Recovery of instrumental conditioned reflexes in rats after pyramidotomy and action of bacterial melanin

Unilateral pyramidotomy at the bulbar level performed after preliminary elaboration of instrumental conditioned reflex (ICR) and without its elaboration caused hemiparesis of extremities in rats. Next day after this operation a half of the operated animals both with elaborated ICE and without its elaboration was injected intramuscularly with bacterial melanin solution (170 mg/kg). Comparison of terms of recovery of ICR and of a balancing movement of the paralyzed hindlimb in the operated rats without administration of melanin and with its injection showed that in the “melanin” animals the time of posttraumatic rehabilitation was much shorter and recovery of movement occurred faster and more completely. It has been found that the difference in terms of recovery of rat motor functions in this case is due to the protective action of bacterial melanin.     

Motor skill recovery in rats with various forelimb preference subjected to lesions in the caudate nucleus

Rats were trained for retrieving a sunflower seed from a tube using a forelimb. In the process of training, all the animals were divided in two groups: with the right- and left-limb preference. The period of learning was divided in two emotional (after the period of anxiety the animal takes a sunflower seed from the flow of a chamber) and three motor stages (different degrees of dexterity and precision in reaching a seed in the tube and grasping it). After a rat learned the skilled movement, the caudate nucleus was lesioned contralaterally to the preferred limb. After surgery, the animals were tested for recovery of the motor skill. The stages of recovery were the same as during learning. "Right-" and "left-handed" rats were compared in the percentage of animals performing each stage, duration of each stage, the depth of the tube reached by the animal's forelimb, and time required for obtaining the best results. The "right-handed" rats dominated in the percentage of animals performing the emotional stages and later began to recover the skilled movement (the crucial motor stage) but faster acquired their individual maximal depth of reaching a seed in the tube than the "left-handed" animals. The recovery of the skilled movement in the "left-handed" animals was a more gradual process than that in the "right-handed" ones.     

Different recovery of a skilled movement after destruction of the caudate nucleus in rats with right- or left-forelimb preference]

Recovery of the motor skill of taking a sunflower seed from a tube after the electrolytic destruction of the caudate nucleus was studied in Wistar rats with left- (n = 27) and right-forelimb (n = 22) preference. The reaching performance was divided in 5 stages. The first two stages characterize the emotional condition of an animal, the following three were immediately related with the skilled movement. After preliminary testing, the head of the caudate nucleus contralateral to the preferred limb was electrolytically damaged. After surgery, rats were tested during 20 weeks, and in every test the following indices were measured: percent of rats which perform the movement characteristic for the given stage, stage duration, depth of reaching a flower seed from the tube, time necessary to obtain the best results etc. It was found that at the "emotional" stages, higher percent of the right-handed animals perform the characteristic movements, but they spend more time to complete these stages than the left-handed rats. In rats with the left-limb dominance the recovery of the specialized movement is positively correlated with the time of the recovery onset at all the stages, while there is no such correlation in rats with right-limb dominance. Both groups of animals reach the same depth of the tube in the course of recovery, but the left-handed rats attain these results later and gradually, while the right-handed animals attain the maximal extent of recovery spontaneously and earlier than left-handed ones.     

Effect of adaptation to altitude hypoxia in early ontogeny on the indices of higher nervous activity]

Newborn male and female Wistar rats were adapted to hypoxia in a pressure chamber. Adaptation was started from the "altitude" of 1000 m, 1 hour daily, and then the time and the intensity of hypoxic actions were gradually increased: beginning from the 17th day the animals were subjected to adaptation to the "altitude" of 5000 m, for 5 hours, 5 days a week. The defence conditioned reflex of active avoidance was provoked in the animals after 2-month adaptation. A tendency to a more rapid elaboration of the reflex and a markedly enhanced degree of its retention in comparison with control was revealed in the adapted males. No significant changes in the elaboration and rentention of the reflex were found in the female animals adapted to hypoxia under analogous conditions.     

Influence of neurotensin on behavior of rats with lesions in serotonergic neurons

After serotonergic lesion by administration of 5,7-dihydroxytryptamine into the dorsalis raphe nucleus, effects of neurotensin microinjections into the caudate nucleus and substantia nigra on rat behavior were compared. Serotonergic lesions resulted in motivated excitement of rats manifested as an increase in the number of intersignal motor reactions during realization and, particularly, extinction of thirst conditioned reflex. Neurotensin microinjections into the caudate nucleus facilitated extinction of the conditioned reflex both in operated and control rats, but such microinjection into the substantia nigra facilitated this process only in operated animals. Neurotensin did not change conditioned reflex realization in both groups of animals but decreased emotional excitement of rats in the "open field". The behavioral effects of neurotensin in operated rats are connected with normalization of motivational and emotional states of animals and may be explained by recovery of interaction between the dopamine- and serotonergic systems. It is suggested that the mechanisms of this normalizing effects of neurotensin at the levels of the caudate nucleus and substantia nigra are different and are associated preferentially with its action either on dopamine- or serotonergic structures.     

Hair analysis for drugs of abuse XXI. Effect of para-substituents on benzene ring of methamphetamine on drug incorporation into rat hair

In order to study the effect of para-substituents on the benzene ring of methamphetamine on drug incorporation into hair from blood, the plasma AUCs and hair concentrations of 7 methamphetamines [methamphetamine(MA), p-hydroxymethamphetamine(OHMA), p-bromomethamphetamine (BMA), p-aminomethamphetamine (AMA), p-nitromethamphetamine (NMA), p-methoxymethamphetamine (MOMA) and 3,4-methylenedioxymethamphetamine (MDMA)] plus propylhexedrine(PHX) in DA rats was determined after intraperitoneal injection at 5 mg/kg, with single dose for the plasma AUC and 10 doses for the hair concentration. Drug incorporation rates into hair (ICRs) were calculated by dividing each hair concentration by each plasma AUC. Comparing the highest value (NMA) to the lowest one (OHMA), the ICR of NMA was 31.7 times larger than that of OHMA. Using the ICR of MA which has no substitute on the benzene ring as a base point, nitro, bromo, methylenedioxy, methoxy and amino groups raised the drug incorporation into rat hair in this order. On the other hand, hydroxy substitution showed a negative effect on the ICR. In comparison between the ICRs of MA and PHX, it was found that the benzene ring shows higher affinity to melanin and less lipophilicity than the cyclohexyl ring. Our results showed that there is a relatively strong effect of the functional groups on drug incorporation into hair. The combination of melanin affinity and lipophilicity are clearly correlated with their ICR.     

Piracetam correction of deficient GABA-ergic inhibition due to early postnatal exposure to cycloheximide

Cycloheximide, administered to 7-day-old rats, caused the delay of CNS activity changes in grown-up rats. The animals showed impaired memory function, expressed in the decrease of habituation in the "open field" test, and the alteration of passive avoidance reflex. The increased number of intersignal reactions due to hypermotility was found during elaboration of avoidance reflex. The analysis of evoked potential recovery revealed the deficiency of GABA-ergic inhibition in the neocortex. Piracetam was shown to prevent completely behavioural disturbances and deficiency of GABA-ergic processes in grown-up animals.     

Glyoxalase-1 overexpression partially prevents diabetes-induced impaired arteriogenesis in a rat hindlimb ligation model

We hypothesize that diabetes-induced impaired collateral formation after a hindlimb ligation in rats is in part caused by intracellular glycation and that overexpression of glyoxalase-I (GLO-I), i.e. the major detoxifying enzyme for advanced-glycation-endproduct (AGE) precursors, can prevent this. Wild-type and GLO-I transgenic rats with or without diabetes (induced by 55 mg/kg streptozotocin) were subjected to ligation of the right femoral artery. Laser Doppler perfusion imaging showed a significantly decreased blood perfusion recovery after 6 days in the diabetic animals compared with control animals, without any effect of Glo1 overexpression. In vivo time-of-flight magnetic resonance angiography at 7-Tesla showed a significant decrease in the number and volume of collaterals in the wild-type diabetic animals compared with the control animals. Glo1 overexpression partially prevented this decrease in the diabetic animals. Diabetes-induced impairment of arteriogenic adaptation can be partially rescued by overexpressing of GLO-I, indicating a role of AGEs in diabetes-induced impaired collateral formation.     

Bacterial overgrowth in the jejunum of ICR mice and Wistar rats orally administered with a single lethal dose of fusarenon-X, a trichothecene mycotoxin

A single oral dose of fusarenon-X (F-X), a trichothecene mycotoxin, resulted in abnormal microflora in the jejunum in ICR mice and Wistar rats with some differences in dose response between the species. In the acute phase, enterobacteria, streptococci, Clostridium perfringens and bacteroides showed remarkably increased counts in the jejunum of mice and rats dosed with F-X while lactobacilli showed a decrease in count. F-X brought an invasion of Pseudomonas aeruginosa into the livers, lungs, kidneys and spleens of ICR mice. Changes in the jejunal microflora appeared after 7 h in ICR mice and after 24 h in Wistar rats after a single oral dose of F-X of 7.5 and 4.0 mg/kg b.w., respectively, and the microflora returned to its normal state at 72 h in mice and 96 h in rats. The changes of intestinal microflora were followed by alterations in the growth curves of both animal species. The pH in the glandular stomach was also greatly enhanced before changes in the jejunal microflora. Acute F-X intoxication may be an involved manifestation of essential cytotoxicity of F-X mycotoxin alone and secondary bacterial overgrowth in the bowel.     

Bacterial overgrowth in the jejunum of ICR mice and Wistar rats orally administered with a single lethal dose of fusarenon-X, a trichothecene mycotoxin

A single oral dose of fusarenon-X (F-X), a trichothecene mycotoxin, resulted in abnormal microflora in the jejunum in ICR mice and Wistar rats with some differences in dose response between the species. In the acute phase, enterobacteria, streptococci, Clostridium perfringens and bacteroides showed remarkably increased counts in the jejunum of mice and rats dosed with F-X while lactobacilli showed a decrease in count. F-X brought an invasion of Pseudomonas aeruginosa into the livers, lungs, kidneys and spleens of ICR mice. Changes in the jejunal microflora appeared after 7 h in ICR mice and after 24 h in Wistar rats after a single oral dose of F-X of 7–5 and 4–0 mg/kg b.w., respectively, and the microflora returned to its normal state at 72 h in mice and 96 h in rats. The changes of intestinal microflora were followed by alterations in the growth curves of both animal species. The pH in the glandular stomach was also greatly enhanced before changes in the jejunal microflora. Acute F-X intoxication may be an involved manifestation of essential cytotoxicity of F-X mycotoxin alone and secondary bacterial overgrowth in the bowel.     

Effect of an analog of vasopressin on the fixation of temporary connections in the intact rat and after destruction of the locus coeruleus

Subcutaneous injection to intact rats and rats with destroyed locus coeruleus of 10 mcg of triglycide-glycinamide-lysinevasopressin (TGDGA-LVP) immediately after elaboration of labyrinth defensive conditioned reflex improves consolidation of temporary connections and raises its resistance to extinction. Improved consolidation of temporary connection is retained in animals with destroyed locus coeruleus for a shorter time than in intact animals. At the same time according to some characteristics, TGDGA-LVP has a more expressed influence of positive character on animals with destroyed locus coeruleus than on intact rats.     

The long-term effect of acute hypoxic hypoxia on shuttlebox avoidance learning in rats

Long-term influence was studied of the acute hypoxic hypoxia seance on rats behaviour in situation of elaboration of the conditioned reaction of active avoidance of electric shocks in shuttle chamber. It was found that in 2.5-3 months after the hypoxia seance, the experimental animals significantly differed from the intact controls by dynamics of CR elaboration (rats which had hypoxia were ahead of the control ones) and by distribution of the conditioned reactions latencies (for experimental animals this distribution was shifted to minor values). The character of these behavioural shifts coincided with that observed in the group of rats with local unilateral hippocampus lesion. The obtained results and numerous data presented in literature on the influence of the hippocampus lesion on animals shuttle avoidance learning, allow to conclude that the seance of hypoxic hypoxia leads to the disturbance of the hippocampus function. This conclusion conforms to the data on diffusive death of the hippocampal and neocortical neurones as a result of hypoxia action.     

Interhemispheric asymmetry of "emotional resonance" in the rat

Hemispheric asymmetry of "emotional resonance" elaborated by the method of P.V. Simonov was studied in Wistar rats. Inactivation of hemispheres was carried out by means of spreading depression. When using as "victims" and "recipients" the animals of the same sex, lateralization of emotional resonance was found to depend upon the velocity of reaction elaboration. In rats rapidly elaborating avoidance reaction the right hemisphere dominated during its performance and so did the left one in animals learning after additional training. When using the animals of different sex as "victims" and "recipients", the right hemisphere dominated in "emotional resonance" performance. Hemispheric asymmetry of "emotional" resonance was more expressed in males than in females.     

Norepinephrine turnover in the heart and blood vessels of rats subjected to bilateral renal infarction

The total norepinephrine (NE) content, the uptake of [3H]NE, the turnover rate and the synthesis rate of the neurotransmitter at the heart and blood vessels have been studied during the development of hypertension in rats subjected to bilateral renal infarction. Normal and sham-operated rats were used as controls. Fifty percent of the rats with renal infarction became hypertensive. The weight of the hearts and blood vessels of the experimental animals was significantly increased 15 days after renal infarction. Changes were greater in hypertensive animals. NE concentration in the heart was slightly decreased without achieving statistical significance, while total NE content was unchanged. In the artery wall NE concentration was significantly decreased in normotensive and hypertensive operated rats. [3H]NE uptake in the heart and blood vessels was similar in experimental and control animals. In relation to NE turnover, in both the heart and blood vessels, normal and sham-operated animals behaved as one population while normotensive and hypertensive rats behaved as another population. The rate constant of NE turnover was increased in both tissues of operated experimental animals without achieving statistical significance in the case of the heart. NE synthesis rate was unchanged in the cardiac muscle but was significantly increased in the blood vessels of operated animals. Present data indicate that results describing NE dynamics in the heart cannot be extrapolated for the blood vessels level; on the other hand changes in the neurotransmitter do not seem to be related to the development of high blood pressure after renal infarction in the rat.     

Strain differences in avoidance reactions of septal laboratory rats

The effect of a lesion of the dorsal septum on active and passive type of avoidance reactions of adult male Wistar (W) and Long-Evans (LE) rats was studied. The rate of acquisition and extinction of the reaction was studied by 3 different testing methods. The animals were operated on when juvenile (30 days) or adult (90 days). The experiments were always started 50 days after the operation, when the "rage syndrome" was no longer present. In the three different tests we found a single common variable--the strain factor. It was this that determined whether the operation was effective, as well as the direction of deviations and the operation age which led to manifest changes. In W males the lesion did not impair either the acquisition or the extinction of an active avoidance reaction (AAR); the passive avoidance reaction (PAR) was acquired and extinguished more slowly after a lesion in adulthood. The spontaneous passive avoidance reaction (i.e. preference of a small, dark space) was likewise negatively affected by the operation. In intact LE animals the AAR was extinguished more slowly than in intact W males; after a lesion produced in juvenile or adult age extinction was speeded up, so that there were no differences compared either with intact or with septal W animals. Intact LE rats also acquired a PAR more slowly than W rats; a septal lesion led to faster acquisition, irrespective of the age at which the operation was performed, so that the rate for septal LE rats were the same as for intact W animals. The extinction of this reaction took longer after an operation at juvenile age and the rate for these septal LE rats were the same as for those of septal W individuals operated on at 90 days. The spontaneous PAR was qualitatively poorer in intact LE animals than in intact W rats, since only 40% of them preferred the small, dark space. A lesion was followed by improvement, which was especially marked after operation on the 30th day, when all the animals preferred this space, and in a very short time, so that they were equal to intact W males. Correlation of the acquisition and the extinction rate showed that there was imbalance of excitation and inhibition processes in the AAR of the W controls, that they were balanced in the PAR and that the lesion reversed these relationships. Both processes were balanced in the LE control and a septal lesion did not alter the situation.     

Divergence of the Response Induced by Xenogenic Immunization in the Sepsis Survival of Rats

We have previously described that boosted natural xenoantibodies in rats cross-react to bacteria by targeting carbohydrate antigens. This type of immunization is associated with reduced survival after cecal ligation and puncture (CLP). In the present study, we investigated further this phenomenon by immunizing Lewis rats with three intraperitoneal injections, every other day, of hamster blood compared to saline-injected control animals. One day after the last injection, CLP was performed to produce a low-grade sepsis. Induction of xenoantibodies was associated with a reduction in animal survival after CLP relative to controls (45% vs. 90%, p<0.01). No bacterial blood load was observed after CLP in this model either with or without xenoantibody enhancement, indicating that the augmented mortality was not mediated by a direct effect of boosted xenoantibodies over blood bacteria. Nevertheless, the xenoimmunization produced a systemic inflammatory response in all rats. Additionally, a lack of weight gain at the time of CLP was present in animals that died after the procedure, which was not observed in surviving rats and controls. The cytokine profile at the time of CLP in animals that died after the procedure was characterized by an increase in the serum level of several cytokines, particularly adipokines. In contrast, the cytokine profile at CLP of xenoimmunized rats that survived the procedure was characterized by a reduction in the level of cytokines. In conclusion, this study failed to show a direct effect of boosted xenoantibodies over blood bacterial isolates as cause for the decreased survival after CLP. However, it evidenced that non-infectious systemic inflammation may lead to a pattern of augmented cytokines, particularly adipokines, which impairs survival after subsequent CLP. Therefore, the profile of cytokines existing before the infectious insult appears more crucial than that resulting from the condition for the outcome of sepsis.     

Albumin synthesis and catabolism following partial hepatectomy in the rat. The effects of amino acids and adrenocortical steroids on albumin synthesis after partial hepatectomy.

Plasma albumin levels were measured in partially hepatectomized, sham operated and control rats. The levels fell in both the partially hepatectomized and sham operated groups; while the latter group returned to normal within a few days, the low plasma albumin in the partially hepatectomized animals was sustained. Albumin synthesis rates in the isolated perfused rat liver were measured in the three groups of animals at varying intervals after partial hepatectomy. There was a significant depression of albumin synthesis rate in terms of both liver and whole animal weights when compared to the sham operated and control animals. This depression was almost completely reversed by the addition of arginine, asparagine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, threonine, tryptophan and valine added together to 10 times their normal plasma concentrations. The addition of hydrocortisone had no effect on the albumin synthesis rate after partial hepatectomy. Studies in vivo in the three groups of animals (partially hepatectomized, sham operated and control animals) revealed a fall in the albumin catabolic rate after partial hepatectomy coinciding with the fall in the albumin synthesis rate. An hypothesis whereby the amino acids may have their stimulatory effect is proposed.     

Melanin protects choroidal blood vessels against light toxicity

Low ocular pigmentation and high long-term exposure to bright light are believed to increase the risk of developing age-related macular degeneration (ARMD). To investigate the role of pigmentation during bright light exposure, cell damage in retinae and choroids of pigmented and non-pigmented rats were compared. Pigmented Long Evans (LE) rats and non-pigmented (albino) Wistar rats were exposed to high intensity visible light from a cold light source with 140,000 lux for 30 min. Control animals of both strains were not irradiated. The animals had their pupils dilated to prevent light absorbance by iris pigmentation. 22 h after irradiation, the rats were sacrificed and their eyes enucleated. Posterior segments, containing retina and choroid, were prepared for light and electron microscopy. Twenty different sections of specified and equal areas were examined in every eye. In albino rats severe retinal damage was observed after light exposure, rod outer segments (ROS) were shortened and the thickness of the outer nuclear layer (ONL) was significantly diminished. Choriocapillaris blood vessels were obstructed. In wide areas the retinal pigment epithelium (RPE) was absent in albino rats after irradiation. In contrast, LE rats presented much less cell damage in the RPE and retina after bright light exposure, although intra-individual differences were observed. The thickness of the ONL was almost unchanged compared to controls. ROS were shortened in LE rats, but the effect was considerably less than that seen in the albinos. Only minimal changes were found in choroidal blood vessels of pigmented rats. The RPE showed certain toxic damage, but cells were not destroyed as in the non-pigmented animals. The number of melanin granules in the RPE of LE rats was reduced after irradiation. Ocular melanin protects the retina and choroid of pigmented eyes against light-induced cell toxicity. Physical protection of iris melanin, as possible in eyes with non-dilated pupils, does not seem to play a major role in our setup. Biochemical mechanisms, like reducing oxidative intracellular stress, are more likely to be responsible for melanin-related light protection in eyes with dilated lens aperture.     

Cardiopulmonary bypass and long-term neurocognitive dysfunction in the rat

Neurologic and neurocognitive complications after cardiac surgery with cardiopulmonary bypass (CPB) have been reported repeatedly. To better understand its etiology and design protective strategies, an appropriate animal model may prove useful. Although impaired short-term neurocognitive function has been recently demonstrated after CPB in rats, the demonstration of persistent long-term neurocognitive changes would be more relevant from a clinical perspective. We hypothesized that CPB results in long-term impairment of neurocognitive performance in rats. Male rats were exposed to either 60 min of normothermic non-pulsatile CPB, using a roller-pump and a neonatal membrane oxygenator, or to cannulation only (sham animals). Long-term neurocognitive function was assessed at 4 to 7 weeks after CPB (Can test), and again after 12 weeks (Morris water maze) in both operated groups and in a non-operated control group, followed by histologic evaluation of the hippocampus. In separate groups of CPB and sham animals, we also measured TNF-alpha and IL-6 in plasma. There were no significant differences in long-term neurocognitive performance or histological outcome between the three groups. Cytokine patterns were also similar in both operated groups. We conclude that CPB did not appear to cause long-term neurocognitive dysfunction in this model of CPB in young healthy rats. The lack of long-term deficits may be due to the absence of clinically important etiologic factors such as atheromatous and gaseous embolization in this model. Similar cytokine patterns in both operated groups suggest that surgical trauma rather than exposure of blood to extra-corporeal circuit was probably responsible for the inflammatory response.     

Testing the efficacy of antimicrobial peptides in the topical treatment of induced osteomyelitis in rats

Joint replacement infections and osteomyelitis are among the most serious complications in orthopaedics and traumatology. The risk factors for these infections are often bacterial resistance to antimicrobials. One of the few solutions available to control bacterial resistance involves antimicrobials, which have a different mechanism of action from traditional antibiotics. Antimicrobial peptides (AMP) appear to be highly promising candidates in the treatment of resistant infections. We have identified several AMP in the venom of various wild bees and designed analogues that show potent antimicrobial activity and low toxicity against eukaryotic cells. The aim of the present study was to test the efficacy of one of those synthetic peptide analogues for the treatment of acute osteomyelitis invoked in laboratory rats. Femoral cavities of 20 laboratory Wistar rats were infected with Staphylococcus aureus. After 1 week, eight rats received an injectable calcium phosphate carrier alone, another eight rats were treated with a calcium phosphate mixed with AMP, and four rats were left without any further treatment. After another week, all rats were euthanized and radiographs were made of both the operated and healthy limbs. The animals with the carrier alone exhibited more severe acute osteomyelitis on radiographs in comparison to the recipients of the calcium phosphate carrier loaded AMP and untreated infected individuals. Based on the results of the above mentioned experiment, it was concluded that when injected directly into the site of femoral acute osteomyelitis, the calcium phosphate carrier mixed with AMP reduced osteomyelitis signs visible on radiographs.