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1

Comparison of miRNA profiling during airway epithelial repair in undifferentiated and differentiated cells in vitro

100%
Langwinski W., Narozna B., Lackie P. M., Holloway J. W., Szczepankiewicz A.
Journal of Applied Genetics
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2017
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tom 58
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nr 2
2

Role of ADRB2 gene polymorphism in asthma and response to beta2-agonists in Polish children

100%
Szczepankiewicz A., Breborowicz A., Sobkowiak P., Kramer L., Popiel A.
Journal of Applied Genetics
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2009
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tom 50
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nr 3
275-281
The aims of this study were: (1) to find associations of asthma with single-nucleotide polymorphisms (SNPs) within the ADRB2 gene: Arg16Gly, Gln27Glu, -1023 G/A, -367 T/C, -47 C/T ; (2) to define linkage disequilibrium in the gene region, basing on the analyzed SNPs; and (3) to analyze the importance of ADRB2 polymorphism for response to bronchodilator drugs in children diagnosed with bronchial asthma. We compared 113 asthmatic children and 123 healthy subjects from the Polish population. Genotyping was performed by PCR-RFLP. We found an association of the A allele of -1023A/G ADRB2 polymorphism with asthma (P = 0.024). No significant associations with other SNPs were detected. Moderate linkage was found between Gln27Glu and -47C/T polymorphisms in linkage disequilibrium analysis (D’ = 0.85, r² = 0.429, LOD = 31.97). No significant differences were found in haplotype frequencies in comparison to the control group, implicating that they are not associated with susceptibility to asthma in the analyzed population. There was no significant correlation between the analyzed SNPs of the ADRB2 gene and the response to β₂ -agonists. This is the first report providing suggestive evidence for association of -1023A/G ADRB2 polymorphism with an increased risk of asthma. The analyzed SNPs may not play a major role in response to β₂-agonists in asthmatic children.
3

Polymorphic variants of MIF gene and prognosis in steroid therapy in children with idiopathic nephrotic syndrome

100%
Swierczewska M., Ostalska-Nowicka D., Kempisty B., Szczepankiewicz A., Nowicki M.
Acta Biochimica Polonica
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2014
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tom 61
|
nr 1
 Nephrotic syndrome (NS) is the most common reason of proteinuria in children and can be caused by the pathology of renal glomeruli. Steroid therapy is typically used in this disorder. It has been shown that MIF is a cytokine which counteracts the immunosuppressive properties of glucocorticoids. The aim of this study was looking for a correlation between MIF polymorphisms and genetic susceptibility to steroid resistance in children with INS (Idiopathic NS). Methods: The study was performed in 71 patients with INS including SRNS (steroid resistance nephrotic syndrome) (41) and SSNS (steroid sensitive nephrotic syndrome) (30) and in 30 control subjects. We employed Sanger sequencing and capillary electrophoresis. Linkage disequilibrium was made using Haploview and PHASE. Results: We didn't observe a statistical significance between SNPs detected in patients with INS and controls. Our studies revealed statistical significance for two polymorphisms: rs2070767C>T and rs2000466T>G between patients with SRNS and SSNS. The results for rs34383331T>A are close to being statistically significant. Statistical significance was revealed for CATT5/CATT6 genotype in SRNS group vs SSNS group (OR=4.604, 95%CI=1.356-15.632, p=0.0168). We found that the frequency of 5/X-CATT genotype compared with X/X-CATT genotype was significantly higher in SRNS patients vs SSNS (OR=3.167, 95%CI=1.046-9.585, p=0.0426). In linkage disequilibrium analysis we didn't show involvement in susceptibility to INS and steroid sensitive phenotype. Conclusions: Our results suggest that the role of MIF polymorphisms in the susceptibility to positive response to steroid therapy is still unresolved. It indicates that MIF may be involved in indirect and complex molecular mechanisms of steroid activity in hormone-dependent metabolic pathways in children with INS. Because of ambiguous findings, pleiotropic features of this cytokine require that more research should be undertaken
4

A nouvel method to visualize tissue molecular architecture at the level of synapse

86%
Szepesi Z., Szczepankiewicz A., Walczak A., Wilczynski G., Wlodarczyk J., Kaczmarek L.
Acta Neurobiologiae Experimentalis
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2009
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tom 69
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nr 3
To understand various biological functions of CNS, visualization of the fi ne details of tissue molecular architectures and synaptic structures is of the major importance. Herein, we use a method for immunfl uorescence microscopy called Array Tomography which allows for imaging details of tissue structure. To this end, we cut resin-embedded specimen into serial sections (50–250 nm) by ultramicrotome. Hydrophilic resin permits the antibody penetration into the tissue, thus providing an opportunity for post-embedding immunostaining. The ribbon of serial sections represents tissue volume. Reconstruction of three dimensional distributions of antigens is obtained by imaging and post-processing of consecutive images. The array can be eluted and re-stained, thus allowing for detection of several antigens in the same sample. The above methodology provides visualization at the spatial resolution that is higher than the one obtained using conventional confocal microscopy. Furthermore, we use this method together with the super-resolution (70 nm) stimulated emission depletion (STED) microscopy. Thus, combining of these two methods provides very detailed image of tissue architecture and offers a novel opportunity to reconstruct the molecular architecture with sub-micrometer resolution in three dimensions.
5

Influence of dopaminergic and serotoninergic genes on working memory in healthy subjects

86%
Wilkosc M., Hauser J., Tomaszewska M., Dmitrzak-Weglarz M., Skibinska M., Szczepankiewicz A., Borkowska A.
Acta Neurobiologiae Experimentalis
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2010
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tom 70
|
nr 1
Working memory is an ability to keep information in short-term memory and manipulate them “on line”. Working memory is also involved in complex frontal executive functions. The role of dopaminergic system in modulating working memory processes in prefrontal cortex is well established. Also the role of serotoninergic receptors is postulated. The purpose of this study was to assess the association between the polymorphisms of dopaminergic (DRD1, DRD3, DRD4, COMT) and serotoninergic (SERT – serotonin transporter, 5HT2A, 5HT2C) genes’ polymorphisms and performance on WCST in 200 volunteers from the Polish population. We found the association between DRD1, DRD4, COMT and SERT genes polymorphisms and the performance on WCST. The results obtained in the study indicate that dopaminergic and serotoninergic genes may play a role in modulating the executive function and working memory processes in healthy subjects. The pattern of this influence may be different in males and females. Moreover, the relationship between the efficacy of prefrontal cognitive function and genes polymorphisms may differ between healthy subjects and schizophrenic patients.
6

Higher-order nuclear structure in epileptogenesis

86%
Szczepankiewicz A., Walczak A., Szczepinska T., Broszkiewicz M., Pawlowski K., Wilczynski G.
Acta Neurobiologiae Experimentalis
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2009
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tom 69
|
nr 3
Although the molecular mechanisms of gene-expression in neurons are comprehensively described in the literature, little is known about the relationship between these processes and the architecture of the neuronal cell nucleus. For example, it has never been examined whether bursts of transcriptional activity associated with seizures involve any regulation at the level of higher-order nuclear structure. Accordingly, we have performed studies on the structure of neuronal nucleus in epileptic animals. Using electron microscopy, we found the striking appearance of large interchromatin granule clusters (IGCs) in epileptic nuclei. We confi rmed this observation by immunofl uorescence-confocal analysis of IGC marker, a spliceosome assembly factor SC-35. Interestingly, there was also aggregation of spots immunoreactive for phophorylated and acetylated Histone H3, a marker of transcriptionally active chromatin. The fi ndings are consistent with the results of bioinformatic analysis of transcription profi ling in the rat kainate-induced status epilepticus (public gene expression data), showing tendency for coordinated expression of positional gene-clusters along chromosomes. Taken together, our results suggest that upon epileptogenesis there is prominent reorganization of neuronal nucleus, putatively involving formation of molecular factories, where transcription, splicing, and export of pre-mRNA are orchestrated.
7

Neuronal PML bodies are modulated by activity in adult murine brain

86%
Broszkiewicz M., Szczepankiewicz A., Yeghiazaryan M., Patel S., Ruszczycki B., Pawlak R., Wilczynski G.
Acta Neurobiologiae Experimentalis
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2011
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tom 71
|
nr S
PML is a tumor-suppresor protein involved in the pathogenesis of promyelocytic leukemia. In proliferating mammalian cells PML is a principal component of characteristic nuclear bodies, which contain other proteins but do not contain nucleic acids. There are several PML bodies per nucleus. The molecular function of PML protein is unclear, yet the majority of data points to its involvement in regulation of gene-expression and/or intranuclear protein storage and degradation. In the brain PML has been implicated in the pathogenesis of neurodegenerative disorders, glioma and in the control of embryonic neurogenesis. It is not clear whether the protein is expressed, and has a function, in the normal adult brain. Therefore we have investigated the expression and localization of PML at the cellular and subcellular levels, in the adult mouse brain. By immunofluorescence, typical PML bodies were found in a subset of neurons in the cerebral cortex, hippocampus and amygdala. In the cortex, the protein was present predominantly in layer II/III. Stimulation of neuronal activity by a) pentylenetetrazole seizures or b) immobilization stress, greatly increased the size and signal-intensity of the neuronal PML bodies. Our results suggest that function of PML protein in the brain can be associated with neuronal activity and plasticity.
8

Repositioning of selected genes in neuronal cell nucleus upon seizures

59%
Walczak A., Szczepankiewicz A., Szczepinska T., Ruszczycki B., Dabrowski M., Broszkiewicz M., Siudek M., Pyskaty M., Wilczek E., Szczerbal I., Switonski M., Pawlowski K., Wilczynski G.
Acta Neurobiologiae Experimentalis
|
2011
|
tom 71
|
nr S
Even though the molecular mechanisms of gene expression in neurons are quite broadly described in the literature, little is known about the relationship between these processes and the architecture of the neuronal cell nucleus. For example, it is firmly established that waves of gene expression occurring after neuronal stimulation comprise plenty of genes playing important roles in cognitive and epileptic phenomena. However, it has never been examined whether these bursts of transcriptional activity involve any regulation at the level of higher-order nuclear structure. Accordingly, we have performed studies on the structure of neuronal nucleus in epileptic animals. Based on the literature we chose the genes for BDNF and TRKB that play key roles in synaptic plasticity in the brain. We have investigated clustering of these genes in the nucleus by means of fluorescent in situ hybridization. We found that in hippocampal neurons the bdnf-trkb distance decreases 2 hours after seizure induction compared to control. Moreover, using bioinformatical approach we selected 2 linear clusters of the genes that are upregulated after seizures. One of them was selected on the chromosome 1 and the other one on chromosome 20. Those linear gene clusters appeared to associate 4 weeks after seizure compared to control. Our results suggest that upon epileptogenesis there is a reorganization of neuronal nucleus involving clustering of the genes in three-dimensional space. The phenomenon could occur also in other forms of synaptic plasticity, and should be a topic to follow up in the future.
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