated fatty acids in phosphatidylcholine between rats and mice. The treatment of guinea-pigs with either of the two peroxisome proliferators caused no change in acyl composition of phosphatidylcholine. The possible role of stearoyl-CoA desaturation in the regulation of acyl composition of phosphatidylcholine was discussed.     

Plasma xanthine oxidase activity correlates with the resistance to severe hypoxia in different species

A rapid polarographic method of measuring the O-form of xanthine oxidase (XOD) is described. Activities of this enzyme and oxypurine concentrations were measured in the plasma and cerebrospinal fluid (CSF) of 1, 12, 25 and 60-day-old rats. In the CSF, the highest oxypurine concentration was found in the group of 12-day-old rats (34.07 +/- 12.37 mumol.l-1), and it remained nearly at the same level in groups of older animals. Oxypurines in the plasma, on the contrary, are polarographically measurable on the first day of life; in groups of older rats their concentration is below the sensitivity of the method (5 mumol.l-1). This was explained by the development of XOD activity in the plasma, which increased from 8.17 +/- 2.80 to 99.46 +/- 13.85 nkat.l-1 during the first 60 days of postnatal life. The higher the normal plasma XOD activity, the shorter was the survival time of the species during interrupted hypobaric hypoxia. Adult guinea-pigs and hamsters have no measurable XOD activity in the plasma; adult rats and mice have 99.46 +/- 13.85 and 259.69 +/- 58.23 nkat.l-1, respectively. The survival time of these animals at 10,500 m was measured following exposure to an altitude of 8,000 m for 30 min and 15 minute normoxia. Guinea-pigs survived 100.8 +/- 13.84 min, hamsters 54.25 +/- 11.33, rats 25.2 +/- 5.37 and mice 3.33 +/- 1.00 min.     

Species variations in the biliary and urinary excretion of arsenate, arsenite and their metabolites

In most mammalian species, inorganic arsenicals are extensively biotransformed and excreted both in unchanged form and as metabolites. In the bile of rats receiving arsenate (AsV) or arsenite (AsIII) we have identified monomethylarsonous acid (MMAsIII), purportedly the most toxic metabolite of inorganic arsenic. As rats are not commonly accepted for studying arsenic metabolism, we carried out a comparative investigation on the excretion of AsV, AsIII and their metabolites in five animal species in order to determine whether they also form MMAsIII from AsV and AsIII. Anaesthetised bile duct-cannulated rats, mice, hamsters, rabbits, and guinea pigs were injected with AsV or AsIII (50 micromol/kg, i.v.) and their bile and urine was collected for 2 h. Arsenic in bile and urine was speciated by HPLC-hydride generation-atomic fluorescence spectrometry and the excretion rates of AsV, AsIII, monomethylarsonic acid (MMAsV), MMAsIII and dimethylarsinic acid (DMAsV) were quantified. All species injected with AsV excreted arsenic preferentially into urine, whereas all animals receiving AsIII, except rabbits, delivered more arsenic into bile than urine. Bile contained almost exclusively trivalent arsenic (i.e. AsIII and/or MMAsIII), whereas AsV, AsIII and DMAsV appeared in urine. Except for guinea pigs, which do not methylate arsenic, the other species formed MMAsIII and excreted it into bile. Having excreted as much as 8% of the dose of AsIII or AsV in 2 h as MMAsIII, rats were by far the most efficient producers of this supertoxic metabolite. Thus, although the rat is not a good model for studying long-term arsenic disposition, this species appears especially valuable in studies on AsIII methyltransferase and in vivo formation of MMAsIII.     

A survey of laboratory animal disease in Kenya.

Diseases and infections diagnosed in laboratory mice, rats, guinea-pigs, golden hamsters and rabbits at the Veterinary Research Laboratory, Kabete, Kenya, are listed and discussed. Zoonoses encountered included salmonellosis and lymphocytic choriomeningitis. A number of traditionally recognised conditions were recorded but there were some notable omissions. The changing picture in laboratory animal science in East Africa is outlined and attention is drawn to the need for work on related diseases and infections.