TY  - CONF
AU  - Milošević, Katarina
AU  - Milošević, Ana
AU  - Živković, Anica
AU  - Stevanović, Ivana
AU  - Laketa, Danijela
AU  - Božić, Iva
AU  - Janjić, Marija
AU  - Bjelobaba, Ivana
AU  - Lavrnja, Irena
AU  - Savić, Danijela
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5875
AB  - Oxidative burst is a component of neuroinflammation whereby microglia-generated reactive oxygen species (ROS) either target pathogens or act as secondary messengers for microglial activation. In response to increased ROS during microglial activation, cytoprotective mechanisms are initiated primarily via Nrf2 activation and HO-1 expression. Agmatine is known to exert neuroprotective effect in vivo due to Nrf2 induction. While agmatine has been shown to activate the Nrf2/HO-1 signaling and protect macrophages from Lps-induced inflammation in vitro, its interaction with this pathway in activated microglia remains unexplored. Therefore, we sought to examine the potential of 100 μM agmatine as a pretreatment of Lps to activate Nrf2 in BV-2 microglia. In addition to cell viability, we analyzed the nuclear level of Nrf2 by Western blot and the expression of Hmox1 by PCR, as well as the protein level of HO-1. We also measured indicators of prooxidant and antioxidant activity: 4-HNE and total glutathione, respectively. Agmatine induces oxidative stress in non-stimulated microglia, as confirmed by the increase in the lipid peroxidation marker — 4-HNE, while cell viability stays preserved. Moreover, agmatine alone causes delayed Nrf2 nuclear overexpression and an increase in total glutathione content, eventually leading to an adaptive stress response. On the other hand, in Lps-stimulated microglia, agmatine prevents lipid peroxidation, readily upregulates the nuclear protein levels of Nrf2, which increases gene and protein expression of HO-1, and maintains delayed Nrf2 nuclear overexpression, resulting in increased total glutathione content associated with cytoprotection. Overall, we interpret agmatine-induced oxidative stress in non-activated microglia as triggering the adaptive response via Nrf2 and total glutathione, enabling them to cope with subsequent stressors, ie, Lps.
PB  - Belgrade: Serbian Neuroscience Society
C3  - Book of abstracts: 8th Congress of Serbian neuroscience society with international participation; 2023 May 31 - Jun 2; Belgrade, Serbia
T1  - Agmatine upregulates Nrf2/HO-1 pathway in Lps-stimulated microglia
SP  - 106
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5875
ER  - 

@conference{
author = "Milošević, Katarina and Milošević, Ana and Živković, Anica and Stevanović, Ivana and Laketa, Danijela and Božić, Iva and Janjić, Marija and Bjelobaba, Ivana and Lavrnja, Irena and Savić, Danijela",
year = "2023",
abstract = "Oxidative burst is a component of neuroinflammation whereby microglia-generated reactive oxygen species (ROS) either target pathogens or act as secondary messengers for microglial activation. In response to increased ROS during microglial activation, cytoprotective mechanisms are initiated primarily via Nrf2 activation and HO-1 expression. Agmatine is known to exert neuroprotective effect in vivo due to Nrf2 induction. While agmatine has been shown to activate the Nrf2/HO-1 signaling and protect macrophages from Lps-induced inflammation in vitro, its interaction with this pathway in activated microglia remains unexplored. Therefore, we sought to examine the potential of 100 μM agmatine as a pretreatment of Lps to activate Nrf2 in BV-2 microglia. In addition to cell viability, we analyzed the nuclear level of Nrf2 by Western blot and the expression of Hmox1 by PCR, as well as the protein level of HO-1. We also measured indicators of prooxidant and antioxidant activity: 4-HNE and total glutathione, respectively. Agmatine induces oxidative stress in non-stimulated microglia, as confirmed by the increase in the lipid peroxidation marker — 4-HNE, while cell viability stays preserved. Moreover, agmatine alone causes delayed Nrf2 nuclear overexpression and an increase in total glutathione content, eventually leading to an adaptive stress response. On the other hand, in Lps-stimulated microglia, agmatine prevents lipid peroxidation, readily upregulates the nuclear protein levels of Nrf2, which increases gene and protein expression of HO-1, and maintains delayed Nrf2 nuclear overexpression, resulting in increased total glutathione content associated with cytoprotection. Overall, we interpret agmatine-induced oxidative stress in non-activated microglia as triggering the adaptive response via Nrf2 and total glutathione, enabling them to cope with subsequent stressors, ie, Lps.",
publisher = "Belgrade: Serbian Neuroscience Society",
journal = "Book of abstracts: 8th Congress of Serbian neuroscience society with international participation; 2023 May 31 - Jun 2; Belgrade, Serbia",
title = "Agmatine upregulates Nrf2/HO-1 pathway in Lps-stimulated microglia",
pages = "106",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5875"
}

Milošević, K., Milošević, A., Živković, A., Stevanović, I., Laketa, D., Božić, I., Janjić, M., Bjelobaba, I., Lavrnja, I.,& Savić, D.. (2023). Agmatine upregulates Nrf2/HO-1 pathway in Lps-stimulated microglia. in Book of abstracts: 8th Congress of Serbian neuroscience society with international participation; 2023 May 31 - Jun 2; Belgrade, Serbia
Belgrade: Serbian Neuroscience Society., 106.
https://hdl.handle.net/21.15107/rcub_ibiss_5875

Milošević K, Milošević A, Živković A, Stevanović I, Laketa D, Božić I, Janjić M, Bjelobaba I, Lavrnja I, Savić D. Agmatine upregulates Nrf2/HO-1 pathway in Lps-stimulated microglia. in Book of abstracts: 8th Congress of Serbian neuroscience society with international participation; 2023 May 31 - Jun 2; Belgrade, Serbia. 2023;:106.
https://hdl.handle.net/21.15107/rcub_ibiss_5875 .

Milošević, Katarina, Milošević, Ana, Živković, Anica, Stevanović, Ivana, Laketa, Danijela, Božić, Iva, Janjić, Marija, Bjelobaba, Ivana, Lavrnja, Irena, Savić, Danijela, "Agmatine upregulates Nrf2/HO-1 pathway in Lps-stimulated microglia" in Book of abstracts: 8th Congress of Serbian neuroscience society with international participation; 2023 May 31 - Jun 2; Belgrade, Serbia (2023):106,
https://hdl.handle.net/21.15107/rcub_ibiss_5875 .

TY  - CONF
AU  - Dimitrijević, Dunja
AU  - Boranijašević, Sanja
AU  - Lavrnja, Irena
AU  - Adžić, Marija
AU  - Dragić, Milorad
AU  - Stekić, Anđela
AU  - Mihajlović, Katarina
AU  - Milenković, Ivan
AU  - Laketa, Danijela
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5983
AB  - In preterm infants, insufficient exposure to endogenous glucocorticoids often leads to fatal complications. Therefore, synthetic glucocorticoids (sGC) are commonly applied to pregnant women at risk of preterm delivery between the 24th and 34th week of gestation. Despite the risk of adverse neurodevelopmental effects, repeat courses are frequently given. In the auditory system, the repeated sGC treatment prolonged neural transmission time and increased auditory thresholds in Wistar rats. Purinergic signaling plays an important role in the development of the auditory system.
We investigated the effects of repeated antenatal treatment with sGC on the components of the purinergic system in the developing auditory brainstem, at postnatal days (PD) 8,14, and 20 (pre-, post-hearing onset, and juvenile stage, respectively). Pregnant C57BL/6 dams received 0.4 mg/kg dexamethasone (DEX) s.c., at gestation days (GD) 15-17 (repeated course - 3DEX), mimicking clinical treatment for three consecutive weeks. In a single treatment (1 DEX), dams received DEX at GD 15, then saline at GD16 and 17. The control group (Sh) received saline. After treatment with 3DEX, a sharp decrease in immunoreactivity for A1 receptors and P2Y1 mRNA expression was observed (in PD8-20 and PD8, respectively). Although treatment effects were not detected for P2X2 receptor, we observed a developmental increase in its mRNA expression. P2X3 receptor, as well as CD73, CD39, and NTPDase2, exhibited stable expression.
In conclusion, repeated antenatal DEX treatment induced changes in A1 and P2Y1 receptors expression in the developing auditory brainstem, suggesting adverse neurodevelopmental effects, urging for evaluation of the current protocols for antenatal sGC treatment.
PB  - Belgrade: Serbian Neuroscience Society
C3  - Book of abstracts: 8th Congress of Serbian neuroscience society with international participation; 2023 May 31 - Jun 2; Belgrade, Serbia
T1  - Developmental effects of repeated antenatal synthetic glucocorticoid treatment on purinergic signaling in the auditory brainstem
SP  - 64
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5983
ER  - 

@conference{
author = "Dimitrijević, Dunja and Boranijašević, Sanja and Lavrnja, Irena and Adžić, Marija and Dragić, Milorad and Stekić, Anđela and Mihajlović, Katarina and Milenković, Ivan and Laketa, Danijela",
year = "2023",
abstract = "In preterm infants, insufficient exposure to endogenous glucocorticoids often leads to fatal complications. Therefore, synthetic glucocorticoids (sGC) are commonly applied to pregnant women at risk of preterm delivery between the 24th and 34th week of gestation. Despite the risk of adverse neurodevelopmental effects, repeat courses are frequently given. In the auditory system, the repeated sGC treatment prolonged neural transmission time and increased auditory thresholds in Wistar rats. Purinergic signaling plays an important role in the development of the auditory system.
We investigated the effects of repeated antenatal treatment with sGC on the components of the purinergic system in the developing auditory brainstem, at postnatal days (PD) 8,14, and 20 (pre-, post-hearing onset, and juvenile stage, respectively). Pregnant C57BL/6 dams received 0.4 mg/kg dexamethasone (DEX) s.c., at gestation days (GD) 15-17 (repeated course - 3DEX), mimicking clinical treatment for three consecutive weeks. In a single treatment (1 DEX), dams received DEX at GD 15, then saline at GD16 and 17. The control group (Sh) received saline. After treatment with 3DEX, a sharp decrease in immunoreactivity for A1 receptors and P2Y1 mRNA expression was observed (in PD8-20 and PD8, respectively). Although treatment effects were not detected for P2X2 receptor, we observed a developmental increase in its mRNA expression. P2X3 receptor, as well as CD73, CD39, and NTPDase2, exhibited stable expression.
In conclusion, repeated antenatal DEX treatment induced changes in A1 and P2Y1 receptors expression in the developing auditory brainstem, suggesting adverse neurodevelopmental effects, urging for evaluation of the current protocols for antenatal sGC treatment.",
publisher = "Belgrade: Serbian Neuroscience Society",
journal = "Book of abstracts: 8th Congress of Serbian neuroscience society with international participation; 2023 May 31 - Jun 2; Belgrade, Serbia",
title = "Developmental effects of repeated antenatal synthetic glucocorticoid treatment on purinergic signaling in the auditory brainstem",
pages = "64",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5983"
}

Dimitrijević, D., Boranijašević, S., Lavrnja, I., Adžić, M., Dragić, M., Stekić, A., Mihajlović, K., Milenković, I.,& Laketa, D.. (2023). Developmental effects of repeated antenatal synthetic glucocorticoid treatment on purinergic signaling in the auditory brainstem. in Book of abstracts: 8th Congress of Serbian neuroscience society with international participation; 2023 May 31 - Jun 2; Belgrade, Serbia
Belgrade: Serbian Neuroscience Society., 64.
https://hdl.handle.net/21.15107/rcub_ibiss_5983

Dimitrijević D, Boranijašević S, Lavrnja I, Adžić M, Dragić M, Stekić A, Mihajlović K, Milenković I, Laketa D. Developmental effects of repeated antenatal synthetic glucocorticoid treatment on purinergic signaling in the auditory brainstem. in Book of abstracts: 8th Congress of Serbian neuroscience society with international participation; 2023 May 31 - Jun 2; Belgrade, Serbia. 2023;:64.
https://hdl.handle.net/21.15107/rcub_ibiss_5983 .

Dimitrijević, Dunja, Boranijašević, Sanja, Lavrnja, Irena, Adžić, Marija, Dragić, Milorad, Stekić, Anđela, Mihajlović, Katarina, Milenković, Ivan, Laketa, Danijela, "Developmental effects of repeated antenatal synthetic glucocorticoid treatment on purinergic signaling in the auditory brainstem" in Book of abstracts: 8th Congress of Serbian neuroscience society with international participation; 2023 May 31 - Jun 2; Belgrade, Serbia (2023):64,
https://hdl.handle.net/21.15107/rcub_ibiss_5983 .

TY  - JOUR
AU  - Dragić, Milorad
AU  - Mihajlovic, Katarina
AU  - Adžić, Marija
AU  - Jakovljević, Marija
AU  - Zarić Kontić, Marina
AU  - Mitrović, Nataša
AU  - Laketa, Danijela
AU  - Lavrnja, Irena
AU  - Kipp, Markus
AU  - Grković, Ivana
AU  - Nedeljkovic, Nadezda
PY  - 2022
UR  - http://journals.sagepub.com/doi/10.1177/17590914221102068
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4984
AB  - Ectonucleoside triphosphate diphosphohydrolase 2 (NTPDase2) hydrolyzes extracellular ATP to ADP, which is the ligand for P2Y1,12,13 receptors. The present study describes the distribution of NTPDase2 in adult rat brains in physiological conditions, and in hippocampal neurodegeneration induced by trimethyltin (TMT). The study also describes the regulation of NTPDase2 by inflammatory mediators in primary astrocytes and oligodendroglial cell line OLN93. In physiological conditions, NTPDase2 protein was most abundant in the hippocampus, where it was found in fibrous astrocytes and synaptic endings in the synaptic-rich hippocampal layers. In TMT-induced neurodegeneration, NTPDase2-mRNA acutely decreased at 2-dpi and then gradually recovered to the control level at 7-dpi and 21-dpi. As determined by immunohistochemistry and double immunofluorescence, the decrease was most pronounced in the dentate gyrus (DG), where NTPDase2 withdrew from the synaptic boutons in the polymorphic layer of DG, whereas the recovery of the expression was most profound in the subgranular layer. Concerning the regulation of NTPDase2 gene expression, proinflammatory cytokines IL-6, IL-1β, TNFα, and IFNγ negatively regulated the expression of NTPDase2 in OLN93 cells, while did not altering the expression in primary astrocytes. Different cell-intrinsic stressors, such as depletion of intracellular energy store, oxidative stress, endoplasmic reticulum stress, and activation of protein kinase C, also massively disturbed the expression of the NTPDase2 gene. Together, our results suggest that the expression and the activity of NTPDase2 transiently cease in neurodegeneration and brain injury, most likely as a part of the acute adaptive response designed to promote cell defense, survival, and recovery.
T2  - ASN Neuro
T1  - Expression of Ectonucleoside Triphosphate Diphosphohydrolase 2 (NTPDase2) Is Negatively Regulated Under Neuroinflammatory Conditions In Vivo and In Vitro.
VL  - 14
DO  - 10.1177/17590914221102068
SP  - 17590914221102068
ER  - 

@article{
author = "Dragić, Milorad and Mihajlovic, Katarina and Adžić, Marija and Jakovljević, Marija and Zarić Kontić, Marina and Mitrović, Nataša and Laketa, Danijela and Lavrnja, Irena and Kipp, Markus and Grković, Ivana and Nedeljkovic, Nadezda",
year = "2022",
abstract = "Ectonucleoside triphosphate diphosphohydrolase 2 (NTPDase2) hydrolyzes extracellular ATP to ADP, which is the ligand for P2Y1,12,13 receptors. The present study describes the distribution of NTPDase2 in adult rat brains in physiological conditions, and in hippocampal neurodegeneration induced by trimethyltin (TMT). The study also describes the regulation of NTPDase2 by inflammatory mediators in primary astrocytes and oligodendroglial cell line OLN93. In physiological conditions, NTPDase2 protein was most abundant in the hippocampus, where it was found in fibrous astrocytes and synaptic endings in the synaptic-rich hippocampal layers. In TMT-induced neurodegeneration, NTPDase2-mRNA acutely decreased at 2-dpi and then gradually recovered to the control level at 7-dpi and 21-dpi. As determined by immunohistochemistry and double immunofluorescence, the decrease was most pronounced in the dentate gyrus (DG), where NTPDase2 withdrew from the synaptic boutons in the polymorphic layer of DG, whereas the recovery of the expression was most profound in the subgranular layer. Concerning the regulation of NTPDase2 gene expression, proinflammatory cytokines IL-6, IL-1β, TNFα, and IFNγ negatively regulated the expression of NTPDase2 in OLN93 cells, while did not altering the expression in primary astrocytes. Different cell-intrinsic stressors, such as depletion of intracellular energy store, oxidative stress, endoplasmic reticulum stress, and activation of protein kinase C, also massively disturbed the expression of the NTPDase2 gene. Together, our results suggest that the expression and the activity of NTPDase2 transiently cease in neurodegeneration and brain injury, most likely as a part of the acute adaptive response designed to promote cell defense, survival, and recovery.",
journal = "ASN Neuro",
title = "Expression of Ectonucleoside Triphosphate Diphosphohydrolase 2 (NTPDase2) Is Negatively Regulated Under Neuroinflammatory Conditions In Vivo and In Vitro.",
volume = "14",
doi = "10.1177/17590914221102068",
pages = "17590914221102068"
}

Dragić, M., Mihajlovic, K., Adžić, M., Jakovljević, M., Zarić Kontić, M., Mitrović, N., Laketa, D., Lavrnja, I., Kipp, M., Grković, I.,& Nedeljkovic, N.. (2022). Expression of Ectonucleoside Triphosphate Diphosphohydrolase 2 (NTPDase2) Is Negatively Regulated Under Neuroinflammatory Conditions In Vivo and In Vitro.. in ASN Neuro, 14, 17590914221102068.
https://doi.org/10.1177/17590914221102068

Dragić M, Mihajlovic K, Adžić M, Jakovljević M, Zarić Kontić M, Mitrović N, Laketa D, Lavrnja I, Kipp M, Grković I, Nedeljkovic N. Expression of Ectonucleoside Triphosphate Diphosphohydrolase 2 (NTPDase2) Is Negatively Regulated Under Neuroinflammatory Conditions In Vivo and In Vitro.. in ASN Neuro. 2022;14:17590914221102068.
doi:10.1177/17590914221102068 .

Dragić, Milorad, Mihajlovic, Katarina, Adžić, Marija, Jakovljević, Marija, Zarić Kontić, Marina, Mitrović, Nataša, Laketa, Danijela, Lavrnja, Irena, Kipp, Markus, Grković, Ivana, Nedeljkovic, Nadezda, "Expression of Ectonucleoside Triphosphate Diphosphohydrolase 2 (NTPDase2) Is Negatively Regulated Under Neuroinflammatory Conditions In Vivo and In Vitro." in ASN Neuro, 14 (2022):17590914221102068,
https://doi.org/10.1177/17590914221102068 . .

TY  - JOUR
AU  - Trifunović, Svetlana
AU  - Stevanović, Ivana
AU  - Milošević, Ana
AU  - Ristić, Nataša
AU  - Janjić, Marija
AU  - Bjelobaba, Ivana
AU  - Savić, Danijela
AU  - Božić, Iva
AU  - Jakovljević, Marija
AU  - Milošević, Katarina
AU  - Laketa, Danijela
AU  - Lavrnja, Irena
PY  - 2021
UR  - https://www.frontiersin.org/articles/10.3389/fnins.2021.649485/full
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4436
AB  - Multiple sclerosis (MS) is an inflammatory, demyelinating disease with an unknown origin. Previous studies showed the involvement of the hypothalamic-pituitary-adrenal (HPA) axis to susceptibility to autoimmune diseases, including MS, and its best-characterized animal model, experimental autoimmune encephalomyelitis (EAE). During MS/EAE, innate immune cells are activated and release cytokines and other inflammatory mediators, leading to a vicious cycle of inflammation. In response to inflammation, the activated HPA axis modulates immune responses via glucocorticoid activity. Because the mechanisms involving oxidative stress to the HPA axis are relatively unrevealed, in this study, we investigate the inflammatory and oxidative stress status of HPA axis during EAE. Our results reveal an upregulation of Pomc gene expression, followed by POMC and ACTH protein increase at the peak of the EAE in the pituitary. Also, prostaglandins are well-known contributors of HPA axis activation, which increases during EAE at the periphery. The upregulated Tnf expression in the pituitary during the peak of EAE occurred. This leads to the activation of oxidative pathways, followed by upregulation of inducible NO synthase expression. The reactive oxidant/nitrosative species (ROS/RNS), such as superoxide anion and NO, increase their levels at the onset and peak of the disease in the pituitary and adrenal glands, returning to control levels at the end of EAE. The corticotrophs in the pituitary increased in number and volume at the peak of EAE that coincides with high lipid peroxidation levels. The expression of MC2R in the adrenal glands increases at the peak of EAE, where strong induction of superoxide anion and malondialdehyde (MDA), reduced total glutathione (GSH) content, and catalase activity occurred at the peak and end of EAE compared with controls. The results obtained from this study may help in understanding the mechanisms and possible pharmacological modulation in MS and demonstrate an effect of oxidative stress exposure in the HPA activation during the course of EAE.
PB  - Lausanne: Frontiers Media SA
T2  - Frontiers in Neuroscience
T1  - The Function of the Hypothalamic-Pituitary-Adrenal Axis During Experimental Autoimmune Encephalomyelitis: Involvement of Oxidative Stress Mediators.
VL  - 15
DO  - 10.3389/fnins.2021.649485
SP  - 649485
ER  - 

@article{
author = "Trifunović, Svetlana and Stevanović, Ivana and Milošević, Ana and Ristić, Nataša and Janjić, Marija and Bjelobaba, Ivana and Savić, Danijela and Božić, Iva and Jakovljević, Marija and Milošević, Katarina and Laketa, Danijela and Lavrnja, Irena",
year = "2021",
abstract = "Multiple sclerosis (MS) is an inflammatory, demyelinating disease with an unknown origin. Previous studies showed the involvement of the hypothalamic-pituitary-adrenal (HPA) axis to susceptibility to autoimmune diseases, including MS, and its best-characterized animal model, experimental autoimmune encephalomyelitis (EAE). During MS/EAE, innate immune cells are activated and release cytokines and other inflammatory mediators, leading to a vicious cycle of inflammation. In response to inflammation, the activated HPA axis modulates immune responses via glucocorticoid activity. Because the mechanisms involving oxidative stress to the HPA axis are relatively unrevealed, in this study, we investigate the inflammatory and oxidative stress status of HPA axis during EAE. Our results reveal an upregulation of Pomc gene expression, followed by POMC and ACTH protein increase at the peak of the EAE in the pituitary. Also, prostaglandins are well-known contributors of HPA axis activation, which increases during EAE at the periphery. The upregulated Tnf expression in the pituitary during the peak of EAE occurred. This leads to the activation of oxidative pathways, followed by upregulation of inducible NO synthase expression. The reactive oxidant/nitrosative species (ROS/RNS), such as superoxide anion and NO, increase their levels at the onset and peak of the disease in the pituitary and adrenal glands, returning to control levels at the end of EAE. The corticotrophs in the pituitary increased in number and volume at the peak of EAE that coincides with high lipid peroxidation levels. The expression of MC2R in the adrenal glands increases at the peak of EAE, where strong induction of superoxide anion and malondialdehyde (MDA), reduced total glutathione (GSH) content, and catalase activity occurred at the peak and end of EAE compared with controls. The results obtained from this study may help in understanding the mechanisms and possible pharmacological modulation in MS and demonstrate an effect of oxidative stress exposure in the HPA activation during the course of EAE.",
publisher = "Lausanne: Frontiers Media SA",
journal = "Frontiers in Neuroscience",
title = "The Function of the Hypothalamic-Pituitary-Adrenal Axis During Experimental Autoimmune Encephalomyelitis: Involvement of Oxidative Stress Mediators.",
volume = "15",
doi = "10.3389/fnins.2021.649485",
pages = "649485"
}

Trifunović, S., Stevanović, I., Milošević, A., Ristić, N., Janjić, M., Bjelobaba, I., Savić, D., Božić, I., Jakovljević, M., Milošević, K., Laketa, D.,& Lavrnja, I.. (2021). The Function of the Hypothalamic-Pituitary-Adrenal Axis During Experimental Autoimmune Encephalomyelitis: Involvement of Oxidative Stress Mediators.. in Frontiers in Neuroscience
Lausanne: Frontiers Media SA., 15, 649485.
https://doi.org/10.3389/fnins.2021.649485

Trifunović S, Stevanović I, Milošević A, Ristić N, Janjić M, Bjelobaba I, Savić D, Božić I, Jakovljević M, Milošević K, Laketa D, Lavrnja I. The Function of the Hypothalamic-Pituitary-Adrenal Axis During Experimental Autoimmune Encephalomyelitis: Involvement of Oxidative Stress Mediators.. in Frontiers in Neuroscience. 2021;15:649485.
doi:10.3389/fnins.2021.649485 .

Trifunović, Svetlana, Stevanović, Ivana, Milošević, Ana, Ristić, Nataša, Janjić, Marija, Bjelobaba, Ivana, Savić, Danijela, Božić, Iva, Jakovljević, Marija, Milošević, Katarina, Laketa, Danijela, Lavrnja, Irena, "The Function of the Hypothalamic-Pituitary-Adrenal Axis During Experimental Autoimmune Encephalomyelitis: Involvement of Oxidative Stress Mediators." in Frontiers in Neuroscience, 15 (2021):649485,
https://doi.org/10.3389/fnins.2021.649485 . .

TY  - CONF
AU  - Jakovljević, Marija
AU  - Lavrnja, Irena
AU  - Božić, Iva
AU  - Milošević, Ana
AU  - Bjelobaba, Ivana
AU  - Savić, Danijela
AU  - Peković, Sanja
AU  - Nedeljković, Nadežda
AU  - Laketa, Danijela
PY  - 2019
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5982
AB  - Considering neuroinflammatory paradigm, increased extracellular levels of ATP have adverse effects, while adenosine is predominantly anti-inflammatory. In the CNS, NTPDase1/CD39 is the main enzyme that initiates the degradation pathway of extracellular ATP to adenosine. The aim of the study was to explore the activation state of the cells that express NTPDase1/CD39 – microglia and macrophages, during experimental autoimmune encephalomyelitis (EAE). Acute monophasic EAE was induced in female Dark Agouti rats. Animals were sacrificed at the disease onset (Eo), peak (Ep) and end (Ee). The lumbosacral parts of spinal cords were isolated for gene (qRT-PCR and in situ hybridization) and protein expression analysis (Western Blot, immunofluorescence and flow cytometry). Activation state of microglia/macrophages was assessed by colocalization analysis of NTPDase1/Iba1 and NTPDase1/CD68 with iNOS or Arg1 as specific markers of pro- and antiinflammatory state, respectively. During EAE, NTPDase1/CD39 was significantly increased both at mRNA and protein level at Ep. Immunofluorescence combined with flow cytometry showed that reactive microglia and mononuclear infiltrates accounted for the most of the observed increase. Both Iba1 and CD68 microglia/macrophage markers showed higher co-occurrence with iNOS at Eo and Arg1 at Ep, suggesting prevalence of M1-like at Eo and M2-like at Ep. In addition, NTPDase1 showed about three-times higher colocalization with Arg1 compared to iNOS at Ep, suggesting its higher association with M2-like activation state of microglia/ macrophages. Together, our data suggest an association between NTPDase1 up-regulation by reactive microglia and infiltrated macrophages and their transition toward anti-inflammatory phenotype in EAE.
PB  - Belgrade: Serbian Neuroscience Society
C3  - Book of Abstract: Federation of European Neuroscience Societies (FENS) Regional Meeting; 2019 Jul 10-13; Belgrade, Serbia
T1  - NTPDase1/CD39 Expression Increased During EAE in Association with Number and Activation State of Microglia/Macrophages
SP  - 492
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5982
ER  - 

@conference{
author = "Jakovljević, Marija and Lavrnja, Irena and Božić, Iva and Milošević, Ana and Bjelobaba, Ivana and Savić, Danijela and Peković, Sanja and Nedeljković, Nadežda and Laketa, Danijela",
year = "2019",
abstract = "Considering neuroinflammatory paradigm, increased extracellular levels of ATP have adverse effects, while adenosine is predominantly anti-inflammatory. In the CNS, NTPDase1/CD39 is the main enzyme that initiates the degradation pathway of extracellular ATP to adenosine. The aim of the study was to explore the activation state of the cells that express NTPDase1/CD39 – microglia and macrophages, during experimental autoimmune encephalomyelitis (EAE). Acute monophasic EAE was induced in female Dark Agouti rats. Animals were sacrificed at the disease onset (Eo), peak (Ep) and end (Ee). The lumbosacral parts of spinal cords were isolated for gene (qRT-PCR and in situ hybridization) and protein expression analysis (Western Blot, immunofluorescence and flow cytometry). Activation state of microglia/macrophages was assessed by colocalization analysis of NTPDase1/Iba1 and NTPDase1/CD68 with iNOS or Arg1 as specific markers of pro- and antiinflammatory state, respectively. During EAE, NTPDase1/CD39 was significantly increased both at mRNA and protein level at Ep. Immunofluorescence combined with flow cytometry showed that reactive microglia and mononuclear infiltrates accounted for the most of the observed increase. Both Iba1 and CD68 microglia/macrophage markers showed higher co-occurrence with iNOS at Eo and Arg1 at Ep, suggesting prevalence of M1-like at Eo and M2-like at Ep. In addition, NTPDase1 showed about three-times higher colocalization with Arg1 compared to iNOS at Ep, suggesting its higher association with M2-like activation state of microglia/ macrophages. Together, our data suggest an association between NTPDase1 up-regulation by reactive microglia and infiltrated macrophages and their transition toward anti-inflammatory phenotype in EAE.",
publisher = "Belgrade: Serbian Neuroscience Society",
journal = "Book of Abstract: Federation of European Neuroscience Societies (FENS) Regional Meeting; 2019 Jul 10-13; Belgrade, Serbia",
title = "NTPDase1/CD39 Expression Increased During EAE in Association with Number and Activation State of Microglia/Macrophages",
pages = "492",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5982"
}

Jakovljević, M., Lavrnja, I., Božić, I., Milošević, A., Bjelobaba, I., Savić, D., Peković, S., Nedeljković, N.,& Laketa, D.. (2019). NTPDase1/CD39 Expression Increased During EAE in Association with Number and Activation State of Microglia/Macrophages. in Book of Abstract: Federation of European Neuroscience Societies (FENS) Regional Meeting; 2019 Jul 10-13; Belgrade, Serbia
Belgrade: Serbian Neuroscience Society., 492.
https://hdl.handle.net/21.15107/rcub_ibiss_5982

Jakovljević M, Lavrnja I, Božić I, Milošević A, Bjelobaba I, Savić D, Peković S, Nedeljković N, Laketa D. NTPDase1/CD39 Expression Increased During EAE in Association with Number and Activation State of Microglia/Macrophages. in Book of Abstract: Federation of European Neuroscience Societies (FENS) Regional Meeting; 2019 Jul 10-13; Belgrade, Serbia. 2019;:492.
https://hdl.handle.net/21.15107/rcub_ibiss_5982 .

Jakovljević, Marija, Lavrnja, Irena, Božić, Iva, Milošević, Ana, Bjelobaba, Ivana, Savić, Danijela, Peković, Sanja, Nedeljković, Nadežda, Laketa, Danijela, "NTPDase1/CD39 Expression Increased During EAE in Association with Number and Activation State of Microglia/Macrophages" in Book of Abstract: Federation of European Neuroscience Societies (FENS) Regional Meeting; 2019 Jul 10-13; Belgrade, Serbia (2019):492,
https://hdl.handle.net/21.15107/rcub_ibiss_5982 .

TY  - CONF
AU  - Laketa, Danijela
AU  - Jakovljević, Marija
AU  - Božić, Iva
AU  - Bjelobaba, Ivana
AU  - Savić, Danijela
AU  - Peković, Sanja
AU  - Nedeljković, Nadežda
AU  - Lavrnja, Irena
PY  - 2019
UR  - https://biore.bio.bg.ac.rs/handle/123456789/2265
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5893
AB  - Ectonucleoside triphosphate diphosphohydrolase 1 (NTPDase1/CD39) is the main ATP- and ADPdegrading
enzyme in extracellular fluid of the central nervous system. In the hydrolysis cascade
NTPDase1 removes ATP and ADP and produces AMP, which is hydrolysed by ecto-5'-nucleotidase to
adenosine. During neuroinflammation, increased extracellular ATP levels exert proinflammatory effects
at microglia as resident immune cells, while adenosine effects are antiinflammatory. Literature data
indicate involvement of purinergic signaling in experimental autoimmune encephalomyelitis (EAE), while
decreased number of NTPDase1/CD39+ regulatory T-cells was evidenced in multiple sclerosis.
Downregulation of NTPDase1 expression was observed in proinflammatory activation phenotype of
macrophages. However, data on the role of NTPDase1 on glial cells in neuroinflammation are still
scarce. We have shown increase in ATP-, ADP- and AMP-hydrolysis, together with upregulated mRNA
and protein expression of NTPDase1 in lumbar spinal cord, correlated to the disease course during EAE.
In this study we aimed to explore contribution of particular cell subsets to the observed changes in
NTPDase1 expression.
Acute monophasic EAE was induced in female rats of Dark Agouti strain by active immunization with a
mixture of spinal cord homogenate in complete Freund’s adjuvant. Immunized animals were sacrificed at
the onset, peak and end of symptoms, while naïve animals were used as control. Significant increase of
NTPDase1 immunofluorescence in lumbar spinal cord cross-sections was related to prominent infiltrates
at the peak of EAE and increased expression of NTPDase1 among isolated mononuclear cells. Analysis
of triple-labeled Arginase1/NTPDase1/Iba1 and iNOS/NTPDase1/Iba1 immunofluorescent micrographs
showed prevalent contribution of Arginase1+ microglia in comparison to iNOS+ microglia in NTPDase1
immunofluorescence, at the peak of EAE. Further studies are needed to reveal possible association of
NTPDase1 with antiinflammatory phenotype in microglia.
PB  - German Neuroscience Society
C3  - Proceedings: 13th Göttingen Meeting of the German Neuroscience Society 2019 and 37th Göttingen Neurobiology Conference; 2019 Mar 20-23; Göttingen, Germany
T1  - Microglia-related increase in NTPDase1 expression during EAE
SP  - T12-5B
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5893
ER  - 

@conference{
author = "Laketa, Danijela and Jakovljević, Marija and Božić, Iva and Bjelobaba, Ivana and Savić, Danijela and Peković, Sanja and Nedeljković, Nadežda and Lavrnja, Irena",
year = "2019",
abstract = "Ectonucleoside triphosphate diphosphohydrolase 1 (NTPDase1/CD39) is the main ATP- and ADPdegrading
enzyme in extracellular fluid of the central nervous system. In the hydrolysis cascade
NTPDase1 removes ATP and ADP and produces AMP, which is hydrolysed by ecto-5'-nucleotidase to
adenosine. During neuroinflammation, increased extracellular ATP levels exert proinflammatory effects
at microglia as resident immune cells, while adenosine effects are antiinflammatory. Literature data
indicate involvement of purinergic signaling in experimental autoimmune encephalomyelitis (EAE), while
decreased number of NTPDase1/CD39+ regulatory T-cells was evidenced in multiple sclerosis.
Downregulation of NTPDase1 expression was observed in proinflammatory activation phenotype of
macrophages. However, data on the role of NTPDase1 on glial cells in neuroinflammation are still
scarce. We have shown increase in ATP-, ADP- and AMP-hydrolysis, together with upregulated mRNA
and protein expression of NTPDase1 in lumbar spinal cord, correlated to the disease course during EAE.
In this study we aimed to explore contribution of particular cell subsets to the observed changes in
NTPDase1 expression.
Acute monophasic EAE was induced in female rats of Dark Agouti strain by active immunization with a
mixture of spinal cord homogenate in complete Freund’s adjuvant. Immunized animals were sacrificed at
the onset, peak and end of symptoms, while naïve animals were used as control. Significant increase of
NTPDase1 immunofluorescence in lumbar spinal cord cross-sections was related to prominent infiltrates
at the peak of EAE and increased expression of NTPDase1 among isolated mononuclear cells. Analysis
of triple-labeled Arginase1/NTPDase1/Iba1 and iNOS/NTPDase1/Iba1 immunofluorescent micrographs
showed prevalent contribution of Arginase1+ microglia in comparison to iNOS+ microglia in NTPDase1
immunofluorescence, at the peak of EAE. Further studies are needed to reveal possible association of
NTPDase1 with antiinflammatory phenotype in microglia.",
publisher = "German Neuroscience Society",
journal = "Proceedings: 13th Göttingen Meeting of the German Neuroscience Society 2019 and 37th Göttingen Neurobiology Conference; 2019 Mar 20-23; Göttingen, Germany",
title = "Microglia-related increase in NTPDase1 expression during EAE",
pages = "T12-5B",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5893"
}

Laketa, D., Jakovljević, M., Božić, I., Bjelobaba, I., Savić, D., Peković, S., Nedeljković, N.,& Lavrnja, I.. (2019). Microglia-related increase in NTPDase1 expression during EAE. in Proceedings: 13th Göttingen Meeting of the German Neuroscience Society 2019 and 37th Göttingen Neurobiology Conference; 2019 Mar 20-23; Göttingen, Germany
German Neuroscience Society., T12-5B.
https://hdl.handle.net/21.15107/rcub_ibiss_5893

Laketa D, Jakovljević M, Božić I, Bjelobaba I, Savić D, Peković S, Nedeljković N, Lavrnja I. Microglia-related increase in NTPDase1 expression during EAE. in Proceedings: 13th Göttingen Meeting of the German Neuroscience Society 2019 and 37th Göttingen Neurobiology Conference; 2019 Mar 20-23; Göttingen, Germany. 2019;:T12-5B.
https://hdl.handle.net/21.15107/rcub_ibiss_5893 .

Laketa, Danijela, Jakovljević, Marija, Božić, Iva, Bjelobaba, Ivana, Savić, Danijela, Peković, Sanja, Nedeljković, Nadežda, Lavrnja, Irena, "Microglia-related increase in NTPDase1 expression during EAE" in Proceedings: 13th Göttingen Meeting of the German Neuroscience Society 2019 and 37th Göttingen Neurobiology Conference; 2019 Mar 20-23; Göttingen, Germany (2019):T12-5B,
https://hdl.handle.net/21.15107/rcub_ibiss_5893 .

TY  - CONF
AU  - Laketa, Danijela
AU  - Josipović, Nataša
AU  - Lavrnja, Irena
AU  - Bjelobaba, Ivana
AU  - Jakovljević, Marija
AU  - Božić, Iva
AU  - Savić, Danijela
AU  - Dacić, Sanja
AU  - Peković, Sanja
AU  - Nedeljković, Nadežda
PY  - 2017
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5988
AB  - Introduction. Ecto-5'–nucleotidase (eN) catalyzes terminal step of extracellular ATP hydrolysis, producing anti-inflammatory adenosine. We reported significantly increased eN activity in lumbar spinal cord during experimental autoimmune encephalomyelitis (EAE), together with increased protein expression connected mainly with reactive astrocytes and appearance of new isoform at ~75kDa at the peak of the disease, besides usual ~71kDa isoform. Since eN is glycoprotein with five potential N-glycosylation sites and 
redicted molecular weight of 57-59 kDa, we hypothesized that occurrence of second isoform during EAE is due to changes in glycosylation pattern, possibly affecting kinetic properties of the enzyme. Methods. Lumbar parts of the spinal cords were obtained from Dark Agouti rats at the onset (Eo), peak (Ep) and the end of symptoms (Er) during EAE and from naïve control animals (C). Results. We here report significant changes of kinetic properties regarding AMP-hydrolysis during EAE, with almost 50% increase of maximal velocity at Ep (92.35±1.86nmolPi/min/mg) and Er (90.68±2.17nmolPi/min/mg), compared to C, whilst Km increased double at Ep (0.041±0.003mmol/l). Enzymatic deglycosylation caused triple decrease of Vmax (33.6±1.8nmolPi/mg/min) at Ep, and double decrease of Km (0.022±0.008mmol/l), whilst immunoblot
probed with anti-eN antibody revealed triple protein band at ~60kDa at all investigated time-points. Conclusion. Our results show that changes of kinetic properties during EAE, at least partially, are governed by modification of glycosylation pattern. Also, appearance of new isoform at the peak of EAE is direct consequence of glycosylation changes. In summary, besides gene and protein expression changes of eN, glycosylation might be additional route of inflammation control conducted by astrocytes.
PB  - Belgrade: Serbian Neuroscience Society
C3  - Book of Abstract: 7th Congress of Serbian Neuroscience Society with international participation; 2017 Oct 25-27; Belgrade, Serbia
T1  - Appearance of second ecto-5'-nucleotidase isoform during experimental autoimmune encephalomyelitis is caused by changes in glycosylation pattern
SP  - 70
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5988
ER  - 

@conference{
author = "Laketa, Danijela and Josipović, Nataša and Lavrnja, Irena and Bjelobaba, Ivana and Jakovljević, Marija and Božić, Iva and Savić, Danijela and Dacić, Sanja and Peković, Sanja and Nedeljković, Nadežda",
year = "2017",
abstract = "Introduction. Ecto-5'–nucleotidase (eN) catalyzes terminal step of extracellular ATP hydrolysis, producing anti-inflammatory adenosine. We reported significantly increased eN activity in lumbar spinal cord during experimental autoimmune encephalomyelitis (EAE), together with increased protein expression connected mainly with reactive astrocytes and appearance of new isoform at ~75kDa at the peak of the disease, besides usual ~71kDa isoform. Since eN is glycoprotein with five potential N-glycosylation sites and 
redicted molecular weight of 57-59 kDa, we hypothesized that occurrence of second isoform during EAE is due to changes in glycosylation pattern, possibly affecting kinetic properties of the enzyme. Methods. Lumbar parts of the spinal cords were obtained from Dark Agouti rats at the onset (Eo), peak (Ep) and the end of symptoms (Er) during EAE and from naïve control animals (C). Results. We here report significant changes of kinetic properties regarding AMP-hydrolysis during EAE, with almost 50% increase of maximal velocity at Ep (92.35±1.86nmolPi/min/mg) and Er (90.68±2.17nmolPi/min/mg), compared to C, whilst Km increased double at Ep (0.041±0.003mmol/l). Enzymatic deglycosylation caused triple decrease of Vmax (33.6±1.8nmolPi/mg/min) at Ep, and double decrease of Km (0.022±0.008mmol/l), whilst immunoblot
probed with anti-eN antibody revealed triple protein band at ~60kDa at all investigated time-points. Conclusion. Our results show that changes of kinetic properties during EAE, at least partially, are governed by modification of glycosylation pattern. Also, appearance of new isoform at the peak of EAE is direct consequence of glycosylation changes. In summary, besides gene and protein expression changes of eN, glycosylation might be additional route of inflammation control conducted by astrocytes.",
publisher = "Belgrade: Serbian Neuroscience Society",
journal = "Book of Abstract: 7th Congress of Serbian Neuroscience Society with international participation; 2017 Oct 25-27; Belgrade, Serbia",
title = "Appearance of second ecto-5'-nucleotidase isoform during experimental autoimmune encephalomyelitis is caused by changes in glycosylation pattern",
pages = "70",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5988"
}

Laketa, D., Josipović, N., Lavrnja, I., Bjelobaba, I., Jakovljević, M., Božić, I., Savić, D., Dacić, S., Peković, S.,& Nedeljković, N.. (2017). Appearance of second ecto-5'-nucleotidase isoform during experimental autoimmune encephalomyelitis is caused by changes in glycosylation pattern. in Book of Abstract: 7th Congress of Serbian Neuroscience Society with international participation; 2017 Oct 25-27; Belgrade, Serbia
Belgrade: Serbian Neuroscience Society., 70.
https://hdl.handle.net/21.15107/rcub_ibiss_5988

Laketa D, Josipović N, Lavrnja I, Bjelobaba I, Jakovljević M, Božić I, Savić D, Dacić S, Peković S, Nedeljković N. Appearance of second ecto-5'-nucleotidase isoform during experimental autoimmune encephalomyelitis is caused by changes in glycosylation pattern. in Book of Abstract: 7th Congress of Serbian Neuroscience Society with international participation; 2017 Oct 25-27; Belgrade, Serbia. 2017;:70.
https://hdl.handle.net/21.15107/rcub_ibiss_5988 .

Laketa, Danijela, Josipović, Nataša, Lavrnja, Irena, Bjelobaba, Ivana, Jakovljević, Marija, Božić, Iva, Savić, Danijela, Dacić, Sanja, Peković, Sanja, Nedeljković, Nadežda, "Appearance of second ecto-5'-nucleotidase isoform during experimental autoimmune encephalomyelitis is caused by changes in glycosylation pattern" in Book of Abstract: 7th Congress of Serbian Neuroscience Society with international participation; 2017 Oct 25-27; Belgrade, Serbia (2017):70,
https://hdl.handle.net/21.15107/rcub_ibiss_5988 .

TY  - CONF
AU  - Jakovljević, Marija
AU  - Lavrnja, Irena
AU  - Božić, Iva
AU  - Adžić, Marija
AU  - Bjelobaba, Ivana
AU  - Savić, Danijela
AU  - Peković, Sanja
AU  - Nedeljković, Nadežda
AU  - Laketa, Danijela
PY  - 2017
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5985
AB  - Introduction. Multiple sclerosis (MS) is a chronic disease of central nervous system (CNS), characterized by neuroinflammation, demyelination and neurodegeneration. Despite well-established role of purinergic signaling in MS pathology, the role of ADP as ectonucleotidase inter-product between proinflammatory ATP and anti-inflammatory adenosine is still obscure. Among ADP-sensitive receptors P2Y1, P2Y12 and P2Y13, there are few data indicating involvement of P2Y12 receptor in MS. The aim of this study was to elucidate a potential impact of ADP on CNS pathology in animal model of MS - experimental autoimmune encephalomyelitis (EAE). Material and Methods. EAE was induced in 8-week old female rats of Dark Agouti strain. The abundance and localization of ADP receptors – P2Y1, P2Y12 and P2Y13 was analyzed in lumbosacral spinal cord tissue by real-time PCR, Western Blot and immunohistochemistry at different disease stages – onset (Eo), peak (Ep) and recovery (Er). Results. Results of this study show that ADP-sensing purinergic receptors are differentially regulated during EAE. Namely, mRNA and protein expression of P2Y1 and P2Y12was decreased at Eo, while significantly increased for P2Y12and P2Y13 at Ep and/or Er. In addition, immunohistochemical labeling showed similar pattern of changes during EAE for investigated receptors, thus providing novel insight into their spinal cord tissue distribution. Conclusion. Our results strongly indicate involvement of ADP-sensitive purinergic receptors P2Y1, P2Y12 and P2Y13 in pathophysiology of EAE. Their differential regulation and tissue distribution implies diverse effects in the course of the disease during EAE. Further studies would be necessary to elucidate their mechanisms of action.
PB  - Belgrade: Serbian Neuroscience Society
C3  - Book of Abstract: 7th Congress of Serbian Neuroscience Society with international participation; 2017 Oct 25-27; Belgrade, Serbia
T1  - ADP receptors P2Y1, P2Y12 and P2Y13 are differentially regulated in a rat model of multiple sclerosis
SP  - 68
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5985
ER  - 

@conference{
author = "Jakovljević, Marija and Lavrnja, Irena and Božić, Iva and Adžić, Marija and Bjelobaba, Ivana and Savić, Danijela and Peković, Sanja and Nedeljković, Nadežda and Laketa, Danijela",
year = "2017",
abstract = "Introduction. Multiple sclerosis (MS) is a chronic disease of central nervous system (CNS), characterized by neuroinflammation, demyelination and neurodegeneration. Despite well-established role of purinergic signaling in MS pathology, the role of ADP as ectonucleotidase inter-product between proinflammatory ATP and anti-inflammatory adenosine is still obscure. Among ADP-sensitive receptors P2Y1, P2Y12 and P2Y13, there are few data indicating involvement of P2Y12 receptor in MS. The aim of this study was to elucidate a potential impact of ADP on CNS pathology in animal model of MS - experimental autoimmune encephalomyelitis (EAE). Material and Methods. EAE was induced in 8-week old female rats of Dark Agouti strain. The abundance and localization of ADP receptors – P2Y1, P2Y12 and P2Y13 was analyzed in lumbosacral spinal cord tissue by real-time PCR, Western Blot and immunohistochemistry at different disease stages – onset (Eo), peak (Ep) and recovery (Er). Results. Results of this study show that ADP-sensing purinergic receptors are differentially regulated during EAE. Namely, mRNA and protein expression of P2Y1 and P2Y12was decreased at Eo, while significantly increased for P2Y12and P2Y13 at Ep and/or Er. In addition, immunohistochemical labeling showed similar pattern of changes during EAE for investigated receptors, thus providing novel insight into their spinal cord tissue distribution. Conclusion. Our results strongly indicate involvement of ADP-sensitive purinergic receptors P2Y1, P2Y12 and P2Y13 in pathophysiology of EAE. Their differential regulation and tissue distribution implies diverse effects in the course of the disease during EAE. Further studies would be necessary to elucidate their mechanisms of action.",
publisher = "Belgrade: Serbian Neuroscience Society",
journal = "Book of Abstract: 7th Congress of Serbian Neuroscience Society with international participation; 2017 Oct 25-27; Belgrade, Serbia",
title = "ADP receptors P2Y1, P2Y12 and P2Y13 are differentially regulated in a rat model of multiple sclerosis",
pages = "68",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5985"
}

Jakovljević, M., Lavrnja, I., Božić, I., Adžić, M., Bjelobaba, I., Savić, D., Peković, S., Nedeljković, N.,& Laketa, D.. (2017). ADP receptors P2Y1, P2Y12 and P2Y13 are differentially regulated in a rat model of multiple sclerosis. in Book of Abstract: 7th Congress of Serbian Neuroscience Society with international participation; 2017 Oct 25-27; Belgrade, Serbia
Belgrade: Serbian Neuroscience Society., 68.
https://hdl.handle.net/21.15107/rcub_ibiss_5985

Jakovljević M, Lavrnja I, Božić I, Adžić M, Bjelobaba I, Savić D, Peković S, Nedeljković N, Laketa D. ADP receptors P2Y1, P2Y12 and P2Y13 are differentially regulated in a rat model of multiple sclerosis. in Book of Abstract: 7th Congress of Serbian Neuroscience Society with international participation; 2017 Oct 25-27; Belgrade, Serbia. 2017;:68.
https://hdl.handle.net/21.15107/rcub_ibiss_5985 .

Jakovljević, Marija, Lavrnja, Irena, Božić, Iva, Adžić, Marija, Bjelobaba, Ivana, Savić, Danijela, Peković, Sanja, Nedeljković, Nadežda, Laketa, Danijela, "ADP receptors P2Y1, P2Y12 and P2Y13 are differentially regulated in a rat model of multiple sclerosis" in Book of Abstract: 7th Congress of Serbian Neuroscience Society with international participation; 2017 Oct 25-27; Belgrade, Serbia (2017):68,
https://hdl.handle.net/21.15107/rcub_ibiss_5985 .

TY  - JOUR
AU  - Adžić, Marija
AU  - Stevanović, Ivana
AU  - Josipović, Nataša
AU  - Laketa, Danijela
AU  - Lavrnja, Irena
AU  - Bjelobaba, Ivana
AU  - Božić, Iva
AU  - Jovanović, Marija
AU  - Milošević, Milena
AU  - Nedeljković, Nadežda
PY  - 2017
UR  - http://doi.wiley.com/10.1002/jnr.23950
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2560
AB  - It is widely accepted that adenosine triphosphate (ATP) acts as a universal danger-associated molecular pattern with several known mechanisms for immune cell activation. In the central nervous system, ATP activates microglia and astrocytes and induces a neuroinflammatory response. The aim of the present study was to describe responses of isolated astrocytes to increasing concentrations of ATP (5 µM to 1 mM), which were intended to mimic graded intensity of the extracellular stimulus. The results show that ATP induces graded activation response of astrocytes in terms of the cell proliferation, stellation, shape remodeling, and underlying actin and GFAP filament rearrangement, although the changes occurred without an apparent increase in GFAP and actin protein expression. On the other hand, ATP in the range of applied concentrations did not evoke IL-1β release from cultured astrocytes, nor did it modify the release from LPS and LPS+IFN-γ–primed astrocytes. ATP did not promote astrocyte migration in the wound-healing assay, nor did it increase production of reactive oxygen and nitrogen species and lipid peroxidation. Instead, ATP strengthened the antioxidative defense of astrocytes by inducing Cu/ZnSOD and MnSOD activities and by increasing their glutathione content. Our current results suggest that although ATP triggers several attributes of activated astrocytic phenotype with a magnitude that increases with the concentration, it is not sufficient to induce full-blown reactive phenotype of astrocytes in vitro. © 2016 Wiley Periodicals, Inc.
T2  - Journal of Neuroscience Research
T1  - Extracellular ATP induces graded reactive response of astrocytes and strengthens their antioxidative defense in vitro
IS  - 4
VL  - 95
DO  - 10.1002/jnr.23950
SP  - 1053
EP  - 1066
ER  - 

@article{
author = "Adžić, Marija and Stevanović, Ivana and Josipović, Nataša and Laketa, Danijela and Lavrnja, Irena and Bjelobaba, Ivana and Božić, Iva and Jovanović, Marija and Milošević, Milena and Nedeljković, Nadežda",
year = "2017",
abstract = "It is widely accepted that adenosine triphosphate (ATP) acts as a universal danger-associated molecular pattern with several known mechanisms for immune cell activation. In the central nervous system, ATP activates microglia and astrocytes and induces a neuroinflammatory response. The aim of the present study was to describe responses of isolated astrocytes to increasing concentrations of ATP (5 µM to 1 mM), which were intended to mimic graded intensity of the extracellular stimulus. The results show that ATP induces graded activation response of astrocytes in terms of the cell proliferation, stellation, shape remodeling, and underlying actin and GFAP filament rearrangement, although the changes occurred without an apparent increase in GFAP and actin protein expression. On the other hand, ATP in the range of applied concentrations did not evoke IL-1β release from cultured astrocytes, nor did it modify the release from LPS and LPS+IFN-γ–primed astrocytes. ATP did not promote astrocyte migration in the wound-healing assay, nor did it increase production of reactive oxygen and nitrogen species and lipid peroxidation. Instead, ATP strengthened the antioxidative defense of astrocytes by inducing Cu/ZnSOD and MnSOD activities and by increasing their glutathione content. Our current results suggest that although ATP triggers several attributes of activated astrocytic phenotype with a magnitude that increases with the concentration, it is not sufficient to induce full-blown reactive phenotype of astrocytes in vitro. © 2016 Wiley Periodicals, Inc.",
journal = "Journal of Neuroscience Research",
title = "Extracellular ATP induces graded reactive response of astrocytes and strengthens their antioxidative defense in vitro",
number = "4",
volume = "95",
doi = "10.1002/jnr.23950",
pages = "1053-1066"
}

Adžić, M., Stevanović, I., Josipović, N., Laketa, D., Lavrnja, I., Bjelobaba, I., Božić, I., Jovanović, M., Milošević, M.,& Nedeljković, N.. (2017). Extracellular ATP induces graded reactive response of astrocytes and strengthens their antioxidative defense in vitro. in Journal of Neuroscience Research, 95(4), 1053-1066.
https://doi.org/10.1002/jnr.23950

Adžić M, Stevanović I, Josipović N, Laketa D, Lavrnja I, Bjelobaba I, Božić I, Jovanović M, Milošević M, Nedeljković N. Extracellular ATP induces graded reactive response of astrocytes and strengthens their antioxidative defense in vitro. in Journal of Neuroscience Research. 2017;95(4):1053-1066.
doi:10.1002/jnr.23950 .

Adžić, Marija, Stevanović, Ivana, Josipović, Nataša, Laketa, Danijela, Lavrnja, Irena, Bjelobaba, Ivana, Božić, Iva, Jovanović, Marija, Milošević, Milena, Nedeljković, Nadežda, "Extracellular ATP induces graded reactive response of astrocytes and strengthens their antioxidative defense in vitro" in Journal of Neuroscience Research, 95, no. 4 (2017):1053-1066,
https://doi.org/10.1002/jnr.23950 . .

TY  - CONF
AU  - Božić, Iva
AU  - Savić, Danijela
AU  - Laketa, Danijela
AU  - Nedeljković, Nadežda
AU  - Peković, Sanja
AU  - Lavrnja, Irena
PY  - 2015
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6059
AB  - Microglial cells are immune cells of the central nervous system (CNS) that
play a major role in its surveillance. Changes in CNS homeostasis, invading
pathogens or neuron impairment, lead to activation of microglial cells that
quickly proliferate, acquire amoeboid morphology and produce toxic
mediators such as nitric oxide (NO) and pro-inflammatory cytokines. These
changes are regulated by transcription factors, most importantly NF-κB.
Although microglial activation is important for maintaining tissue
homeostasis, excessive activation leads to chronic inflammation that can
damage healthy neurons. Substances that suppress microglial activation are
potential therapeutics for neurodegenerative diseases. Benfotiamine (Sbenzoylthiamine-
O-monophosphate) is a synthetic derivative of vitamin B1
that has anti-inflammatory properties. In this study we investigated antiinflammatory
properties of benfotiamine on activated microglia in vitro.
BV-2 microglia were pre-treated with benfotiamine, stimulated with LPS
and their viability and morphology were evaluated. LPS induced prominent
alterations in cell morphology, enlargement of cell bodies and spreading of
multiple processes. Pre-treatment with benfotiamine before LPS
stimulation suppressed these morphological changes. Additionally,
benfotiamine diminished LPS induced NO production, without altering cell
viability. Furthermore, benfotiamine decreased LPS induced production of
pro-inflammatory cytokines TNF- α and IL-6, while increasing production
of anti-inflammatory cytokine IL-10. Analysis of NF-κB activation
revealed that benfotiamine's effects were mediated by this transcription
factor. In conclusion, benfotiamine exerts anti-inflammatory properties in LPS
activated microglia in vitro and should be further investigated as a potential
therapeutic for neurodegenerative diseases.
PB  - Belgrade: Immunological Society of Serbia
C3  - 3rd Belgrade EFIS symposium on Immunoregulation: Immunity, Infection, Autoimmunity and Aging; 2015 May 24-27; Arandjelovac, Serbia
T1  - Anti-inflammatory effects of benfotiamine in LPS stimulated microglia: role of NF-κB signaling
SP  - 71
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6059
ER  - 

@conference{
editor = "Lukić, Miodrag, Jonjic, Stipan, Nikolich-Zugich, Janko",
author = "Božić, Iva and Savić, Danijela and Laketa, Danijela and Nedeljković, Nadežda and Peković, Sanja and Lavrnja, Irena",
year = "2015",
abstract = "Microglial cells are immune cells of the central nervous system (CNS) that
play a major role in its surveillance. Changes in CNS homeostasis, invading
pathogens or neuron impairment, lead to activation of microglial cells that
quickly proliferate, acquire amoeboid morphology and produce toxic
mediators such as nitric oxide (NO) and pro-inflammatory cytokines. These
changes are regulated by transcription factors, most importantly NF-κB.
Although microglial activation is important for maintaining tissue
homeostasis, excessive activation leads to chronic inflammation that can
damage healthy neurons. Substances that suppress microglial activation are
potential therapeutics for neurodegenerative diseases. Benfotiamine (Sbenzoylthiamine-
O-monophosphate) is a synthetic derivative of vitamin B1
that has anti-inflammatory properties. In this study we investigated antiinflammatory
properties of benfotiamine on activated microglia in vitro.
BV-2 microglia were pre-treated with benfotiamine, stimulated with LPS
and their viability and morphology were evaluated. LPS induced prominent
alterations in cell morphology, enlargement of cell bodies and spreading of
multiple processes. Pre-treatment with benfotiamine before LPS
stimulation suppressed these morphological changes. Additionally,
benfotiamine diminished LPS induced NO production, without altering cell
viability. Furthermore, benfotiamine decreased LPS induced production of
pro-inflammatory cytokines TNF- α and IL-6, while increasing production
of anti-inflammatory cytokine IL-10. Analysis of NF-κB activation
revealed that benfotiamine's effects were mediated by this transcription
factor. In conclusion, benfotiamine exerts anti-inflammatory properties in LPS
activated microglia in vitro and should be further investigated as a potential
therapeutic for neurodegenerative diseases.",
publisher = "Belgrade: Immunological Society of Serbia",
journal = "3rd Belgrade EFIS symposium on Immunoregulation: Immunity, Infection, Autoimmunity and Aging; 2015 May 24-27; Arandjelovac, Serbia",
title = "Anti-inflammatory effects of benfotiamine in LPS stimulated microglia: role of NF-κB signaling",
pages = "71",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6059"
}

Lukić, M., Jonjic, S., Nikolich-Zugich, J., Božić, I., Savić, D., Laketa, D., Nedeljković, N., Peković, S.,& Lavrnja, I.. (2015). Anti-inflammatory effects of benfotiamine in LPS stimulated microglia: role of NF-κB signaling. in 3rd Belgrade EFIS symposium on Immunoregulation: Immunity, Infection, Autoimmunity and Aging; 2015 May 24-27; Arandjelovac, Serbia
Belgrade: Immunological Society of Serbia., 71.
https://hdl.handle.net/21.15107/rcub_ibiss_6059

Lukić M, Jonjic S, Nikolich-Zugich J, Božić I, Savić D, Laketa D, Nedeljković N, Peković S, Lavrnja I. Anti-inflammatory effects of benfotiamine in LPS stimulated microglia: role of NF-κB signaling. in 3rd Belgrade EFIS symposium on Immunoregulation: Immunity, Infection, Autoimmunity and Aging; 2015 May 24-27; Arandjelovac, Serbia. 2015;:71.
https://hdl.handle.net/21.15107/rcub_ibiss_6059 .

Lukić, Miodrag, Jonjic, Stipan, Nikolich-Zugich, Janko, Božić, Iva, Savić, Danijela, Laketa, Danijela, Nedeljković, Nadežda, Peković, Sanja, Lavrnja, Irena, "Anti-inflammatory effects of benfotiamine in LPS stimulated microglia: role of NF-κB signaling" in 3rd Belgrade EFIS symposium on Immunoregulation: Immunity, Infection, Autoimmunity and Aging; 2015 May 24-27; Arandjelovac, Serbia (2015):71,
https://hdl.handle.net/21.15107/rcub_ibiss_6059 .

TY  - JOUR
AU  - Lavrnja, Irena
AU  - Laketa, Danijela
AU  - Savić, Danijela
AU  - Božić, Iva
AU  - Bjelobaba, Ivana
AU  - Peković, Sanja
AU  - Nedeljkovic, Nadezda
PY  - 2015
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1981
AB  - Ecto-5'-nucleotidase/cluster of differentiation 73 (CD73) (eN) is a
   70-kDa glycoprotein expressed in several different mammalian tissues and
   cell types. It is the rate-limiting enzyme of the purine catabolic
   pathway, which catalyzes the hydrolysis of AMP to produce adenosine with
   known anti-inflammatory and immunosuppressive actions. There is strong
   evidence for lymphocyte and endothelial cell eN having a role in
   experimental autoimmune encephalomyelitis (EAE), but the role of eN in
   cell types within the central nervous system is less clear. We have
   previously shown that eN activity significantly increased in the lumbar
   spinal cord during EAE. The present study is aimed to explore molecular
   pattern of the eN upregulation over the course of the disease and cell
   type(s) accountable for the induction. EAE was induced in Dark Agouti
   (DA) rats by immunization with the spinal cord tissue homogenate and
   adjuvant. Animals were sacrificed 8, 15, and 28 days following
   immunization (D8, D15, and D28), i.e., at time points which corresponded
   to the presymptomatic, symptomatic, and postsymptomatic phases of the
   disease, respectively. Significant increase in eN activity and its
   upregulation at the gene and the protein levels were demonstrated at D15
   and less prominently at D28 in comparison to control. Additionally,
   reactive astrocytes abundantly present in the lumbar spinal cord
   parenchyma were identified as principal cell type with significantly
   elevated eN expression. In all experimental groups, eN was expressed as
   a 71-kDa protein band of uniform abundance, whereas the overexpression
   of eN at D15 and D28 was associated with the expression of a second
   75-kDa eN variant. The possible outcome of eN upregulation during EAE as
   a part of protective astrocyte repertoire contributing to the resolution
   of the disease is discussed.
T2  - Journal of Molecular Neuroscience
T1  - Expression of a Second Ecto-5'-Nucleotidase Variant Besides the Usual
 Protein in Symptomatic Phase of Experimental Autoimmune
 Encephalomyelitis
IS  - 4
VL  - 55
DO  - 10.1007/s12031-014-0445-x
SP  - 898
EP  - 911
ER  - 

@article{
author = "Lavrnja, Irena and Laketa, Danijela and Savić, Danijela and Božić, Iva and Bjelobaba, Ivana and Peković, Sanja and Nedeljkovic, Nadezda",
year = "2015",
abstract = "Ecto-5'-nucleotidase/cluster of differentiation 73 (CD73) (eN) is a
   70-kDa glycoprotein expressed in several different mammalian tissues and
   cell types. It is the rate-limiting enzyme of the purine catabolic
   pathway, which catalyzes the hydrolysis of AMP to produce adenosine with
   known anti-inflammatory and immunosuppressive actions. There is strong
   evidence for lymphocyte and endothelial cell eN having a role in
   experimental autoimmune encephalomyelitis (EAE), but the role of eN in
   cell types within the central nervous system is less clear. We have
   previously shown that eN activity significantly increased in the lumbar
   spinal cord during EAE. The present study is aimed to explore molecular
   pattern of the eN upregulation over the course of the disease and cell
   type(s) accountable for the induction. EAE was induced in Dark Agouti
   (DA) rats by immunization with the spinal cord tissue homogenate and
   adjuvant. Animals were sacrificed 8, 15, and 28 days following
   immunization (D8, D15, and D28), i.e., at time points which corresponded
   to the presymptomatic, symptomatic, and postsymptomatic phases of the
   disease, respectively. Significant increase in eN activity and its
   upregulation at the gene and the protein levels were demonstrated at D15
   and less prominently at D28 in comparison to control. Additionally,
   reactive astrocytes abundantly present in the lumbar spinal cord
   parenchyma were identified as principal cell type with significantly
   elevated eN expression. In all experimental groups, eN was expressed as
   a 71-kDa protein band of uniform abundance, whereas the overexpression
   of eN at D15 and D28 was associated with the expression of a second
   75-kDa eN variant. The possible outcome of eN upregulation during EAE as
   a part of protective astrocyte repertoire contributing to the resolution
   of the disease is discussed.",
journal = "Journal of Molecular Neuroscience",
title = "Expression of a Second Ecto-5'-Nucleotidase Variant Besides the Usual
 Protein in Symptomatic Phase of Experimental Autoimmune
 Encephalomyelitis",
number = "4",
volume = "55",
doi = "10.1007/s12031-014-0445-x",
pages = "898-911"
}

Lavrnja, I., Laketa, D., Savić, D., Božić, I., Bjelobaba, I., Peković, S.,& Nedeljkovic, N.. (2015). Expression of a Second Ecto-5'-Nucleotidase Variant Besides the Usual
 Protein in Symptomatic Phase of Experimental Autoimmune
 Encephalomyelitis. in Journal of Molecular Neuroscience, 55(4), 898-911.
https://doi.org/10.1007/s12031-014-0445-x

Lavrnja I, Laketa D, Savić D, Božić I, Bjelobaba I, Peković S, Nedeljkovic N. Expression of a Second Ecto-5'-Nucleotidase Variant Besides the Usual
 Protein in Symptomatic Phase of Experimental Autoimmune
 Encephalomyelitis. in Journal of Molecular Neuroscience. 2015;55(4):898-911.
doi:10.1007/s12031-014-0445-x .

Lavrnja, Irena, Laketa, Danijela, Savić, Danijela, Božić, Iva, Bjelobaba, Ivana, Peković, Sanja, Nedeljkovic, Nadezda, "Expression of a Second Ecto-5'-Nucleotidase Variant Besides the Usual
 Protein in Symptomatic Phase of Experimental Autoimmune
 Encephalomyelitis" in Journal of Molecular Neuroscience, 55, no. 4 (2015):898-911,
https://doi.org/10.1007/s12031-014-0445-x . .

TY  - JOUR
AU  - Božić, Iva
AU  - Savić, Danijela
AU  - Laketa, Danijela
AU  - Bjelobaba, Ivana
AU  - Milenkovic, Ivan
AU  - Peković, Sanja
AU  - Nedeljkovic, Nadezda
AU  - Lavrnja, Irena
PY  - 2015
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2001
AB  - Microglial cells are resident immune cells of the central nervous system
   (CNS), recognized as key elements in the regulation of neural
   homeostasis and the response to injury and repair. As excessive
   activation of microglia may lead to neurodegeneration, therapeutic
   strategies targeting its inhibition were shown to improve treatment of
   most neurodegenerative diseases. Benfotiamine is a synthetic vitamin B1
   (thiamine) derivate exerting potentially anti-inflammatory effects.
   Despite the encouraging results regarding benfotiamine potential to
   alleviate diabetic microangiopathy, neuropathy and other oxidative
   stress-induced pathological conditions, its activities and cellular
   mechanisms during microglial activation have yet to be elucidated. In
   the present study, the anti-inflammatory effects of benfotiamine were
   investigated in lipopolysaccharide (LPS)-stimulated murine BV-2
   microglia. We determined that benfotiamine remodels activated microglia
   to acquire the shape that is characteristic of non-stimulated BV-2
   cells. In addition, benfotiamine significantly decreased production of
   pro-inflammatory mediators such as inducible form of nitric oxide
   synthase (iNOS) and NO; cyclooxygenase-2 (COX-2), heat-shock protein 70
   (Hsp70), tumor necrosis factor alpha a (TNF-alpha), interleukin-6
   (IL-6), whereas it increased anti-inflammatory interleukin-10 (IL-10)
   production in LPS stimulated BV-2 microglia. Moreover, benfotiamine
   suppressed the phosphorylation of extracellular signal-regulated kinases
   1/2 (ERK1/2), c-Jun N-terminal kinases (JNK) and protein kinase B
   Akt/PKB. Treatment with specific inhibitors revealed that
   benfotiamine-mediated suppression of NO production was via JNK1/2 and
   Akt pathway, while the cytokine suppression includes ERK1/2, JNK1/2 and
   Akt pathways. Finally, the potentially protective effect is mediated by
   the suppression of translocation of nuclear factor
   kappa-light-chain-enhancer of activated B cells (NF-kappa B) in the
   nucleus. Therefore, benfotiamine may have therapeutic potential for
   neurodegenerative diseases by inhibiting inflammatory mediators and
   enhancing anti-inflammatory factor production in activated microglia.
T2  - Plos One
T1  - Benfotiamine Attenuates Inflammatory Response in LPS Stimulated BV-2
 Microglia
IS  - e0118372
VL  - 10
DO  - 10.1371/journal.pone.0118372
ER  - 

@article{
author = "Božić, Iva and Savić, Danijela and Laketa, Danijela and Bjelobaba, Ivana and Milenkovic, Ivan and Peković, Sanja and Nedeljkovic, Nadezda and Lavrnja, Irena",
year = "2015",
abstract = "Microglial cells are resident immune cells of the central nervous system
   (CNS), recognized as key elements in the regulation of neural
   homeostasis and the response to injury and repair. As excessive
   activation of microglia may lead to neurodegeneration, therapeutic
   strategies targeting its inhibition were shown to improve treatment of
   most neurodegenerative diseases. Benfotiamine is a synthetic vitamin B1
   (thiamine) derivate exerting potentially anti-inflammatory effects.
   Despite the encouraging results regarding benfotiamine potential to
   alleviate diabetic microangiopathy, neuropathy and other oxidative
   stress-induced pathological conditions, its activities and cellular
   mechanisms during microglial activation have yet to be elucidated. In
   the present study, the anti-inflammatory effects of benfotiamine were
   investigated in lipopolysaccharide (LPS)-stimulated murine BV-2
   microglia. We determined that benfotiamine remodels activated microglia
   to acquire the shape that is characteristic of non-stimulated BV-2
   cells. In addition, benfotiamine significantly decreased production of
   pro-inflammatory mediators such as inducible form of nitric oxide
   synthase (iNOS) and NO; cyclooxygenase-2 (COX-2), heat-shock protein 70
   (Hsp70), tumor necrosis factor alpha a (TNF-alpha), interleukin-6
   (IL-6), whereas it increased anti-inflammatory interleukin-10 (IL-10)
   production in LPS stimulated BV-2 microglia. Moreover, benfotiamine
   suppressed the phosphorylation of extracellular signal-regulated kinases
   1/2 (ERK1/2), c-Jun N-terminal kinases (JNK) and protein kinase B
   Akt/PKB. Treatment with specific inhibitors revealed that
   benfotiamine-mediated suppression of NO production was via JNK1/2 and
   Akt pathway, while the cytokine suppression includes ERK1/2, JNK1/2 and
   Akt pathways. Finally, the potentially protective effect is mediated by
   the suppression of translocation of nuclear factor
   kappa-light-chain-enhancer of activated B cells (NF-kappa B) in the
   nucleus. Therefore, benfotiamine may have therapeutic potential for
   neurodegenerative diseases by inhibiting inflammatory mediators and
   enhancing anti-inflammatory factor production in activated microglia.",
journal = "Plos One",
title = "Benfotiamine Attenuates Inflammatory Response in LPS Stimulated BV-2
 Microglia",
number = "e0118372",
volume = "10",
doi = "10.1371/journal.pone.0118372"
}

Božić, I., Savić, D., Laketa, D., Bjelobaba, I., Milenkovic, I., Peković, S., Nedeljkovic, N.,& Lavrnja, I.. (2015). Benfotiamine Attenuates Inflammatory Response in LPS Stimulated BV-2
 Microglia. in Plos One, 10(e0118372).
https://doi.org/10.1371/journal.pone.0118372

Božić I, Savić D, Laketa D, Bjelobaba I, Milenkovic I, Peković S, Nedeljkovic N, Lavrnja I. Benfotiamine Attenuates Inflammatory Response in LPS Stimulated BV-2
 Microglia. in Plos One. 2015;10(e0118372).
doi:10.1371/journal.pone.0118372 .

Božić, Iva, Savić, Danijela, Laketa, Danijela, Bjelobaba, Ivana, Milenkovic, Ivan, Peković, Sanja, Nedeljkovic, Nadezda, Lavrnja, Irena, "Benfotiamine Attenuates Inflammatory Response in LPS Stimulated BV-2
 Microglia" in Plos One, 10, no. e0118372 (2015),
https://doi.org/10.1371/journal.pone.0118372 . .

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  - Laketa, Danijela
AU  - Savić, Jasmina
AU  - Bjelobaba, Ivana
AU  - Lavrnja, Irena
AU  - Vasić, Vesna
AU  - Stojiljković, Mirjana
AU  - Nedeljković, Nadežda
PY  - 2015
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2093
AB  - Background: Cortical stab injury (CSI) induces changes in the activity,
   expression and cellular distribution of specific ectonucleotidases at
   the injury site. Also, several experimentally induced neuropathologies
   are associated with changes in soluble ectonucleotidase activities in
   the plasma and serum, whilst various insults to the brain alter purine
   compounds levels in cerebrospinal fluid, but also in serum, indicating
   that insults to the brain may induce alterations in nucleotides release
   and rate of their hydrolysis in the vascular system. Since adenine
   nucleotides and adenosine regulate diverse cellular functions in the
   vascular system, including vascular tone, platelet aggregation and
   inflammatory responses of lymphocytes and macrophages, alterations of
   ectonucleotidase activities in the vascular system may be relevant for
   the clinical outcome of the primary insult.
   Methods: We explored ectonucleotidase activities using specific enzyme
   assays and determined adenine nucleotides concentrations by the UPLC
   method in the rat serum after cortical stab injury.
   Results: At 4-h post-injury, ATP and AMP hydrolysis increased by about
   60\% and 40\%, respectively, while phosphodiesterase activity remained
   unchanged. Also, at 4-h postinjury a marked decrease in ATP
   concentration and more than 2-fold increase in AMP concentration were
   recorded.
   Conclusions: CSI induces rapid up-regulation of nucleotide catabolizing
   soluble ectonucleotidases in rat serum, which leads to the observed
   shift in serum nucleotide levels. The results obtained imply that
   ectonucleotidases and adenine nucleotides participate in the
   communication between the brain and the vascular system in physiological
   and pathological conditions and thereby may be involved in the
   development of various human neuropathologies.
T2  - Journal of Medical Biochemistry
T1  - BRAIN INJURY ALTERS ECTONUCLEOTIDASE ACTIVITIES AND ADENINE NUCLEOTIDE
 LEVELS IN RAT SERUM
IS  - 2
VL  - 34
DO  - 10.2478/jomb-2014-0025
SP  - 215
EP  - 222
ER  - 

@article{
author = "Laketa, Danijela and Savić, Jasmina and Bjelobaba, Ivana and Lavrnja, Irena and Vasić, Vesna and Stojiljković, Mirjana and Nedeljković, Nadežda",
year = "2015",
abstract = "Background: Cortical stab injury (CSI) induces changes in the activity,
   expression and cellular distribution of specific ectonucleotidases at
   the injury site. Also, several experimentally induced neuropathologies
   are associated with changes in soluble ectonucleotidase activities in
   the plasma and serum, whilst various insults to the brain alter purine
   compounds levels in cerebrospinal fluid, but also in serum, indicating
   that insults to the brain may induce alterations in nucleotides release
   and rate of their hydrolysis in the vascular system. Since adenine
   nucleotides and adenosine regulate diverse cellular functions in the
   vascular system, including vascular tone, platelet aggregation and
   inflammatory responses of lymphocytes and macrophages, alterations of
   ectonucleotidase activities in the vascular system may be relevant for
   the clinical outcome of the primary insult.
   Methods: We explored ectonucleotidase activities using specific enzyme
   assays and determined adenine nucleotides concentrations by the UPLC
   method in the rat serum after cortical stab injury.
   Results: At 4-h post-injury, ATP and AMP hydrolysis increased by about
   60\% and 40\%, respectively, while phosphodiesterase activity remained
   unchanged. Also, at 4-h postinjury a marked decrease in ATP
   concentration and more than 2-fold increase in AMP concentration were
   recorded.
   Conclusions: CSI induces rapid up-regulation of nucleotide catabolizing
   soluble ectonucleotidases in rat serum, which leads to the observed
   shift in serum nucleotide levels. The results obtained imply that
   ectonucleotidases and adenine nucleotides participate in the
   communication between the brain and the vascular system in physiological
   and pathological conditions and thereby may be involved in the
   development of various human neuropathologies.",
journal = "Journal of Medical Biochemistry",
title = "BRAIN INJURY ALTERS ECTONUCLEOTIDASE ACTIVITIES AND ADENINE NUCLEOTIDE
 LEVELS IN RAT SERUM",
number = "2",
volume = "34",
doi = "10.2478/jomb-2014-0025",
pages = "215-222"
}

Laketa, D., Savić, J., Bjelobaba, I., Lavrnja, I., Vasić, V., Stojiljković, M.,& Nedeljković, N.. (2015). BRAIN INJURY ALTERS ECTONUCLEOTIDASE ACTIVITIES AND ADENINE NUCLEOTIDE
 LEVELS IN RAT SERUM. in Journal of Medical Biochemistry, 34(2), 215-222.
https://doi.org/10.2478/jomb-2014-0025

Laketa D, Savić J, Bjelobaba I, Lavrnja I, Vasić V, Stojiljković M, Nedeljković N. BRAIN INJURY ALTERS ECTONUCLEOTIDASE ACTIVITIES AND ADENINE NUCLEOTIDE
 LEVELS IN RAT SERUM. in Journal of Medical Biochemistry. 2015;34(2):215-222.
doi:10.2478/jomb-2014-0025 .

Laketa, Danijela, Savić, Jasmina, Bjelobaba, Ivana, Lavrnja, Irena, Vasić, Vesna, Stojiljković, Mirjana, Nedeljković, Nadežda, "BRAIN INJURY ALTERS ECTONUCLEOTIDASE ACTIVITIES AND ADENINE NUCLEOTIDE
 LEVELS IN RAT SERUM" in Journal of Medical Biochemistry, 34, no. 2 (2015):215-222,
https://doi.org/10.2478/jomb-2014-0025 . .

TY  - JOUR
AU  - Božić, Iva
AU  - Savić, Danijela
AU  - Jovanović, Marija
AU  - Bjelobaba, Ivana
AU  - Laketa, Danijela
AU  - Nedeljković, Nadežda
AU  - Stojiljković, Mirjana
AU  - Peković, Sanja
AU  - Lavrnja, Irena
PY  - 2015
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2047
AB  - Microglia play a key role in defending central nervous system from
   various internal and external threats. However, their excessive and/or
   chronic activation is associated with deleterious effects in a variety
   of neurodegenerative diseases. Previously, we have shown that ribavirin
   when applied in clinically relevant dosage (10 mu M) modulates activated
   microglia in complex fashion inducing both anti-and proinflammatory
   effects, simultaneously causing cytotoxicity. Here, we examined
   potential of low-dose ribavirin (0.1 and 1 mu M) to modulate activated
   BV-2 microglia. Morphological and functional activation of BV-2 cells
   was achieved with lipopolysaccharide (LPS) stimulation. Our results
   demonstrated that low-dose ribavirin did not induce cell death, while 10
   mu M ribavirin promoted LPS induced apoptosis. We determined that 1 mu M
   ribavirin was equally efficient in deactivation of LPS induced
   morphological changes as 10 mu M ribavirin treatment. Ribavirin showed
   halfway success in reducing markers of functional activation of
   microglia. Namely, none of the doses had effect on LPS triggered
   production of proinflammatory cytokine tumor necrosis factor alpha. On
   the other hand, low-dose ribavirin proved its effectiveness in reduction
   of another inflammatory mediator, nitric oxide, by inhibiting inducible
   form of nitric oxide synthase. Our results imply that low-dose ribavirin
   may alleviate nitrosative stress during neuroinflammation.
T2  - Analytical Cellular Pathology
T1  - Low-Dose Ribavirin Treatments Attenuate Neuroinflammatory Activation of
 BV-2 Cells by Interfering with Inducible Nitric Oxide Synthase
IS  - 923614
DO  - 10.1155/2015/923614
ER  - 

@article{
author = "Božić, Iva and Savić, Danijela and Jovanović, Marija and Bjelobaba, Ivana and Laketa, Danijela and Nedeljković, Nadežda and Stojiljković, Mirjana and Peković, Sanja and Lavrnja, Irena",
year = "2015",
abstract = "Microglia play a key role in defending central nervous system from
   various internal and external threats. However, their excessive and/or
   chronic activation is associated with deleterious effects in a variety
   of neurodegenerative diseases. Previously, we have shown that ribavirin
   when applied in clinically relevant dosage (10 mu M) modulates activated
   microglia in complex fashion inducing both anti-and proinflammatory
   effects, simultaneously causing cytotoxicity. Here, we examined
   potential of low-dose ribavirin (0.1 and 1 mu M) to modulate activated
   BV-2 microglia. Morphological and functional activation of BV-2 cells
   was achieved with lipopolysaccharide (LPS) stimulation. Our results
   demonstrated that low-dose ribavirin did not induce cell death, while 10
   mu M ribavirin promoted LPS induced apoptosis. We determined that 1 mu M
   ribavirin was equally efficient in deactivation of LPS induced
   morphological changes as 10 mu M ribavirin treatment. Ribavirin showed
   halfway success in reducing markers of functional activation of
   microglia. Namely, none of the doses had effect on LPS triggered
   production of proinflammatory cytokine tumor necrosis factor alpha. On
   the other hand, low-dose ribavirin proved its effectiveness in reduction
   of another inflammatory mediator, nitric oxide, by inhibiting inducible
   form of nitric oxide synthase. Our results imply that low-dose ribavirin
   may alleviate nitrosative stress during neuroinflammation.",
journal = "Analytical Cellular Pathology",
title = "Low-Dose Ribavirin Treatments Attenuate Neuroinflammatory Activation of
 BV-2 Cells by Interfering with Inducible Nitric Oxide Synthase",
number = "923614",
doi = "10.1155/2015/923614"
}

Božić, I., Savić, D., Jovanović, M., Bjelobaba, I., Laketa, D., Nedeljković, N., Stojiljković, M., Peković, S.,& Lavrnja, I.. (2015). Low-Dose Ribavirin Treatments Attenuate Neuroinflammatory Activation of
 BV-2 Cells by Interfering with Inducible Nitric Oxide Synthase. in Analytical Cellular Pathology(923614).
https://doi.org/10.1155/2015/923614

Božić I, Savić D, Jovanović M, Bjelobaba I, Laketa D, Nedeljković N, Stojiljković M, Peković S, Lavrnja I. Low-Dose Ribavirin Treatments Attenuate Neuroinflammatory Activation of
 BV-2 Cells by Interfering with Inducible Nitric Oxide Synthase. in Analytical Cellular Pathology. 2015;(923614).
doi:10.1155/2015/923614 .

Božić, Iva, Savić, Danijela, Jovanović, Marija, Bjelobaba, Ivana, Laketa, Danijela, Nedeljković, Nadežda, Stojiljković, Mirjana, Peković, Sanja, Lavrnja, Irena, "Low-Dose Ribavirin Treatments Attenuate Neuroinflammatory Activation of
 BV-2 Cells by Interfering with Inducible Nitric Oxide Synthase" in Analytical Cellular Pathology, no. 923614 (2015),
https://doi.org/10.1155/2015/923614 . .

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  - 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  - CONF
AU  - Dačić, Sanja
AU  - Stojiljković, Mirjana B
AU  - Lavrnja, Irena
AU  - Savić, Danijela
AU  - Parabučki, Ana B.
AU  - Božić, Iva D.
AU  - Bjelobaba, Ivana
AU  - Laketa, Danijela
AU  - Nedeljković, Nadezda N
AU  - Rakić, Ljubisa M
AU  - Peković, Sanja
PY  - 2011
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1262
C3  - Glia
T1  - Treatment With Combination of B Vitamins Reduces Neurocan Expression After Cortical Ablation
IS  - null
VL  - 59
EP  - S52
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_1262
ER  - 

@conference{
author = "Dačić, Sanja and Stojiljković, Mirjana B and Lavrnja, Irena and Savić, Danijela and Parabučki, Ana B. and Božić, Iva D. and Bjelobaba, Ivana and Laketa, Danijela and Nedeljković, Nadezda N and Rakić, Ljubisa M and Peković, Sanja",
year = "2011",
journal = "Glia",
title = "Treatment With Combination of B Vitamins Reduces Neurocan Expression After Cortical Ablation",
number = "null",
volume = "59",
pages = "S52",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_1262"
}

Dačić, S., Stojiljković, M. B., Lavrnja, I., Savić, D., Parabučki, A. B., Božić, I. D., Bjelobaba, I., Laketa, D., Nedeljković, N. N., Rakić, L. M.,& Peković, S.. (2011). Treatment With Combination of B Vitamins Reduces Neurocan Expression After Cortical Ablation. in Glia, 59(null).
https://hdl.handle.net/21.15107/rcub_ibiss_1262

Dačić S, Stojiljković MB, Lavrnja I, Savić D, Parabučki AB, Božić ID, Bjelobaba I, Laketa D, Nedeljković NN, Rakić LM, Peković S. Treatment With Combination of B Vitamins Reduces Neurocan Expression After Cortical Ablation. in Glia. 2011;59(null):null-S52.
https://hdl.handle.net/21.15107/rcub_ibiss_1262 .

Dačić, Sanja, Stojiljković, Mirjana B, Lavrnja, Irena, Savić, Danijela, Parabučki, Ana B., Božić, Iva D., Bjelobaba, Ivana, Laketa, Danijela, Nedeljković, Nadezda N, Rakić, Ljubisa M, Peković, Sanja, "Treatment With Combination of B Vitamins Reduces Neurocan Expression After Cortical Ablation" in Glia, 59, no. null (2011),
https://hdl.handle.net/21.15107/rcub_ibiss_1262 .

TY  - JOUR
AU  - Laketa, Danijela
AU  - Bjelobaba, Ivana
AU  - Savić, Jasmina Z
AU  - Lavrnja, Irena
AU  - Stojiljković, Mirjana B
AU  - Rakić, Ljubisav
AU  - Nedeljković, Nadezda N
PY  - 2010
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1372
AB  - Biochemical properties of nucleotide pyrophosphatase/phosphodiesterase (NPP) in rat serum have been described by assessing its nucleotide phosphodiesterase activity, using p-nitrophenyl-5'-thymidine monophosphate (p-Nph-5'-TMP) as a substrate. It was demonstrated that NPP activity shares some typical characteristics described for other soluble NPP, such as divalent cation dependence, strong alkaline pH optimum (pH 10.5), inhibition by glycosaminoglycans, and K (m) for p-Nph-5'-TMP hydrolysis of 61.8 +/- A 5.2 mu M. In order to characterize the relation between phosphodiesterase and pyrophosphatase activities of NPP, we have analyzed the effects of different natural nucleotides and nucleotide analogs. ATP, ADP, and AMP competitively inhibited p-Nph-5'-TMP hydrolysis with K (i) values ranging 13-43 mu M. Nucleotide analogs, alpha,beta-metATP, BzATP, 2-MeSATP, and dialATP behaved as competitive inhibitors, whereas alpha,beta-metADP induced mixed inhibition, with K (i) ranging from 2 to 20 mu M. Chromatographic analysis revealed that alpha,beta-metATP, BzATP, and 2-MeSATP were catalytically degraded in the serum, whereas dialATP and alpha,beta-metADP resisted hydrolysis, implying that the former act as substrates and the latter as true competitive inhibitors of serum NPP activity. Since NPP activity is involved in generation, breakdown, and recycling of extracellular adenine nucleotides in the vascular compartment, the results suggest that both hydrolyzable and non-hydrolyzable nucleotide analogs could alter the amplitude and direction of ATP actions and could have potential therapeutic application.
T2  - Molecular and Cellular Biochemistry
T1  - Biochemical characterization of soluble nucleotide pyrophosphatase/phosphodiesterase activity in rat serum
IS  - 1-2
VL  - 339
EP  - 106
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_1372
ER  - 

@article{
author = "Laketa, Danijela and Bjelobaba, Ivana and Savić, Jasmina Z and Lavrnja, Irena and Stojiljković, Mirjana B and Rakić, Ljubisav and Nedeljković, Nadezda N",
year = "2010",
abstract = "Biochemical properties of nucleotide pyrophosphatase/phosphodiesterase (NPP) in rat serum have been described by assessing its nucleotide phosphodiesterase activity, using p-nitrophenyl-5'-thymidine monophosphate (p-Nph-5'-TMP) as a substrate. It was demonstrated that NPP activity shares some typical characteristics described for other soluble NPP, such as divalent cation dependence, strong alkaline pH optimum (pH 10.5), inhibition by glycosaminoglycans, and K (m) for p-Nph-5'-TMP hydrolysis of 61.8 +/- A 5.2 mu M. In order to characterize the relation between phosphodiesterase and pyrophosphatase activities of NPP, we have analyzed the effects of different natural nucleotides and nucleotide analogs. ATP, ADP, and AMP competitively inhibited p-Nph-5'-TMP hydrolysis with K (i) values ranging 13-43 mu M. Nucleotide analogs, alpha,beta-metATP, BzATP, 2-MeSATP, and dialATP behaved as competitive inhibitors, whereas alpha,beta-metADP induced mixed inhibition, with K (i) ranging from 2 to 20 mu M. Chromatographic analysis revealed that alpha,beta-metATP, BzATP, and 2-MeSATP were catalytically degraded in the serum, whereas dialATP and alpha,beta-metADP resisted hydrolysis, implying that the former act as substrates and the latter as true competitive inhibitors of serum NPP activity. Since NPP activity is involved in generation, breakdown, and recycling of extracellular adenine nucleotides in the vascular compartment, the results suggest that both hydrolyzable and non-hydrolyzable nucleotide analogs could alter the amplitude and direction of ATP actions and could have potential therapeutic application.",
journal = "Molecular and Cellular Biochemistry",
title = "Biochemical characterization of soluble nucleotide pyrophosphatase/phosphodiesterase activity in rat serum",
number = "1-2",
volume = "339",
pages = "106",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_1372"
}

Laketa, D., Bjelobaba, I., Savić, J. Z., Lavrnja, I., Stojiljković, M. B., Rakić, L.,& Nedeljković, N. N.. (2010). Biochemical characterization of soluble nucleotide pyrophosphatase/phosphodiesterase activity in rat serum. in Molecular and Cellular Biochemistry, 339(1-2).
https://hdl.handle.net/21.15107/rcub_ibiss_1372

Laketa D, Bjelobaba I, Savić JZ, Lavrnja I, Stojiljković MB, Rakić L, Nedeljković NN. Biochemical characterization of soluble nucleotide pyrophosphatase/phosphodiesterase activity in rat serum. in Molecular and Cellular Biochemistry. 2010;339(1-2):null-106.
https://hdl.handle.net/21.15107/rcub_ibiss_1372 .

Laketa, Danijela, Bjelobaba, Ivana, Savić, Jasmina Z, Lavrnja, Irena, Stojiljković, Mirjana B, Rakić, Ljubisav, Nedeljković, Nadezda N, "Biochemical characterization of soluble nucleotide pyrophosphatase/phosphodiesterase activity in rat serum" in Molecular and Cellular Biochemistry, 339, no. 1-2 (2010),
https://hdl.handle.net/21.15107/rcub_ibiss_1372 .

TY  - JOUR
AU  - Bjelobaba, Ivana
AU  - Stojiljković, Mirjana B
AU  - Lavrnja, Irena
AU  - Savić, Danijela
AU  - Peković, Sanja
AU  - Dačić, Sanja
AU  - Laketa, Danijela
AU  - Rakić, Ljubisav
AU  - Nedeljković, Nadezda N
PY  - 2009
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1473
AB  - During a variety of insults to the brain adenine nucleotides are released in large quantities from damaged cells, triggering local cellular and biochemical responses to injury. Different models of brain injury reveal that the local increase in adenine nucleotides levels is followed by a compensatory up-regulation of ectonucleotidase enzymes that catalyze sequential hydrolysis of ATP to ADP, AMP and adenosine. However, recent studies imply that changes in adenine nucleotides release may also occur in the areas distant from the site of direct damage. Therefore, in the present study we have used the model of cortical stab injury to analyze extracellular ATP, ADP and AMP hydrolysis in the membrane preparations obtained from the brain regions that were not subjected to direct tissue damage. The brain regions analyzed were contralateral cortex, hippocampus, caudate nucleus, thalamus and hypothalamus. It was evidenced that cortical stab injury induced early widespread decrease in AMP hydrolysis in all brain areas tested, except in the hypothalamus, without changes in ATP hydrolysis. These findings imply that brain injury affects global extracellular adenine nucleotide and nucleoside levels, consequently affecting neuronal function in the regions distant to the primary damage.
T2  - General Physiology and Biophysics
T1  - Regional changes in ectonucleotidase activity after cortical stab injury in rat
IS  - null
VL  - 28
EP  - 68
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_1473
ER  - 

@article{
author = "Bjelobaba, Ivana and Stojiljković, Mirjana B and Lavrnja, Irena and Savić, Danijela and Peković, Sanja and Dačić, Sanja and Laketa, Danijela and Rakić, Ljubisav and Nedeljković, Nadezda N",
year = "2009",
abstract = "During a variety of insults to the brain adenine nucleotides are released in large quantities from damaged cells, triggering local cellular and biochemical responses to injury. Different models of brain injury reveal that the local increase in adenine nucleotides levels is followed by a compensatory up-regulation of ectonucleotidase enzymes that catalyze sequential hydrolysis of ATP to ADP, AMP and adenosine. However, recent studies imply that changes in adenine nucleotides release may also occur in the areas distant from the site of direct damage. Therefore, in the present study we have used the model of cortical stab injury to analyze extracellular ATP, ADP and AMP hydrolysis in the membrane preparations obtained from the brain regions that were not subjected to direct tissue damage. The brain regions analyzed were contralateral cortex, hippocampus, caudate nucleus, thalamus and hypothalamus. It was evidenced that cortical stab injury induced early widespread decrease in AMP hydrolysis in all brain areas tested, except in the hypothalamus, without changes in ATP hydrolysis. These findings imply that brain injury affects global extracellular adenine nucleotide and nucleoside levels, consequently affecting neuronal function in the regions distant to the primary damage.",
journal = "General Physiology and Biophysics",
title = "Regional changes in ectonucleotidase activity after cortical stab injury in rat",
number = "null",
volume = "28",
pages = "68",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_1473"
}

Bjelobaba, I., Stojiljković, M. B., Lavrnja, I., Savić, D., Peković, S., Dačić, S., Laketa, D., Rakić, L.,& Nedeljković, N. N.. (2009). Regional changes in ectonucleotidase activity after cortical stab injury in rat. in General Physiology and Biophysics, 28(null).
https://hdl.handle.net/21.15107/rcub_ibiss_1473

Bjelobaba I, Stojiljković MB, Lavrnja I, Savić D, Peković S, Dačić S, Laketa D, Rakić L, Nedeljković NN. Regional changes in ectonucleotidase activity after cortical stab injury in rat. in General Physiology and Biophysics. 2009;28(null):null-68.
https://hdl.handle.net/21.15107/rcub_ibiss_1473 .

Bjelobaba, Ivana, Stojiljković, Mirjana B, Lavrnja, Irena, Savić, Danijela, Peković, Sanja, Dačić, Sanja, Laketa, Danijela, Rakić, Ljubisav, Nedeljković, Nadezda N, "Regional changes in ectonucleotidase activity after cortical stab injury in rat" in General Physiology and Biophysics, 28, no. null (2009),
https://hdl.handle.net/21.15107/rcub_ibiss_1473 .