Kidney Of Wistar Rats Research Articles

Decrease in kidney calbindin-d 28kda as a possible mechanism mediating cyclosporine A- and FK-506-induced calciuria and tubular mineralization

The use of the immunosuppressant cyclosporine A (CsA) is limited by its adverse renal effects. Most recently, we reported that the drug markedly decreases the levels of the calcium-binding protein calbindin-D 28kDa in kidneys of male Wistar rats. In the present study, the potential relationship between drug-induced nephrotoxicity and the decrease in kidney calbindin-D 28kDa was investigated. Four groups of male Wistar rats were treated for 10 or 31 days with either the immunosuppressant CsA (50 mg/kg/day), FK-506 (5 mg/kg/day), rapamycin (5 mg/kg/day) or with the nonimmunosuppressive cyclosporine derivative 3'keto-[Bmt 1]-[Val 2]-CsA (SDZ PSC-833) (50 mg/kg/day), and the effects on calcium homeostasis, kidney histology and renal calbindin-D 28kDa were examined. Similar effects were found with CsA and FK-506; both drugs strongly reduced kidney calbindin-D 28kDa protein levels, increased urine calcium excretion, caused intratubular calcification, and induced basophilic tubules. In contrast, rapamycin and SDZ PSC-833 caused no decrease in renal calbindin-D 28kDa levels, no noticeable alterations in calcium metabolism, and no renal calcification. The results provide evidence for a link between decreased renal calbindin, increased calcium urine excretion, and intratubular kidney calcification. The present data show no correlation between the decrease in renal calbindin and the induction of basophilic tubules; however, it needs to be investigated if these apparently independent kidney effects may have a common origin upstream of calbindin expression.

Overexpression of integrin-associated protein (CD47) in rat kidney treated with a renal carcinogen, ferric nitrilotriacetate.

An iron chelate, ferric nitrilotriacetate (Fe–NTA), induces renal proximal tubular necrosis, a consequence of free radical–associated damage, that ultimately leads to a polycystic change of the renal cortex and a high incidence of renal cell carcinoma (RCC) in rodents. The differential display technique was used to search for inducible genes in the kidney of male Wistar rats treated with Fe–NTA and in the induced RCCs. Six fragments were selected that showed specific quantitative changes in mRNA. Two of them exhibited similar patterns in northern blots as well. One fragment showed a high limnology (89%) to murine integrin–associated protein (IAP; CD47). We thus cloned rat IAP cDNA including the entire coding region for use in further analysis. Rat IAP cDNA showed a 21–amino–acid deletion that was also observed in human, but not in mouse. Northern blots revealed that IAP was consistently overexpressed in non–tumorous parts of the kidney (2.4‐fold increase, n=9, P< 0.0001) as compared with matched controls 1to 2 years after Fe–NTA treatment. IAP Overexpression of more than 2.9–fold was found in 25% (2/8) of RCCs studied, and was limited to cases of a high histological grade and lung metastasis. Unexpectedly, IAP expression was higher in the non–tumorous part of the kidney after Fe–NTA treatment (2.8–fold) than in RCC (1.5–fold) in each case (n=4, P<0.05). Abundant expression of IAP mRNA in the renal tubular epithelium after Fe‐ ‐NTA treatment and RCC cells was observed by in situ hybridization. The results suggest that IAP Overexpression may he associated with Fe–NTA–induced renal cortical tubular damage and regeneration that lead to a polycystic state, and with tumor progression and metastasis of the induced RCCs.

Induction of a Wide Range of C 2–12 Aldehydes and C 7–12 Acyloins in the Kidney of Wistar Rats After Treatment With a Renal Carcinogen, Ferric Nitrilotriacetate

An iron chelate, ferric nitrilotriacetate (Fe-NTA), induces renal proximal tubular necrosis associated with lipid peroxidation and oxidative DNA damage that finally leads to a high incidence of renal cell carcinoma in rodents. In the present study, we investigated what kinds of C 2–12 saturated and unsaturated aldehydes and C 7–12 acyloins, metabolites of saturated aldehydes, are produced in the kidney and liver within 24 h after single ip administration of 15 mg Fe/kg of Fe-NTA, or after repeated (1 or 3 wk) ip administration of 5–10 mg Fe/kg of Fe-NTA. Amounts of twenty one aldehydes and five acyloins were determined by capillary column gas chromatography-negative-ion chemical ionization mass spectrometry with ammonia as reagent gas. Most of the aldehydes and all the acyloins measured revealed a significant dose-dependent increase 1 to 3 h after single administration in the kidney, among which 4-hydroxy-2-nonenal (HNE) showed the highest increase (27.3-fold) and malondialdehyde (MDA) was the most abundant aldehyde (2.40 nmol/100 mg wet tissue). In the liver, however, the increase in aldehydes and acyloins was less prominent. After repeated administration of Fe-NTA, only 9 aldehydes (ethanal; furfural; trans,trans-2,4-heptadienal; nonanal; trans-2,cis-6-nonadienal; HNE; decanal; trans-4,cis-4-decenal; MDA) and 4 acyloins (3-hydroxyheptan-2-one; 3-hydroxyoctan-2-one; 3-hydroxynonan-2-one; 3-hydroxydodecan-2-one) showed a significant increase. Immunohistochemistry further demonstrated an increased amount of HNE-modified and MDA-modified proteins in the renal proximal tubules after repeated Fe-NTA administration. Some of the aldehydes measured such as HNE and MDA are reportedly cytotoxic, genotoxic and mutagenic. Accumulation of these aldehydes may play a role in this renal carcinogenesis model. Copyright © 1997 Elsevier Science Inc.

Ultrastructural study of glomerular capillary loops at different perfusion pressures as revealed by quick-freezing, freeze-substitution and conventional fixation methods.

Wistar rat kidneys were perfused with some fixatives at different pressures of 100, 150, 200 or 250 cm H2O via the aorta and were studied by the quick-freezing and freeze-substitution (QF-FS) or deep-etching (QF-DE) methods, in addition to the conventional immersion or perfusion fixation method. Different parts of glomerular capillary walls were selected for morphometric analyses. It was demonstrated that the widths of glomerular slit diaphragms and the spaces between foot processes were more widely dilated at higher perfusion pressures (200 and 250 cm H2O) than those seen at both normal perfusion pressure (150 cm H2O) and lower perfusion pressure (100 cm H2O). On the other hand, the glomerular basement membranes were thinner at higher perfusion pressures. By the QF-FS and QF-DE methods, the foot processes showed different shapes from those revealed by the conventional preparation methods, even at the same perfusion pressure. It is concluded that the widths of glomerular slit diaphragms and glomerular basement membranes and the spaces between foot processes may be significantly changed in vivo, depending on the hemodynamics in the glomerular capillary.

In vitro macro- and microautoradiographic localization of V1 and V2 receptors in the rat kidney using OPC-21268 and OPC-31260.

To elucidate the precise localization of vasopressin (VP) V1 and V2 receptors in the kidney, we utilized in vitro macroautoradiography (macro-ARG) and microautoradiography (micro-ARG) of these receptors in Wistar rat kidneys. This was done by using OPC-21268 and OPC-31260, two newly developed selective V1 (OPC-21268) and V2 (OPC-31260) receptor antagonists. For macro-ARG, 10-microm kidney sections were incubated with Tris-HCl buffer containing [3H]-VP with or without unlabeled ligand (VP, OPC-21268, or OPC-31260) at 20 degrees C for 40 min. These sections were then loaded into X-ray cassettes with Hyperfilm-[3H] and exposed in the dark for 2 months. The autoradiograms were quantitatively analyzed by using the research analysis system RAS 1,000; the V1 and V2 receptors were quantitated by subtracting the nonspecific binding (incubated with OPC-21268 and OPC-31260, respectively) from the total binding. To assess a more precise localization of the V1 and V2 receptors, we also investigated the micro-ARG of the renal V1 and V2 receptors by dipping the kidney section slides used for macro-ARG into a photographic emulsion and observing the receptors under light microscopy. [3H]-VP binding to the rat kidney was completely displaced by unlabeled excess VP, but not by unlabeled angiotensin II, indicating that [3H]-VP binding was specific for VP receptors. Computerized quantification showed that V2 receptors, visualized by OPC-31260, were the predominant type of VP receptor in the kidney. Conversely, V1 receptors, visualized by OPC-21268, were fewer in number. V1 receptors were partly localized to the glomerulus, cortical vessels, interstitial cells, and the medullary vessels. The V2 receptors localized to the collecting ducts and medullary tubules. Our findings indicated that renal V1 and V2 receptors can be detected by in vitro macro- and micro-ARG by using OPC-21268 and OPC-31260.

Effect of radon exposure on superoxide dismutase (SOD) activity in rats.

Effect of radon exposure on the activities of superoxide dismutase (SOD) in blood, kidney, liver and spleen of male Wistar rats were investigated. Radon exposure was carried out in a newly developed chamber with 226Ra as the radon source. SOD activity, measured by the spin trap method using an electron spin resonance spectrometer, increased greatly in liver and kidney after 4 hour exposure whereas it decreased significantly after 16 hour exposure. Despite approximately same total doses by the radon exposure, the stimulated SOD activity after 4 hour exposure was not observed after 16 hour exposure in kidney, liver and spleen except blood, suggesting that the stimulating effect could last for a short period.

Alteration of antioxidant status following sympathectomy: Differential effects of modified plasma levels of adrenaline and noradrenaline

The influence of altered levels of endogenous catecholamines following adrenalectomy or 6-hydroxydopamine (6-OH) treatment (alone or in combination) on enzymatic (glutathione reductase, catalase, glutathione peroxidase and Cu, Zn superoxide dismutase) and non-enzymatic (glutathione) antioxidant components of heart, liver, kidney, lung and erythrocytes in male Wistar rats was investigated. Functional antioxidant status was assessed in terms of susceptibility to t-butylhydroperoxide-induced sulfhydryl group oxidation (an indirect measure of glutathione depletion) and lipid peroxidation, as measured by thiobarbituric acid-reactive substance (TBARS) formation. Reduced levels of adrenaline and noradrenaline resulted from adrenalectomy and 6-OH treatment, respectively, while a combination of these treatments led to a reduction in the levels of both catecholamines. Adrenalectomy was associated with alterations in glutathione reductase activity in the heart and liver (increased). 6-OH treatment alone produced an elevation in glutathione reductase activity only in the heart. In adrenalectomized animals, 6-OH treatment produced no further increases in glutathione reductase activities of heart or liver. In lung, however, the combination of adrenalectomy and 6-OH treatment caused an elevation in both glutathione peroxidase and glutathione reductase activities. Glutathione levels of liver alone were elevated following adrenalectomy, while those of erythrocytes and liver (but not other tissues investigated) were increased by the combination of adrenalectomy and 6-OH treatment. The kidney was relatively resistant to the effects of sympathectomy and showed no changes in any of the antioxidant components measured. Adrenalectomy alone or in combination with 6-OH produced an increased in susceptibility to peroxide-induced sulfhydryl group oxidation only in the heart. 6-OH treatment caused a reduction in peroxide-induced TBARS formation only in the kidney. Both adrenalectomy and the combination of adrenalectomy and 6-OH treatment were associated with reduced TBARS formation in the liver, lung and kidney, but not heart. Results from this study demonstrate that the effects of sympathectomy on antioxidant status vary among tissues. Differences between adrenalectomy and 6-OH treatment on antioxidant components are suggestive of differential actions of adrenaline and noradrenaline on tissue antioxidant status which may have important implications under conditions associated with elevations in levels of these catecholamines including chronic stress and myocardial infarction.

Cyclosporine enhances the expression of TGF-β in the juxtaglomerular cells of the rat kidney

The mediators of cyclosporine (CsA) nephrotoxicity remain ill defined. In this study, we describe evidence of increased amounts of transforming growth factor-beta (TGF-beta) in the kidneys of adult male Wistar rats treated with CsA (5 to 25 mg/kg/day) for four weeks. Localization of TGF-beta was undertaken immunocytochemically at both light and electron microscope levels and Northern blot analysis was applied to detect changes in transcription of TGF-beta. In control rats, weak to moderate immunostaining for TGF-beta was observed, in the juxtaglomerular arterioles. CsA treatment resulted in a dose-dependent increase in the number of stained afferent and interlobular arterioles and in the intensity of staining. The number of stained afferent arterioles increased from a control value of 0.21 +/- 0.08/mm2 cortex to 0.84 +/- 0.15/mm2 cortex, P < 0.01, and to 1.12 +/- 0.10/mm2 cortex, P < 0.01, in rats treated with CsA 12.5 mg/kg/day and 25 mg/kg/day, respectively. The number of interlobular arterioles stained for TGF-beta increased from a control value of 0.07 +/- 0.05/mm2 to 0.31 +/- 0.02/mm2, P < 0.05, and 0.39 +/- 0.07/mm2, P < 0.01, in rats treated with CsA, 12.5 mg/kg/day and 25 mg/kg/day, respectively. At the electron microscope level, TGF-beta was localized exclusively within the granular cells of the juxtaglomerular arterioles. Northern blot analysis suggested that this enhanced staining is due to increased transcription of TGF-beta 1. We have therefore observed an association between TGF-beta and CsA-induced nephrotoxicity. While this does not establish a causal link, it leads us to postulate that TGF-beta, alone or in combination with other growth factors, may play a role in the pathogenesis of CsA induced nephrotoxicity.

Alteration of antioxidant status in diabetic rats by chronic exposure to psychological stressors

Antioxidant status was measured in heart, liver, kidney, lung, and erythrocytes of 2-week streptozotocin-diabetic male Wistar rats exposed to chronic intermittent psychological stress consisting of 1 h of restraint twice daily for 14 days. Diabetes reduced erythrocyte and heart and liver susceptibility to hydrogen peroxide-induced glutathione depletion. Susceptibility to peroxide-induced thiobarbituric acid reactive substance (TBARS) formation increased in erythrocytes, liver, kidney, and lung but decreased in heart. Significant changes also occurred in glutathione levels (increased in heart and decreased in liver) and in the activities of catalase (reduced in liver and kidney), glutathione reductase (elevated in heart and liver), and glutathione peroxidase (decreased in liver and lung), but not Cu,Zn-superoxide dismutase. Stress potentiated diabetes-associated hyperglycemia and attenuated diabetesinduced hyperlipidemia. In addition, the reduction in peroxide-induced glutathione depletion in heart and liver and the increased TBARS formation in kidney and lung were reversed. Similarly, the diabetes-induced increase in liver glutathione reductase and decreases in liver and lung glutathione peroxidase activities were abolished by stress. Thus, the relative resistance of antioxidant systems to stress can be modified under pathologic conditions in which antioxidant alterations are present.

Comparison of gentamicin nephrotoxicity between rats and mice

Toxic effects of gentamicin administration (10–80 mg/kg body weight, subcutaneously (s.c.), once daily for 7 days) on several enzyme activities of kidney and duodenal mucosa together with other parameters were compared between male rats and mice. In Wistar rat kidney, tubular brush border Mg 2+-dependent, HCO 3 −-stimulated ATPase (Mg 2+-HCO 3 −-ATPase) activity was inhibited by 40–80 mg/kg gentamicin in an almost dose-dependent manner with no changes in microsomal Mg 2+-Na +-K +-ATPase activity. Cytosol carbonic anhydrase (CA) activity was inhibited only by 80 mg/kg gentamicin. In rat duodenal mucosa, Mg 2+-HCO 3 −-ATPase and CA activities were unchanged by any dose of gentamicin. Rat serum urea nitrogen (UN), GOT and GPT concentrations and urinary N-acetyl- β-D-glucosaminidase (NAG) activity were significantly increased by 80 mg/kg gentamicin. In ddY mice, however, almost all parameters described above were unaffected by gentamicin except for the urinary NAG activity which was increased only by 80 mg/kg gentamicin. The concentration of gentamicin in cytosol of rat whole kidney was approximately 3.4-fold higher compared with that in mouse kidney after 80 mg/kg treatment. In light microscopic analysis, 80 mg/kg gentamicin produced necrosis in the greater part of rat kidney proximal tubuli with no pathological findings in mouse kidney. In conclusion, Mg 2+-HCO 3 −-ATPase activity in brush border membrane of rat proximal tubuli was selectively damaged in gentamicin nephrotoxicity, indicating that the rats are the suitable model for studies of gentamicin nephrotoxicity in humans.

Frequent mutations of Ki-ras codon 12 in N-bis (2-hydroxypropyl)-nitrosamine-initiated thyroid, kidney and lung tumors in Wistar rats.

N-Bis (2-hydroxypropyl) nitrosamine (DHPN) is a very potent mutagen and wide spectrum carcinogen in rodents. In the present study, we investigated mutational activation of the Ki-ras gene in eight thyroid, five kidney (four mesenchymal and one transitional cell lesion) and two lung tumors induced with DHPN. Mutations were identified using single-strand conformation polymorphism, restriction fragment length polymorphism and DNA sequencing analysis. All of the 15 neoplasms could be shown to have mutations in codon 12 (GGT-->GAT). These results suggest that Ki-ras mutations are frequent events during the development of DHPN-induced carcinomas in these organs. In a separate experiment, moreover, we analyzed the presence of Ki-ras mutations in various tissues 8 weeks after DHPN treatment. One of five thyroid tissues treated with DHPN was found to have the same characteristic mutation, suggesting that it may represent an early event during carcinogen-induced tumor formation in the thyroid.

Simultaneous Determinations of Histamine and Nτ-Methylhistamine by High-Performance Liquid Chromatography-Chemiluminescence Coupled with Immobilized Diamine Oxidase

A method for the simultaneous determinations of histamine and its metabolite Nτ-methylhistamine by HPLC-chemiluminescence coupled with immobilized diamine oxidase was developed. The method was based on the determination of chemiluminescence formed by the reaction of a luminol-ferricyanide mixture in alkaline medium with hydrogen peroxide which is one of the metabolic products of histamine and Nτ-methylhistamine formed by diamine oxidase. HPLC with postcolumn derivatization resulted in good separation of the two amines and gave linear relationships between the concentrations of both and their chemiluminescence intensities. The lower limits of chemiluminescent detection of histamine and Nτ-methylhistamine were 5 and 10 pmol, respectively. The immobilized column showed good operational stability for more than 1 month, during which period 200 samples were analyzed. With this system, the histamine contents of the cerebral cortex, forestomach, glandular stomach, and kidney of Wistar rats were found to be 0.30, 58, 396, and 2.4 nmol/g wet wt, respectively. These values are very similar to those determined by HPLC-fluorometry. The Nτ-methylhistamine contents of these tissues were 0.36, 0.40, 0.72, and 3.8 nmol/g wet wt, respectively. This method will be useful for studying the roles of histamine in both brain and peripheral tissues.

Monovalent cation selective channel in the apical membrane of rat inner medullary collecting duct cells in primary culture

Ion channels in the apical membrane of rat inner medullary collecting duct (IMCD) were investigated by the patch clamp technique. Owing to the histological heterogeneity of IMCD, cells were cultured from the lower half of the inner medulla of Wistar rat kidney. Channel activity was rarely seen in cell attached patch, but membrane excision activated multiple units of 28.2 ± 0.7 pS cation selective channel. A Na or K selective channel was not found. The 28 pS channel showed membrane voltage dependency, no rectification, almost equal permeability to monovalent cations (Na/K/Li/Cs/Rb/NH 4 = 1:1.00:0.82:0.97:1.10:1.71) and no significant permeation to anions or divalent cations. Calcium of the cytoplasmic side from 10 −7 M to 10 −4 M affected the mean number of open channels ( nP o ) dose-dependently in excised patch (IC 50 = 5 · 10 −6 M). 1 mM of ATP, ADP, AMP and gadolinium reversibly suppressed nP o to near zero whereas amiloride, cAMP or cGMP had no effect. Multiple conductance substates were frequently observed. These results suggested that this channel belongs to the nonselective cation channels which has been identified in other epithelia and is not responsible for amiloride sensitive Na transport through IMCD cells.

Effect of intake of exogenous vitamins C, E and beta-carotene on the antioxidative status in kidneys of rats with streptozotocin-induced diabetes.

We studied the effect of supplementation with vitamins C, E and beta-carotene (PARABION, produced by Syndipharma) on antioxidative status in kidneys of male Wistar rats with diabetes induced by intravenous application of streptozotocin (45 mg.kg-1 of body weight). The animals received subtherapeutic doses of Insulin Interdep (6 U.kg-1 of body weight). A significant decrease of malondialdehyde (MDA), reduced (GSH) and oxidized (GSSG) glutathione and reduction of the activities of Se-glutathione peroxidase (Se-GSH-PX, EC. 1.11.1.9.) and glutathione S-transferase (GST, EC. 2.5.1.18.) were observed in kidneys of diabetic rats treated with these vitamins. On the contrary, the activity of CuZn-superoxide dismutase (CuZn-SOD, EC. 1.15.1.1) and the level of vitamin C (vit. C) increased significantly. No changes were observed for vitamin E (vit. E), beta-carotene and catalase (CAT, EC. 1.11.1.6). Supplementation with vitamins C, E and beta-carotene resulted in an improvement of antioxidative status of kidneys of rats with streptozotocin-induced diabetes.

The effect of sacrifice on image signal, T1, T2, and in liver, kidney, and brain of the Wistar rat

Signal intensity in SE240/25 and GE240/25 images, T*2, T2, and T1 were measured prior to and after sacrifice at 1.5 T in liver, kidney, and brain of Wistar rats. Presacrifice, animals were maintained on 2-3% halothane, ensuring a high saturation of venous haemoglobin with oxygen, which emphasised blood oxygenation level dependent (BOLD) effects upon deoxygenation of haemoglobin postsacrifice. T*2 values (in ms) changed on sacrifice from 27.8 +/- 4.1 to 15.0 +/- 1.1 in liver (p << .001), from 57.4 +/- 6.5 to 52.8 +/- 5.7 in brain (p < .02), and from 52.3 +/- 10.3 to 37.6 +/- 11.9 in kidney cortex (p < .01). T*2 was also reduced in kidney medulla and papilla. T2 reduced on sacrifice (by a lesser amount than T*2), except in brain, which showed a small increase. T1 (ms) changed only in liver, showing a significant increase from 670 +/- 31 to 834 +/- 69 (p << .001). The ratio of signal postsacrifice to presacrifice was lowest in the SE240/25 and GE240/25 images of liver (0.73 +/- 0.03 and 0.52 +/- 0.05, respectively), but increased T1 may account for some of these changes. Expected signal ratios at sacrifice calculated from measured relaxation times agreed well with those observed experimentally. Relaxation changes observed on sacrifice were consistent with deoxygenation of haemoglobin. These results give an estimate of the maximum effects which changes of blood oxygen level might produce at 1.5 T and suggest that contrast due to variation of endogenous deoxyhaemoglobin may be exploitable in the liver.

Toxicity of neomycin on enzyme activities of kidney and duodenal mucosa in vivo: organ specificity and species difference between rats and mice

Inhibitory effects of neomycin administration (10–80 mg/kg body weight/day, s.c. for 7 days) on several enzyme activities of kidney and duodenal mucosa were compared between male rats and mice. In Wistar rat kidney, tubular brush border Mg 2+ -dependent, HCO 3 − -stimulated ATPase activity was inhibited by neomycin in a dose-dependent manner, while microsomal Mg 2+ -Na + -K + -ATPase activity and cytosol carbonic anhydrase (CA) activity were inhibited only by larger doses. In rat duodenal mucosa, Mg 2+ -HCO 3 − -ATPase and CA activities were also inhibited only by larger doses. Serum urea nitrogen (UN) concentration and urinary N -acetyl- β -D-glucosaminidase (NAG) activity were increased in a dose-dependent manner. In ddY and C57black mice, however, all enzyme activities in kidney and duodenal mucosa were almost unaffected by any dose of neomycin and showed no changes in serum UN concentration and urinary NAG activity except for ddY mice in which NAG activity was only increased by the largest dose of neomycin. In light microscopic analysis, 80 mg neomycin/kg produced necrosis in the greater part of rat proximal tubuli with no changes in duodenal brush border. On the other hand, no histological changes were produced in the renal cortex or duodenal mucosa of mice by any dosage. In conclusion, there are organ-specific and species differences in the effects of neomycin between rats and mice.

Toxicity of neomycin on enzyme activities of kidney and duodenal mucosa in vivo: organ specificity and species difference between rats and mice

Inhibitory effects of neomycin administration (10-80 mg/kg body weight/day, s.c. for 7 days) on several enzyme activities of kidney and duodenal mucosa were compared between male rats and mice. In Wistar rat kidney, tubular brush border Mg(2+)-dependent, HCO3(-)-stimulated ATPase activity was inhibited by neomycin in a dose-dependent manner, while microsomal Mg(2+)-Na(+)-K(+)-ATPase activity and cytosol carbonic anhydrase (CA) activity were inhibited only by larger doses. In rat duodenal mucosa, Mg(2+)-HCO3(-)-ATPase and CA activities were also inhibited only by larger doses. Serum urea nitrogen (UN) concentration and urinary N-acetyl-beta-D-glucosaminidase (NAG) activity were increased in a dose-dependent manner. In ddY and C57black mice, however, all enzyme activities in kidney and duodenal mucosa were almost unaffected by any dose of neomycin and showed no changes in serum UN concentration and urinary NAG activity except for ddY mice in which NAG activity was only increased by the largest dose of neomycin. In light microscopic analysis, 80 mg neomycin/kg produced necrosis in the greater part of rat proximal tubuli with no changes in duodenal brush border. On the other hand, no histological changes were produced in the renal cortex or duodenal mucosa of mice by any dosage. In conclusion, there are organ-specific and species differences in the effects of neomycin between rats and mice.

Changes in bioavailability and tissue distribution of zinc caused by magnesium deficiency in rats.

The effect of a Mg-deficient diet (200 mg Mg/kg feed) on the bioavailability of dietary Zn and the concentration of this cation in plasma, whole blood, skeletal muscle, kidney, heart and brain of Wistar rats was studied after 7, 35, 42, 49, 56, 63 and 70 d. Mg deficiency significantly decreased Zn in whole blood on day 42 of the experiment, but there was no significant change in plasma Zn throughout the 70 d study period. The Mg-deficient diet significantly increased intestinal absorption of Zn, Zn balance, and Zn concentration in femur and kidney, but decreased Zn concentration in the heart despite the increase in dry weight of this organ. No change was found in brain Zn concentration.

Accumulation of autofluorescent yellow lipofuscin in rat tissues estimated by sodium dodecylsulfate extraction

Autofluorescent yellow lipofuscin accumulated in the older Wistar rat brain, kidney, spleen and testis was successfully extracted by use of 0.5% sodium dodecylsulfate (SDS)/0.05 M phosphate buffered saline (pH 7.4). The extracts contained three fluorophores, one of which corresponded to yellow lipofuscin and showed an excitation maximum at 400 nm and an emission maximum at 620 nm. The SDS extraction method was found suitable for the accurate quantification of lipofuscin in rat tissues. Lipofuscin thus determined accumulated in brain, kidney, spleen and testis with the increase of age of rats. It was found that kidney was the most susceptible tissue in lipofuscin accumulation, and accumulation in male rat kidney was much higher than that in female rat kidney. Accumulation of lipofuscin in rat tissues was slightly dependent on the lipid peroxidation state of the tissues and the dietary conditions: a vitamin E-adequate, vitamin E-deficient, vitamin E-excessive and oil-rich diet, except that the accumulation in male and female rat kidney was significantly enhanced when fed a vitamin E-deficient diet for 7–10 weeks.

Cetaben and fibrates both influence the activities of peroxisomal enzymes in different ways

The effects of cetaben and clofibric acid were compared on the activities of peroxisomal enzymes in the liver and kidney of male Wistar rats. Cetaben at 200 mg kg body wt increased the activities of all of the enzymes in the liver that were studied two to eight times, whereas the changes induced by the same dose of clofibric acid increased some of the enzymes and decreased others. In the kidney, cetaben increased the activities of all investigated peroxisomal enzymes, while clofibric acid only increased the activity of palmitoyl-CoA oxidase. The data obtained in the dose-response study of cetaben revealed a significant rise in the activities of peroxisomal enzymes in both the liver and kidney at doses of 50–100 mg/kg body wt administered over 10 days, but the maximal effect was observed at 250 mg/kg. Palmitoyl-CoA oxidase and d-amino acid oxidase respond most markedly to cetaben. Cetaben could represent an atypical peroxisomal proliferator, since it increased the activities of all peroxisomal enzymes investigated. The fact that the individual components localized in the peroxisomes do not change markedly could be of importance with respect to the function and physical properties of peroxisomes.