A study of the action of clonidine on secretion from the adrenal medulla in dogs.

The effects of clonidine on adrenal catecholamine (adrenaline and noradrenaline) secretion were investigated in chloralose-anaesthetized dogs. Intravenous administration of clonidine (10 and 20 micrograms kg-1) induced a decrease in both adrenal catecholamine secretion rates and cardiovascular parameters (blood pressure and heart rate). In contrast, a dose of 5 micrograms kg-1 was ineffective. Intracisternal clonidine (in a lower dose of 3 micrograms kg-1) also decreased adrenaline and noradrenaline release from the adrenal gland. Clonidine failed to modify adrenal catecholamine release evoked by electrical stimulation of the splanchnic nerve. These results demonstrate that clonidine decreases adrenaline release from the adrenal gland through a central and not a peripheral mechanism in dogs. This action might contribute to its antihypertensive effects.

Olfactory memory in rats, cholinergic agents and benzodiazepine receptor ligands.

Drugs and their effects on olfactory learning processes in rats were tested using a modified version of the runway apparatus developed by Ades. Rats were first exposed to a conspecific urine sample and 24 h later were exposed to the same stimulus in the runway. Observations recorded the time spent investigating the urine and the number of sniffs at the site, these being considered to be indices of memory. Diazepam-treated rats (4 or 6 mg/kg) and scopolamine-treated rats (0.5 or 1 mg/kg) showed increases for both parameters. When both drugs were administered simultaneously, the impairing effect was potentiated. However, no changes in learning responses were observed in rats treated with physostigmine (0.125, 0.25, 0.5 mg/kg) or methyl beta-carboline-3-carboxylate (0.3, 0.5, 1 mg/kg), although the administration of physostigmine or methyl beta-carboline-3-carboxylate was shown to antagonize the impairing effect of diazepam or scopolamine respectively. These observations support the hypothesis of interactions existing between cholinergic agents and benzodiazepine receptor ligands and of such interactions affecting olfactory acquisition processes. The runway apparatus appears to be a valid candidate model to be used for the assessment of pharmacological influences on olfactory learning in rats.

Opposite effects of cholinergic agents and benzodiazepine receptor ligands in a passive avoidance task in rats.

Benzodiazepine (Bzd) agonist, diazepam (Dzp) and inverse agonist methyl beta-carboline-3-carboxylate (beta-CCM); acetylcholinesterase inhibitor, physostigmine (Physo) and muscarinic antagonist, scopolamine (Scopo), were investigated for their mnesic effect in a passive avoidance (PA) task in rats. Impairments were observed after Dzp- and/or Scopo-pretraining treatments. Physo was without effect but antagonized the Dzp-induced impairments. beta-CCM enhanced acquisition and antagonized the Scopo-induced impairing effect. All these drugs had no effect in posttraining administration.

Septal scopolamine-systemic diazepam interaction in passive avoidance learning in rats.

The role of the septal region in passive avoidance (step-through) learning was studied by administering 15µg/0.5µl of scopolamine intraseptally to rats, together with an i.p. injection of 2mg/kg of diazepam to the same animals. The combined treatment provoked a severe impairment in retention of the learning situation, whereas animals which received either scopolamine or diazepam alone did not show any deficit.

Intraseptal injection of scopolamine increases the effect of systemic diazepam on passive avoidance learning and emotionality in rats.

This experiment was designed to assess the role of the septo-hippocampal cholinergic (ACh) system in the deleterious effects produced by systemic benzodiazepine injection on learning processes in rats. Retention of a step through passive avoidance task was analysed after systemic injection of increasing doses of either scopolamine or diazepam applied alone 30 min before the acquisition phase. Results indicated a dose related impairment of retention by each drug: in addition, sub-threshold doses of scopolamine and diazepam applied in combination (diazepam: 2mg/kg plus scopolamine: 0.3mg/kg) produced a decrease of retention latencies, thus showing an additive effect of the combined treatment. Secondly, a sub-threshold dose of scopolamine (15microg/0.5microl) was also administered into the medial septal area, together with an i.p. injection of 2mg/kg of diazepam. This combined treatment produced a severe impairment of retention, in parallel with a large reduction in emotionality (number of faeces). The data are consistent with the hypothesis that peripheral administration of behaviorally effective doses of diazepam on passive avoidance learning might act partially via a septal ACh-GABA/benzodiazepine mechanism. It is also suggested that this mechanism subserves both anxiety and the memorisation of contextual stimuli associated with passive avoidance acquisition, through the modification of the septo-hippocampal activity.

Paraquat potentiates iron-induced microsomal lipid peroxidation: modulation by the diet lipid composition.

The study concerns the role of two combined factors-lipid composition of the microsomal membranes and the iron concentration in the incubation medium-in lipid peroxidation catalysed by paraquat (P(++)). Rats were subjected to diets containing 5% lipids composed of either tripalmitin (T), peanut oil/rapeseed oil (v/v) (C) or fish oil (F). The level of polyunsaturated fatty acids in the microsomal membranes was higher in C and F than in T. The level of vitamin E was lowest in F. The activity of the system 'Cyt P450-NADPH cyt c reductase' increased in the order T

Phospholipid fatty acid composition affects enzymatic antioxidant defenses in cultured Swiss 3T3 fibroblasts.

The aim of this work was to study the adaptation of enzymatic antioxidant cell defense to the nature of the membrane polyunsaturated fatty acids (PUFA). 3T3 Swiss fibroblasts were grown for 5 days in a medium supplemented with 50 microM linoleic acid (LA) or eicosapentaenoic acid (EPA) and compared to control cells (C). The phospholipid fatty acid content was evaluated: LA were enriched in n-6 PUFA (27.8%) in comparison to C (6.7%) or EPA (5.6%); EPA were enriched in n-3 PUFA (26.2%) in comparison to LA (4.4%) or C (4.6%). The fatty acid double bond index (DBI) increased from C to LA and EPA. The activities of the three key enzymatic antioxidant defenses, SOD, GPx and GST, increased with the degree of unsaturation of the phospholipid fatty acids. In the cells with fatty acids that are very sensitive to oxidative stress, the higher activities of SOD and GPx might act to limit the initiation of lipid peroxidation and the higher activities of GST and GPx to decrease the toxic effects of the various species produced from lipid degradation.

Potentiation of iron-induced lipid peroxidation by a series of bipyridyls in relation to their ability to reduce iron.

We determine the relative abilities of three bipyridyls (Paraquat P + +, Benzylviologen B + + and Diquat D + +) to stimulate iron-induced lipid peroxidation in relation to their ability to redox cycle and to reduce iron. The ability to redox cycle increases with the redox potential, in the order: P + + < B + + < D + +. The initial rates of Fe3+ reduction increased from P + + to D + + and B + +. NADPH-dependent microsomal lipid peroxidation was measured in different conditions (1) increasing incubation time, 100 microM bipyridyl, 10 microM FeCl3; (2) increasing concentrations of bipyridyls, 10 microM FeCl3, 10 min incubation time; (3) increasing concentrations of FeCl3, 100 microM bipyridyl, 10 min incubation time. Bipyridyls potentiated the production of malondialdehyde (MDA), particularly with the shortest incubation times and the lowest concentrations of FeCl3, in the order of activity P + + < D + + < B + +. The strong correlation between the initial rate of lipid peroxidation and Fe3+ reduction indicates that the rate of Fe3+ reduction determines the intensity of the potentiation effect of bipyridyls on lipid peroxidation--on the condition that the amount of peroxidizable substrate is not the limiting factor.

Total parenteral nutrition decreases liver oxidative metabolism and antioxidant defenses in healthy rats: comparative effect of dietary olive and soybean oil.

BACKGROUND: Total parenteral nutrition (TPN) is used for critically ill patients undergoing surgery, after trauma, or during disease conditions that favor oxidative stress. We studied the effect of TPN on liver oxidative metabolism and antioxidant defenses in rats, and we compared the effect of soybean oil- and olive oil-based diets. METHODS: Seven-week-old rats (n = 28) were divided into four groups. Two experimental groups received a TPN solution containing soybean oil (TPN-S) or a mixture of olive/soybean oil, 80/20 (TPN-O), IV for 6 days. Orally fed animals received a solid diet including soybean oil (Oral-S) or olive/soybean oil, 80/20 (Oral-O). The following parameters were measured: DL-alpha-tocopherol, vitamin A, malondialdehyde and thiobarbituric acid reactive substances (MDA-TBARS), and total radical-trapping antioxidant parameter (TRAP) in serum; DL-alpha-tocopherol, vitamin A, glutathione (GSH), and catalase (Cat) activity in liver homogenate; fatty acids from phospholipid, cytochrome P-450 content, NADPH-cytochrome c2 reductase activity in liver microsomes; superoxide dismutase (SOD), glutathione peroxidase (Gpx), glutathione reductase (GR), glutathione transferase (GST), and glucose-6-phosphate dehydrogenase (G6PD) in liver cytosol. RESULTS: The soybean or olive oil diets modified the liver microsomal fatty acid phospholipid composition, but the unsaturation index remained unchanged. TPN specifically increased the saturation of the membrane. The cytochrome P-450 level and the NADPH-cytochrome c2 reductase, SOD, Gpx, Cat, and GST activities were unchanged by soybean oil or olive oil diet but decreased receiving TPN. CONCLUSIONS: In rats, TPN decreased the liver oxidative metabolism and enzymatic antioxidant defenses. This may be related to saturation of the liver microsomal fatty acids.

Paraquat and iron-dependent lipid peroxidation. NADPH versus NADPH-generating systems.

The aim of this work was to study the effect of paraquat (P2+) on NADPH iron-dependent lipid peroxidation (basal peroxidation) either in the presence of NADPH or in the presence of NADPH-generating systems. When NADPH is present, P2+ potentiates NADPH iron-dependent lipid peroxidation, but use of NADPH-generating systems cancels this effect. This may be attributed to certain components in NADPH-generating systems such as glucose-6-phosphate and sodium isocitrate, which act as iron chelators. The binding of iron by these molecules facilitates its reduction and enhances its reactivity toward dioxygen molecules, leading to the formation of reactive species capable of initiating lipid peroxidation, such as Fe3+-O2-*. Under these conditions of rapid basal peroxidation, any additional reduction of iron(III) by a reduced form of P2+ (P+*) has no apparent effect on the peroxidation itself, probably because the initial reaction between iron(II) and O2 followed by initiation of the peroxidation are both rate-limiting steps in the process. Consequently, any alteration of the composition of the reacting mixture (e.g., buffers or the generating system) must be taken into consideration because the formation of new iron chelates can change the rate of basal peroxidation and will modify the effect of redoxcycling molecules.

Microsomal membrane peroxidation by an Fe3+/paraquat system. Consequences of phenobarbital induction.

Descriptions of the effects of paraquat (P2+) on the peroxidation of liver microsomes are very divergent. Therefore, the presence of ferric iron in the medium and the activity of microsomal mixed-function oxidase system are two factors that we have taken into consideration to explain the discrepancies. The results showed that 100 microM P2+ potentializes the slight production of MDA induced by low concentrations of Fe3+ (< or = 15 microM). In these conditions, P+., arising from the one-step reduction of P2+ by NADPH-cytochrome C reductase, could reduce Fe3+ and cause the formation of species that initiate peroxidation. However, unlike the results obtained with CBrCl3, for animals induced by phenobarbital (Ph), the production of MDA in the presence of FeCl3 and of P2+ was weaker than for the controls. The establishment of a new Fe3+/Fe2+ equilibrium owing to increased production of P+. could be responsible.

[Analysis of the behavior of rats in the Morris water-basin: new methodology and pharmacological application]. / Analyse du comportement du rat dans la piscine de Morris: nouvelle méthodologie et application pharmacologique.

The Morris water-maze has been designed to test spatial orientation ability, learning and memory processes. In order to improve the analyse of the organization of the trajectory of rats, during the training phase, a computer program was elaborated. The study of the effect of a benzodiazepine, diazepam, was chosen to illustrate and validate this methodological approach. Results showed that rats pre-treated with diazepam (2 mg/kg) presented an impairment of spatial learning associated with the occurrence of a stereotyped circular swimming behaviour.

Plasma and hepatic antioxidant control and vitamin A nutritional status.

Twenty-seven rats were divided into three groups and fed on diets containing 0.3, 6 or 60 RE (retinol equivalent) retinyl palmitate/g food. After 7 weeks, hepatic vitamin A uptake was found to be more efficient in vitamin A-deficient rats than in rats given adequate vitamin A. We showed that during the metabolic adaptation of the animals to the level of vitamin A in the diet, extensive modifications occur in the antioxidant defences of the organism. In parallel with the increase in the level of vitamin A, the decrease in the level of alpha-tocopherol in the plasma can bring about a greater susceptibility of the lipoproteins to oxidative stress. Similarly, the decrease in the hepatic alpha-tocopherol level and in glutathione peroxidase activity leads to the weakening of the liver's antioxidant defences.

[Nephroepithelioma of children manifested by isolated recurrent hematuria. Diagnostic problems apropos of 3 cases]. / Le nephroepitheliome de l'enfant à forme d'hematurie recidivante isolee. Problemes diagnostiques a propos de 3 observations

In children, nephroepithelioma represent less than 10% of the malignant tumors of the kidney. Reporting 3 cases, the authors emphasize that the clinical expression of nephroepithelioma, often made only of recurrent hematuria occuring over years, may mislead adequate diagnostic approach. Thus, a kidney arteriogram should be included in the investigations of nephroepithelioma. Earlier diagnosis may improve the rate of recovery from nephroepithelioma which, in children, is about 30%.

Rilmenidine (S 3341) and the sympathoadrenal system: adrenoreceptors, plasma and adrenal catecholamines in dogs.

1. The effects of rilmenidine, a new alpha 2-adrenoreceptor agonist with antihypertensive properties, were investigated on plasma catecholamines, blood cell adrenoreceptors and adrenal medullary function. 2. In conscious sino-aortic denervated (SAD) dogs, rilmenidine (1 mg kg-1 orally for 2 weeks) significantly reduced both blood pressure and heart rate when compared with placebo treatment. The drug decreased plasma noradrenaline and adrenaline levels and corrected the decrease in leucocyte beta-adrenoreceptors observed in placebo-treated SAD dogs. There was no change in platelet alpha 2-adrenoreceptors. 3. In anaesthetized normotensive dogs, rilmenidine (0.1 and 0.3 mg kg-1 i.v.) induced a dose-dependent decrease in both cardiovascular parameters (blood pressure and heart rate) and catecholamine release from the adrenal medulla. 4. The present study shows that rilmenidine decreases sympathetic tone mainly by an action on the adrenal medulla. In addition, its ability to lower blood pressure in SAD dogs, i.e. a model of hypertension in which high sympathetic tone is present, indicates that rilmenidine may also depress other parts of the sympathetic nervous system.

Is adrenal medulla involved in the antihypertensive effect of nicardipine?

The effects of nicardipine, a dihydropyridine calcium channel blocker, on adrenal medulla were investigated in chloralose-anesthetized dogs. For analysis of the adrenal medullary function, adrenal venous catecholamine rates were determined. Intravenous administration of nicardipine (50 micrograms/kg) induced a marked increase in both adrenal catecholamine rates and heart rate and a simultaneous decrease in blood pressure. This effect on epinephrine and norepinephrine release is probably explained by the involvement of baroreflex mechanisms. After acute splanchnicectomy, nicardipine (50 and 100 micrograms/kg i.v.) failed to modify catecholamine secretion rates from the denervated adrenal medulla during electrical stimulation of the splanchnic nerve at low (2 Hz) and high (5 Hz) frequencies. In conclusion, these results suggest that in vivo a functional dihydropyridine-sensitive calcium channel is not required for calcium entry mechanisms into dog chromaffin cells. Moreover, adrenal medulla is not involved in the antihypertensive action of nicardipine.