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S Hynie

Publications and source records attributed to S Hynie.

At least 19 recordsLinked to original sources

Effects of stress and of amphetamine on passive avoidance conditioning in rats.

This study examined the effects of immobilization stress combined with water immersion (ICS) and/or amphetamine (AM) on different memory phases in the passive avoidance task in rats. The performance of rats was evaluated in the retention tests 24 and 48 h after a single acquisition trial. ICS exposure lasting 1 h impaired retention of the learned avoidance response if applied 2 to 4 h before or immediately after training. The stressor did not affect retrieval if presented 5 or 2 h before the retention test. AM was used i.p. at the dose of 8 or 1 mg/kg. Neither 8 mg AM administered 4 h before nor 8 or 1 mg doses given after training did not impair the retention performance in unstressed rats. The 1 mg AM prevented the impairment of retention in animals exposed to the stressor 3 or 4 h before training but had no effect when the stronger impairment was induced by ICS 2 h before training. However, when given 1 h before retention testing, 1 mg AM attenuated even the severe impairment induced by the pre-training stressor exposure. Our results suggest that ICS impairs primarily the early phase of memory consolidation and a low dose of AM can prevent this effect.

Amphetamines↗

In situ detection of cyclic AMP-phosphodiesterase activity in the heart of Lewis and Sprague-Dawley rats: the effect of restraint stress or amphetamine application.

Cyclic AMP plays an important role in heart functions under normal as well as pathological conditions. Since phosphodiesterase (PDE), responsible for the hydrolysis of cAMP, is equally important as synthesizing adenylyl cyclase, we decided to determine its activity by cytochemical procedure after exposure of rats to restraint stress or an acute dose of amphetamine. Sprague-Dawley (S-D) and Lewis (LE) rats, the latter known to have a deficient hypothalamo-pituitary-adrenal axis activity, were used in order to disclose the possible significance of rat strain on PDE activity. Animals were divided into 3 groups: controls, rats treated with an acute dose of amphetamine (8 mg/kg, i.p., for 60 min) and rats under restraint stress for 60 min. Control hearts of both strains revealed PDE activity on sarcolemma of cardiomyocytes and plasmalemma of endothelial cells of microvessels. In LE rats we observed an additional enzyme reaction in junctional sarcoplasmic reticulum. In addition, cardiomyocytes of LE rats revealed a higher PDE activity when compared to S-D rats. Restraint stress decreased PDE activity in cardiomyocytes of LE rats while amphetamine markedly inhibited enzyme activity in cardiomyocytes of S-D rats. Endothelial PDE was more resistant to stress. Our results indicate differences in PDE localization and variations in sensitivity of myocardial cAMP-PDE of LE and S-D rat strains to restraint stress and amphetamine application.

Amphetamines↗

Exposure to intermittent high altitude induces different changes in adenylyl cyclase activity in hearts of young and adult Wistar rats.

This study investigates changes of adenylyl cyclase activity in the heart of young and adult Wistar rats exposed to experimental conditions simulating high altitude hypoxia as a model for interpretation of some adaptive changes of adenylyl cyclase observed in human. The exposure of rats to intermittent high altitude (IHA) hypoxia (5000 m) showed significant adaptive changes. The right ventricular weight and the ratio of right/left ventricular weights of adult rats exposed to IHA were significantly increased when compared to appropriate controls; adaptive changes of cardiac adenylyl cyclase being dependent on the age of the animals. The isoprenaline-stimulated activity was higher in the left than in the right ventricle, and in both ventricles it was higher in young rats than in adult rats. When compared to controls, isoprenaline stimulation was decreased in the right ventricles of adapted young rats and, by contrast, it was increased in the left ventricles of adapted adult rats. This decrease and increase of adenylyl cyclase activity evoked by isoprenaline was paralleled by forskolin-induced adenylyl cyclase activity in these experimental groups. It seems therefore that the changes in the pattern of total adenylyl cyclase activity observed under IHA hypoxia may at least be partially explained by the changes of beta-adrenergic receptor susceptibility following IHA hypoxia.

Adaptation, Physiological↗

Impaired passive avoidance acquisition in Wistar rats after restraint/cold stress and/or stresscopin administration.

Stresscopin (SCP) and related peptides are new members of the corticotropin-releasing factor (CRF) peptide family that are selective ligands for CRF type 2 receptor; these ligands are essential for maintaining homeostasis after stress. SCP (i.p. injections) was tested on the passive avoidance learning task in stressed Wistar rats; it impaired the formation of memory trace. The retention performance deficit induced by SCP was comparable with the deficit induced by the stressor of restraint/cold. More profound impairment of avoidance response occurred following combined application of SCP and stressor. More specific actions of SCP can be expected from its studies with targeted intracerebral applications.

Animals↗

Differences in the behaviour of Sprague--Dawley and Lewis rats during repeated passive avoidance procedure: effect of amphetamine.

The present paper investigated the differences in passive avoidance learning between Sprague--Dawley and Lewis rats. After initial habituation (experimental Part 1), measured as latencies to enter the dark, preferable compartment, the effect of treatment with amphetamine (8 mg kg(-1)b.w.), the retention performance compared with controls (saline) was tested in both rat strains in Parts 2--4. The intervals between Parts 2--4 were 24 or 49 days. Each experimental part consisted of testing lasting 6 days. On the 7th day the rats received drug treatment 1 h before the application of foot shock. The differences between rat strains were already detectable at the beginning of the study. During the repeated exposures of rats in Part 1, only Lewis rats, in contrast to Sprague--Dawley rats, exhibited the habituation. The repeated testing of rats in Parts 2--4, due to previous experience with an aversive stimulus, was considered as the retention test. In Parts 2--3 we observed only minor differences in the responses of both rat strains tested. Also no significant differences were observed between rat strains after amphetamine treatment that induced an amnesia-like effect in all retention trials. However, data shown in Part 4 revealed the largest differences between both strains. Control Lewis rats exhibited significantly higher retention responses than Sprague--Dawley rats. In the latter strain we observed no differences in avoidance latencies between controls and amphetamine treated rats. In Lewis rats the difference in avoidance performance between controls and amphetamine treated animals was highly significant due to their enhanced retention performance. In conclusion, the results presented in this study extend the known behavioural differences in tested rat strains to the passive avoidance procedure that, in addition, was performed for a total period of 4 months. Due to a known deficiency of hypothalamo-pituitary-adrenal axis activity in Lewis rats it can be hypothesized that the behavioural dissociation of this strain from Sprague--Dawley rats could be related to the different activity of this regulatory axis in the rat strains tested.

Amphetamine↗

Rat strain differences in responses of plasma prolactin and PRL mRNA expression after acute amphetamine treatment or restraint stress.

1. The aim of this study was to compare the effects of acute amphetamine (AMPH) treatment and restraint stress on plasma level of prolactin (PRL) and PRL mRNA expression in the adenohypophysis in Sprague-Dawley and Lewis male rats, the latter known to have a deficient hypothalamo-pituitary-adrenal (HPA) axis. 2. Both restraint stress and AMPH treatment (i.p. in a dose of 8 mg/kg of b.w.) were applied 15 or 30 min before termination of the experiment. Plasma PRL and corticosterone (CORT) were determined by radioimmunoassay. PRL mRNA expression was estimated by a dot-blot hybridization. 3. Restraint stress and AMPH treatment induced a significant increase in the CORT plasma level, as an indicator of stress response. Compared to Sprague-Dawley rats, the magnitude of CORT increase after both stimuli was significantly lower in Lewis rats. 4. Although restraint stress significantly increased the PRL plasma levels in both rat strains, AMPH treatment reduced the PRL levels in both rat strains. However, the changes of PRL plasma levels had another pattern in Lewis rats than in Sprague-Dawley rats. Control plasma PRL levels were significantly higher in Lewis rats, and in this rat strain AMPH treatment for 30 min increased the PRL levels as compared to the values obtained after AMPH treatment for 15 min. 5. Expression of PRL mRNA in adenohypophysis by restraint stress and AMPH treatment had a similar pattern. After a 15-min lasting restraint stress, the expression of PRL mRNA was decreased insignificantly in both rat strains. AMPH treatment induced in Sprague-Dawley rats a significant decrease of PRL mRNA after a 15-min interval while after 30 min there was a significant increase. However, in Lewis rats AMPH failed to significantly change PRL mRNA. 6. The results from the present study indicate that the mechanisms mediating the effects of acute restraint stress and acute AMPH treatment differ in PRL response in Sprague-Dawley and Lewis male rat strains. Differences in the observed responses in Lewis rats could be related to the deficient activity of HPA axis in this rat strain.

Acute Disease↗

[Nobel Prize winning laureates in physiology or medicine for the year 2000--a few comments on discoveries related to signal transduction].

The Nobel Assembly awarded The Nobel Prize in Physiology or Medicine for 2000 jointly to Arvid Carlsson, Paul Greengard and Eric Kandel for their discoveries concerning signal transduction in the nervous system. On the examples of their predecessors we attempted to demonstrate how results of basic research serve as building blocks for new discoveries and for the application of research results into the praxis. We presented not only the basic discoveries of laureates of Nobel Prize for year 2000 (biological role of dopamine, regulation of cell functions by phosphorylation of proteins, changes in transduction of signals during processes of memory), but we also mentioned previous discoveries that helped in the research of the last laureates. These discoveries concerned not only the storage and metabolism of transmitters, formulation of the concept of cyclic AMP as a second messenger of hormonal action, the role of G-proteins in transduction processes in receptor-effector complexes, processes of phosphorylation of proteins as regulators of cell functions, but we also mentioned the discovery of other second messengers and substances functioning as local hormones (prostaglandins and related compounds). Most of the described discoveries have not only the value as stones that can help to fill still incomplete mosaic of our present knowledge, but they also represent the immediate basis for the development and use of very important remedies, such as are antiparkinsonics, antidepressive drugs, nonsteroidal antiinflammatory drugs, etc.

History, 20th Century↗

Silymarin effects on intracellular calcuim and cytotoxicity: a study in perfused rat hepatocytes after oxidative stress injury.

The re-emergence of silymarin as a natural remedy for diseases of the liver and biliary tract necessitates reevaluation of the efficiency of this compound and its possible mode of action. The aim of this study was to investigate the potentials of silymarin on the amelioration of hepatic injuries. The possible mechanism(s) that contribute to the hepatoprotective effect of silymarin and the role played by intracellular calcium (Ca(2+)(i)) was investigated using tert-butyl hydroperoxide (TBH) and D-galactosamine (D-Gal) intoxication in a model of the isolated immobilized and perfused hepatocytes. Silymarin decreased lactate dehydrogenase (LDH) leakage, increased oxygen consumption, reduced the formation of lipid peroxides (malondialdehyde, MDA) in hepatocytes that were altered by TBH and increased urea synthesis in the perfusion medium. TBH treatment increased Ca(2+)(i) in hepatocytes significantly to a value of more than 600 nM and silymarin pre-treatment reduced the TBH-induced rise in Ca(2+)(i) and brought Ca(2+)(i) level to below 300 nM. Silymarin did not affect LD leakage or urea synthesis in D-Gal-injured cells. It is concluded that silymarin hepatoprotective effect under the present experimental conditions is due to the inhibition of lipid peroxidation and that the modulation of hepatocyte Ca(2+)(i) plays a pivotal role in a protective effect.

Animals↗

Cyclosporin A modifies cytoplasmic calcium levels in isolated hepatocytes exposed to oxidative stress due to tert-butyl hydroperoxide.

Within the framework of our studies on hypertension in various rat strains, we have examined the effect of cyclosporin A (CsA) on intracellular calcium signaling under conditions of oxidative stress. For these preliminary experiments, we have chosen isolated hepatocytes of normotensive rats as a model system for the study of the role of intracellular calcium. We used tert-butyl hydroperoxide (t-BHP, 1 mmol x l(-1)) as an prooxidant agent. When compared to the controls, we found increased levels of cytosolic free calcium concentration (Ca2+i) during 120 min incubation. The preincubation of hepatocytes with CsA in the concentration of 0.5 micromol x l(-1)] did not change the physiological level of cytosolic calcium. However, a dual action of CsA on elevated Ca2+i was observed during oxidative injury of hepatocytes: while in the first period of incubation CsA increased Ca2+i, CsA reduced the effect of t-BHP on Ca2+i during the next period of incubation. This indicates the ability of CsA to modify oxidative stress, but further studies are necessary to explain these findings.

Animals↗

Effects of immobilization stress combined with water immersion and chronic amphetamine treatment on the adenylyl cyclase activity in rat neurohypophysis.

Several papers have indicated the participation of cyclic AMP as a second messenger for the release of neurohypophysial hormones. Since very little is known about the effects of stress and drugs of abuse on this process, we studied the activity of adenylyl cyclase in the neurohypophyses after immobilization stress and chronic amphetamine treatment. Our findings indicate the involvement of cyclic AMP in the regulation of neurohypophysis as well as the increase in total adenylyl cyclase both after application of immobilization stress combined with water immersion and after chronic amphetamine treatment.

Adenylyl Cyclases↗

Phase I and phase II xenobiotic biotransformation in different inbred strains of rats: study in immobilized perfused hepatocytes.

The present study was designed to compare phase I and phase II biotransformation reactions in immobilized perfused hepatocytes as a cellular system obtained from inbred rat strains which represent models for some cardiovascular diseases, namely, spontaneously hypertensive rats (SHR), rats sensitive and resistant to isoprenaline-induced myocardial lesions (IS and IR, respectively) as compared to Wistar rats (W). The biotransformation kinetics for hexobarbital (HX), 7-ethoxycoumarin (7-EC), 1-chloro-2,4-dinitrobenzene (CDNB) and 4-nitrophenol (4-NP) were followed up in the hepatocyte perfusate. W and SHR rat hepatocytes have metabolized HX at a higher rate than those of the IR and IS strains. Hepatocytes from the W strain exhibited a higher rate of 7-EC deethylation activity compared to hepatocytes obtained from the IR or IS strains. Hepatocytes obtained from SHR and IR rats showed the highest glutathione-S-transferase (GST) activity towards CDNB compared to the IS or W strain. 4-NP disappearance was higher in the perfusion medium of hepatocytes obtained from the W and IS strains compared to the IR strain. These significant differences in drug biotransformation between various studied strains, which may be genetically determined, can be well demonstrated by using an efficient drug metabolizing model of the immobilized perfused hepatocytes. The importance of these differences should be considered during the study of the experimental therapy of the relevant disease as obtained from the specific experimental strain, where it may be expected that the pharmacokinetic profile of a drug in vivo and consequently its pharmacodynamic or toxic effects will be strain dependent.

Animals↗

Different activity of adenylyl cyclase in prefrontal cortex in three rat strains. The effect of amphetamine.

Since the literature data do not provide enough information on the effects of amphetamine on the prefrontal cortex and since many controversial findings were reported in various rat strains we decided to compare adenylyl cyclase activity in the prefrontal cortex in various rat strains and test the effects of chronic amphetamine treatment (for 14 days) on the activity of this enzyme. Basal adenylyl cyclase activity was lower in Wistar rats than in Sprague-Dawley and Lewis rat strains. Amphetamine treatment produced in Wistar rats a substantial decrease in basal adenylyl cyclase activity. In Sprague-Dawley rats, we observed the highest enzyme activity which was slightly reduced after amphetamine treatment. In Lewis rats which had basal activity close to the activity of Wistar rats, amphetamine produced an increase in enzyme activity. The total adenylyl cyclase activity, estimated in the presence of forskolin, was the lowest in Wistar rats. The highest stimulation was observed in Lewis rats. Amphetamine treatment caused a very significant inhibition of total adenylyl activity in Wistar rats and a smaller inhibition in Sprague-Dawley rats. However, in Lewis rats amphetamine treatment increased the dose-response curve of forskolin stimulation. These results show that Lewis rats, compared to the other two strains, develop not only quantitatively but also qualitatively different responses.

Adenylyl Cyclases↗

Differences in heart phospholipids in two inbred rat strains differing in sensitivity to the development of heart lesions.

The content of phospholipids and their fatty acid composition were followed in the hearts of two inbred strains of rats: IR, resistant against the development of isoprenaline-induced myocardial lesions and IS, sensitive to their development. In the hearts of rats of the resistant strain, a lower content of phosphatidylcholine and its plasmalogen fraction was found compared to IS rats. The total amount of phospholipids was only insignificantly lower in IR rats. Greater differences were found in individual fatty acids. The most important finding concerned lower arachidonic acid and higher linoleic acid content in heart phospholipids of IR rats. These differences were exactly opposite to changes reported in the literature in animals known to have a higher resistance against myocardial damage due to various interventions. Our results do not support the hypothesis claiming the importance of changes in phospholipids and their FA composition for the resistance of the heart against the development of necrotic lesions.

Animals↗

The effects of adrenergic agonists on intraocular pressure and on adenylyl cyclase activity of ciliary processes in pigmented rabbits.

The effects of beta-adrenergic agonists isoprenaline, fenoterol and clenbuterol on the activity of adenylyl cyclase from ciliary processes and on intraocular pressure were examined in pigmented rabbits. Isoprenaline, fenoterol and clenbuterol stimulated adenylyl cyclase activity in vitro, but clenbuterol behaved as a partial agonist. Preincubation of ciliary processes with any of these three drugs led to the heterologous desensitization of adenylyl cyclase to the stimulatory effects of beta-adrenergic agonists or vasoactive intestinal peptide (VIP). This desensitization was dose-dependent and was expressed mainly as a decrease of the highest effects of stimulatory drugs. The exact mechanism of this phenomenon is not yet known. After topical administration, all three tested beta-adrenergic agonists decreased intraocular pressure with approximately the same intensity. The relationship between ocular hypotensive effects of beta-adrenergic agonists and their effects on adenylyl cyclase of ciliary processes is discussed. It is concluded that ocular hypotensive effects of adrenergic agonists and other drugs stimulating adenylyl cyclase cannot be explained simply by stimulation or desensitization of adenylyl cyclase of ciliary processes.

Adenylyl Cyclases↗

Repeated topical administration of fenoterol in rabbit reverses its initial ocular hypotensive effect and decreases sensitivity of adenylyl cyclase in ciliary processes to stimulatory agents.

PURPOSE: The effects of repeated topical administration of the selective beta 2-adrenergic agonist fenoterol on the intraocular pressure and on the adenylyl cyclase activity in ciliary processes in rabbit were examined in order to detect their possible causal relationship. METHODS: Intraocular pressure was measured by pneumatonometry. Adenylyl cyclase activity in homogenates of ciliary processes was assayed ex vivo by measurement of conversion of 32P-alpha-ATP to 32P-cyclic AMP. RESULTS: A single topical dose of 1% solution of fenoterol elicited a clear-cut decrease of the intraocular pressure lasting for several h. Repeated administration of fenoterol for 2-5 days led to a significant increase of intraocular pressure, observable from the second to the fifth day. The stimulation of adenylyl cyclase activity ex vivo by isoproterenol, vasoactive intestinal polypeptide or forskolin was significantly decreased on the fifth day (24 h after the administration of the last dose of fenoterol). CONCLUSIONS: Our data showed that repeated topical administration of the selective beta 2-adrenergic agonist increased intraocular pressure and desensitized adenylyl cyclase in ciliary processes; if these two effects are related then they would support the idea of direct relationship of decreased cAMP production in ciliary processes to the increase of intraocular pressure, and vice versa. However, conclusive evidence of this suggestion and of its possible significance in another animal species or man would require further study.

Adenylyl Cyclases↗

Effects of nitroprusside as a nitric oxide donor on anoxia/reoxygenation and D-galactosamine hepatic injuries: a study in perfused hepatocytes.

At present, the physiological role of NO. synthesis in the liver is ambiguous. Studies directed to reveal the role of NO. in relation to liver function were primarily initiated by an interest in the hepatic response to infections and the consequent modulation of liver function. The purpose of the present investigation was to use perfused rat hepatocytes to test the ability of the latter to produce NO. and to delineate the relationship between exogenously delivered NO. and any alteration in the degree of injury as produced by anoxia/reoxygenation (AR) or D-galactosamine (GalN, 5 mM) intoxication. NO. production in rats was stimulated by a single dose of lipopolysaccharide (LPS, 20 mg/kg i.p.) from which hepatocytes were isolated and perfused. Exogenous NO. was delivered to the perfusate of hepatocytes that were isolated from untreated rats, by the addition of sodium nitroprusside (SNP, 2 mM and 0.2 mM). AR and GalN hepatocyte injury was followed after the addition of SNP. Rat hepatocytes were immobilized in low-gelling agarose and perfused with Williams E medium. Endogenous synthesis of NO. and exogenous NO. as produced by SNP was evaluated by estimating the end products of NO. (NO2- + NO3-) in the perfusion medium. The functional and structural integrity of hepatocytes was evaluated from lactate dehydrogenase (LD) leakage and urea synthesis in the perfusion medium. Normal, AR- and GalN-injured hepatocytes did not exhibit measurable NO. while LPS-treated hepatocytes produced NO. (80 microM NO2- + NO3-). SNP-produced NO. significantly increased or decreased LD leakage in AR at 2 mM or 0.2 mM, respectively, and also reduced or increased the rate of urea synthesis, respectively. 0.2 mM SNP increased trypan blue exclusion by hepatocytes. On the other hand, GalN toxicity was not significantly altered by SNP as demonstrated by LD leakage and the rate of urea synthesis was increased by SNP addition. The present data suggest both deleterious and beneficial role of NO. in AR liver injury model depending on the level of NO. generated.

Animals↗

Effects of drugs acting on adrenergic and adenosine receptors on the intraocular pressure and the activity of adenylyl cyclase in ciliary processes and their sensitivity to pertussis toxin.

The effects of the selective alpha2-adrenergic agonist p-aminoclonidine, the nonselective adrenergic agonist epinephrine, the selective beta2-adrenergic agonist fenoterol and the adenosine A1 agonist R-PIA on intraocular pressure were studied in control and pertussis toxin-pretreated rabbits. Pretreatment of rabbits with pertussis toxin decreased the ocular hypotensive effects of p-aminoclonidine and epinephrine, did not influence the same effects of fenoterol or R-PIA and markedly potentiated the initial ocular hypertensive effects of epinephrine and R-PIA. As far as the action on adenylyl cyclase in ciliary processes is concerned, isoproterenol stimulated its activity in control rabbits and epinephrine exerted dual, i.e. stimulatory and inhibitory effects on the activity of this enzyme. The data obtained with epinephrine and p-aminoclonidine confirm the view that their ocular hypotensive effects are associated with their inhibitory action on adenylyl cyclase and contradict the opinion that the hypotensive action of adrenergic drugs depends on adenylyl cyclase activation.

Adenosine↗

Response of immobilized hepatocytes in a perfusion system to anoxia/reoxygenation: effect of cyclosporine A pretreatment.

The present study was designed to investigate the ameliorative effect of cyclosporine A (CsA) pretreatment on an anoxia/reoxygenation injury model by using immobilized perfused hepatocytes. Rats received an i.p. injection of two successive doses of CsA (5 mg/kg/day). Twenty-four hours later hepatocytes were isolated from CsA-treated and control rats. After hepatocyte isolation, immobilization, perfusion, induction of anoxia/reoxygenation, the structural and functional integrity of the hepatocytes was followed in a perfusion medium by measuring the leakage of lactate dehydrogenase (LD) and the time course of urea biosynthesis. CsA pretreatment reduced the initial rate of urea synthesis during normoxia but reduced the drop in the relative percentage rate of urea synthesis during the period of anoxia. LD leakage was increased threefold by anoxia and sevenfold by reoxygenation in cells of untreated animals. After CsA pretreatment in vivo, hepatocytes showed no increase in LD leakage into the medium. These findings demonstrate that the perfused immobilized hepatocytes can be used as a cellular model to assess the effects of liver insults such as anoxia/reoxygenation injury and that CsA modulates the injury. The mechanisms of CsA beneficial effects at the experimental level remain to be elucidated.

Animals↗