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Application of real-time PCR technique for validation of microarray experiment

100%
Ciesielska A., Sikorski M. M.
Biotechnologia
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2008
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issue 4
88-100
EN
Microarray technology has recently reached the mainstream. It is widely applied in many biological fields of studies. Despite of its uncontested advantages microarray results have to be validated using independent methodology. For this purpose real-time PCR platform has become the most popular method of choice. The ability of PCR product measurement after each cycle is connected here with wide dynamic range of detection. This review is a summary of our knowledge on the technical issue of quantitative PCR utilization in microarray results confirmation.
2
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New insights into the signaling and function of cytokinins in higher plants

61%
Ciesielska A., Ruszkowski M., Kasperska A., Femiak I., Michalski Z., Sikorski M. M.
BioTechnologia
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2012
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issue 4
3
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Dopamine, serotonin and noradrenaline changes in the striatum of C57BL mice following myelin oligodendrocyte glycoprotein (MOG) 35-55 and complete Freund adjuvant (CFA) administration

61%
Balkowiec-Iskra E., Kurkowska-Jastrzebska I., Joniec I., Ciesielska A., Czlonkowska A., Czlonkowski A.
Acta Neurobiologiae Experimentalis
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2007
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vol. 67
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issue 4
379-388
EN
Many data suggest involvement of inflammation in neurodegeneration. However, the exact mechanisms of this cooperation are poorly understood. We have previously shown that induction of inflammatory reaction, both before and after injury of the striatum, affects regeneration of dopaminergic neurons. In the present research we studied the role of inflammatory reaction in non-injured striatum. We used myelin oligodendrocyte glycoprotein (MOG) 35-55 in complete Freund's adjuvant (CFA) to elicit experimental autoimmune encephalomyelitis (EAE) mice model. As determined by HPLC, striatal dopamine (DA) and serotonin levels in mice treated with either MOG 35-55 in CFA or CFA alone were significantly higher compared to vehicle-treated controls on 13th day after induction. The ratio of homovanilic acid/dopamine (HVA/DA) and 3, 4 dihydroxyphenylacetic acid/dopamine (DOPAC/DA) were significantly lower in the MOG and CFA groups on 13th day, indicating decreased DA metabolism. Noradrenaline (NA) concentration did not differ between groups. Moreover, the striatal mRNA IL-1beta and TNF-alpha levels were elevated during induction phase of EAE in both groups, as determined by RT-PCR. Our data indicate regulatory connection between dopaminergic and immune systems.
4
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Dynamics of expression of the mRNA for cytokines and inducible nitric synthase in a murine model of the Parkinson disease

61%
Ciesielska A., Joniec I., Przybylkowski A., Gromadzka G., Czlonkowska A., Czlonkowski A.
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EN
The inflammatory reaction and oxidative stress has been linked with PD. Proinflammatory cytokines promote neurodegeneration or neuroprotection in different animal models. In addition, these cytokines have been reported to increase iNOS expression. With the RT-PCR method we evaluated mRNA levels for IL1beta, IL6, TNF, IFN gamma, IL-10 and iNOS in the striatum of C57BL/6 mice after MPTP intoxication. The IL1beta mRNA expression rapidly increased, nad peaked at 6 h. The first increase of mRNA for TNFalpha and INFgamma was noticed at 6-24 h and the second at the 7th day after MPTP intoxication. Two peaks of IL10 mRNA were seen, immediately (6 h) and at the 3rd day post MPTP injection. The peak of mRNA level for IL6 was observed at the 7th day. Expression of mRNA for iNOS peaked at 24 h, started decreasing on the 3rd day, but was still present till the 14th day Those findings suggest that cytokine network and iNOS may be involved in the development of immune changes accompanying degeneration of the nigrostriatal system.
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