The ecophysiological significance of calcium bicarbonate in the urine of subterranean rodents: testing a hypothesis

1. A comparative study of calcium and bicarbonate in the urine was carried out on the subterranean mole rat Cryptomys hottenttus and the terrestrial vlei rat Otomys irroratus. 2. The two species were kept on two different diets; carrots, a high calcium diet (41 mg/ 100 kg) or potatoes, a low calcium diet (14 mg/ 100g). 3. The results show that the urine of the mole rat contained high values of calcium bicarbonate on either diet. 4. The urine of the vlei rat showed high values of calcium bicarbonate only when kept on the high calcium diet. 5. From these results we assume that in subterranean rodents excretion of calcium bicarbonate is an adaptive mechanism to unload CO2 without increasing its concentration in the hypercapnic environment.     

Urine analysis of three rodent species with emphasis on calcium and magnesium bicarbonate

The urine composition of three rodent species, mole rat Spalax ehrenbergi, the golden hamster Mesocricetus auratus and the white rat Rattus norvegicus was studied. These three species represent different degrees of fossoriality. The results show that the urine of the species that show a higher degree of fossoriality, the mole rat and the hamster, contain high values of calcium and magnesium bicarbonates when compared with the white rat. From these results it may be assumed that the mole rat as well as the hamster can use the kidney as a pathway for releasing bicarbonate and carbonate. This mechanism may reduce the CO2 concentration in their hypercapnic environment.     

The importance of sodium and chloride ions for the development of DOCA-NaCl hypertension: a haemodynamic study

The role of sodium and its accompanying anion for the development of DOCA-salt hypertension was studied in uninephrectomized DOCA-treated weanling Wistar rats which were fed a diet containing either sodium chloride or sodium bicarbonate (170 mmol/kg). The blood pressure was increased in both groups of rats with sodium overload as compared to rats fed a low-salt diet only. A decreased cardiac output and substantially elevated systemic resistance were demonstrated in both groups of rats with high sodium intake in comparison with rats kept on a low-salt diet. However, these haemodynamic changes were more pronounced in rats with sodium chloride overload than in animals with a high sodium bicarbonate intake. On the other hand, the rigidity of major arteries which was estimated as the pulse pressure/stroke volume ratio, was increased only in rats fed a diet with sodium chloride but not in rats with sodium bicarbonate overload. Thus high sodium intake was responsible for the changes of systemic resistance in DOCA-treated animals and its action was only slightly augmented by a high chloride intake. In contrast to this, the chloride overload seemed to be essential for the induction of increased arterial rigidity.     

Effect of glutathione depletion on urinary acidification in the rat

Glutathione (GSH) depletion by diethyl maleate (DEM) administration and its rapid repletion were associated with the development of a moderate acidosis in the rat. The acidosis observed after DEM treatment could be a consequence of an impairment of lactate metabolism. GSH-depleted rats also showed an increased urine pH and a higher bicarbonate fractional excretion compared with control rats. Renal bicarbonate excretion was magnified when blood bicarbonate levels were normalized by means of a bicarbonate infusion in GSH-depleted rats; however, the amount of bicarbonate excreted in the urine was a very small fraction (less than 5%) of the calculated filtered load. GSH-depleted rats failed to elevate the relation urine minus blood (U-B) pCO2 as compared with control rats when they were subjected to a high bicarbonate load to the distal portions of the nephron. All these data were consistent with a distal renal tubular acidosis due to GSH depletion which could participate in the maintenance of the systemic acidosis, although it is unlikely that it is the primary cause of the acidosis.     

The role of the gut flora in the metabolism of cyclamate

1. [(14)C]Cyclamate was not metabolized when incubated with the liver, spleen, kidney or blood of rats of rabbits kept on a cyclamate-containing diet, and that had become converters of cyclamate into cyclohexylamine. 2. [(14)C]Cyclamate was converted into cyclohexylamine when incubated under anaerobic conditions with the contents of the caecum, colon or rectum or with the faeces of cyclamate-pretreated rats. Similar results were obtained with cyclamate-pretreated rabbits. With cyclamate-pretreated guinea pigs, which did not readily convert cyclamate into cyclohexylamine, the colon contents showed only low activity in this respect. 3. The faeces of a human converter of [(14)C]cyclamate into cyclohexylamine were also very active, but became less active when cyclamate was removed from his diet. 4. On subculturing the organisms from the contents of the colon and rectum of rats, the ability to convert cyclamate into cyclohexylamine was lost during three subcultures, but the loss of the activity was considerably decreased by subculturing in the presence of cyclamate. 5. Incubation of rat faeces in broths containing cyclamate increased their ability to metabolize cyclamate, but similar treatment of rabbit and human faeces suppressed this activity. 6. When rats are kept on a cyclamate diet the number of clostridia in the faeces increased considerably. In human dietary cyclamate did not appear to alter the counts of various faecal micro-organisms. 7. The gut organisms that appear to develop the ability to convert cyclamate into cyclohexylamine are clostridia in rats, enterobacteria in rabbits and enterococci in man. 8. [(14)C]Cyclohexylamine injected into the caecum or colon of rats is readily absorbed and excreted in the urine. 9. It appears that on continued intake of cyclamate the gut flora develop the ability to convert cyclamate into cyclohexylamine, which is then absorbed and excreted mainly in the urine, although a small proportion is metabolized to other compounds.     

Bicarbonaturic effect of acetazolamide in the dog: the influence of graded volume expansion

Studies were performed in anesthetized dogs to characterize the effect of a progressive volume expansion on the acetazolamide-induced bicarbonaturia. A closed system with urine reinfusion was used in all these experiments. In normovolemic dogs, 24% of the filtered bicarbonate was excreted into the urine while this value reached 62% when a 10% expansion was superimposed on a continuous infusion of acetazolamide. When a single dose of acetazolamide was given, fractional bicarbonate excretion increased from 21% in normovolemic dogs to 46% during 10% expansion. Without acetazolamide administration, 13% of the filtered bicarbonate was excreted during a 10% expansion. The continuous infusion of acetazolamide in normovolemic dogs increased fractional bicarbonate excretion in a progressive fashion, from 25 to 40%. This study shows that an acute volume expansion potentiates markedly the bicarbonaturic effect of acetazolamide, fractional bicarbonate excretion exceeding by far the simple additive effect of acetazolamide and expansion. We speculate that volume expansion might prevent a compensatory rise in acetazolamide-insensitive bicarbonate reabsorption in sites other than the superficial proximal convoluted tubules.     

LABELLING OF ACETYLCHOLINE IN THE BRAIN OF MICE FED ON A DIET CONTAINING DEUTERIUM LABELLED CHOLINE: STUDIES UTILIZING GAS CHROMATOGRAPHY—MASS SPECTROMETRY

—A method to achieve labelling of the acetylcholine stores of the brain under ideal physiological conditions is described. To this end, mice fed on a choline free diet were supplied with deuterium labelled choline in the drinking water. Labelled and unlabelled choline in plasma and in the brain as well as labelled and unlabelled acetyicholine in the brain were measured by a gas chromatographic-mass spectrometric method. It was found that after 1–25 days on the deuterium choline diet, substantial amounts of the plasma choline and brain acetylcholine were displaced by deuterium choline and deuterium acetylcholine, respectively. Already on the first day, the mole ratio of deuterium choline/total choline in plasma was 0·22, and it approached a maximum of 0·57 on the 14th day. The mole ratios of deuterium acetylcholine/total acetylcholine in the brain were slightly but significantly lower than those of deuterium choline/total choline in plasma 1–14 days, but asymptotically approached the mole ratios of deuterium Ch/total Ch in plasma by 25 days. Intact brains submitted to incubation at room temperature for 10 min increased their total choline content by about 500 per cent. Concurrently, in brains from animals kept on a deuterium choline diet for 1–2 days, the level of deuterium choline rose only by 50 per cent after incubation. Deuterium choline levels increased, however, by 200–300 per cent in the brains from animals kept on the deuterium diet for longer time periods. On the basis of these data it is suggested that: (a) choline in plasma is partly supplied from the food and partly from endogenous sources; (b) plasma choline rapidly equilibrates (less than one day) with a pool of Ch in the brain which is responsible for biosynthesis of acetylcholine; (c) the size of this choline pool is in the order of 34–40 nmol/g.     

Influence of dietary curcumin and cholesterol on the progression of experimentally induced diabetes in albino rat

Effect of feeding 0.5% curcumin diet or 1% cholesterol diet was examined in albino rats rendered diabetic with streptozotocin injection. Diabetic rats maintained on curcumin diet for 8 weeks excreted comparatively less amounts of albumin, urea, creatinine and inorganic phosphorus. Urinary excretion of the electrolytes sodium and potassium were also significantly lowored under curcumin treatment. Dietary curcumin also partially reversed the abnormalities in plasma albumin, urea, creatinine and inorganic phosphorus in diabetic animals. On the other hand, glucose excretion or the fasting sugar level was unaffected by dietary curcumin and so also the body weights were not improved to any significant extent. Diabetic rats fed curcumin diet had a lowered relative liver weight at the end of the study compared to other diabetic rat groups. Diabetic rats fed a curcumin diet also showed lowered lipid peroxidation in plasma and urine when compared to other diabetic groups. The extent of lipid peroxidation on the other hand, was still higher in cholesterol fed diabetic groups compared to diabetic rats fed with control diet.Thus, the study reveals that curcumin feeding improves the metabolic status in diabetic condition, despite no effect on hyperglycemic status or the body weights. The mechanism by which curcumin improves this situation is probably by virtue of its hypocholesterolemic influence, antioxidant nature and free radical scavenging property.     

The bile flow and biliary excretion of ursocholate in the rat

Several studies reported that ursodeoxycholate (but not its conjugates), when administered intravenously, increased the biliary bicarbonate concentration in the rat (1–3). At the same time, a complete dissociation between bile flow and the bile salt excretion rate was produced in the second hr of infusion (2). In order to examine whether this property was due to the 7β-hydroxy group in its molecular structure, the choleretic property of ursocholate (3α, 7β, 12α-trihydroxy-5β-cholanoic acid) was investigated in male Wistar rats. Immediately after the start of iv infusion of ursocholate at a rate of 1.2 μmole/min/100 g b. wt., both the bile flow and bile salt excretion rate began to increase. However, unlike with ursodeoxycholate, the bile salt excretion rate continued to be high in the second and third hr of infusion, while the bile flow rate gradually increased. Furthermore, the bicarbonate concentration in the bile fell slightly 10 min after the start of ursocholate infusion. Although the concentration tended to return to the baseline value before the bile salt infusion in the later period of observation, no significant increase in bicarbonate concentration was observed during the whole observation period. These properties were quite similar to those of cholate rather than those of ursodeoxycholate. However, a cholate infusion at the same rate of 1.2 μmole/min/100 g b.wt. caused a cholestasis as early as 20 to 30 min after the start of an infusion. These results suggest that the previously reported properties of ursodeoxycholate (that it causes a complete dissociation between the bile flow and bile salt excretion rate in the second hr and that it increases the biliary bicarbonate concentration) were not due to the 7β-hydroxy group in its steroidal structure, and that the choleretic property of ursocholate is similar to its 7α-hydroxy epimar, cholate. However, the much lower cytotoxicity of ursocholate compared to cholate appears to be due to the 7β-hydroxy group that ursocholate has.     

Foraging strategy in a subterranean rodent,Spalax ehrenbergi: a test case for optimal foraging theory

Summary The foraging behavior of the subterranean mole rat Spalax ehrenbergi (Rodentia, Spalacidae) was tested according to the framework of optimal foraging theory. We compared the frequencies of food species hoarded in storage chambers of mole rats with the frequencies of these species occurring in the vicinity of the mole rats' nest mounds during the winter and spring seasons. In addition, we examined the food composition of several summer nest mounds. Laboratory observations were conducted in order to test the foraging behavior of mole rats under simulated subterranean conditions. The mole rat is a generalist and collects a variety of food species. Out of 33 plant species that were hoarded by mole rats in the 21 studied nest mounds, 61% (n=20) were geophytes, 21% (n=7) perennial herbs, 15% (n=5) annual herbs and 3% (n=1) dwarf shrubs. The frequency of each collected species in the 16 winter and spring nest mounds is in general accordance with its frequency in the mole rat's territory. This implies that the mole rat randomly samples the food reserve of its territory without special preference or directed search for a particular species. The collection or avoidance of any food item is not dependent on the presence or absence of any other food item. We suggest that the foragin generalism of the mole rat is a product of the constraints of a subterranean niche — the necessity to hoard food as much as possible in a limited time period and the high energetic investment of tunneling to the food items.     

r-Galactonolactone in experimental galactosemic animals

An accumulation of galactose-1,4-lactone, an oxidation product of galactose, was observed in various tissues of galactosemic guinea pigs fed a 40% galactose diet for 6 weeks. In addition, an accumulation of the two galactose metabolites varied among organs. The highest content of the lactone was observed in the liver and the content of the lactone exceeded that of the reduced counterpart. The lens gave the highest galactitol content. In the serum the level of the lactone was very low. A trace amount of the lactone was detected in the kidney while it was mostly excreted into urine within 54 h upon withdrawal of the diet. On the other hand, in the animals kept on a high galactose diet for only 2 days, urinary lactone rapidly decreased. These observations indicated that a high galactose level in the circulation was associated with the production of the lactone in various tissues and that the accumulated lactone was released into the circulation very slowly and then excreted into the urine. Suppression of galactitol production by administration of an aldose reductase inhibitor resulted in the accumulation of the lactone in the lens, the testis, and the muscle, as well as in the circulation. The lactone thus produced was excreted exclusively into the urine. This observation indicates a close relationship between the oxidative and reductive metabolisms of galactose at a toxic level.     

A Simple Bioassay for Chemicals Active to the Photosynthetic or Respiratory Systems of Plants

After male rats of the Sprague Dawley strain, 5 weeks old, were fed a 20% casein diet with or without 0.5% nicotinamide for 13 days, 180 mg/kg body weight of alloxan was injected in- traperitoneally into the rats. The rats were kept for 18 days with the same diet. The level of blood glucose was increased 6-fold in the group on a 20% casein diet by the injection of alloxan, while there was only a 2-foid increase in the group on a nicotinamide-containing diet and the decreased body weight was also lower in the group on the nicotinamide diet than the group on the casein diet. The body weight was indirectly related to the concentration of blood glucose. A marked increase was observed in the activities of tryptophan oxygenase, aminocarboxymuconate-semialdehyde decarboxylase, and nicotinamide methyltransferase upon the injection of alloxan with both diets; on the other hand, the activities of kynureninase and NAD⁺ synthetase were decreased by the injection of alloxan. The activity of kynurenine aminotransferase increased in the group on the 20% casein diet by the injection of alloxan, while in the group on the nicotinamide-containing diet its activity was not increased by the injection. These changes in the above enzyme activities mean that the conversion ratio from tryptophan to niacin is lower in the alloxan diabetic rat than normal rat. It was found that the activities of tryptophan oxygenase, aminocarboxymuconate-semialdehyde decarboxylase, and nicotinamide methyltransferase were directly related to the concentration of blood glucose, and that the activities of kynureninase and NAD⁺ synthetase were inversely related. There was no difference in the activities of 3-hydroxyanthranilic acid oxygenase and nicotinamide mononucleotide adenylyltransferase upon the injection of alloxan with both diets.     

Safety assessment of transgenic Bacillus thuringiensis rice T1c-19 in Sprague-Dawley rats from metabonomics and bacterial profile perspectives

Bacillus thuringiensis rice is facing commercialization as the main food source in the near future. The unintended effects of genetically modified (GM) organisms are the most important barriers to their promotion. We aimed to establish a new in vivo evaluation model for genetically modified foods by using metabonomics and bacterial profile approaches. T1c-19 rice flour or its transgenic parent MH63 was used at 70% wt/wt to produce diets that were fed to rats for ～ 90 days. Urine metabolite changes were detected using (1)H NMR. Denaturing gradient gel electrophoresis and real-time polymerase chain reaction (RT-PCR) were used to detect the bacterial profiles between the two groups. The metabonomics was analyzed for metabolite changes in rat urine, when compared with the non-GM rice group, where rats were fed a GM rice diet. Several metabolites correlated with rat age and sex but not with GM rice diet. Significant biological differences were not identified between the GM rice diet and the non-GM rice diet. The bacteria related to rat urine metabolites were also discussed. The results from metabonomics and bacterial profile analyses were comparable with the results attained using the traditional method. Because metabonomics and bacterial profiling offer noninvasive, dynamic approaches for monitoring food safety, they provide a novel process for assessing the safety of GM foods.     

Benzophenone metabolism. I. Isolation of p-hydroxybenzophenone from rat urine

Two sets of male Sprague-Dawley rats were administered 6.2 and 5.0 mmole of benzophenone by stomach tube. Ethyl acetate extracts obtained from 24 hour β-glucuronidase/aryl sulfatase hydrolyzed urine samples led to the isolation of 44.4 and 55.4 μmole, respectively, of p-hydroxybenzophenone. The phenol was identified from its melting point, elemental analysis, infrared, proton magnetic resonance, and mass spectral characteristics. Thin-layer chromatographic and mass spectral analyses of unhydrolyzed 24 hour urine and fecal samples showed no detectable quantities of p-hydroxybenzophenone. The identification of p-hydroxybenzophenone in rat urine may prove useful as a trapping agent for detecting benzophenone as an intermediate in the metabolism of (AR)₂- CHX type diphenylmethyl drugs.     

The effect of melatonin administration and short exposures to cold on body temperature of the blind subterranean mole rat (Rodentia, Spalax ehrenberghi, Nehring)

Improvement of the cold resistance capacity of the mole rat (Spalax ehrenbergi, Nehring), a blind subterranean rodent, was achieved following subcutaneous melatonin administration. Melatonin-treated mole rats exhibited reduction both in initial body temperature (Tbo) and in the decrease of body temperature (delta Tb) after 6 hours of exposure to cold. The tested mole rats were acclimated to and kept during the experiment, under long (16L/8D) photoperiod schedule while the effects of melatonin administration on body temperature simulated the effects of short photoperiod conditions. These results suggest that in mole rats, melatonin biochemically mediates photoperiodic information into body temperature physiological response. As a byproduct of the original experiment, it was found that frequently repeated short (6 hours) exposure to cold (Ta = 5 +/- 1 degrees C) can in itself initiate an improvement of cold resistance capacity similar to the effect of melatonin administration. This supports the idea that mole rats may improve their cold resistance in response to either photoperiodic or climatic changes.     

A high cholesterol diet ameliorates renal tubular lesions in diabetic rats

These studies examine the effect of cholesterol feeding in normal rats and in rats with streptozotocin-induced diabetes mellitus. Four groups were studied: normal rats fed either a standard rat chow or a standard rat chow supplemented with cholesterol and diabetic rats fed standard chow or standard chow plus cholesterol. Diabetic rats fed a standard diet excreted more creatinine and urea in the urine, had higher levels of blood urea nitrogen, and lower serum albumin levels than rats fed standard diet plus cholesterol. Blood glucose levels were similar in the two groups; however, diabetic rats given cholesterol had a greater body weight at the end of the study than diabetic rats eating standard chow. Urine volumes and sodium and potassium excretion in the urine were greater in diabetic rats fed a standard diet than in those fed a high cholesterol diet. Diabetic rats fed a standard diet had distinctive renal lesions characterized by swelling of tubular epithelial cells with clearing of cytoplasm. The nephron segments involved by this striking vacuolar change were the distal convoluted tubule and the thick limbs of Henle's loop. These lesions were identical to those described by Armanni-Ebstein in severely glycosuric patients. These lesions were not observed in any of the animals of the other three groups (including diabetic rats fed a high cholesterol diet). Glomeruli were normal in animals of all groups. Thus, cholesterol administration prevents the development of the Armanni-Ebstein lesions in diabetic rats despite persistent hyperglycemia. The mechanism by which cholesterol administration prevents the accumulation of glycogen in distal tubule cells has not been elucidated. It is suggested that glycogen accumulation in distal tubular segments may explain the greater urine volumes, natriuresis, kaliuresis, and proteinuria observed in diabetic animals fed a standard diet when compared with rats fed the same diet plus cholesterol.     

Hypoxic survival differs between two mole rat species (Spalax ehrenbergi) of humid and arid habitats.

1. Two chromosomal species, 2n = 52 and 2n = 60 of the mole rat superspecies (Spalax ehrenbergi), occupy humid (2n = 52) and arid (2n = 60) habitats in Israel. 2. Gas conductivity of the soil of the 2n = 52 mole rat is lower than that of the 2n = 60 mole rat, and the 2n = 52 mole rat is better adapted to hypoxia. 3. The hypothesis that the 2n = 52 mole rat can survive to a lower pO2 than the 2n = 60 mole rat was tested. 4. Terminal pO2 (Torr) of four females 2n = 52 was lower, 18.0 +/- 2.9 (SD), than the terminal pO2 of five females 2n = 60, 28.2 +/- 5.1 (SD). 5. The hypoxic survival of the 2n = 52 mole rat as compared to that of the 2n = 60 mole rat correlates with other physiological traits: breathing and heart frequencies, blood hemoglobin and tissue gas tensions.     

Urinary aldehydes as indicators of lipid peroxidation in vivo

The excretion of malondialdehyde (MDA), lipophilic aldehydes and related carbonyl compounds in rat and human urine was investigated. MDA was found to be excreted mainly in the form of two adducts with lysine, indicating that its predominant reaction in vivo is with the lysine residues of proteins. Adducts with the phospholipid bases serine and ethanolamine and the nucleic acid bases guanine and deoxyguanosine also were found. Except for the adduct with deoxyguanosine (dG-MDA), the excretion of these compounds increased with peroxidative stress imposed in the form of vitamin E deficiency or the administration of iron or carbon tetrachloride. Marked differences in the concentration of dG-MDA in different tissues were correlated with their content of fatty acids having three or more double bonds, the putative source of MDA. Fourteen nonpolar and eleven polar lipophilic aldehydes and other carbonyl compounds were identified as their 2,4-diphenylhydrazine derivatives in rat urine. The excretion of five nonpolar and nine polar compounds was increased under conditions of peroxidative stress. The profile of lipophilic aldehydes obtained for human urine resembled that for rat urine. Except for a reported 4-hydroxynon-2-enal conjugate with mercapturic acid, the conjugated forms of the lipophilic aldehydes excreted in urine remain unidentified. Aldehyde excretion is influenced by numerous factors that affect the formation of lipid peroxides in vivo such as energy status, physical activity and environmental temperature, as well as by wide variations in the intake of peroxides in the diet. Consequently, urinalysis for aldehydic products of lipid peroxidation is an unreliable indicator of the general state of peroxidative stress in vivo.     

Regulation of glucose utilization in chick embryo fibroblasts by bicarbonate ion

The amount of glucose consumed by chick embryo fibroblasts in primary culture is strongly influenced by the presence of bicarbonate ion in the culture medmum. Cells grown on glucose at physiologic concentration (5.5 mm) and in the absence of bicarbonate ion have a reduced rate of glucose utilization when compared to their counterparts cultivated in medium containing the usual 25 mM bicarbonate. The presence or absence of bicarbonate is without effect on chick embryo fibroblast proliferation over a 6-day growth period. Both lactic acid accumulation per mole of glucose consumed and the utilization of glutamine increase as a function of bicarbonate ion in the growth medium.     

Tissue metabolism and enzyme activities in the rodent Heterocephalus glaber, a poor temperature regulator.

1. Tissue oxygen uptake and enzyme activities were investigated in the naked mole rat, Heterocephalus glaber, a mammal notable for its low body temperature and metabolism and poor temperature regulating ability. 2. Q10 for O2 uptake of Heterocephalus crude liver homogenates ranged from 1.91 for the temperature interval 25-30 degrees C to 1.76 within the range 30-38 degrees C, values similar to those reported for typical homoiotherms. 3. Km pyruvate of lactate dehydrogenase in heart muscle had the same temperature dependence in the mole rat and mouse. 4. O2 uptake and cytochrome oxidase activity of skeletal muscle were higher for mole rat than mouse. The reverse was true for heart muscle. Brain and liver O2 uptake showed similar values for both species, while kidney O2 uptake was highest in the mouse. 5. Pyruvate kinase activity in heart and skeletal muscle was higher in mouse than mole rat, suggesting a greater reliance on glycolysis in the former. 6. Na+, K+ -ATPase activity of liver and kidney was 60% higher in mouse than mole rat, while brain was 30% higher in mouse. 7. The results indicate that the effects of temperature on tissue metabolism in the mole rat conform to those in typical homoiotherms. The low body temperature and O2 uptake in the mole rat find no expression in the tissue respiratory capacity.