TY  - JOUR
AU  - Lavrnja, Irena
AU  - Savić, Danijela
AU  - Parabucki, Ana
AU  - Dacic, Sanja
AU  - Laketa, Danijela
AU  - Peković, Sanja
AU  - Stojiljkovic, Mirjana
PY  - 2015
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2062
AB  - Compelling evidence now points to the critical role of the cytoskeleton
   in neurodegeneration. In the present study, using an immunohistochemical
   approach, we have shown that cortical stab injury (CSI) in adult Wistar
   rats significantly affects temporal pattern of expression of
   neurofilament proteins (NFs). a major cytoskeleton components of
   neurons, and microtubule-associated proteins (MAP2). At 3 days
   post-injury (dpi) most of the NFs immunoreactivity was found in pyknotic
   neurons and in fragmentized axonal processes in the perilesioned cortex.
   These cytoskeletal alterations became more pronounced by 10 dpi. At the
   subcellular level CSI also showed significant impact on NFs and MAP-2
   expression. Thus, at 3 dpi most of the dendrites disappeared, while
   large neuronal somata appeared like open circles pointing to membrane
   disintegration. Conversely, at 10 dpi neuronal perikarya and a few new
   apical dendrites were strongly labeled. Since aberrant NF
   phosphorylation is a pathological hallmark of many human
   neurodegenerative disorders, as well as is found after stressor stimuli,
   the present results shed light into the expression of neurofilaments
   after the stab brain injury. (C) 2014 Elsevier GmbH. All rights
   reserved.
T2  - Acta Histochemica
T1  - Effect of stab injury in the rat cerebral cortex on temporal pattern of
 expression of neuronal cytoskeletal proteins: An immunohistochemical
 study
IS  - 2
VL  - 117
DO  - 10.1016/j.acthis.2014.12.004
SP  - 155
EP  - 162
ER  - 

@article{
author = "Lavrnja, Irena and Savić, Danijela and Parabucki, Ana and Dacic, Sanja and Laketa, Danijela and Peković, Sanja and Stojiljkovic, Mirjana",
year = "2015",
abstract = "Compelling evidence now points to the critical role of the cytoskeleton
   in neurodegeneration. In the present study, using an immunohistochemical
   approach, we have shown that cortical stab injury (CSI) in adult Wistar
   rats significantly affects temporal pattern of expression of
   neurofilament proteins (NFs). a major cytoskeleton components of
   neurons, and microtubule-associated proteins (MAP2). At 3 days
   post-injury (dpi) most of the NFs immunoreactivity was found in pyknotic
   neurons and in fragmentized axonal processes in the perilesioned cortex.
   These cytoskeletal alterations became more pronounced by 10 dpi. At the
   subcellular level CSI also showed significant impact on NFs and MAP-2
   expression. Thus, at 3 dpi most of the dendrites disappeared, while
   large neuronal somata appeared like open circles pointing to membrane
   disintegration. Conversely, at 10 dpi neuronal perikarya and a few new
   apical dendrites were strongly labeled. Since aberrant NF
   phosphorylation is a pathological hallmark of many human
   neurodegenerative disorders, as well as is found after stressor stimuli,
   the present results shed light into the expression of neurofilaments
   after the stab brain injury. (C) 2014 Elsevier GmbH. All rights
   reserved.",
journal = "Acta Histochemica",
title = "Effect of stab injury in the rat cerebral cortex on temporal pattern of
 expression of neuronal cytoskeletal proteins: An immunohistochemical
 study",
number = "2",
volume = "117",
doi = "10.1016/j.acthis.2014.12.004",
pages = "155-162"
}

Lavrnja, I., Savić, D., Parabucki, A., Dacic, S., Laketa, D., Peković, S.,& Stojiljkovic, M.. (2015). Effect of stab injury in the rat cerebral cortex on temporal pattern of
 expression of neuronal cytoskeletal proteins: An immunohistochemical
 study. in Acta Histochemica, 117(2), 155-162.
https://doi.org/10.1016/j.acthis.2014.12.004

Lavrnja I, Savić D, Parabucki A, Dacic S, Laketa D, Peković S, Stojiljkovic M. Effect of stab injury in the rat cerebral cortex on temporal pattern of
 expression of neuronal cytoskeletal proteins: An immunohistochemical
 study. in Acta Histochemica. 2015;117(2):155-162.
doi:10.1016/j.acthis.2014.12.004 .

Lavrnja, Irena, Savić, Danijela, Parabucki, Ana, Dacic, Sanja, Laketa, Danijela, Peković, Sanja, Stojiljkovic, Mirjana, "Effect of stab injury in the rat cerebral cortex on temporal pattern of
 expression of neuronal cytoskeletal proteins: An immunohistochemical
 study" in Acta Histochemica, 117, no. 2 (2015):155-162,
https://doi.org/10.1016/j.acthis.2014.12.004 . .

TY  - JOUR
AU  - Lavrnja, Irena
AU  - Parabucki, Ana
AU  - Brkić, Predrag
AU  - Jovanović, Tomislav
AU  - Dacic, Sanja
AU  - Savić, Danijela
AU  - Pantić, Igor
AU  - Stojiljković, Mirjana
AU  - Peković, Sanja
PY  - 2015
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2070
AB  - The exact mechanisms by which treatment with hyperbaric oxygen (HBOT)
   exerts its beneficial effects on recovery after brain injury are still
   unrevealed. Therefore, in this study we investigated the influence of
   repetitive HBOT on the reactive astrogliosis and expression of mediators
   of inflammation after cortical stab injury (CSI). CSI was performed on
   male Wistar rats, divided into control, sham, and lesioned groups with
   appropriate HBO. The HBOT protocol was as follows: 10 minutes of slow
   compression, 2.5 atmospheres absolute (ATA) for 60 minutes, and 10
   minutes of slow decompression, once a day for 10 consecutive days. Data
   obtained using real-time polymerase chain reaction, Western blot, and
   immunohistochemical and immunofluorescence analyses revealed that
   repetitive HBOT applied after the CSI attenuates reactive astrogliosis
   and glial scarring, and reduces expression of GFAP (glial fibrillary
   acidic protein), vimentin, and ICAM-1 (intercellular adhesion
   molecule-1) both at gene and tissue levels. In addition, HBOT prevents
   expression of CD40 and its ligand CD40L on microglia, neutrophils,
   cortical neurons, and reactive astrocytes. Accordingly, repetitive HBOT,
   by prevention of glial scarring and limiting of expression of
   inflammatory mediators, supports formation of more permissive
   environment for repair and regeneration.
T2  - Mediators of Inflammation
T1  - Repetitive Hyperbaric Oxygenation Attenuates Reactive Astrogliosis and
 Suppresses Expression of Inflammatory Mediators in the Rat Model of
 Brain Injury
IS  - 498405
DO  - 10.1155/2015/498405
ER  - 

@article{
author = "Lavrnja, Irena and Parabucki, Ana and Brkić, Predrag and Jovanović, Tomislav and Dacic, Sanja and Savić, Danijela and Pantić, Igor and Stojiljković, Mirjana and Peković, Sanja",
year = "2015",
abstract = "The exact mechanisms by which treatment with hyperbaric oxygen (HBOT)
   exerts its beneficial effects on recovery after brain injury are still
   unrevealed. Therefore, in this study we investigated the influence of
   repetitive HBOT on the reactive astrogliosis and expression of mediators
   of inflammation after cortical stab injury (CSI). CSI was performed on
   male Wistar rats, divided into control, sham, and lesioned groups with
   appropriate HBO. The HBOT protocol was as follows: 10 minutes of slow
   compression, 2.5 atmospheres absolute (ATA) for 60 minutes, and 10
   minutes of slow decompression, once a day for 10 consecutive days. Data
   obtained using real-time polymerase chain reaction, Western blot, and
   immunohistochemical and immunofluorescence analyses revealed that
   repetitive HBOT applied after the CSI attenuates reactive astrogliosis
   and glial scarring, and reduces expression of GFAP (glial fibrillary
   acidic protein), vimentin, and ICAM-1 (intercellular adhesion
   molecule-1) both at gene and tissue levels. In addition, HBOT prevents
   expression of CD40 and its ligand CD40L on microglia, neutrophils,
   cortical neurons, and reactive astrocytes. Accordingly, repetitive HBOT,
   by prevention of glial scarring and limiting of expression of
   inflammatory mediators, supports formation of more permissive
   environment for repair and regeneration.",
journal = "Mediators of Inflammation",
title = "Repetitive Hyperbaric Oxygenation Attenuates Reactive Astrogliosis and
 Suppresses Expression of Inflammatory Mediators in the Rat Model of
 Brain Injury",
number = "498405",
doi = "10.1155/2015/498405"
}

Lavrnja, I., Parabucki, A., Brkić, P., Jovanović, T., Dacic, S., Savić, D., Pantić, I., Stojiljković, M.,& Peković, S.. (2015). Repetitive Hyperbaric Oxygenation Attenuates Reactive Astrogliosis and
 Suppresses Expression of Inflammatory Mediators in the Rat Model of
 Brain Injury. in Mediators of Inflammation(498405).
https://doi.org/10.1155/2015/498405

Lavrnja I, Parabucki A, Brkić P, Jovanović T, Dacic S, Savić D, Pantić I, Stojiljković M, Peković S. Repetitive Hyperbaric Oxygenation Attenuates Reactive Astrogliosis and
 Suppresses Expression of Inflammatory Mediators in the Rat Model of
 Brain Injury. in Mediators of Inflammation. 2015;(498405).
doi:10.1155/2015/498405 .

Lavrnja, Irena, Parabucki, Ana, Brkić, Predrag, Jovanović, Tomislav, Dacic, Sanja, Savić, Danijela, Pantić, Igor, Stojiljković, Mirjana, Peković, Sanja, "Repetitive Hyperbaric Oxygenation Attenuates Reactive Astrogliosis and
 Suppresses Expression of Inflammatory Mediators in the Rat Model of
 Brain Injury" in Mediators of Inflammation, no. 498405 (2015),
https://doi.org/10.1155/2015/498405 . .

TY  - JOUR
AU  - Brisevac, Dusica
AU  - Adzic, Marija
AU  - Laketa, Danijela
AU  - Parabucki, Ana
AU  - Milosevic, Milena
AU  - Lavrnja, Irena
AU  - Bjelobaba, Ivana
AU  - Sevigny, Jean
AU  - Kipp, Markus
AU  - Nedeljkovic, Nadezda
PY  - 2015
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2349
AB  - Extracellular ATP (eATP) acts as a danger-associated molecular pattern
   which induces reactive response of astrocytes after brain insult,
   including morphological remodeling of astrocytes, proliferation,
   chemotaxis, and release of proinflammatory cytokines. The responses
   induced by eATP are under control of ecto-nucleotidases, which catalyze
   sequential hydrolysis of ATP to adenosine. In the mammalian brain,
   ecto-nucleotidases comprise three enzyme families: ecto-nucleoside
   triphosphate diphosphohydrolases 1-3 (NTPDase1-3), ecto-nucleotide
   pyrophosphatase/phospodiesterases 1-3 (NPP1-3), and ecto-5'-nucleotidase
   (eN), which crucially determine ATP/adenosine ratio in the pericellular
   milieu. Altered expression of ecto-nucleotidases has been demonstrated
   in several experimental models of human brain dysfunctions. In the
   present study, we have explored the pattern of NTPDase1-3, NPP1-3, and
   eN expression by cultured cortical astrocytes challenged with 1 mmol/L
   ATP (eATP). At the transcriptional level, eATP upregulated expression of
   NTPDase1, NTPDase2, NPP2, and eN, while, at translational and functional
   levels, these were paralleled only by the induction of NTPDase2 and eN.
   Additionally, eATP altered membrane topology of eN, from clusters
   localized in membrane domains to continuous distribution along the cell
   membrane. Our results suggest that eATP, by upregulating NTPDase2 and eN
   and altering the enzyme membrane topology, affects local kinetics of ATP
   metabolism and signal transduction that may have important roles in the
   process related to inflammation and reactive gliosis.
T2  - Journal of Molecular Neuroscience
T1  - Extracellular ATP Selectively Upregulates Ecto-Nucleoside Triphosphate
 Diphosphohydrolase 2 and Ecto-5'-Nucleotidase by Rat Cortical Astrocytes
 In Vitro
IS  - 3
VL  - 57
DO  - 10.1007/s12031-015-0601-y
SP  - 452
EP  - 462
ER  - 

@article{
author = "Brisevac, Dusica and Adzic, Marija and Laketa, Danijela and Parabucki, Ana and Milosevic, Milena and Lavrnja, Irena and Bjelobaba, Ivana and Sevigny, Jean and Kipp, Markus and Nedeljkovic, Nadezda",
year = "2015",
abstract = "Extracellular ATP (eATP) acts as a danger-associated molecular pattern
   which induces reactive response of astrocytes after brain insult,
   including morphological remodeling of astrocytes, proliferation,
   chemotaxis, and release of proinflammatory cytokines. The responses
   induced by eATP are under control of ecto-nucleotidases, which catalyze
   sequential hydrolysis of ATP to adenosine. In the mammalian brain,
   ecto-nucleotidases comprise three enzyme families: ecto-nucleoside
   triphosphate diphosphohydrolases 1-3 (NTPDase1-3), ecto-nucleotide
   pyrophosphatase/phospodiesterases 1-3 (NPP1-3), and ecto-5'-nucleotidase
   (eN), which crucially determine ATP/adenosine ratio in the pericellular
   milieu. Altered expression of ecto-nucleotidases has been demonstrated
   in several experimental models of human brain dysfunctions. In the
   present study, we have explored the pattern of NTPDase1-3, NPP1-3, and
   eN expression by cultured cortical astrocytes challenged with 1 mmol/L
   ATP (eATP). At the transcriptional level, eATP upregulated expression of
   NTPDase1, NTPDase2, NPP2, and eN, while, at translational and functional
   levels, these were paralleled only by the induction of NTPDase2 and eN.
   Additionally, eATP altered membrane topology of eN, from clusters
   localized in membrane domains to continuous distribution along the cell
   membrane. Our results suggest that eATP, by upregulating NTPDase2 and eN
   and altering the enzyme membrane topology, affects local kinetics of ATP
   metabolism and signal transduction that may have important roles in the
   process related to inflammation and reactive gliosis.",
journal = "Journal of Molecular Neuroscience",
title = "Extracellular ATP Selectively Upregulates Ecto-Nucleoside Triphosphate
 Diphosphohydrolase 2 and Ecto-5'-Nucleotidase by Rat Cortical Astrocytes
 In Vitro",
number = "3",
volume = "57",
doi = "10.1007/s12031-015-0601-y",
pages = "452-462"
}

Brisevac, D., Adzic, M., Laketa, D., Parabucki, A., Milosevic, M., Lavrnja, I., Bjelobaba, I., Sevigny, J., Kipp, M.,& Nedeljkovic, N.. (2015). Extracellular ATP Selectively Upregulates Ecto-Nucleoside Triphosphate
 Diphosphohydrolase 2 and Ecto-5'-Nucleotidase by Rat Cortical Astrocytes
 In Vitro. in Journal of Molecular Neuroscience, 57(3), 452-462.
https://doi.org/10.1007/s12031-015-0601-y

Brisevac D, Adzic M, Laketa D, Parabucki A, Milosevic M, Lavrnja I, Bjelobaba I, Sevigny J, Kipp M, Nedeljkovic N. Extracellular ATP Selectively Upregulates Ecto-Nucleoside Triphosphate
 Diphosphohydrolase 2 and Ecto-5'-Nucleotidase by Rat Cortical Astrocytes
 In Vitro. in Journal of Molecular Neuroscience. 2015;57(3):452-462.
doi:10.1007/s12031-015-0601-y .

Brisevac, Dusica, Adzic, Marija, Laketa, Danijela, Parabucki, Ana, Milosevic, Milena, Lavrnja, Irena, Bjelobaba, Ivana, Sevigny, Jean, Kipp, Markus, Nedeljkovic, Nadezda, "Extracellular ATP Selectively Upregulates Ecto-Nucleoside Triphosphate
 Diphosphohydrolase 2 and Ecto-5'-Nucleotidase by Rat Cortical Astrocytes
 In Vitro" in Journal of Molecular Neuroscience, 57, no. 3 (2015):452-462,
https://doi.org/10.1007/s12031-015-0601-y . .

TY  - JOUR
AU  - Parabucki, Ana
AU  - Savić, Danijela
AU  - Laketa, Danijela
AU  - Peković, Sanja
AU  - Stojiljković, Mirjana
AU  - Nedeljković, Nadežda
AU  - Bjelobaba, Ivana
PY  - 2014
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5864
AB  - Ectonucleotidases are cell surface-located enzymes responsible for the extracellular degradation of nucleotides. They are comprised of several protein families: ectonucleoside triphosphate diphosphohydrolases (E-NTPDase), ectonucleotide pyrophosphatase/phosphodiesterases (E-NPPases) and ecto-5'-nucleotidase. Previously we showed that cortical stab injury alters ectonucleotidase activities in the rat brain, but that the specific enzymes responsible for these changes were not identified. In this study we investigated the gene expression of the specific ectonucleotidase enzymes, NTP-Dase1-3, NPP1-3 and ecto-5'-nucleotidase, two and seven days after cortical stab injury in rats, using real-time PCR. Two days after the injury we observed only one significant change: the downregulation in NTPDase2 mRNA expression. Our results indicate that traumatic brain injury induces significant upregulation of NTPDasel, NTPDase2 and ecto-5'-nucleotidase transcripts, and the downregulation of NPP1, seven days after the injury. Thus, traumatic brain injury has diverse impacts on ectonucleotidases gene expression, which may be reflected in the enzyme activities and extracellular nucleotide concentrations in the perilesional tissue.
PB  - Belgrade: Serbian Biological Society
T2  - Archives of Biological Sciences
T1  - Expression of major ectonucleotidases after cortical stab brain injury in rats: A real-time PCR study
IS  - 1
VL  - 66
DO  - 10.2298/ABS1401148P
SP  - 149
EP  - 155
ER  - 

@article{
author = "Parabucki, Ana and Savić, Danijela and Laketa, Danijela and Peković, Sanja and Stojiljković, Mirjana and Nedeljković, Nadežda and Bjelobaba, Ivana",
year = "2014",
abstract = "Ectonucleotidases are cell surface-located enzymes responsible for the extracellular degradation of nucleotides. They are comprised of several protein families: ectonucleoside triphosphate diphosphohydrolases (E-NTPDase), ectonucleotide pyrophosphatase/phosphodiesterases (E-NPPases) and ecto-5'-nucleotidase. Previously we showed that cortical stab injury alters ectonucleotidase activities in the rat brain, but that the specific enzymes responsible for these changes were not identified. In this study we investigated the gene expression of the specific ectonucleotidase enzymes, NTP-Dase1-3, NPP1-3 and ecto-5'-nucleotidase, two and seven days after cortical stab injury in rats, using real-time PCR. Two days after the injury we observed only one significant change: the downregulation in NTPDase2 mRNA expression. Our results indicate that traumatic brain injury induces significant upregulation of NTPDasel, NTPDase2 and ecto-5'-nucleotidase transcripts, and the downregulation of NPP1, seven days after the injury. Thus, traumatic brain injury has diverse impacts on ectonucleotidases gene expression, which may be reflected in the enzyme activities and extracellular nucleotide concentrations in the perilesional tissue.",
publisher = "Belgrade: Serbian Biological Society",
journal = "Archives of Biological Sciences",
title = "Expression of major ectonucleotidases after cortical stab brain injury in rats: A real-time PCR study",
number = "1",
volume = "66",
doi = "10.2298/ABS1401148P",
pages = "149-155"
}

Parabucki, A., Savić, D., Laketa, D., Peković, S., Stojiljković, M., Nedeljković, N.,& Bjelobaba, I.. (2014). Expression of major ectonucleotidases after cortical stab brain injury in rats: A real-time PCR study. in Archives of Biological Sciences
Belgrade: Serbian Biological Society., 66(1), 149-155.
https://doi.org/10.2298/ABS1401148P

Parabucki A, Savić D, Laketa D, Peković S, Stojiljković M, Nedeljković N, Bjelobaba I. Expression of major ectonucleotidases after cortical stab brain injury in rats: A real-time PCR study. in Archives of Biological Sciences. 2014;66(1):149-155.
doi:10.2298/ABS1401148P .

Parabucki, Ana, Savić, Danijela, Laketa, Danijela, Peković, Sanja, Stojiljković, Mirjana, Nedeljković, Nadežda, Bjelobaba, Ivana, "Expression of major ectonucleotidases after cortical stab brain injury in rats: A real-time PCR study" in Archives of Biological Sciences, 66, no. 1 (2014):149-155,
https://doi.org/10.2298/ABS1401148P . .

TY  - JOUR
AU  - Savić, Danijela
AU  - Stojiljkovic, Mirjana
AU  - Lavrnja, Irena
AU  - Parabucki, Ana
AU  - Bjelobaba, Ivana
AU  - Nedeljkovic, Nadezda
AU  - Herdegen, Thomas
AU  - Peković, Sanja
PY  - 2014
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2120
AB  - Ribavirin (RBV) is synthetic purine nucleoside analogue, licensed as
   anti-viral drug that displays immunomodulatory actions on various immune
   cells. Our previous ex vivo studies have demonstrated immunosuppressive
   effects of RBV on reactive T-lymphocytes in experimental autoimmune
   encephalomyelitis. Here, we examined the effects of RBV on inflammatory
   response of microglia. RBV potency to down-regulate microglia
   inflammatory response was assessed by measuring microglia cell body
   size, and the production of nitric oxide (NO) and pro-and
   anti-inflammatory cytokines. RBV exerted cytotoxic effects on
   LPS-stimulated microglia, leaving non-stimulated microglia unaffected.
   The exposure of activated microglia to RBV led to: decrease in the level
   of NO as a result of decreased cell number, lower average cell surface,
   the reduction of membrane ruffling, the suppression of interleukin-6
   release and promoted interleukin-10 production. On the other hand, RBV
   promoted LPS-induced interleukin-1 beta release. Our results imply that
   RBV is a complex immunomodulator showing both anti-and pro-inflammatory
   effects on activated microglia.
T2  - Immunopharmacology and Immunotoxicology
T1  - Ribavirin shows immunomodulatory effects on activated microglia
IS  - 6
VL  - 36
DO  - 10.3109/08923973.2014.971962
SP  - 433
EP  - 441
ER  - 

@article{
author = "Savić, Danijela and Stojiljkovic, Mirjana and Lavrnja, Irena and Parabucki, Ana and Bjelobaba, Ivana and Nedeljkovic, Nadezda and Herdegen, Thomas and Peković, Sanja",
year = "2014",
abstract = "Ribavirin (RBV) is synthetic purine nucleoside analogue, licensed as
   anti-viral drug that displays immunomodulatory actions on various immune
   cells. Our previous ex vivo studies have demonstrated immunosuppressive
   effects of RBV on reactive T-lymphocytes in experimental autoimmune
   encephalomyelitis. Here, we examined the effects of RBV on inflammatory
   response of microglia. RBV potency to down-regulate microglia
   inflammatory response was assessed by measuring microglia cell body
   size, and the production of nitric oxide (NO) and pro-and
   anti-inflammatory cytokines. RBV exerted cytotoxic effects on
   LPS-stimulated microglia, leaving non-stimulated microglia unaffected.
   The exposure of activated microglia to RBV led to: decrease in the level
   of NO as a result of decreased cell number, lower average cell surface,
   the reduction of membrane ruffling, the suppression of interleukin-6
   release and promoted interleukin-10 production. On the other hand, RBV
   promoted LPS-induced interleukin-1 beta release. Our results imply that
   RBV is a complex immunomodulator showing both anti-and pro-inflammatory
   effects on activated microglia.",
journal = "Immunopharmacology and Immunotoxicology",
title = "Ribavirin shows immunomodulatory effects on activated microglia",
number = "6",
volume = "36",
doi = "10.3109/08923973.2014.971962",
pages = "433-441"
}

Savić, D., Stojiljkovic, M., Lavrnja, I., Parabucki, A., Bjelobaba, I., Nedeljkovic, N., Herdegen, T.,& Peković, S.. (2014). Ribavirin shows immunomodulatory effects on activated microglia. in Immunopharmacology and Immunotoxicology, 36(6), 433-441.
https://doi.org/10.3109/08923973.2014.971962

Savić D, Stojiljkovic M, Lavrnja I, Parabucki A, Bjelobaba I, Nedeljkovic N, Herdegen T, Peković S. Ribavirin shows immunomodulatory effects on activated microglia. in Immunopharmacology and Immunotoxicology. 2014;36(6):433-441.
doi:10.3109/08923973.2014.971962 .

Savić, Danijela, Stojiljkovic, Mirjana, Lavrnja, Irena, Parabucki, Ana, Bjelobaba, Ivana, Nedeljkovic, Nadezda, Herdegen, Thomas, Peković, Sanja, "Ribavirin shows immunomodulatory effects on activated microglia" in Immunopharmacology and Immunotoxicology, 36, no. 6 (2014):433-441,
https://doi.org/10.3109/08923973.2014.971962 . .

TY  - JOUR
AU  - Laketa, Danijela
AU  - Bjelobaba, Ivana
AU  - Savić, Danijela
AU  - Lavrnja, Irena
AU  - Parabucki, Ana
AU  - Stojiljković, Mirjana
AU  - Nedeljković, Nadežda
PY  - 2013
PY  - 2013
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/326
AB  - Injury and other pathological conditions induce a massive release of ATP and ADP that initiate an immune response. Extracellular nucleotides are degraded by ectonucleotidases: enzymes from E-NTPDase and E-NPP families sequentially hydrolyze ATP and ADP to AMP, which is further hydrolyzed by ecto-5'-nucleotidase to adenosine that exerts suppressive effects on immune cells. We investigated the ectonucleotidase activities of peripheral lymphocytes at different post-injury times after an unilateral brain injury in the rat. Significant and dynamic changes in the lymphocytic ectonucleotidase activities were obtained. ATP- and ADP-hydrolysis changes, together with their calculated ratios, indicate the major contribution of E-NTPDase 1 and its comparable upregulation between sham operation and injury. AMP hydrolysis changes were more brain-injury specific, with a longer-lasting lymphocytic response induced by cortical stab injury (CSI). In summary, CSI and sham operation induce the upregulation of the whole enzyme chain for adenine nucleotide hydrolysis in lymphocytes, suggesting an important roles of ectonucleotidases in the course of recovery after brain injury.
T2  - Archives of Biological Sciences
T1  - Brain cortical injury induces changes in peripheral lymphocyte ectonucleotidase activities
IS  - 1
VL  - 65
SP  - 33
EP  - 42
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_326
ER  - 

@article{
author = "Laketa, Danijela and Bjelobaba, Ivana and Savić, Danijela and Lavrnja, Irena and Parabucki, Ana and Stojiljković, Mirjana and Nedeljković, Nadežda",
year = "2013, 2013",
abstract = "Injury and other pathological conditions induce a massive release of ATP and ADP that initiate an immune response. Extracellular nucleotides are degraded by ectonucleotidases: enzymes from E-NTPDase and E-NPP families sequentially hydrolyze ATP and ADP to AMP, which is further hydrolyzed by ecto-5'-nucleotidase to adenosine that exerts suppressive effects on immune cells. We investigated the ectonucleotidase activities of peripheral lymphocytes at different post-injury times after an unilateral brain injury in the rat. Significant and dynamic changes in the lymphocytic ectonucleotidase activities were obtained. ATP- and ADP-hydrolysis changes, together with their calculated ratios, indicate the major contribution of E-NTPDase 1 and its comparable upregulation between sham operation and injury. AMP hydrolysis changes were more brain-injury specific, with a longer-lasting lymphocytic response induced by cortical stab injury (CSI). In summary, CSI and sham operation induce the upregulation of the whole enzyme chain for adenine nucleotide hydrolysis in lymphocytes, suggesting an important roles of ectonucleotidases in the course of recovery after brain injury.",
journal = "Archives of Biological Sciences",
title = "Brain cortical injury induces changes in peripheral lymphocyte ectonucleotidase activities",
number = "1",
volume = "65",
pages = "33-42",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_326"
}

Laketa, D., Bjelobaba, I., Savić, D., Lavrnja, I., Parabucki, A., Stojiljković, M.,& Nedeljković, N.. (2013). Brain cortical injury induces changes in peripheral lymphocyte ectonucleotidase activities. in Archives of Biological Sciences, 65(1), 33-42.
https://hdl.handle.net/21.15107/rcub_ibiss_326

Laketa D, Bjelobaba I, Savić D, Lavrnja I, Parabucki A, Stojiljković M, Nedeljković N. Brain cortical injury induces changes in peripheral lymphocyte ectonucleotidase activities. in Archives of Biological Sciences. 2013;65(1):33-42.
https://hdl.handle.net/21.15107/rcub_ibiss_326 .

Laketa, Danijela, Bjelobaba, Ivana, Savić, Danijela, Lavrnja, Irena, Parabucki, Ana, Stojiljković, Mirjana, Nedeljković, Nadežda, "Brain cortical injury induces changes in peripheral lymphocyte ectonucleotidase activities" in Archives of Biological Sciences, 65, no. 1 (2013):33-42,
https://hdl.handle.net/21.15107/rcub_ibiss_326 .

TY  - THES
AU  - Parabucki, Ana
PY  - 2013
UR  - http://eteze.bg.ac.rs/application/showtheses?thesesId=763
UR  - https://fedorabg.bg.ac.rs/fedora/get/o:7070/bdef:Content/download
UR  - http://vbs.rs/scripts/cobiss?command=DISPLAY&base=70036&RID=1024629426
UR  - http://nardus.mpn.gov.rs/123456789/2104
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2403
AB  - Glavno obeležje traumatske povrede mozga (TPM) je reaktivna astroglioza koja, između ostalog, uzrokuje i promene u signalizaciji purinima. Posebno važan aspekt purinske signalizacije u patološkim procesima centralnog nervnog sistema predstavlja dinamika promena vanćelijskih koncentracija neuroprotektora adenozina. Stoga je cilj ove doktorske teze bio ispitivanje ekspresije i funkcije komponenti adenozinskog signalnog sistema astrocita u in vivo i in vitro modelu moždane povrede, sa posebnim osvrtom na ulogu ekvilibrišućih nukleozidnih transportera (ENT).
U in vivo studiji, izvedenoj na modelu ubodne lezije kore prednjeg mozga pacova, je pokazano da povreda dovodi do dinamičnih promena u ekspresiji ENT, ektonukleotidaza i adenozinskog A1 receptora. Sem toga, povreda uzrokuje i ćelijsku re-distribuciju ENT1/2 i ushodnu regulaciju transportera na reaktivnim astrocitima, što je posebno izraženo sedmog dana nakon ozlede.
Uloga astrocita u orkestraciji adenozinskog signalnog sistema nakon povrede je detaljnije ispitana in vitro, nakon skarifikacije astrocitnog jednosloja. Rezultati su pokazali da skarifikacija povećava ekspresiju ENT1 i ENT2 tek u kasnijim vremenima. Bifazna promena u ekspresiji ekto-5`-nukleotidaze (e-5NT) je iskazana prvobitnim smanjenjem i potom povećanjem ekspresije u kasnijim vremenima nakon skarifikacije. Pored toga, skarifikacija astrocitnog jednosloja vodi promenama u koncentracijama adenozina i njegovih metabolita u ćelijskom medijumu. Naime, porast koncentracija adenozina u ranim vremenima nakon povrede, bio je praćen padom u kasnijim vremenima. Blokiranje ENT dipiridamolom (DPM) je dovelo do promena u koncentracijama adenozina nakon
skarifikacije, ukazavši na ulogu ENT1/2 u kontroli vanćelijskih koncentracija ovog nukleozida.
Kako bi se nagovestili mogući putevi regulacije ENT, kultura astrocita je podvrgnuta tretmanima adenozinom, ATP, DPM i blokatorom A1 receptora DPCPX. Rezultati su pokazali da ATP ushodno reguliše oba transportera i e-5NT, dok adenozin povećava ekspresiju ENT2 a smanjuje ekspresiju ENT1. Takođe, drugačiji efekat na ekspresiju ENT1/2 su imali i DPM i DPCPX, potvrdivši da su ova dva transportera regulisana drugačijim putevima.
Na kraju smo želeli da uporedimo efekte mehaničke povrede i CoCl2 izazvane hipoksije nakon četvoročasovnog na aktivnost komponenti adenozinskog sistema. Za razliku od skarifikacije, CoCl2 vodi ranom povećanju ekspresije ENT2, e-5NT i HIF-1α, kao i daleko većem povećanju vanćelijskih koncentracija adenozina.
Dobijeni rezultati predstavljaju pregled eskpresije i aktivnosti najvažnijih komponenti adenozinskog sistema astrocita nakon povrede, dajući dobru osnovu za dalja ispitivanja uloge astrocita u kontroli kruženja adenozina u fiziološkim i patološkim stanjima, kao i potencijalne kandidate za buduće terapije TPM.
AB  - Reactive astrogliosis is a hallmark of traumatic brain injury (TBI), which, among the others alterations, causes changes in purinergic signaling. Due to its neuroprotective features, fluctuations of adenosine extracellular concentration are particularly important aspect of purinergic signaling in brain pathology. Hence, herein given thesis aimed to investigate expression and function of astrocytes’ adenosine signaling system components after brain injury in vitro and in vivo, with special regard to the role of equilibrative nucleoside transporters (ENT).
In vivo study, performed on a model of cortical stub injury of rat forebrain, showed that injury caused dynamic changes in expression of ENTs, ectonucleotidases, and adenosine A1 receptor. Moreover, injury induced cell redistribution of ENT1/2 and upregulation of transporters on reactive astrocytes, which is especially pronounced seven day after the impact.
The role of astrocytes in orchestration of adenosine signaling system after the injury was examined in more details in vitro, after scratch wound injury of astrocytic monolayer. Results have shown that scarification induced upregulation of ENT1 and ENT2 in later time points. Biphasic alteration in expression of e-5NT was shown in early downregulation followed by upregulation of the enzyme in later time points after the induction of scratch wound. Beside, scarification of astrocytic monolayer caused changes in concentration of adenosine and its metabolites in extracellular medium. The rise of adenosine concentration was noted early after the injury, which was followed by drop of the concentration in later time points examined. Blocking of ENT with dipyridamole (DPM) resulted in changes of observed adenosine concentration after the scarification, pointing out
that ENT1/2 have significant role in controlling extracellular concentration of this nucleoside.
In order to identify possible regulation pathways for ENT, astrocytes where treated with adenosine, ATP, DPM and DPCPX, antagonist of A1 receptor. Results have shown that ATP upregulates both transporters as well as e-5NT, while adenosine enhances expression of ENT2 and diminishes expression of ENT1. Moreover, DPM and DPCPX have shown different effect on ENT1/2 expression, confirming that regulation of these two transporters depends on different pathways.
Toward the end, we wanted to compare effects of mechanical injury to those caused by chemically induced hypoxia after 4-hour CoCl2 treatment, on activity of the components of adenosine system. Unlike the sratch wound injury, CoCl2 causes early upregulation of ENT2, e-5NT and HIF-1α, as well as much greater increase in extracellular adenosine concentration.
Herein given results represent an overview of activity and expression the most important components of adenosine signaling system after the injury, therefore giving good foundation for future research of astrocyte role in control of adenosine in both physiological and pathological conditions, as well as for investigation of potential candidates for future TBI therapies.
PB  - Belgrade: University of Belgrade, Faculty of Biology
T2  - University of Belgrade, Faculty of Biology
T1  - Aktivnost komponenti adenozinskog signalnog sistema astrocita u modelu moždane povrede in vivo i in vitro
T1  - Activity of the astrocytes' adenosine signaling system components in model of traumatic brain injury in vivo and in vitro
SP  - 1
EP  - 182
UR  - https://hdl.handle.net/21.15107/rcub_nardus_2104
ER  - 

@phdthesis{
author = "Parabucki, Ana",
year = "2013",
abstract = "Glavno obeležje traumatske povrede mozga (TPM) je reaktivna astroglioza koja, između ostalog, uzrokuje i promene u signalizaciji purinima. Posebno važan aspekt purinske signalizacije u patološkim procesima centralnog nervnog sistema predstavlja dinamika promena vanćelijskih koncentracija neuroprotektora adenozina. Stoga je cilj ove doktorske teze bio ispitivanje ekspresije i funkcije komponenti adenozinskog signalnog sistema astrocita u in vivo i in vitro modelu moždane povrede, sa posebnim osvrtom na ulogu ekvilibrišućih nukleozidnih transportera (ENT).
U in vivo studiji, izvedenoj na modelu ubodne lezije kore prednjeg mozga pacova, je pokazano da povreda dovodi do dinamičnih promena u ekspresiji ENT, ektonukleotidaza i adenozinskog A1 receptora. Sem toga, povreda uzrokuje i ćelijsku re-distribuciju ENT1/2 i ushodnu regulaciju transportera na reaktivnim astrocitima, što je posebno izraženo sedmog dana nakon ozlede.
Uloga astrocita u orkestraciji adenozinskog signalnog sistema nakon povrede je detaljnije ispitana in vitro, nakon skarifikacije astrocitnog jednosloja. Rezultati su pokazali da skarifikacija povećava ekspresiju ENT1 i ENT2 tek u kasnijim vremenima. Bifazna promena u ekspresiji ekto-5`-nukleotidaze (e-5NT) je iskazana prvobitnim smanjenjem i potom povećanjem ekspresije u kasnijim vremenima nakon skarifikacije. Pored toga, skarifikacija astrocitnog jednosloja vodi promenama u koncentracijama adenozina i njegovih metabolita u ćelijskom medijumu. Naime, porast koncentracija adenozina u ranim vremenima nakon povrede, bio je praćen padom u kasnijim vremenima. Blokiranje ENT dipiridamolom (DPM) je dovelo do promena u koncentracijama adenozina nakon
skarifikacije, ukazavši na ulogu ENT1/2 u kontroli vanćelijskih koncentracija ovog nukleozida.
Kako bi se nagovestili mogući putevi regulacije ENT, kultura astrocita je podvrgnuta tretmanima adenozinom, ATP, DPM i blokatorom A1 receptora DPCPX. Rezultati su pokazali da ATP ushodno reguliše oba transportera i e-5NT, dok adenozin povećava ekspresiju ENT2 a smanjuje ekspresiju ENT1. Takođe, drugačiji efekat na ekspresiju ENT1/2 su imali i DPM i DPCPX, potvrdivši da su ova dva transportera regulisana drugačijim putevima.
Na kraju smo želeli da uporedimo efekte mehaničke povrede i CoCl2 izazvane hipoksije nakon četvoročasovnog na aktivnost komponenti adenozinskog sistema. Za razliku od skarifikacije, CoCl2 vodi ranom povećanju ekspresije ENT2, e-5NT i HIF-1α, kao i daleko većem povećanju vanćelijskih koncentracija adenozina.
Dobijeni rezultati predstavljaju pregled eskpresije i aktivnosti najvažnijih komponenti adenozinskog sistema astrocita nakon povrede, dajući dobru osnovu za dalja ispitivanja uloge astrocita u kontroli kruženja adenozina u fiziološkim i patološkim stanjima, kao i potencijalne kandidate za buduće terapije TPM., Reactive astrogliosis is a hallmark of traumatic brain injury (TBI), which, among the others alterations, causes changes in purinergic signaling. Due to its neuroprotective features, fluctuations of adenosine extracellular concentration are particularly important aspect of purinergic signaling in brain pathology. Hence, herein given thesis aimed to investigate expression and function of astrocytes’ adenosine signaling system components after brain injury in vitro and in vivo, with special regard to the role of equilibrative nucleoside transporters (ENT).
In vivo study, performed on a model of cortical stub injury of rat forebrain, showed that injury caused dynamic changes in expression of ENTs, ectonucleotidases, and adenosine A1 receptor. Moreover, injury induced cell redistribution of ENT1/2 and upregulation of transporters on reactive astrocytes, which is especially pronounced seven day after the impact.
The role of astrocytes in orchestration of adenosine signaling system after the injury was examined in more details in vitro, after scratch wound injury of astrocytic monolayer. Results have shown that scarification induced upregulation of ENT1 and ENT2 in later time points. Biphasic alteration in expression of e-5NT was shown in early downregulation followed by upregulation of the enzyme in later time points after the induction of scratch wound. Beside, scarification of astrocytic monolayer caused changes in concentration of adenosine and its metabolites in extracellular medium. The rise of adenosine concentration was noted early after the injury, which was followed by drop of the concentration in later time points examined. Blocking of ENT with dipyridamole (DPM) resulted in changes of observed adenosine concentration after the scarification, pointing out
that ENT1/2 have significant role in controlling extracellular concentration of this nucleoside.
In order to identify possible regulation pathways for ENT, astrocytes where treated with adenosine, ATP, DPM and DPCPX, antagonist of A1 receptor. Results have shown that ATP upregulates both transporters as well as e-5NT, while adenosine enhances expression of ENT2 and diminishes expression of ENT1. Moreover, DPM and DPCPX have shown different effect on ENT1/2 expression, confirming that regulation of these two transporters depends on different pathways.
Toward the end, we wanted to compare effects of mechanical injury to those caused by chemically induced hypoxia after 4-hour CoCl2 treatment, on activity of the components of adenosine system. Unlike the sratch wound injury, CoCl2 causes early upregulation of ENT2, e-5NT and HIF-1α, as well as much greater increase in extracellular adenosine concentration.
Herein given results represent an overview of activity and expression the most important components of adenosine signaling system after the injury, therefore giving good foundation for future research of astrocyte role in control of adenosine in both physiological and pathological conditions, as well as for investigation of potential candidates for future TBI therapies.",
publisher = "Belgrade: University of Belgrade, Faculty of Biology",
journal = "University of Belgrade, Faculty of Biology",
title = "Aktivnost komponenti adenozinskog signalnog sistema astrocita u modelu moždane povrede in vivo i in vitro, Activity of the astrocytes' adenosine signaling system components in model of traumatic brain injury in vivo and in vitro",
pages = "1-182",
url = "https://hdl.handle.net/21.15107/rcub_nardus_2104"
}

Parabucki, A.. (2013). Aktivnost komponenti adenozinskog signalnog sistema astrocita u modelu moždane povrede in vivo i in vitro. in University of Belgrade, Faculty of Biology
Belgrade: University of Belgrade, Faculty of Biology., 1-182.
https://hdl.handle.net/21.15107/rcub_nardus_2104

Parabucki A. Aktivnost komponenti adenozinskog signalnog sistema astrocita u modelu moždane povrede in vivo i in vitro. in University of Belgrade, Faculty of Biology. 2013;:1-182.
https://hdl.handle.net/21.15107/rcub_nardus_2104 .

Parabucki, Ana, "Aktivnost komponenti adenozinskog signalnog sistema astrocita u modelu moždane povrede in vivo i in vitro" in University of Belgrade, Faculty of Biology (2013):1-182,
https://hdl.handle.net/21.15107/rcub_nardus_2104 .

TY  - JOUR
AU  - Savić, Danijela
AU  - Parabucki, Ana
AU  - Bjelobaba, Ivana
AU  - Santrač, Anja
AU  - Dacić, Sanja
AU  - Peković, Sanja
AU  - Stojiljković, Mirjana
PY  - 2013
PY  - 2013
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/507
AB  - Background: Various in vivo and in vitro models have been described in order to elucidate the pathobiology underlying the traumatic brain injury (TBI) and test potentially suitable treatments. Since TBI is a complex disease, models differ in regard to the aspect of TBI that is being investigated. One of the used in vitro models is the scratch wound assay, first established as a reproducible, low-cost assay for the analysis of cell migration in vitro. The aim of the present study was to further investigate the relevancy of this model as a counter­part of in vivo TBI models. Methods: We have examined the astrocytic response to a mechanical injury in terms of expression of chondroitin sulfate proteoglycans (CSPGs) - phosphacan, neurocan and brevican, using real-time PCR and immunocytochemistry. Results: Our results indicate that in vitro scratch wounding alters the expression profile of examined CSPGs. Four hours after the scratch injury of the astrocytic monolayer, real-time PCR analysis revealed upregulation of mRNA levels for phosphacan (3-fold) and neurocan (2-fold), whereas brevican mRNA was downregulated (2-fold). Immunofluorescent signal for phosphacan and neurocan was more intense in astrocytes close to the injury site, while brevican was scarcely present in cultured astrocytes. Conclusions: Obtained results indicate that CSPGs are differentially expressed by astrocytes after scratch wounding, demonstrating that the scratch wound model might be suitable for investigation of astrocyte-derived response to injury.
AB  - Uvod: Brojni in vivo i in vitro modeli opisani su sa ciljem da se rasvetle patobiološki procesi koji su osnova traumatske povrede mozga (TPM) i testiraju potencijalni tretmani. Imajući u vidu da je TPM kompleksno oboljenje, ovi modeli se međusobno razlikuju shodno aspektu TPM koji se ispituje. Jedan od in vitro modela je i povreda ćelijskog jednosloja grebanjem (engl. 'scratch wound' assay), isprva ustanovljen kao ponovljiv, jeftin test za analizu celijske migracije in vitro. Cilj ove studije je da se bliže ispita relevantnost ovog modela u odnosu na in vivo modele TPM. Metode: Da bi se istražio odgovor astrocita na mehaničku povredu, praćena je ekspresija odabranih hondroitin-sulfatnih proteoglikana (CSPG) - fosfakana, neurokana i brevikana, korišćenjem PCR u realnom vremenu i imunocitohemije. Rezultati: Dobijeni rezultati su pokazali da in vitro povreda astrocitnog jednosloja menja profile ekspresije ispitivanih CSPG. Četiri sata nakon povrede, primena PCR u realnom vremenu analize pokazala je povećanje nivoa iRNK za fosfakan (trostruko) i neurokan (dvostruko), dok je iRNK za brevikan bila smanjena na polovinu kontrolne vrednosti. Imunofluorescentni signal poreklom od fosfakana i neurokana je bio intenzivniji u astrocitima bližim mestu povrede, dok je signal za brevikan bio slab kako u kontrolnoj, tako i u ozleđenoj grupi. Zaključak: Dobijeni rezultati pokazuju da povreda izazvana grebanjem različito utiče na ekspresiju ispitivanih CSPG u astrocitima, sto ukazuju da ovaj model može biti pogodan za ispitivanje odgovora astrocita na povredu.
T2  - Journal of Medical Biochemistry
T1  - PCR i imunocitohemijska studija ekspresije hondroitin-sulfatnih proteoglikana nakon povrede astrocita u kulturi
T1  - Real-time PCR and immunocytochemical study of chondroitin sulfate proteoglycans after scratch wounding in cultured astrocytes
IS  - 4
VL  - 32
SP  - 398
EP  - 405
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_507
ER  - 

@article{
author = "Savić, Danijela and Parabucki, Ana and Bjelobaba, Ivana and Santrač, Anja and Dacić, Sanja and Peković, Sanja and Stojiljković, Mirjana",
year = "2013, 2013",
abstract = "Background: Various in vivo and in vitro models have been described in order to elucidate the pathobiology underlying the traumatic brain injury (TBI) and test potentially suitable treatments. Since TBI is a complex disease, models differ in regard to the aspect of TBI that is being investigated. One of the used in vitro models is the scratch wound assay, first established as a reproducible, low-cost assay for the analysis of cell migration in vitro. The aim of the present study was to further investigate the relevancy of this model as a counter­part of in vivo TBI models. Methods: We have examined the astrocytic response to a mechanical injury in terms of expression of chondroitin sulfate proteoglycans (CSPGs) - phosphacan, neurocan and brevican, using real-time PCR and immunocytochemistry. Results: Our results indicate that in vitro scratch wounding alters the expression profile of examined CSPGs. Four hours after the scratch injury of the astrocytic monolayer, real-time PCR analysis revealed upregulation of mRNA levels for phosphacan (3-fold) and neurocan (2-fold), whereas brevican mRNA was downregulated (2-fold). Immunofluorescent signal for phosphacan and neurocan was more intense in astrocytes close to the injury site, while brevican was scarcely present in cultured astrocytes. Conclusions: Obtained results indicate that CSPGs are differentially expressed by astrocytes after scratch wounding, demonstrating that the scratch wound model might be suitable for investigation of astrocyte-derived response to injury., Uvod: Brojni in vivo i in vitro modeli opisani su sa ciljem da se rasvetle patobiološki procesi koji su osnova traumatske povrede mozga (TPM) i testiraju potencijalni tretmani. Imajući u vidu da je TPM kompleksno oboljenje, ovi modeli se međusobno razlikuju shodno aspektu TPM koji se ispituje. Jedan od in vitro modela je i povreda ćelijskog jednosloja grebanjem (engl. 'scratch wound' assay), isprva ustanovljen kao ponovljiv, jeftin test za analizu celijske migracije in vitro. Cilj ove studije je da se bliže ispita relevantnost ovog modela u odnosu na in vivo modele TPM. Metode: Da bi se istražio odgovor astrocita na mehaničku povredu, praćena je ekspresija odabranih hondroitin-sulfatnih proteoglikana (CSPG) - fosfakana, neurokana i brevikana, korišćenjem PCR u realnom vremenu i imunocitohemije. Rezultati: Dobijeni rezultati su pokazali da in vitro povreda astrocitnog jednosloja menja profile ekspresije ispitivanih CSPG. Četiri sata nakon povrede, primena PCR u realnom vremenu analize pokazala je povećanje nivoa iRNK za fosfakan (trostruko) i neurokan (dvostruko), dok je iRNK za brevikan bila smanjena na polovinu kontrolne vrednosti. Imunofluorescentni signal poreklom od fosfakana i neurokana je bio intenzivniji u astrocitima bližim mestu povrede, dok je signal za brevikan bio slab kako u kontrolnoj, tako i u ozleđenoj grupi. Zaključak: Dobijeni rezultati pokazuju da povreda izazvana grebanjem različito utiče na ekspresiju ispitivanih CSPG u astrocitima, sto ukazuju da ovaj model može biti pogodan za ispitivanje odgovora astrocita na povredu.",
journal = "Journal of Medical Biochemistry",
title = "PCR i imunocitohemijska studija ekspresije hondroitin-sulfatnih proteoglikana nakon povrede astrocita u kulturi, Real-time PCR and immunocytochemical study of chondroitin sulfate proteoglycans after scratch wounding in cultured astrocytes",
number = "4",
volume = "32",
pages = "398-405",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_507"
}

Savić, D., Parabucki, A., Bjelobaba, I., Santrač, A., Dacić, S., Peković, S.,& Stojiljković, M.. (2013). PCR i imunocitohemijska studija ekspresije hondroitin-sulfatnih proteoglikana nakon povrede astrocita u kulturi. in Journal of Medical Biochemistry, 32(4), 398-405.
https://hdl.handle.net/21.15107/rcub_ibiss_507

Savić D, Parabucki A, Bjelobaba I, Santrač A, Dacić S, Peković S, Stojiljković M. PCR i imunocitohemijska studija ekspresije hondroitin-sulfatnih proteoglikana nakon povrede astrocita u kulturi. in Journal of Medical Biochemistry. 2013;32(4):398-405.
https://hdl.handle.net/21.15107/rcub_ibiss_507 .

Savić, Danijela, Parabucki, Ana, Bjelobaba, Ivana, Santrač, Anja, Dacić, Sanja, Peković, Sanja, Stojiljković, Mirjana, "PCR i imunocitohemijska studija ekspresije hondroitin-sulfatnih proteoglikana nakon povrede astrocita u kulturi" in Journal of Medical Biochemistry, 32, no. 4 (2013):398-405,
https://hdl.handle.net/21.15107/rcub_ibiss_507 .

TY  - CONF
AU  - Parabucki, Ana
AU  - Korać, Bato
AU  - Otašević, Vesna
AU  - Bjelobaba, Ivana
AU  - Lavrnja, Irena
AU  - Stojiljković, Mirjana
AU  - Nedeljković, Nadežda
PY  - 2009
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6064
AB  - Povreda mozga obuhvata primarnu povredu, koja nastaje kao rezultat
neposrednog mehaničkog oštećenja tkiva i sekundarnu povredu koja se javlja tokom
perioda od narednih nekoliko dana. Iako još uvek nisu poznati svi detalji mehanizama
koji dovode do sekundarne povrede, nekoliko metaboličkih promena, uključujući i
povećanje produkcije reaktivnih kiseoničnih vrsta (ROS) dovedeno je u vezu sa
patofiziologijom sekundarne povrede. Budući da su mitohondrije primarno
unutarćelijsko mesto nastanka ROS, pretpostavlja se da mangan superoksid dismutaza
(MnSOD, SOD2) ima presudnu zaštitnu ulogu u antioksidativnim mehanizmima
odbrane i preživljavanju nervnih ćelija nakon povrede. Stoga je u ovom radu praćena
ekspresija MnSOD u modelu unilateralne ablacije korteksa pacova, sa ciljem da se
rasvetli njena uloga u ranim događajima nakon povrede mozga.
Svi eksperimenti izvedeni su na pacovima soja Wistar starim tri meseca.
Životinjama je pod anestezijom Zoletilom, uklonjen senzomotorni korteks na levoj
strani, pažljivim usisavanjem tkiva kroz polipropilenski vrh. Životinje se žrtvovane 0, 4,
24 i 72 časa nakon povrede i tkivo levog (LCtx) i desnog (RCtx) korteksa disecirano je i
iskorišćeno za pripremu tkivnih homogenata. Za svaku eksprimentalnu tačku, korišćena
je grupa lažno operisanih životinja, dok su neoperisane, intaktne životinje primenjene
kao fiziološka kontrola. Intenzitet signala ekspresije iRNK i proteina kod povređenih
životinja izražen je u odnosu na lažno-operisane životinje, dok je intenzitet dobijen kod
intaktnih životinja arbitrarno definisan kao 1.00.
RtPCR analiza demonstrirala je da u LCtx dolazi do rane indukcije iRNK za
MnSOD između 4 i 24 h nakon povrede. Uočeno vremenski-zavisno povećanje iRNK za
MnSOD bilo je najveće 4 sata nakon povrede. Saglasno tome, imunoblot analiza
pokazala je da je ekspresija MnSOD proteina u LCtx značajno povećana tokom prva
četiri sata nakon povrede. Ekspresija iRNK i MnSOD proteina vraća se na fiziološki
nivo 72 sata nakon povrede. S druge strane, ekspresija MnSOD, kako na nivou iRNK,
tako i na nivou proteina, ne menja se u RCtx tokom celog eksperimentalnog perioda.
Rezultati ovog rada ukazuju da povreda mozga dovodi do brzog i značajnog
povećanja ekspresije MnSOD na mestu povrede, najverovatnije kao deo odgovora na
oksidativni stres uzrokovan primarnom povredom. Budući da MnSOD predstavlja prvu
liniju odbrane od superoksid anjon radikala koji nastaju u mitohondrijama, ovi nalazi
mogu doprineti boljem razumevanju uloge MnSOD u procesu oporavka nakon povrede.
AB  - Brain injury consists of primary injury that is the result of immediate
mechanical damage and secondary injury that evolves over a period of minutes and
days. The precise mechanisms underlying secondary injury are not well understood,
however several metabolic dearangements, including increased generation of reactive
oxygen species (ROS) have been implicated in the pathophysiology following brain
damage. Since mitochondria are the major subcellular site of ROS generation,
manganese superoxid dismutase (MnSOD, SOD2), a potent scavanger of superoxide
radicals could have critical cytoprotective role in the antioxidant defence mechanism and
neuronal survival after brain damage. Thus, in the present study we have evaluated
expression of MnSOD to address its role during early events of brain injury using a
model of unilateral cortical ablation in rat.
Experiments were performed on three-month old Wistar male rats. The
sensomotory cortex was unilaterally removed on the left side by gentle suction
aspiration through polypropilene tip under the Zoletil anesthesia. Animals were
sacrificed 0, 4, 24 and 72 hours after the surgery and left (LCtx) and right (RCtx)
cortical tissues were immediately isolated for tissue homogenate preparations. For each
time point, another group of aged-matched animals was used as sham-operated controls,
whereas non-operated, intact animals were used as a physiological control. Signal
intensities obtained for MnSOD mRNA and protein expression in injured animals were
expressed relative to that obtained for sham-operated animals at each time point after the
surgery, whereas signal intensity obtained for intact control was arbitrarily defined as
1.00.
RtPCR analysis showed a rapid induction of MnSOD mRNA in LCtx between 4
and 24 h after the injury. Observed time-dependent increase in MnSOD mRNA was
maximal 4 hours after the injury compared to the level induced by sham operation alone.
Accordingly, immunoblot analysis demonstrated increased expression of MnSOD
protein in LCtx up to 4 h after the injury. 72 hours after the injury MnSOD mRNA and
protein expression return to the level of the intact control. On the other hand, MnSOD
mRNA and protein expression remained unaffected in the RCtx at all time points after
the surgery.
In conclusion, the result of this study demonstrate that brain injury induce rapid
and marked increase in MnSOD expression at the site of injury, most likely as a
protective response after oxidative stress initiated by primary brain damage. Since
MnSOD provides the first line of defense against superoxide generated in mitochondria,
these findings may contribute to a better understanding of role MnSOD in the recovery
process following brain injury.
PB  - Belgrade : Serbian Society for Mitochondrial and Free-Radical Physiology
C3  - Knjiga sažetaka: Naučni simpozijum: Mitohondrije i slobodni radikali - nov izazov; 2009 Sep 21; Belgrade, Serbia
T1  - Rano povećanje ekspresije mangan superoksid dismutaze nakon eksperimentalne povrede mozga pacova
SP  - 58
EP  - 59
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6064
ER  - 

@conference{
author = "Parabucki, Ana and Korać, Bato and Otašević, Vesna and Bjelobaba, Ivana and Lavrnja, Irena and Stojiljković, Mirjana and Nedeljković, Nadežda",
year = "2009",
abstract = "Povreda mozga obuhvata primarnu povredu, koja nastaje kao rezultat
neposrednog mehaničkog oštećenja tkiva i sekundarnu povredu koja se javlja tokom
perioda od narednih nekoliko dana. Iako još uvek nisu poznati svi detalji mehanizama
koji dovode do sekundarne povrede, nekoliko metaboličkih promena, uključujući i
povećanje produkcije reaktivnih kiseoničnih vrsta (ROS) dovedeno je u vezu sa
patofiziologijom sekundarne povrede. Budući da su mitohondrije primarno
unutarćelijsko mesto nastanka ROS, pretpostavlja se da mangan superoksid dismutaza
(MnSOD, SOD2) ima presudnu zaštitnu ulogu u antioksidativnim mehanizmima
odbrane i preživljavanju nervnih ćelija nakon povrede. Stoga je u ovom radu praćena
ekspresija MnSOD u modelu unilateralne ablacije korteksa pacova, sa ciljem da se
rasvetli njena uloga u ranim događajima nakon povrede mozga.
Svi eksperimenti izvedeni su na pacovima soja Wistar starim tri meseca.
Životinjama je pod anestezijom Zoletilom, uklonjen senzomotorni korteks na levoj
strani, pažljivim usisavanjem tkiva kroz polipropilenski vrh. Životinje se žrtvovane 0, 4,
24 i 72 časa nakon povrede i tkivo levog (LCtx) i desnog (RCtx) korteksa disecirano je i
iskorišćeno za pripremu tkivnih homogenata. Za svaku eksprimentalnu tačku, korišćena
je grupa lažno operisanih životinja, dok su neoperisane, intaktne životinje primenjene
kao fiziološka kontrola. Intenzitet signala ekspresije iRNK i proteina kod povređenih
životinja izražen je u odnosu na lažno-operisane životinje, dok je intenzitet dobijen kod
intaktnih životinja arbitrarno definisan kao 1.00.
RtPCR analiza demonstrirala je da u LCtx dolazi do rane indukcije iRNK za
MnSOD između 4 i 24 h nakon povrede. Uočeno vremenski-zavisno povećanje iRNK za
MnSOD bilo je najveće 4 sata nakon povrede. Saglasno tome, imunoblot analiza
pokazala je da je ekspresija MnSOD proteina u LCtx značajno povećana tokom prva
četiri sata nakon povrede. Ekspresija iRNK i MnSOD proteina vraća se na fiziološki
nivo 72 sata nakon povrede. S druge strane, ekspresija MnSOD, kako na nivou iRNK,
tako i na nivou proteina, ne menja se u RCtx tokom celog eksperimentalnog perioda.
Rezultati ovog rada ukazuju da povreda mozga dovodi do brzog i značajnog
povećanja ekspresije MnSOD na mestu povrede, najverovatnije kao deo odgovora na
oksidativni stres uzrokovan primarnom povredom. Budući da MnSOD predstavlja prvu
liniju odbrane od superoksid anjon radikala koji nastaju u mitohondrijama, ovi nalazi
mogu doprineti boljem razumevanju uloge MnSOD u procesu oporavka nakon povrede., Brain injury consists of primary injury that is the result of immediate
mechanical damage and secondary injury that evolves over a period of minutes and
days. The precise mechanisms underlying secondary injury are not well understood,
however several metabolic dearangements, including increased generation of reactive
oxygen species (ROS) have been implicated in the pathophysiology following brain
damage. Since mitochondria are the major subcellular site of ROS generation,
manganese superoxid dismutase (MnSOD, SOD2), a potent scavanger of superoxide
radicals could have critical cytoprotective role in the antioxidant defence mechanism and
neuronal survival after brain damage. Thus, in the present study we have evaluated
expression of MnSOD to address its role during early events of brain injury using a
model of unilateral cortical ablation in rat.
Experiments were performed on three-month old Wistar male rats. The
sensomotory cortex was unilaterally removed on the left side by gentle suction
aspiration through polypropilene tip under the Zoletil anesthesia. Animals were
sacrificed 0, 4, 24 and 72 hours after the surgery and left (LCtx) and right (RCtx)
cortical tissues were immediately isolated for tissue homogenate preparations. For each
time point, another group of aged-matched animals was used as sham-operated controls,
whereas non-operated, intact animals were used as a physiological control. Signal
intensities obtained for MnSOD mRNA and protein expression in injured animals were
expressed relative to that obtained for sham-operated animals at each time point after the
surgery, whereas signal intensity obtained for intact control was arbitrarily defined as
1.00.
RtPCR analysis showed a rapid induction of MnSOD mRNA in LCtx between 4
and 24 h after the injury. Observed time-dependent increase in MnSOD mRNA was
maximal 4 hours after the injury compared to the level induced by sham operation alone.
Accordingly, immunoblot analysis demonstrated increased expression of MnSOD
protein in LCtx up to 4 h after the injury. 72 hours after the injury MnSOD mRNA and
protein expression return to the level of the intact control. On the other hand, MnSOD
mRNA and protein expression remained unaffected in the RCtx at all time points after
the surgery.
In conclusion, the result of this study demonstrate that brain injury induce rapid
and marked increase in MnSOD expression at the site of injury, most likely as a
protective response after oxidative stress initiated by primary brain damage. Since
MnSOD provides the first line of defense against superoxide generated in mitochondria,
these findings may contribute to a better understanding of role MnSOD in the recovery
process following brain injury.",
publisher = "Belgrade : Serbian Society for Mitochondrial and Free-Radical Physiology",
journal = "Knjiga sažetaka: Naučni simpozijum: Mitohondrije i slobodni radikali - nov izazov; 2009 Sep 21; Belgrade, Serbia",
title = "Rano povećanje ekspresije mangan superoksid dismutaze nakon eksperimentalne povrede mozga pacova",
pages = "58-59",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6064"
}

Parabucki, A., Korać, B., Otašević, V., Bjelobaba, I., Lavrnja, I., Stojiljković, M.,& Nedeljković, N.. (2009). Rano povećanje ekspresije mangan superoksid dismutaze nakon eksperimentalne povrede mozga pacova. in Knjiga sažetaka: Naučni simpozijum: Mitohondrije i slobodni radikali - nov izazov; 2009 Sep 21; Belgrade, Serbia
Belgrade : Serbian Society for Mitochondrial and Free-Radical Physiology., 58-59.
https://hdl.handle.net/21.15107/rcub_ibiss_6064

Parabucki A, Korać B, Otašević V, Bjelobaba I, Lavrnja I, Stojiljković M, Nedeljković N. Rano povećanje ekspresije mangan superoksid dismutaze nakon eksperimentalne povrede mozga pacova. in Knjiga sažetaka: Naučni simpozijum: Mitohondrije i slobodni radikali - nov izazov; 2009 Sep 21; Belgrade, Serbia. 2009;:58-59.
https://hdl.handle.net/21.15107/rcub_ibiss_6064 .

Parabucki, Ana, Korać, Bato, Otašević, Vesna, Bjelobaba, Ivana, Lavrnja, Irena, Stojiljković, Mirjana, Nedeljković, Nadežda, "Rano povećanje ekspresije mangan superoksid dismutaze nakon eksperimentalne povrede mozga pacova" in Knjiga sažetaka: Naučni simpozijum: Mitohondrije i slobodni radikali - nov izazov; 2009 Sep 21; Belgrade, Serbia (2009):58-59,
https://hdl.handle.net/21.15107/rcub_ibiss_6064 .