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1

Influence of isopropyl methylphosphonofluoridate intoxication on cytokine levels in chosen brain structures

100%
Kowalczyk M., Zdanowski R., Antkowiak B., Paluch M.
Acta Neurobiologiae Experimentalis
|
2009
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tom 69
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nr 3
Poisoning with organophosphate compounds induces pronounced neurotransmitter disturbances in central nervous system. There are some data indicating that these poisonings initiate changes in neuro-immune response. Our study was aimed at the evaluation of cytokines levels (interleukin 1β – IL-1β, interleukin 2 – IL-2, tumor necrosis factor α – TNF-α and interferon γ – IFNγ) after isopropyl methylphosphonofl uoridate (IMPF) intoxication in chosen brain structures of BALB/c mice. Tests were performed in supernatants of frontal cortex, hippocampus and brain stem homogenates using immunoenzymatic assay after 24 h, 7, 14 and 21 days of IMPF intoxication (1/2 LD50, s.c.). IMPF induced differential changes in cytokines levels (except IL-2) in the examined brain structures. The signifi cant increase of TNF-α and IL-1β levels was shown in frontal cortex after 14 days of poisoning. Raised level of TNF-α in this structure was observed until the end of the experiment (21st day). We also observed pronounced increase of IFNg level after 7 days in hippocampus and after 7 and 14 days in brain stem. Concurrently the level of IL-1β in these structures was signifi cantly decreased, in hippocampus in all measurements, while in the brain stem only after 24 hours following IMPF application.Our results indicate that IMPF intoxication without inducing convulsions can produce marked changes in brain cytokines levels that may lead to immune response disturbances.
2

Changes in GnRH and GnRH receptor biosynthesis in the hypothalamic-pituitary unit of anestrous ewes after infusion of GnRH into the third cerebral ventricle

76%
Lapot M., Przekop F., Paruszewska E., Antkowiak B., Malewski T., Ciechanowska M.
Acta Neurobiologiae Experimentalis
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2017
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tom 77
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nr Suppl.1
INTRODUCTION: Physiological regulation of GnRH secretion in mammals is associated with complex interplay between excitatory and inhibitory neurotransmitter and neurohormone systems within the hypothalamus that mediate oestrogen signals to GnRH cells. Among numerous studies on the neuroendocrine processes controlling GnRH secretion only a few research have revealed that hypothalamic GnRH acting locally may also participate in the regulation of its own release, but the mechanisms involved remain poorly understood. AIM(S): This study was designed to evaluate the effect of prolonged intermittent infusion of small doses of exogenous GnRH on GnRH and GnRH receptor (GnRHR) biosynthesis in the hypothalamus, GnRHR expression in the anterior pituitary gland (AP), and on LH secretion in sexually inactive sheep. METHOD(S): Studies were conducted on 3–4-year-old Polish Merino ewes during the middle of the anestrous season. Infusions of exogenous GnRH (0.2 µg GnRH per animal daily) were performed into the third cerebral ventricle for 3 consecutive days. The control for GnRH-treated group was animals infused with equivalent to GnRH volume of Ringer’s solution. The measurement of GnRH and GnRHR levels was performed using an enzyme-linked immunosorbent assay (ELISA). Plasma LH concentration was analysed by a double-antibody radioimmunoassay (RIA). RESULTS: The results of this study demonstrate that prolonged infusion of small doses of GnRH into the third cerebral ventricle of anestrous ewes increased drastically GnRH and GnRHR levels in the hypothalamus, but decreased GnRHR expression in the AP and also reduced LH secretion. CONCLUSIONS: The study indicates the existence of ultrashort loop feedback mechanism of GnRH release from the hypothalamus in which hypothalamic GnRHR participate. Decreased expression of GnRH in the AP and diminished LH secretion in GnRH-treated ewes provide indirect evidence for suppressive effect of exogenous GnRH on GnRH release from hypothalamic nerve terminals. FINANCIAL SUPPORT: This work was supported by grant National Science Center Poland No UMO-2012/05/B/ NZ4/02443.
3

The influence of dapaminergic system inhibition on kisspeptin and rfamide-related peptide-3 transcripts and gonadotropin releasing-hormone/receptor proteins in anestrous ewes

76%
Lapot M., Ciechanowska M., Kowalczyk M., Mateusiak K., Paluch M., Antkowiak B., Przekop F.
Acta Neurobiologiae Experimentalis
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2015
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tom 75
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nr Supl.
BACKGROUND AND AIMS: The aim of this study was to explain how prolonged inhibition of central dopaminergic activity affected the neuroendocrine processes controlling GnRH/gonadotropins secretion in the hypothalamus and in the anterior pituitary gland compartments of sheep during the non-breeding time. In this purpose, we investigated the effects of prolonged, intermittent infusions of small doses of sulpiride (dopaminergic D2 receptors antagonist) on transcript levels of the two central regulators of the hypothalamic-pituitary reproductive system – kiss 1 and RFRP-3. It was also examined the influence of sulpiride on the GnRH and GnRH receptor (GnRHR) biosynthesis in the hypothalamus-pituitary unit. Additionally, these analyses were complemented by the study of plasma LH concentration. METHODS: The levels of kiss 1 and RFRP-3 transcripts and GnRH and GnRHR proteins were analyzed using Real-time PCR technique or an enzyme-linked immunoabsorbent assay (ELISA), respectively. Plasma LH concentration was measured by a doubleantibody radioimmunoassay. RESULTS: Pharmacological blockade of D2 receptors by sulpiride significantly increased kiss 1 mRNA levels in the preoptic area and in the arcuate nucleus/ventromedial hypothalamus and decreased the RFRP-3 mRNA in the paraventricular nucleus. The abolition of dopaminergic neurons activity also resulted in augmentation of the GnRH and GnRHR protein levels in the entire hypothalamus and led to similar changes in the expression of GnRHR in the anterior pituitary gland. The increase of kiss 1 transcription and GnRH biosynthesis has appeared with a concomitant increase of LH secretion. CONCLUSIONS: Similar direction of changes of kiss 1 transcript and GnRH protein levels with LH secretion strongly suggest that the inhibition of D2 receptors has a stimulatory effect on kiss1/ GnRH biosynthesis and consequently on LH release. It is likely, that dopamine inhibits gonadotropin synthesis and release in anestrous period through complex actions of kiss1/RFRP-3 neurons on GnRH cells.
4

Mechanisms of central neural regulation of GnRH/LH secretion under short and prolonged stress in the hypothalamic-pituitary unit of ewes in different stages of reproduction

76%
Ciechanowska M., Lapot M., Antkowiak B., Mateusiak K., Paruszewska E., Paluch M., Przekop F.
Acta Neurobiologiae Experimentalis
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2015
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tom 75
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nr Supl.
AIM: The aim of this study was (1) to analyze the effects of short and prolonged stress (footshock stimulation) on the biosynthesis of GnRH and GnRHR proteins in the hypothalamus-pituitary region and on luteinizing hormone (LH) secretion in anestrous and follicular phase ewes (2) to determine whether applied models of physical stress influence mRNA expression of kisspeptin (kiss1) and RFamide-related peptide-3 (RFRP-3), two central regulators of the mammalian reproductive axis. METHODS: The levels of the GnRH and GnRHR proteins were analyzed in selected tissue of the hypothalamus-anterior pituitary unit using an enzyme-linked immunoabsorbent assay (ELISA). Plasma LH concentration was measured by a double-antibody radioimmunoassay. To determine the transcript levels of kiss 1 and RFRP-3 in the preoptic area-hypothalamus region, Real-time PCR with SYBR Green dye was applied. RESULTS: Stress changed drastically the biosynthesis of GnRH and GnRHR, as well as the transcriptional activities of genes encoding kiss 1 and RFRP3 neuropeptides. The pattern of these changes was dependent upon physiological state of animal and on the time course of stressor application. The fluctuations of GnRH and GnRH-R protein levels under short or prolonged stress stimuli were associated with similar changes in LH secretion, thus suggesting the existence of a direct relationship between GnRH and GnRH-R biosynthesis and GnRH/LH release. CONCLUSIONS: The results indicate that disturbances of gonadotropin secretion under stress condition in sheep may be due to dysfunction of the hypothalamus/pituitary GnRH/GnRHR system. It can not be excluded that interaction between kiss 1 and RFRP3 neuronal networks with GnRH cells may also play a critical role in the transduction of stress-induced changes in the activity of hypothalamic-pituitary-gonadal axis. However, a detailed explanation of this phenomenon requires further research.
5

The influence of soman on the expression of tp53 gene in the rat brain; distant effect

76%
Brytan M., Kowalczyk M., Lapot M., Paruszewska E., Antkowiak B., Przekop F., Ciechanowska M.
Acta Neurobiologiae Experimentalis
|
2017
|
tom 77
|
nr Suppl.1
INTRODUCTION: The organophosphorus compound soman (GD), an irreversible inhibitor of cholinesterases, produces seizure activity and related brain damage. Studies using various biochemical markers of programmed cell death indicate apoptotic rather than necrotic mechanism of GD-induced acute cell damage in the brain. One of the most important links between the proliferation and cell death machinery is the tumor suppressor p53, which as a guardian of the genome and the element promoting apoptosis makes it a prime target for a prognostic factor. AIM(S): The aim of this study was to examine distant effects of poisoning with a small, repeated dose of GD on the expression of mRNA encoding p53 protein in the rat brain. METHOD(S): The study was performed on maternal generation (F0) and on first filial generation (F1) of Wistar rats. Low clinically asymptomatic dose of GD (0.2×LD50) was administered by subcutaneous repeated injections, first in pregnancy and subsequently during the lactation period. Six months after the end of poisoning the animals were euthanised and brain structures (hippocampus, cerebellum and piriform cortex) were isolated aseptically for evaluation of p53 mRNA. To determine p53 transcript levels Real-Time PCR with SYBR Green dye was applied. RESULTS: GD action resulted in a significant increase of p53 transcript in the cerebellum and in the piriform cortex of both F0 and F1 females as well as in F1 males. The significant elevation of p53 mRNA level in the hippocampus was observed only in F1 females. CONCLUSIONS: The study demonstrates that GD causes distant changes in the expression of p53 mRNA in the rat brain. Increased expression of p53 mRNA provides indirect evidence that GD-induced distant disorders may include DNA damage and cell cycle disturbances leading to cell dysfunction and their elimination via apoptosis. FINANCIAL SUPPORT: This work was supported by Polish Ministry of Science and Higher Education No O R00 0042 08, “Soldier as a precise weapon – individual sets and kits”.
6

Prolonged SOCS3 gene response following soman intoxication in the rat brain

76%
Paruszewska E., Lapot M., Brytan M., Antkowiak B., Kowalczyk M., Ciechanowska M.
Acta Neurobiologiae Experimentalis
|
2017
|
tom 77
|
nr Suppl.1
INTRODUCTION: In comparison with well documented data concerning the mechanisms of acute neurotoxic action of nerve agents such as soman (GD), the immunomodulatory properties of these compounds are still poorly understood, especially considering their long‑term effects. One promising candidate for mediation of GD-induced immunomodulation seems to be a suppressor of cytokine signaling 3 (SOCS 3) – an intracellular protein which exhibits a wide variety of physiological effects on immune cell function. There also exists strong evidence to support SOCS3 as a crucial regulator of many disease processes in the central nervous system. AIM(S): The aim of the present study was to determine whether perinatal exposure to GD exerted distant action on the expression of mRNA encoding SOCS3 in selected tissue of the rats brain. METHOD(S): Studies were conducted on maternal generation (F0) and on first filial generation (F1) of Wistar rats. F0 animals were treated subcutaneously with a low (0.2×LD50) repeated dose of soman (o-pinacolyl methylphosphonofluoridate). GD was administrated first, in pregnancy, and subsequently during the lactation period. Six months after termination of GD exposure animals were anesthetised and immediately hippocampus, cerebellum and piriform cortex were obtained for subsequent analysis. Real-Time PCR with SYBR Green dye was used to evaluate the level of SOCS3 mRNA in selected structures of the brain. RESULTS: Intoxication with GD decreased significantly SOCS3 mRNA levels in the cerebellum and in the piriform cortex in F0 females and in their offspring of both sexes. The analogous tendency, but without statistical significance, was observed in the hippocampus of all experimental animals. CONCLUSIONS: The current data do not clarify distant signs and symptoms of soman exposure; however, a decrease in expression of SOCS3 following intoxication with GD may suggest a functional role of this protein in pathogenesis of GD-induced neurological disorders. FINANCIAL SUPPORT: This work was supported by Polish Ministry of Science and Higher Education No O R00 0042 08, “Soldier as a precise weapon – packages and sets”.
7
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Effects of corticotropin releasing hormone and corticotropin releasing hormone antagonist on biosynthesis of gonadotropin releasing hormone and gonadotropin releasing hormone receptor in the hypothalamic-pituitary unit of follicular-phase ewes and contribution of kisspeptin

64%
Ciechanowska M., Kowalczyk M., Lapot M., Malewski T., Antkowiak B., Brytan M., Winnicka I., Przekop F.
Journal of Physiology and Pharmacology
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2018
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tom 69
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nr 3
8
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Suppressive effect of electromagnetic field on analgesic activity of tramadol in rats

64%
Bodera P., Stankiewicz W., Antkowiak B., Paluch M., Kieliszek J., Sobiech J., Zdanowski R., Wojdas A., Siwicki A. K., Skopinska-Rozewska E.
Polish Journal of Veterinary Sciences
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2012
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tom 15
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nr 1
The electromagnetic fields (EMFs) have been shown to alter animal and human behavior, such as directional orientation, learning, pain perception (nociception or analgesia) and anxiety-related behaviors. The aim of this study was to evaluate the influence of electromagnetic fields of high-frequency microwaves on pain perception and anti-nociceptive activity of tramadol (TRAM) - analgetic effective in the treatment of moderate to severe acute and chronic pain states. Electromagnetic fields exposures of a) 1500 MHz frequency and b) modulated, 1800 MHz (which is identical to that generated by mobile phones) were applied. Paw withdrawal latency (PWL) to thermal stimulus was measured in vehicle or tramadol (TRAM) treated animals before and after 30, 60 and 90 minutes from injections. The differences in the level of pain (PWL) between control group and rats exposed to EMF alone in three measurements, were not observed. Tramadol alone significantly increased PWLs to thermal stimulus in comparison to vehicle results at 30 (p < 0.001) and 60 minutes (p < 0.05) after drug injection. EMF exposure of both frequencies transiently suppressed analgesic effect of tramadol, significantly reducing paw withdrawal latency in animals treated with this drug at 30 minutes from the drug injection.
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