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1

Europejski Program Standaryzacji Biologicznej. Standaryzacja metod serologicznych stosowanych do określania mocy skojarzonych szczepionek przeciwbłoniczych (EDQM BSP-034)

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Walory J., Bieganska K., Piasek A.
Diagnostyka Laboratoryjna
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2006
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tom 42
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nr 2
2

Lentiviral delivered siRNA against ICER protects primary cortical neurons from apoptosis caused by serum depriviation

100%
Klejman A., Bieganska K., Gierej D., Kaczmarek L.
Acta Neurobiologiae Experimentalis
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2009
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tom 69
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nr 3
CREB activation and CREB-dependent signaling pathways are crucial for neuronal survival. The term ICER (inducible cAMP early repressor) refers to four protein isoforms that are all endogenous, inducible antagonists of CREB. It was previously shown, that all 4 ICER isoforms are induced upon pro-apoptotic treatment, and also that each of them separately evokes neuronal cell death in cortical culture transfected with these genes. The ICER proteins are believed to be strong repressors of Immediate Early Genes, which are involved in cell response to inter- and/or intra-cellular signals. Herein, we have applied the siRNA approach to silence ICER expression. Because ICERs are members of CREM family of proteins, sharing with them the gene sequence, only the small unique region for ICER was selected to design ICER-directed, specifi c siRNA. Indeed, we obtained functional siRNA capable of blocking ICERs but not affecting CREM proteins. With this tool, we have investigated if the ICER’s silencing protects neurons from apoptosis caused by either serum deprivation or excitotoxicity. Using the lentiviral vector, as a vehicle to deliver siRNA (shRNA) we have found that silencing of ICER mildly, although signifi cantly, protects primary cortical neurons from apoptosis caused by serum deprivation.
3

Screening of trace organic compounds in municipal wastewater by gas chromatography-mass spectrometry

100%
Kotowska U., Bieganska K., Isidorov V. A.
Polish Journal of Environmental Studies
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2012
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tom 21
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nr 1
This research proposes the simple and reliable procedure of screening a wide range of polarity trace organic pollutants in influent and effluent municipal wastewater. Such a method includes concentrations of analytes on C18 bonded phase cartridges and its subsequent eluting with organic solvents of different polarity: n-hexane, ethyl ether, and methanol. Each eluate is directed to gas chromatography/mass spectrometry (GC/MS) separation and detection. Silylation of compounds eluted with ethyl ether and methanol was done prior to chromatographic analysis. For reliable identification of the unknown organic compounds a combination of three independent parameters: mass spectra, retention indices and partition coefficients in n-hexane/ acetonitrile system was employed. More than 120 compounds were identified and semi-quantitatively determined in municipal wastewater from the Wastewater Treatment Plant in Białystok, mainly from the groups of aliphatic acids, alcohols and polyalcohols, and carbohydrates. Compounds potentially harmful to the environment and found in the analyzed municipal wastewater are: 2,6-ditertbutylhydroxytoluene, phosphoric acid, phthalic acid esters, phenol, and drug remnants (ibuprofen, naproxen, and caffeine).
4

Hipokretyny (oreksyny) i receptory hipokretynowe: struktura, rozmieszczenie i molekularne podłoże działania

100%
Bieganska K., Urbanska A., Woldan-Tambor A., Zawilska J. B.
Postępy Biologii Komórki
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2011
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tom 38
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nr 2
5

Effects of orexins on survival of the primary neuronal and glial cell cultures

88%
Sokolowska P., Urbanska A., Bieganska K., Namiecinska M., Zawilska J.
Acta Neurobiologiae Experimentalis
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2011
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tom 71
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nr S
Orexins (hypocretins) are hypothalamic peptides present in neurons that project throughout the brain. They act through two receptors: OX1 and OX2. Recently, it has been demonstrated that orexins exert potent proapoptotic activity in various cancer cell lines. On the other hand, little is known about the role of these peptides in survival of neurons and their supportive cells in the brain. Therefore, the aim of our study was to investigate whether orexins are implicated in receptor-mediated survival- or deathpromoting effects in cultured neurons and astrocytes derived from rat cerebral cortex. Real-time PCR experiments indicated that both types of orexin receptors were expressed in rat neurons and astrocytes. In cultured neurones OX2R expression was considerably higher than that of OX1R. In astrocytes similar expressions of both types of orexin receptors were identified but they were markedly lower compared to neurons. Incubation of primary neuronal cultures with orexin A, orexin B and [Ala11-D-Leu15]orexin B (a selective agonist of OX2R) resulted in a marked increase of cells viability and a parallel reduction of apoptotic cells as assessed by MTT test and caspase-3 assay kit. In cultured astrocytes the tested neuropeptides increased cell viability (MTT) and stimulated 3 H-thymidine incorporation but had no effect on caspase-3 activity, an observation indicating that orexins may affect astrocytes survival by enhancing cell proliferation. In the next set of experiments cultured neurons were subjected to hypoxia induced chemically by iron chelator cobalt chloride. Orexins A and B, and [Ala11-D-Leu15]orexin B effectively protected neuronal cells, suggesting that the peptides may be endowed with neuroprotective potential in the brain. Supported by MNiSW (grant No 4254/B/ PO1/2010/38) and InterMolMed (grant No POIG.01.01.02-10- 107/09).
6

The orexin-induced release of cytokines in primary microglial cell cultures

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Namiecinska M., Sokolowska P., Bieganska K., Urbanska A., Andrzejczak D., Zawilska J. B.
Acta Neurobiologiae Experimentalis
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2011
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tom 71
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nr S
Orexin A and B (hypocretin 1 and 2) are hypothalamic neuropeptides that have been implicated in a variety of physiological functions, including sleep and arousal, reaction to stress, regulation of energy homeostasis and hypothalamo-pituitary-adrenal axis. The neuropeptides exert their numerous actions by interacting with two specific, membrane-bound, G-protein-coupled receptors, OX1R and OX2R. We have recently demonstrated that orexin receptors are expressed in rat astrocytes and modify their function. Thus, searching for new orexins roles, in this work we investigated whether these peptides can affect the production of cytokines by microglial cells. Studies were performed on primary microglial cultures from rat cerebral cortex. Orexin A and orexin B stimulated the release of IL-6 as well as TNF-α from microglial cells, as measured by rat IL-6 and TNF-α immunoassay ELISA kits. Interestingly, orexins did not affect the release of IL-1. The obtained results might be important in the aspect of interaction between microglia and neurons/astrocytes in the central nervous system. Supported by MNiSW (grant No 4254/B/PO1/2010/38) and InterMolMed (grant No POIG.01.01.02-10-107/09).
7

Effects of orexins on survival of rat C6 glioma cells. A comparison to the primary rat astrocyte cell cultures

76%
Bieganska K., Sokolowska P., Namiecinska M., Urbanska A., Sobczak E., Zawilska J. B.
Acta Neurobiologiae Experimentalis
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2011
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tom 71
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nr S
Orexin-A and orexin-B, also named hypocretin-1 and hypocretin-2, are hypothalamic neuropeptides which are important regulators of sleep-wake cycles, reward-seeking, and body energy balance. These neuropeptides bind to two specific, membranebound receptors: OX1R and OX2R, members of the GPCR superfamily. A potent proapoptotic activity of orexins has recently been demonstrated in colon cancer cell lines and human colorectal tumor. In our studies we investigated effects of orexins on survival of rat C6 glioma cells, an experimental model for studies on glioblastoma multiforme, and compared them with those exerted on cultured astocytes from rat cerebral cortex. Orexins A and B decreased the number of surviving C6 glioma cells after 48 h of treatment (MTT test), and reduced [3H]thymidine incorporation into proliferating C6 cells. On the contrary, 48 h incubation of cultured astrocytes with orexins increased astrocyte viability and moderately stimulated their proliferation. Taken together, our results suggest that effects of orexins on cell survival depend on the cell type (normal versus cancer). It can be speculated that orexins may serve as potential anticancer factors in therapy of brain tumors. Supported by MNiSW (grant No 4254/B/PO1/2010/38) and InterMolMed (grant No POIG.01.01.02-10-107/09).
8

Neurogenesis and behavioral strategies in ICER overexpressing rats

64%
Klejman A., Janusz A., Kiryk-Jaskiewicz A., Gorlewicz A., Bieganska K., Jedynak P., Kaczmarek L., Konopka W.
Acta Neurobiologiae Experimentalis
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2019
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tom 79
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nr Suppl.1
INTRODUCTION: ICER (Inducible cAMP Early Repressor) is an effective endogenous repressor of CREB/ CREM/ATF transcription factors family, including its own expression. We have developed a Syn‑Flag‑ICER II transgenic rat line. In transgenic animals, we have surprisingly detected increased levels of mRNA for CREB or CREM transcription factors. We have also detected upregulation of CREB dependent miR‑132. Nerogenesis is a process of generation and maturation of newborn neurons into neuronal networks in the developing brain. We have found that ICER II overexpressing rats showed reduced hippocampal adult neurogenesis. The number of the SGZ BrdU positive cells was similar, but in the mature granular neurons layer, the number of BrdU positive cells was decreased when compared to control animals. One of the crucial elements enabling the incorporation of newborn neurons into neuronal network of the brain is the active reorganization of the extracellular matrix mostly by action of metalloproteinases. The most known for its activity in the brain is matrix metalloproteinase 9 (MMP-9), which is also one of the known targets of miR‑132. We have examined MMP-9 activity in the ICER overexpressing rat brain lysates, and we observed decreased activity of MMP‑9 in ICER mutants as compared to WT controls. RESULTS: We have also found that ICER rats with affected neurogenesis employ different learning strategies than their control littermates in the Morris Water Maze learning paradigm. The results of this behavioral tests indicate that transgenic rats didn’t differ from controls in their learning and memory capabilities, but they showed differences in strategies of finding the hidden platform. Male ICER rats more often were choosing imprecise strategies to find the platform than control males. CONCLUSIONS: Those results demonstrate that disruption of CREB dependent gene expression in neurons by overexpression of ICER affects adult neurogenesis and causes changes that affect discrete aspects of animal cognitive behavior.
9

Manipulation of CREB related gene expression in neurons influences adult neurogenesis and morris water maze strategies

64%
Janusz A., Klejman A., Kiryk-Jaskiewicz A., Gorlewicz A., Bieganska K., Jedynak P., Kaczmarek L., Konopka W.
Acta Neurobiologiae Experimentalis
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2017
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tom 77
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nr Suppl.1
INTRODUCTION: Active reorganization of extracellular matrix in the brain allows for growth of neuronal dendrites and axons which guarantees successful incorporation of new born neurons into neuronal network during adult neurogenesis in the hippocampus. Activity of surrounding neurons may affect adult neurogenesis. AIM(S): In order to test whether manipulation of CREB dependent gene expression in neurons and hence their activity will influence adult neurogenesis we have developed the Syn-Flag-ICER II transgenic rat line. The ICER (Inducible cAMP Early Repressor) is effective endogenous repressor of CREB/CREM/ATF transcription factors family. METHOD(S): BrdU labeling to asses a level of adult neurogenesis in the hippocampus qPCR for changes in transcription of CREB/CREM and related genes gelatin zymography to measure MMP9 activity Morris Water Maze spatial learning tests Patch Clamp RESULTS: ICER II overexpressing rats showed diminished hippocampal neurogenesis. We have observed a reduced number of mature BrdU positive cells in granular zone of hippocampus of transgenic rats, in comparison to control group. We have observed also that neurons of dentate gyrus demonstrate increased excitability. Paradoxically, we have detected increased levels of mRNA for CREB or CREM factors. Also CREB dependent miR-132 expression was upregulated in transgenic rats, which regulates expression of MMP-9 – extracellular matrix metalloproteinase. We have found the decreased activity of MMP9 in ICER overexpressing rats. Morris Water Maze tests didn’t show overall differences in rats learning and memory capabilities, however male ICER rats chose more often imprecise strategies to find hidden platform than control males. CONCLUSIONS: Obtained results indicate that CREB dependent gene expression in neurons regulates a set of genes e.g. miR-132 that may in turn regulate translation of proteins involved in remodeling of extracellular matrix and affect adult neurogenesis, what changes discrete aspects of animal cognitive behavior.
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