Justyna Rogalska

Perinatal asphyxia, hyperthermia and hyperferremia as factors inducing behavioural disturbances in adulthood : A rat model

Alertness was studied in adult male Wistar rats after neonatal critical anoxia applied under three different thermal conditions: (i) at physiological neonatal body temperature of 33 degrees C, (ii) at body temperature elevated to 37 degrees C, and (iii) at body temperature elevated to 39 degrees C (both during anoxia and for 2 h postanoxia). To elucidate the effect of iron-dependent postanoxic oxidative damage to the brain, half of the group (iii) was injected with deferoxamine, a chelator of iron. Postanoxic behavioural disturbances were recorded in open-field, elevated plus-maze, and sudden silence tests when the rats reached the age of 4 month. Moreover, spontaneous motor activity of the rats was recorded radiotelemetrically in their home-cages. Both open-field stress-induced and spontaneous motor activity were reduced in rats subjected to neonatal anoxia under hyperthermic conditions. In contrast, these rats were hyperactive in the plus-maze test. Both the plus-maze and sudden silence tests revealed that these rats show reduced alertness to external stimuli signalling potential dangers. The behavioural disturbances were prevented by the body temperature of 33 degrees C and by postanoxic administration of deferoxamine. These data support the conclusion that permanent postanoxic behavioural disturbances are due to iron-dependent oxidative damage to the brain, which can be prevented by the reduced neonatal body temperature.

Effect of temperature on postanoxic, potentially neurotoxic changes of plasma pH and free iron level in newborn rats.

In asphyxiated newborns, iron, released from heme and ferritin and deposited in the brain, contributes to neurodegeneration. Because hypothermia provides neuroprotection, newborn mammals, showing reduced body temperature, might avoid iron-mediated neurotoxicity. However, hypothermia leads to acidosis, which induces hyperferremia. Therefore, we decided to study the effects of body temperature on plasma pH and iron levels in newborn rats exposed to a critical anoxia. Rectal temperature was kept at 33 degrees C (typical of neonates), reduced by 2 degrees C, or elevated to a level typical of healthy (37 degrees C) or febrile (39 degrees C) adults. Arterial blood samples were collected at 0, 10, 20, 30, and 120 min postanoxia. Control samples were obtained from normoxic, temperature-matched neonates. Anoxia tolerance time decreased progressively at rectal temperatures exceeding 33 degrees C. Neither pH nor plasma iron were significantly affected by anoxia at 33 degrees C. Although hypothermia (31 degrees C) resulted in acidosis in normoxic rats, both pH and iron levels were hardly influenced by anoxia. However, acidosis and hyperferremia, proportional to body temperature, developed at 37 and 39 degrees C. In conclusion, reduced body temperature is likely to protect asphyxiated newborns against iron-mediated brain injury.

Effect of neonatal body temperature on postanoxic, potentially neurotoxic iron accumulation in the rat brain

In asphyxiated newborns iron, released from heme and ferritin and deposited in the brain, contributes to neurodegeneration. Because hypothermia provides neuroprotection, newborn mammals, showing spontaneously reduced body temperature, might avoid the iron-mediated neurotoxicity. Therefore, we decided to study the effects of body temperature and chelation of iron with deferoxamine on iron accumulation in the brain of three weeks old rats exposed neonatally to a critical anoxia. At the age of two days, newborn rats were exposed to anoxia in 100% nitrogen atmosphere. Rectal temperature was kept at 33 degrees C (typical of the rat neonates), or elevated to a level typical of febrile (39 degrees C) adults. Control rats were exposed to atmospheric air in the respective thermal conditions. Half of the rats exposed to anoxia under hyperthermic conditions were injected with deferoxamine (DF), immediately after anoxia and 24 h later. Regional changes in cerebral iron deposition were examined in the frontal cortex, the hippocampus and the striatum, using iron histochemistry, when the rats reached the age of three weeks. Increased iron staining was found in neurons of each of the three cerebral regions in rats exposed to neonatal anoxia under hyperthermic conditions and the iron accumulation was prevented by postanoxic DF injection. In conclusion, febrile body temperature amplifies cerebral hyperferremia, which might induce neurodegenerative disturbances in the brain. On the other hand, a protection against the brain hyperferremia can be achieved by both the reduced physiological neonatal body temperature and by postasphyxic DF administration.

Stress-induced behaviour in juvenile rats: effects of neonatal asphyxia, body temperature and chelation of iron.

Newborn mammals, showing reduced normal body temperature, might be protected against iron-mediated, delayed neurotoxicity of perinatal asphyxia. Therefore, we investigated the effects of (1) neonatal body temperature and neonatal critical anoxia as well as (2) postanoxic chelation of iron with deferoxamine, on open-field stress-induced behaviour in juvenile rats. The third aim of this study was to compare (after the above-mentioned treatments) circadian changes in spontaneous motor activity and body temperature in juvenile rats permanently protected from any stress. Neonatal anoxia at body temperature adjusted (both during anoxia and 2 h reoxygenation) to a level typical of healthy (37 degrees C) or febrile (39 degrees C) adults led to the stress-induced hyperactivity in juvenile (5-45 days old) rats. Both normal neonatal body temperature of 33 degrees C and chelation of iron prevented the hyperactivity in rats. Neither neonatal body temperature nor neonatal anoxia affected spontaneous motor activity or body temperature of juvenile rats, recorded in their home-cages with implantable transmitters. Circadian rhythmicity was also undisturbed. Presented data support the hypothesis that physiologically reduced neonatal body temperature can provide a protection against iron-mediated postanoxic disturbances of behavioural stress responses in juvenile rats.

Effect of winter torpor upon antioxidative defence in Helix pomatia

Arousal of land snails from torpor is inseparably connected with an increase in oxygen consumption leading to oxidative stress. Therefore, activity of antioxidant defence system (antioxidant enzymes and reduced glutathione) and degree of oxidative damage (concentration of malondialdehyde as an index of lipid peroxidation) in the snail Helix pomatia L., 1758 were tested to check whether torpid snails are able to activate their antioxidative defence against oxidative damage prior to arousal from winter torpor. Snails, which were collected from their natural habitats, were tested at the beginning, in the middle part, and at the end of winter torpor. Active snails collected in autumn and spring were taken as control groups. Snails were immediately killed and their foot, hepatopancreas, and kidney were used for the biochemical assays. Winter torpor induced significant changes in activities of the crucial antioxidant substances. The lowest activities were observed at the beginning of torpor, whereas activity of...

Defence against oxidative stress in two species of land snails (Helix pomatia and Helix aspersa) subjected to estivation.

During summer, land snails are exposed to estivation/arousal cycles that imposes oxidative stress, but they exhibit different patterns of antioxidant defence. To test the ability of two related species, Helix pomatia and Helix aspersa, to modulate their antioxidant defence mechanism during estivation/arousal cycles, we examined activities of catalase and glutathione-related enzymes and concentrations of glutathione and thiobarbituric acid reactive substances (TBARS; as products of lipid peroxidation). In both species, estivation evoked changes in activity of total and selenium-dependent glutathione peroxidase (GPx), but did not affect activity of catalase, glutathione reductase, and glutathione transferase, and had no effect on concentration of glutathione. Activity of catalase in estivating snails, instead of the expected increase, showed a tendency to diminish. Extremely low activities of catalase in the foot were usually associated with extremely high activities of both forms of GPx. In conclusion, maintenance of relatively high activities of the antioxidant enzymes and accumulation of glutathione, resulting in a low and stable concentration of TBARS, plays an important role in scavenging oxygen free radicals from the organism of both species.

Molecular Targets for Components of Essential Oils in the Insect Nervous System—A Review

Essential oils (EOs) are lipophilic secondary metabolites obtained from plants; terpenoids represent the main components of them. A lot of studies showed neurotoxic actions of EOs. In insects, they cause paralysis followed by death. This feature let us consider components of EOs as potential bioinsecticides. The inhibition of acetylcholinesterase (AChE) is the one of the most investigated mechanisms of action in EOs. However, EOs are rather weak inhibitors of AChE. Another proposed mechanism of EO action is a positive allosteric modulation of GABA receptors (GABArs). There are several papers that prove the potentiation of GABA effect on mammalian receptors induced by EOs. In contrast, there is lack of any data concerning the binding of EO components in insects GABArs. In insects, EOs act also via the octopaminergic system. Available data show that EOs can increase the level of both cAMP and calcium in nervous cells. Moreover, some EO components compete with octopamine in binding to its receptor. Electrophysiological experiments performed on Periplaneta americana have shown similarity in the action of EO components and octopamine. This suggests that EOs can modify neuron activity by octopamine receptors. A multitude of potential targets in the insect nervous system makes EO components interesting candidates for bio-insecticides.

Deferoxamine prevents cerebral glutathione and vitamin E depletions in asphyxiated neonatal rats: role of body temperature

Abstract Hypoxic-ischaemic brain injury involves increased oxidative stress. In asphyxiated newborns iron deposited in the brain catalyses formation of reactive oxygen species. Glutathione (GSH) and vitamin E are key factors protecting cells against such agents. Our previous investigation has demonstrated that newborn rats, showing physiological low body temperature as well as their hyperthermic counterparts injected with deferoxamine (DF) are protected against iron-mediated, delayed neurotoxicity of perinatal asphyxia. Therefore, we decided to study the effects of body temperature and DF on the antioxidant status of the brain in rats exposed neonatally to critical anoxia. Two-day-old newborn rats were exposed to anoxia in 100% nitrogen atmosphere for 10 min. Rectal temperature was kept at 33 °C (physiological to rat neonates), or elevated to the level typical of healthy adult rats (37 °C), or of febrile adult rats (39 °C). Half of the rats exposed to anoxia under extremely hyperthermic conditions (39 °C) were injected with DF. Cerebral concentrations of malondialdehyde (MDA, lipid peroxidation marker) and the levels of GSH and vitamin E were determined post-mortem, (1) immediately after anoxia, (2) 3 days, (3) 7 days, and (4) 2 weeks after anoxia. There were no post-anoxic changes in MDA, GSH and vitamin E concentrations in newborn rats kept at body temperature of 33 °C. In contrast, perinatal anoxia at elevated body temperatures intensified oxidative stress and depleted the antioxidant pool in a temperature-dependent manner. Both the depletion of antioxidants and lipid peroxidation were prevented by post-anoxic DF injection. The data support the idea that hyperthermia may extend perinatal anoxia-induced brain lesions.

Neonatal asphyxia under hyperthermic conditions alters HPA axis function in juvenile rats.

Neonatal anoxia is an example of early-life threatening experience that might exert long-lasting behavioral disturbance. One of the consequences of neonatal asphyxia is hyperactivity in open-field test. Changes in open-field activity are coupled with changes in the function of the hypothalamic-pituitary-adrenal (HPA) axis. A critical determinant of the severity of hypoxic-ischemic brain injury in newborn rats is body temperature. Hyperthermia under anoxic conditions increases locomotor activity in the open-field test. Therefore, the aim of the present study was to test whether body temperature during neonatal anoxia can affect basal and stress-induced corticosterone secretion in rats. At the age of 2 days Wistar rat pups were exposed to anoxia in 100% nitrogen atmosphere. Rectal temperature was kept at 33 degrees C (typical for the rat pups), or was elevated to a level typical for febrile adults (39 degrees C). Control rats were exposed to atmospheric air under the respective thermal conditions. Basal and stress-induced corticosterone levels were assessed using sulphuric acid-induced fluorescence, on postnatal day 14. Body temperature during neonatal asphyxia altered the early developmental profile of plasma corticosterone. Hyperthermia under anoxic conditions decreased the corticosterone response to open-field stress. In conclusion, febrile body temperature changes corticosterone release, which might induce neurobehavioral disturbances. On the other hand, a protection against the HPA dysfunction can be achieved by the reduced physiological neonatal body temperature.

Deferoxamine improves antioxidative protection in the brain of neonatal rats: The role of anoxia and body temperature

After hypoxic-ischemic insult iron deposited in the brain catalyzes formation of reactive oxygen species. Newborn rats, showing reduced physiological body temperature and their hyperthermic counterparts injected with deferoxamine (DF), a chelator of iron, are protected both against iron-mediated neurotoxicity and against depletion of low-molecular antioxidants after perinatal asphyxia. Therefore, we decided to study the effects of DF on activity of antioxidant enzymes (superoxide dismutase-SOD, glutathione peroxidase-GPx and catalase-CAT) in the brain of rats exposed neonatally to a critical anoxia at body temperatures elevated to 39°C. Perinatal anoxia under hyperthermic conditions intensified oxidative stress and depleted the pool of antioxidant enzymes. Both the depletion of antioxidants and lipid peroxidation were prevented by post-anoxic DF injection. The present paper evidenced that deferoxamine may act by recovering of SOD, GPx and CAT activity to reduce anoxia-induced oxidative stress.