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INTRODUCTION: The neuronal cell death associated with perinatal anoxia plays a significant role in neonatal morbidity and neurodevelopmental disability. In response to mild stress, a number of compensatory mechanisms are activated, which allows the cells to survive. Beside decreased body temperature, brain-derived neurotrophic factor (BDNF) is also considered to be beneficial to neuronal survival. AIM(S): Therefore, the aim of this study was to determine whether body temperature under anoxic condition affects the ability of BDNF (proBDNF and mBDNF) to decrease caspase‑3 levels in the developing brain. METHOD(S): 2-day-old Wistar rats were divided into 3 temperature groups: i) normothermic ‑33°C (typical body temperature of newborn rats), ii) hyperthermic – 37°C (typical body temperature of adult rat), and iii) extremely hyperthermic – 39°C (typical body temperature of febrile adult rats). The temperature was controlled starting 15 minutes before, and the measurement was continued during 10 minutes of anoxia (pure nitrogen atmosphere), as well as, for 2 hours postanoxia. Levels of BDNF and caspase-3 were determined post mortem, 2 and 72 hours after anoxia using Western blot and ELISA analysis. RESULTS: Body temperature affected the levels of endogenous BDNF, its precursor form (proBDNF), and caspase‑3. In anoxic animals, the levels of proBDNF and caspase-3 increased with increasing neonatal body temperature. In contrast, a significant negative correlation between the total BDNF to proBDNF ratio, and caspase-3 concentrations was observed. CONCLUSIONS: The results suggest that decreased body temperature can not only up-regulate BDNF levels, but also may affect the other functions of this neuropeptide. FINANCIAL SUPPORT: Research on this paper was supported by grant from National Science Centre, Poland, no. 016/21/N/NZ7/00399.
BACKGROUND AND AIMS: Complications after neonatal asphyxia are the most common cause of subsequent neurological disorders. The mechanism involved in brain damage is closely associated with abnormal iron metabolism that is a cofactor of free-radical reactions. There is a number of evidence that one of the endogenous processes that protect the brain from damage due to perinatal hypoxia is decreasing of body temperature. It is also known, that the transcriptional hypoxia-inducible factor1α (HIF-1α) plays the fundamental role in adaptive process in response to hypoxia. HIF-1α upregulates several genes involved in glycolysis, erythropoiesis, and angiogenesis to promote survival. Our experiments aimed at checking the effects of body temperature during simulated perinatal anoxia on the subsequent changes of  HIF-1α expression in brain. Considering the key role of iron as a cofactor of free radical reactions and it’s contribution in proteasomal degradation of α subunit of HIF protein, the second goal of the project was to verify the influence of deferoxamine (iron chelating agent) on the level of expression of HIF-1α in a variety of thermal conditions. METHODS: Two-day-old Wistar rats were divided into 4 temperature groups: (1) hypothermic (31°C), (2) normothermic (33°C), (3) hyperthermic typical to adult rats (37°C) and (4) hyperthermic typical to febrile adults rats (39°C). Within each group, infants were divided into two subgroups: animals with saline injection and animals with deferoxamine injection. The temperature was controlled starting 15 minutes before, and continuing during 10 minutes of anoxia (pure nitrogen atmosphere) as well as for 2 hours postanoxia. Levels of HIF-1α gene expression were analyzed post mortem: immediately, 3 and 7 days after anoxia using Western blot analysis. RESULTS: The results showed, that the body temperature during neonatal anoxia affects the level of HIF-1α expression. Moreover, the use of deferoxamine increases the expression of this gene.
One of the variables oscillating seasonally in Helix pomatia L., described in our previous paper, was their rudimentary cryoprotection provided by modest increases in haemolymph glucose and glycerol concentrations in autumn and early spring, respectively. Because factors governing cryoprotective responses are unknown, we decided to investigate the effects of spring and autumn acclimation of H. pomatia to different thermal and photoperiod conditions on the changes in haemolymph concentrations of glucose and glycerol and on the glycogen level in selected organs. Neither acclimation to short-day photoperiod nor low ambient temperature evoked increase in glucose and glycerol concentration in spring and autumn. Both acclimation variants decreased hepatopancreatic glycogen level. The rudimentary freeze-tolerance in H. pomatia seems to be a combined effect of cold and short-day photoperiod and might also be affected by their nutritional and reproductive status. The effect of exposure to frost-bite is also likely to be involved.
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Seasonal changes in cryoprotectants concentrations in Helix pomatia snails

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Terrestrial snails are often exposed to freezing. Therefore, we investigated seasonal shifts in hemolymph concentrations of cryoprotectants such as glycerol and glucose. We also investigated whether summer acclimation to cold and short-day photoperiod induced synthesis of cryoprotectants in Helix pomatia snails. Concentrations of the both cryoprotectants were elevated in winter and reduced in summer. These changes, however, were not correlated with shifts in liver glycogen content. Summer acclimation to cold (5°C) and short-day photoperiod evoked a selective increase in glycerol concentration. In conclusion, glycerol may play a role in adaptation of the snails to winter cold and glucose is rather unlikely to provide the cryoprotection.
Perinatal asphyxia in mammals leads to iron accumulation in the brain, which results in delayed neurobehavioural disturbances, including impaired learning and abnormal alertness over their entire life span. The aim of this investigation was to verify our hypothesis that newborn rats, showing reduced normal body temperature, are protected against neurotoxicity of the asphyxia up to senescence. Alertness was studied in adult and old male Wistar rats after exposure to critical neonatal anoxia: (i) at physiological neonatal body temperature of 33°C, (ii) at body temperature elevated to 37°C, or (iii) at body temperature elevated to 39°C (the thermal conditions remained unchanged 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 12 and 24 months. Open-field stress-induced motor activity was 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 show reduced alertness of these rats to external stimuli signalling potential dangers. The behavioural disturbances were prevented by body temperature of 33°C and by administration of deferoxamine.
Objective: To study the levels of malonyldialdehyde, marked as the concentration of substances reacting with thiobarbituric acid in liver, brain, serum and kidney in subacute intoxication with low doses of chlorfenvinphos, an organophosphate insecticide. Materials and Methods: The study used male Wistar rats, body weight 250 – 280g. The animals received intragastrically, by a gastric tube, once daily 0.1ml/100g of olive oil (the control group) or oil solution of chlorfenvinphos at a dose of 0.02LD50 (0.3 mg/kg b.w.) (the experimental groups). After 14 and 28 days of the experiment, blood samples were collected by cardiac puncture to obtain serum; liver, kidney and brain sections were taken from the animals in anaesthesia. The level of malonyldialdehyde (as TBA RS) was determined in homogenates of the organs and in serum. Additionally, serum cholinesterase activity was determined. Results: There was an increase in the tissues as well in the serum malonyldialdehyde level. Proportionally the highest increase in TBARS , as compared to the control, was observed in the liver on day 28 of chlorfenvinphos administration and in the kidney on day 14 of the exposure. Conclusion: In subacute chlorfenvinphos intoxication, lipid peroxidation is increased in the rat liver, serum, kidney and brain, which may cause various health effects in the population exposed to its action.
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