TY  - JOUR
AU  - Mitrović-Ajtić, Olivera
AU  - Đikić, Dragoslava
AU  - Subotički, Tijana
AU  - Bižić-Radulović, Sandra
AU  - Beleslin-Čokić, Bojana
AU  - Dragojević, Teodora
AU  - Živković, Emilija
AU  - Miljatović, Sanja
AU  - Vukotić, Milica
AU  - Stanisavljević, Dejana
AU  - Santibanez, Juan F.
AU  - Čokić, Vladan
PY  - 2023
UR  - http://rimi.imi.bg.ac.rs/handle/123456789/1355
AB  - The severity and mortality of coronavirus disease 2019 (COVID-19) are greater in males than in females, though the infection rate is the same in the two sexes. We investigated sex hormone differences associated with the hyperinflammatory immune response to SARS-CoV-2 on the basis of patients’ cytokine profiles and vaccination statuses. Clinical and laboratory data of 117 patients with COVID-19 were collected to examine sex differences associated with oxidative stress markers, neutrophil extracellular traps (NETs), and plasma cytokine levels up to 5 months from hospital admission. The testosterone and free testosterone levels were low in male patients with COVID-19 and returned to normal values after recovery from the disease. The dihydrotestosterone (DHT) levels were transiently reduced, while the sex hormone-binding globulin levels were decreased in post-COVID-19 male patients. The levels of the inflammatory cytokines interleukin-6 (IL-6) and IL-10 appeared generally increased at diagnosis and decreased in post-COVID-19 patients. In females, the concentration of tumor necrosis factor-alpha was increased by four times at diagnosis. The levels of the coagulation markers intercellular adhesion molecule-1 (ICAM-1) and E-selectin were consistently upregulated in post-COVID-19 female patients, in contrast to those of vascular cell adhesion molecule-1 (VCAM-1), P-selectin, and chemokine IL-8. DHT increased the levels of reactive oxygen species in the neutrophils of male patients, while estradiol decreased them in females. Markers for NET, such as circulating DNA and myeloperoxidase, were significantly more abundant in the patients’ plasma. Sex hormones have a potential protective role during SARS-CoV-2 infection, which is weakened by impaired testosterone synthesis in men.
PB  - Multidisciplinary Digital Publishing Institute (MDPI)
T2  - Vaccines
T2  - Vaccines
T1  - Sex Differences and Cytokine Profiles among Patients Hospitalized for COVID-19 and during Their Recovery: The Predominance of Adhesion Molecules in Females and Oxidative Stress in Males
IS  - 10
SP  - 1560
VL  - 11
DO  - 10.3390/vaccines11101560
ER  - 

@article{
author = "Mitrović-Ajtić, Olivera and Đikić, Dragoslava and Subotički, Tijana and Bižić-Radulović, Sandra and Beleslin-Čokić, Bojana and Dragojević, Teodora and Živković, Emilija and Miljatović, Sanja and Vukotić, Milica and Stanisavljević, Dejana and Santibanez, Juan F. and Čokić, Vladan",
year = "2023",
abstract = "The severity and mortality of coronavirus disease 2019 (COVID-19) are greater in males than in females, though the infection rate is the same in the two sexes. We investigated sex hormone differences associated with the hyperinflammatory immune response to SARS-CoV-2 on the basis of patients’ cytokine profiles and vaccination statuses. Clinical and laboratory data of 117 patients with COVID-19 were collected to examine sex differences associated with oxidative stress markers, neutrophil extracellular traps (NETs), and plasma cytokine levels up to 5 months from hospital admission. The testosterone and free testosterone levels were low in male patients with COVID-19 and returned to normal values after recovery from the disease. The dihydrotestosterone (DHT) levels were transiently reduced, while the sex hormone-binding globulin levels were decreased in post-COVID-19 male patients. The levels of the inflammatory cytokines interleukin-6 (IL-6) and IL-10 appeared generally increased at diagnosis and decreased in post-COVID-19 patients. In females, the concentration of tumor necrosis factor-alpha was increased by four times at diagnosis. The levels of the coagulation markers intercellular adhesion molecule-1 (ICAM-1) and E-selectin were consistently upregulated in post-COVID-19 female patients, in contrast to those of vascular cell adhesion molecule-1 (VCAM-1), P-selectin, and chemokine IL-8. DHT increased the levels of reactive oxygen species in the neutrophils of male patients, while estradiol decreased them in females. Markers for NET, such as circulating DNA and myeloperoxidase, were significantly more abundant in the patients’ plasma. Sex hormones have a potential protective role during SARS-CoV-2 infection, which is weakened by impaired testosterone synthesis in men.",
publisher = "Multidisciplinary Digital Publishing Institute (MDPI)",
journal = "Vaccines, Vaccines",
title = "Sex Differences and Cytokine Profiles among Patients Hospitalized for COVID-19 and during Their Recovery: The Predominance of Adhesion Molecules in Females and Oxidative Stress in Males",
number = "10",
pages = "1560",
volume = "11",
doi = "10.3390/vaccines11101560"
}

Mitrović-Ajtić, O., Đikić, D., Subotički, T., Bižić-Radulović, S., Beleslin-Čokić, B., Dragojević, T., Živković, E., Miljatović, S., Vukotić, M., Stanisavljević, D., Santibanez, J. F.,& Čokić, V.. (2023). Sex Differences and Cytokine Profiles among Patients Hospitalized for COVID-19 and during Their Recovery: The Predominance of Adhesion Molecules in Females and Oxidative Stress in Males. in Vaccines
Multidisciplinary Digital Publishing Institute (MDPI)., 11(10), 1560.
https://doi.org/10.3390/vaccines11101560

Mitrović-Ajtić O, Đikić D, Subotički T, Bižić-Radulović S, Beleslin-Čokić B, Dragojević T, Živković E, Miljatović S, Vukotić M, Stanisavljević D, Santibanez JF, Čokić V. Sex Differences and Cytokine Profiles among Patients Hospitalized for COVID-19 and during Their Recovery: The Predominance of Adhesion Molecules in Females and Oxidative Stress in Males. in Vaccines. 2023;11(10):1560.
doi:10.3390/vaccines11101560 .

Mitrović-Ajtić, Olivera, Đikić, Dragoslava, Subotički, Tijana, Bižić-Radulović, Sandra, Beleslin-Čokić, Bojana, Dragojević, Teodora, Živković, Emilija, Miljatović, Sanja, Vukotić, Milica, Stanisavljević, Dejana, Santibanez, Juan F., Čokić, Vladan, "Sex Differences and Cytokine Profiles among Patients Hospitalized for COVID-19 and during Their Recovery: The Predominance of Adhesion Molecules in Females and Oxidative Stress in Males" in Vaccines, 11, no. 10 (2023):1560,
https://doi.org/10.3390/vaccines11101560 . .

TY  - CONF
AU  - Mitrović-Ajtić, Olivera
AU  - Đikić, Dragoslava
AU  - Dragojević, Teodora
AU  - Otašević, Vladimir
AU  - Živković, Emilija
AU  - Vuković, Vojin
AU  - Vukotić, Milica
AU  - Subotički, Tijana
AU  - Diklić, Miloš
AU  - Suvajdžić-Vuković, Nada
AU  - Mitrović, Mirjana
AU  - Mihaljević, Biljana
AU  - Antić, Darko
AU  - Čokić, Vladan
PY  - 2023
UR  - http://rimi.imi.bg.ac.rs/handle/123456789/1420
AB  - Introduction: Patients with hematological malignancies have an increased risk of thrombotic complications, ranging from 3-5% in patients with lymphoma and acute myeloid leukemia (AML). The presented study observed the onset of thrombus formation to predict risk factors for thrombosis in lymphoid and myeloid malignancies. Methods: Coagulation factors, inflammatory signaling pathways and adhesion molecules have been observed in patients with Hodgkin lymphoma (HL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL) and AML. Their mononuclear cells (MNC) trans-endothelial migration through human microvascular endothelial cells (HMEC-1) monolayer is observed by Boyden chamber.
Results: Thrombin was in positive correlation with tumor necrosis factor alpha (TNF-α) in HL, while with P-selectin (p<0.001), tumor growth factor-beta (TGF-β) and factor VIII (p<0.05) in DLBCL and AML. Transendothelial migration of MNC was increased by TNF-α (p<0.001) in DLBCL regardless of previous thrombosis. Regarding coagulation, factor VIII was increased in HL and AML (p<0.05), while tissue factor in non-Hodgkin lymphomas (DLBCL and FL, p<0.05). Tissue factor was in positive correlation with adhesion molecule P-selectin and factor VIII (p<0.05). P-selectin was increased in non-Hodgkin lymphomas (p<0.0001), while TGF-β only in FL (p<0.001). Fibrinolytic activity was decreased in plasma of patients with HL, DLBCL, and FL (p<0.05), but largely in AML (p<0.01) as measured by tissue-type plasminogen activator. Inflammatory NF-κB signaling has been activated in HL and DLBCL, while p38 signaling only in HL. Conclusion: Coagulation factors and inflammation are increased in hematological malignancies along with the interaction of the endothelium and circulating cells that predispose to thrombus formation
PB  - Belgrade: Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade
C3  - CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
T1  - Activation of coagulation factors and prothrombic properties of endothelium in hematological malignancies
EP  - 134
SP  - 134
UR  - https://hdl.handle.net/21.15107/rcub_rimi_1420
ER  - 

@conference{
author = "Mitrović-Ajtić, Olivera and Đikić, Dragoslava and Dragojević, Teodora and Otašević, Vladimir and Živković, Emilija and Vuković, Vojin and Vukotić, Milica and Subotički, Tijana and Diklić, Miloš and Suvajdžić-Vuković, Nada and Mitrović, Mirjana and Mihaljević, Biljana and Antić, Darko and Čokić, Vladan",
year = "2023",
abstract = "Introduction: Patients with hematological malignancies have an increased risk of thrombotic complications, ranging from 3-5% in patients with lymphoma and acute myeloid leukemia (AML). The presented study observed the onset of thrombus formation to predict risk factors for thrombosis in lymphoid and myeloid malignancies. Methods: Coagulation factors, inflammatory signaling pathways and adhesion molecules have been observed in patients with Hodgkin lymphoma (HL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL) and AML. Their mononuclear cells (MNC) trans-endothelial migration through human microvascular endothelial cells (HMEC-1) monolayer is observed by Boyden chamber.
Results: Thrombin was in positive correlation with tumor necrosis factor alpha (TNF-α) in HL, while with P-selectin (p<0.001), tumor growth factor-beta (TGF-β) and factor VIII (p<0.05) in DLBCL and AML. Transendothelial migration of MNC was increased by TNF-α (p<0.001) in DLBCL regardless of previous thrombosis. Regarding coagulation, factor VIII was increased in HL and AML (p<0.05), while tissue factor in non-Hodgkin lymphomas (DLBCL and FL, p<0.05). Tissue factor was in positive correlation with adhesion molecule P-selectin and factor VIII (p<0.05). P-selectin was increased in non-Hodgkin lymphomas (p<0.0001), while TGF-β only in FL (p<0.001). Fibrinolytic activity was decreased in plasma of patients with HL, DLBCL, and FL (p<0.05), but largely in AML (p<0.01) as measured by tissue-type plasminogen activator. Inflammatory NF-κB signaling has been activated in HL and DLBCL, while p38 signaling only in HL. Conclusion: Coagulation factors and inflammation are increased in hematological malignancies along with the interaction of the endothelium and circulating cells that predispose to thrombus formation",
publisher = "Belgrade: Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade",
journal = "CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia",
title = "Activation of coagulation factors and prothrombic properties of endothelium in hematological malignancies",
pages = "134-134",
url = "https://hdl.handle.net/21.15107/rcub_rimi_1420"
}

Mitrović-Ajtić, O., Đikić, D., Dragojević, T., Otašević, V., Živković, E., Vuković, V., Vukotić, M., Subotički, T., Diklić, M., Suvajdžić-Vuković, N., Mitrović, M., Mihaljević, B., Antić, D.,& Čokić, V.. (2023). Activation of coagulation factors and prothrombic properties of endothelium in hematological malignancies. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
Belgrade: Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 134-134.
https://hdl.handle.net/21.15107/rcub_rimi_1420

Mitrović-Ajtić O, Đikić D, Dragojević T, Otašević V, Živković E, Vuković V, Vukotić M, Subotički T, Diklić M, Suvajdžić-Vuković N, Mitrović M, Mihaljević B, Antić D, Čokić V. Activation of coagulation factors and prothrombic properties of endothelium in hematological malignancies. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia. 2023;:134-134.
https://hdl.handle.net/21.15107/rcub_rimi_1420 .

Mitrović-Ajtić, Olivera, Đikić, Dragoslava, Dragojević, Teodora, Otašević, Vladimir, Živković, Emilija, Vuković, Vojin, Vukotić, Milica, Subotički, Tijana, Diklić, Miloš, Suvajdžić-Vuković, Nada, Mitrović, Mirjana, Mihaljević, Biljana, Antić, Darko, Čokić, Vladan, "Activation of coagulation factors and prothrombic properties of endothelium in hematological malignancies" in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia (2023):134-134,
https://hdl.handle.net/21.15107/rcub_rimi_1420 .

TY  - CONF
AU  - Mitrović Ajtić, Olivera
AU  - Đikić, Dragoslava
AU  - Suvajdžić-Vuković, Nada
AU  - Mitrović, Mirjana
AU  - Subotički, Tijana
AU  - Vukotić, Milica
AU  - Dragojević, Teodora
AU  - Diklić, Miloš
AU  - Pantić, Nikola
AU  - Čokić, Vladan
PY  - 2023
UR  - http://rimi.imi.bg.ac.rs/handle/123456789/1425
AB  - Background: Patients with acute myeloid leukemia (AML) have an increased risk of thrombotic complications in the range of 4.2 - 5.2%.
Aims: Our hypothesis is that inflammation is responsible for deterioration of coagulation in AML.
Methods: Quantification of neutrophil extracellular traps (NETs) from peripheral blood of patients with AML by measurement of circulating cell-free DNA (cfDNA) and myeloperoxidase (MPO) activity. Inflammatory cytokines, coagulation factors and chemokines are measured by enzyme-linked immunosorbent assay (ELISA) and flow cytometry in peripheral blood, while fibrinolytic activity with fluorescent tissue-type plasminogen activator (tPA) and urokinase plasminogen activator (uPA) assays.
Results: The pro-inflammatory cytokines IL-1β and TNF-α were significantly increased in AML, but not the chemokines IL-8 and MCP-1. NETs were increased in the peripheral blood of patients with AML (p<0.05) as measured by cfDNA and MPO activity. Regarding coagulation, factor VIII (p<0.05) and adhesion molecule P-selectin (p<0.001) were increased in plasma. Fibrinolytic activity was 3-fold decreased in the plasma of patients with AML (p<0.01) as measured by tPA. In contrast, uPA levels were increased in patients with AML (p<0.05). Tissue factor (CD142+) inflammatory microparticles derived from monocytes (CD14+: 5.1±0.6, p<0.001), activated monocytes (CD14+/CD16+: 2.89±0.4%, p<0.05) and circulating endothelial cells (CD31+/CD144+: 4.08±0.5%, p<0.05) were increased in AML compared to healthy controls.
Summary/Conclusion: Chronic inflammation is present in AML in parallel with reduced fibrinolysis and increased coagulation provoking the risk of thrombosis. A panel of the applied inflammatory/ procoagulant biomarkers can be used as a predictor of thrombosis in AML.
PB  - Wolters Kluwer Health, Inc.
C3  - HemaSphere - EHA2023 Hybrid Congress Abstract Book
T1  - PB1832: Inflammation induced coagulation in acute myeloid leukemia
IS  - S3
SP  - 68019.6f
VL  - 7
DO  - 10.1097/01.HS9.0000974172.68019.6f
ER  - 

@conference{
author = "Mitrović Ajtić, Olivera and Đikić, Dragoslava and Suvajdžić-Vuković, Nada and Mitrović, Mirjana and Subotički, Tijana and Vukotić, Milica and Dragojević, Teodora and Diklić, Miloš and Pantić, Nikola and Čokić, Vladan",
year = "2023",
abstract = "Background: Patients with acute myeloid leukemia (AML) have an increased risk of thrombotic complications in the range of 4.2 - 5.2%.
Aims: Our hypothesis is that inflammation is responsible for deterioration of coagulation in AML.
Methods: Quantification of neutrophil extracellular traps (NETs) from peripheral blood of patients with AML by measurement of circulating cell-free DNA (cfDNA) and myeloperoxidase (MPO) activity. Inflammatory cytokines, coagulation factors and chemokines are measured by enzyme-linked immunosorbent assay (ELISA) and flow cytometry in peripheral blood, while fibrinolytic activity with fluorescent tissue-type plasminogen activator (tPA) and urokinase plasminogen activator (uPA) assays.
Results: The pro-inflammatory cytokines IL-1β and TNF-α were significantly increased in AML, but not the chemokines IL-8 and MCP-1. NETs were increased in the peripheral blood of patients with AML (p<0.05) as measured by cfDNA and MPO activity. Regarding coagulation, factor VIII (p<0.05) and adhesion molecule P-selectin (p<0.001) were increased in plasma. Fibrinolytic activity was 3-fold decreased in the plasma of patients with AML (p<0.01) as measured by tPA. In contrast, uPA levels were increased in patients with AML (p<0.05). Tissue factor (CD142+) inflammatory microparticles derived from monocytes (CD14+: 5.1±0.6, p<0.001), activated monocytes (CD14+/CD16+: 2.89±0.4%, p<0.05) and circulating endothelial cells (CD31+/CD144+: 4.08±0.5%, p<0.05) were increased in AML compared to healthy controls.
Summary/Conclusion: Chronic inflammation is present in AML in parallel with reduced fibrinolysis and increased coagulation provoking the risk of thrombosis. A panel of the applied inflammatory/ procoagulant biomarkers can be used as a predictor of thrombosis in AML.",
publisher = "Wolters Kluwer Health, Inc.",
journal = "HemaSphere - EHA2023 Hybrid Congress Abstract Book",
title = "PB1832: Inflammation induced coagulation in acute myeloid leukemia",
number = "S3",
pages = "68019.6f",
volume = "7",
doi = "10.1097/01.HS9.0000974172.68019.6f"
}

Mitrović Ajtić, O., Đikić, D., Suvajdžić-Vuković, N., Mitrović, M., Subotički, T., Vukotić, M., Dragojević, T., Diklić, M., Pantić, N.,& Čokić, V.. (2023). PB1832: Inflammation induced coagulation in acute myeloid leukemia. in HemaSphere - EHA2023 Hybrid Congress Abstract Book
Wolters Kluwer Health, Inc.., 7(S3), 68019.6f.
https://doi.org/10.1097/01.HS9.0000974172.68019.6f

Mitrović Ajtić O, Đikić D, Suvajdžić-Vuković N, Mitrović M, Subotički T, Vukotić M, Dragojević T, Diklić M, Pantić N, Čokić V. PB1832: Inflammation induced coagulation in acute myeloid leukemia. in HemaSphere - EHA2023 Hybrid Congress Abstract Book. 2023;7(S3):68019.6f.
doi:10.1097/01.HS9.0000974172.68019.6f .

Mitrović Ajtić, Olivera, Đikić, Dragoslava, Suvajdžić-Vuković, Nada, Mitrović, Mirjana, Subotički, Tijana, Vukotić, Milica, Dragojević, Teodora, Diklić, Miloš, Pantić, Nikola, Čokić, Vladan, "PB1832: Inflammation induced coagulation in acute myeloid leukemia" in HemaSphere - EHA2023 Hybrid Congress Abstract Book, 7, no. S3 (2023):68019.6f,
https://doi.org/10.1097/01.HS9.0000974172.68019.6f . .

TY  - CONF
AU  - Subotički, Tijana
AU  - Živković, Emilija
AU  - Mitrović Ajtić, Olivera
AU  - Vukotić, Milica
AU  - Đikić, Dragoslava
AU  - Dragojević, Teodora
AU  - Šefer, D.
AU  - Bižić, S.
AU  - Santibanez, Juan F.
AU  - Gotić, M.
AU  - Čokić, Vladan P.
PY  - 2022
UR  - http://rimi.imi.bg.ac.rs/handle/123456789/1490
AB  - Background: Myeloproliferative neoplasms (MPN) are clonal hematopoietic disorders that include polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF). Bone marrow fibrosis (BMF) is a shared feature of all MPN, although it is most pronounced in PMF and represents a major diagnostic criteria. Studies suggest a correlation between the grade of BMF and prognosis of MPN, with more fibrosis associated with worse outcome. A large body of evidence has suggested that transforming growth factor beta (TGF-β) is among the most prominent inducers of fibrotic processes. Bone morphogenetic proteins (BMPs) are major regulators of cell fate in tissue homeostasis. In MPN, bone marrow-derived mesenchymal stromal cells (BM-MSC) are identified as a major cellular source of fibrosis, but exact molecular mechanism involved have not been identified so far.  Aims: In addition to apoptosis and prolifration, we analyzed the effect of BMP2 and TGF- β / SMAD signaling pathway on the fibrotic phenotype of BM-MSC isolated from MPN patients and healthy donors.  Methods: Bone marrow aspirates from 5 newly diagnosed MPN patients (3 PMF and 2 PV patients) and 3 healthy donors were analyzed by immunofluorescence expression of fibronectin and alpha smooth muscle Actin (αSMA), after treatment with TGF-β and / or BMP2. Using immunocytochemistry, we analyzed HEL 92.1.7 cells with a homozygous expression of JAK2V617F for proliferation (Ki67) and apoptosis (ssDNA) during exposure to BMP2 and selective BMP signaling inhibitor LDN-193189.  Results: Our results showed that TGF-β significantly increased fibronectin expression, in contrast to BMP2, in BM-MSC of healthy donors. Also, the joint treatment of TGF-β and BMP2 reduced the level of fibronectin expression relative to TGF-β. In addition, TGF-β increased αSMA expression in BM-MSC from healthy donors. In contrast, BMP2 reduces the expression of αSMA and fibronectin in BM-MSC of healthy donors.BMP2 significantly reduced fibronectin expression in BM-MSC compared to untreated cells of patients with MPN. BMP2 dose dependently and significantly (p<0.01) increased the proliferation of HEL 92.1.7 cells, while the BMP signaling inhibitor LDN-193189 also demonstrated dose dependence in stimulation of proliferation (p<0.001). Apoptosis of HEL 92.1.7 cells was slightly affected by BMP2 and LDN-193189.  Summary/Conclusion: Our results show that BMP2 has anti-fibrotic activity that is antagonistic to TGF-β. This indicate that BMP2 may modify the TGF-β signaling pathway.
PB  - Wolters Kluwer Health, Inc.
C3  - HemaSphere - EHA2022 Hybrid Congress Abstract Book
T1  - P1002: Anti-fibrotic activity of BMP2 in bone marrow-derived mesenchymal stromal cells of myeloproliferative neoplasms
EP  - 893
IS  - Suppl (S3)
SP  - 892
VL  - 6
DO  - 10.1097/01.HS9.0000846876.57509.68
ER  - 

@conference{
author = "Subotički, Tijana and Živković, Emilija and Mitrović Ajtić, Olivera and Vukotić, Milica and Đikić, Dragoslava and Dragojević, Teodora and Šefer, D. and Bižić, S. and Santibanez, Juan F. and Gotić, M. and Čokić, Vladan P.",
year = "2022",
abstract = "Background: Myeloproliferative neoplasms (MPN) are clonal hematopoietic disorders that include polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF). Bone marrow fibrosis (BMF) is a shared feature of all MPN, although it is most pronounced in PMF and represents a major diagnostic criteria. Studies suggest a correlation between the grade of BMF and prognosis of MPN, with more fibrosis associated with worse outcome. A large body of evidence has suggested that transforming growth factor beta (TGF-β) is among the most prominent inducers of fibrotic processes. Bone morphogenetic proteins (BMPs) are major regulators of cell fate in tissue homeostasis. In MPN, bone marrow-derived mesenchymal stromal cells (BM-MSC) are identified as a major cellular source of fibrosis, but exact molecular mechanism involved have not been identified so far.  Aims: In addition to apoptosis and prolifration, we analyzed the effect of BMP2 and TGF- β / SMAD signaling pathway on the fibrotic phenotype of BM-MSC isolated from MPN patients and healthy donors.  Methods: Bone marrow aspirates from 5 newly diagnosed MPN patients (3 PMF and 2 PV patients) and 3 healthy donors were analyzed by immunofluorescence expression of fibronectin and alpha smooth muscle Actin (αSMA), after treatment with TGF-β and / or BMP2. Using immunocytochemistry, we analyzed HEL 92.1.7 cells with a homozygous expression of JAK2V617F for proliferation (Ki67) and apoptosis (ssDNA) during exposure to BMP2 and selective BMP signaling inhibitor LDN-193189.  Results: Our results showed that TGF-β significantly increased fibronectin expression, in contrast to BMP2, in BM-MSC of healthy donors. Also, the joint treatment of TGF-β and BMP2 reduced the level of fibronectin expression relative to TGF-β. In addition, TGF-β increased αSMA expression in BM-MSC from healthy donors. In contrast, BMP2 reduces the expression of αSMA and fibronectin in BM-MSC of healthy donors.BMP2 significantly reduced fibronectin expression in BM-MSC compared to untreated cells of patients with MPN. BMP2 dose dependently and significantly (p<0.01) increased the proliferation of HEL 92.1.7 cells, while the BMP signaling inhibitor LDN-193189 also demonstrated dose dependence in stimulation of proliferation (p<0.001). Apoptosis of HEL 92.1.7 cells was slightly affected by BMP2 and LDN-193189.  Summary/Conclusion: Our results show that BMP2 has anti-fibrotic activity that is antagonistic to TGF-β. This indicate that BMP2 may modify the TGF-β signaling pathway.",
publisher = "Wolters Kluwer Health, Inc.",
journal = "HemaSphere - EHA2022 Hybrid Congress Abstract Book",
title = "P1002: Anti-fibrotic activity of BMP2 in bone marrow-derived mesenchymal stromal cells of myeloproliferative neoplasms",
pages = "893-892",
number = "Suppl (S3)",
volume = "6",
doi = "10.1097/01.HS9.0000846876.57509.68"
}

Subotički, T., Živković, E., Mitrović Ajtić, O., Vukotić, M., Đikić, D., Dragojević, T., Šefer, D., Bižić, S., Santibanez, J. F., Gotić, M.,& Čokić, V. P.. (2022). P1002: Anti-fibrotic activity of BMP2 in bone marrow-derived mesenchymal stromal cells of myeloproliferative neoplasms. in HemaSphere - EHA2022 Hybrid Congress Abstract Book
Wolters Kluwer Health, Inc.., 6(Suppl (S3)), 892-893.
https://doi.org/10.1097/01.HS9.0000846876.57509.68

Subotički T, Živković E, Mitrović Ajtić O, Vukotić M, Đikić D, Dragojević T, Šefer D, Bižić S, Santibanez JF, Gotić M, Čokić VP. P1002: Anti-fibrotic activity of BMP2 in bone marrow-derived mesenchymal stromal cells of myeloproliferative neoplasms. in HemaSphere - EHA2022 Hybrid Congress Abstract Book. 2022;6(Suppl (S3)):892-893.
doi:10.1097/01.HS9.0000846876.57509.68 .

Subotički, Tijana, Živković, Emilija, Mitrović Ajtić, Olivera, Vukotić, Milica, Đikić, Dragoslava, Dragojević, Teodora, Šefer, D., Bižić, S., Santibanez, Juan F., Gotić, M., Čokić, Vladan P., "P1002: Anti-fibrotic activity of BMP2 in bone marrow-derived mesenchymal stromal cells of myeloproliferative neoplasms" in HemaSphere - EHA2022 Hybrid Congress Abstract Book, 6, no. Suppl (S3) (2022):892-893,
https://doi.org/10.1097/01.HS9.0000846876.57509.68 . .

TY  - CONF
AU  - Dragojević, Teodora
AU  - Mitrović Ajtić, Olivera
AU  - Diklić, Miloš
AU  - Subotički, Tijana
AU  - Đikić, Dragoslava
AU  - Živković, Emilija
AU  - Čokić, Vladan
AU  - Vukotić, Milica
PY  - 2022
UR  - http://rimi.imi.bg.ac.rs/handle/123456789/1427
AB  - Background:: Hydroxyurea (HU) is a chemotherapeutic agent that reduces ribonucleotide reductase, stops DNA synthesis and repar, and therefore causes cell proliferation inhibition and apoptosis. Due to its cytostatic properties, HU is frequently used for treatment of myeloproliferative neoplasms, ovarian cancer, and sickle cell anemia. Nitric oxide (NO), produced by nitric oxide synthase (NOS) enzymes is a potent signaling molecule involved in blood flow regulation, neutrotransmission, and immunity. Although HU treatment increases NO levels, up to date it is not clear whether it originates from activation of NOS enzymes or HU degradation.

Aims: The aim of this study was to determine the involvement of NOS2 enzyme in the cytostatic effect of HU.
Methods: To examine the involvement of the NOS2 enzyme in the molecular mechanism of HU, we treated erythroleukemic HEL92.7.1 cells with pan-selective NOS inhibitor L-NAME (200µM, 1mM, and 5mM), NOS2 specific inhibitor 1400W (1, 10, and 100µM), or NOS2/NOS3 inhibitor DPI (1, 5, and 10µM), in combination with hydroxyurea (200µM), and monitored their effect on proliferation and cell cycle. Immunocytochemistry for the proliferation marker Ki67 was performed to assess proliferation, while cell distribution in cell cycle phases was determined by flow cytometry after propidium iodide staining. Colony forming assay have been performed with the bone marrow cells of Nos2 null mice after oral HU treatment to corroborate the data obtained by enzymatic inhibition.
Results: In this study, we demonstrated that treatment of HEL92.7.1 cells with HU induces a dose-dependent increase in NOS2 protein levels and two products of the enzyme NOS - NO and citrulline. HU-induced citrulline levels can be reduced by treatment with the NOS inhibitor L-NAME, indicating that NO is produced de novo by the NOS enzyme rather than HU degradation. Inhibition of the NOS2 enzyme by L-NAME, 1400W, or DPI was sufficient to abolish HU-mediated inhibition of proliferation. While HU increased the number of cells in S-phase of the cycle at the expanse of the G0/G1 due to blocked DNA synthesis, combined treatment with HU and L-NAME or DPI inhibitor resulted in decreased G0/G1 phase and increased S and G2/M phases pointing to increased proliferation. These data indicate that the cytostatic properties of HU are mediated by the NOS2 enzyme. A colony formation assay showed that Nos2 deficient bone marrow cells isolated from mice treated orally with HU (200mg/kg) formed significantly more erythroid colonies (BFU-E and CFU-E) and granulocyte/macrophage progenitors (CFU-GM) compared to HU treated wild-type and untreated Nos2 null mice showing the involvement of Nos2 in the molecular mechanism of HU in vivo.
Summary/Conclusion: Our results show that HU induces the enzymatic activity of the NOS2 protein which in turn is involved in the HU regulation of proliferation and cell cycle. Comprehensive knowledge of the molecular mechanism of HU might help to improve its beneficial properties and decrease adverse effects.
PB  - Wolters Kluwer Health, Inc.
C3  - HemaSphere - EHA2022 Hybrid Congress Abstract Book
T1  - P1474: Molecural mechanism of chemotherapeutic hydroxyurea is mediated by NOS2
EP  - 1357
IS  - S3
SP  - 1356
VL  - 6
DO  - 10.1097/01.HS9.0000848752.48359.16
ER  - 

@conference{
author = "Dragojević, Teodora and Mitrović Ajtić, Olivera and Diklić, Miloš and Subotički, Tijana and Đikić, Dragoslava and Živković, Emilija and Čokić, Vladan and Vukotić, Milica",
year = "2022",
abstract = "Background:: Hydroxyurea (HU) is a chemotherapeutic agent that reduces ribonucleotide reductase, stops DNA synthesis and repar, and therefore causes cell proliferation inhibition and apoptosis. Due to its cytostatic properties, HU is frequently used for treatment of myeloproliferative neoplasms, ovarian cancer, and sickle cell anemia. Nitric oxide (NO), produced by nitric oxide synthase (NOS) enzymes is a potent signaling molecule involved in blood flow regulation, neutrotransmission, and immunity. Although HU treatment increases NO levels, up to date it is not clear whether it originates from activation of NOS enzymes or HU degradation.

Aims: The aim of this study was to determine the involvement of NOS2 enzyme in the cytostatic effect of HU.
Methods: To examine the involvement of the NOS2 enzyme in the molecular mechanism of HU, we treated erythroleukemic HEL92.7.1 cells with pan-selective NOS inhibitor L-NAME (200µM, 1mM, and 5mM), NOS2 specific inhibitor 1400W (1, 10, and 100µM), or NOS2/NOS3 inhibitor DPI (1, 5, and 10µM), in combination with hydroxyurea (200µM), and monitored their effect on proliferation and cell cycle. Immunocytochemistry for the proliferation marker Ki67 was performed to assess proliferation, while cell distribution in cell cycle phases was determined by flow cytometry after propidium iodide staining. Colony forming assay have been performed with the bone marrow cells of Nos2 null mice after oral HU treatment to corroborate the data obtained by enzymatic inhibition.
Results: In this study, we demonstrated that treatment of HEL92.7.1 cells with HU induces a dose-dependent increase in NOS2 protein levels and two products of the enzyme NOS - NO and citrulline. HU-induced citrulline levels can be reduced by treatment with the NOS inhibitor L-NAME, indicating that NO is produced de novo by the NOS enzyme rather than HU degradation. Inhibition of the NOS2 enzyme by L-NAME, 1400W, or DPI was sufficient to abolish HU-mediated inhibition of proliferation. While HU increased the number of cells in S-phase of the cycle at the expanse of the G0/G1 due to blocked DNA synthesis, combined treatment with HU and L-NAME or DPI inhibitor resulted in decreased G0/G1 phase and increased S and G2/M phases pointing to increased proliferation. These data indicate that the cytostatic properties of HU are mediated by the NOS2 enzyme. A colony formation assay showed that Nos2 deficient bone marrow cells isolated from mice treated orally with HU (200mg/kg) formed significantly more erythroid colonies (BFU-E and CFU-E) and granulocyte/macrophage progenitors (CFU-GM) compared to HU treated wild-type and untreated Nos2 null mice showing the involvement of Nos2 in the molecular mechanism of HU in vivo.
Summary/Conclusion: Our results show that HU induces the enzymatic activity of the NOS2 protein which in turn is involved in the HU regulation of proliferation and cell cycle. Comprehensive knowledge of the molecular mechanism of HU might help to improve its beneficial properties and decrease adverse effects.",
publisher = "Wolters Kluwer Health, Inc.",
journal = "HemaSphere - EHA2022 Hybrid Congress Abstract Book",
title = "P1474: Molecural mechanism of chemotherapeutic hydroxyurea is mediated by NOS2",
pages = "1357-1356",
number = "S3",
volume = "6",
doi = "10.1097/01.HS9.0000848752.48359.16"
}

Dragojević, T., Mitrović Ajtić, O., Diklić, M., Subotički, T., Đikić, D., Živković, E., Čokić, V.,& Vukotić, M.. (2022). P1474: Molecural mechanism of chemotherapeutic hydroxyurea is mediated by NOS2. in HemaSphere - EHA2022 Hybrid Congress Abstract Book
Wolters Kluwer Health, Inc.., 6(S3), 1356-1357.
https://doi.org/10.1097/01.HS9.0000848752.48359.16

Dragojević T, Mitrović Ajtić O, Diklić M, Subotički T, Đikić D, Živković E, Čokić V, Vukotić M. P1474: Molecural mechanism of chemotherapeutic hydroxyurea is mediated by NOS2. in HemaSphere - EHA2022 Hybrid Congress Abstract Book. 2022;6(S3):1356-1357.
doi:10.1097/01.HS9.0000848752.48359.16 .

Dragojević, Teodora, Mitrović Ajtić, Olivera, Diklić, Miloš, Subotički, Tijana, Đikić, Dragoslava, Živković, Emilija, Čokić, Vladan, Vukotić, Milica, "P1474: Molecural mechanism of chemotherapeutic hydroxyurea is mediated by NOS2" in HemaSphere - EHA2022 Hybrid Congress Abstract Book, 6, no. S3 (2022):1356-1357,
https://doi.org/10.1097/01.HS9.0000848752.48359.16 . .

TY  - JOUR
AU  - Đikić, Dragoslava
AU  - Bogdanović, Andrija
AU  - Marković, Dragana
AU  - Mitrović-Ajtić, Olivera
AU  - Subotički, Tijana
AU  - Diklić, Miloš
AU  - Vukotić, Milica
AU  - Dragojević, Teodora
AU  - Živković, Emilija
AU  - Santibanez, Juan F.
AU  - Čokić, Vladan
PY  - 2022
UR  - http://rimi.imi.bg.ac.rs/handle/123456789/1202
AB  - Chronic inflammation is characterized by the production of reactive oxygen species (ROS), reactive nitrogen species, and inflammatory cytokines in myeloproliferative neoplasms (MPNs). In addition to these parameters, the aim of this study was to analyze the influence of ROS on the pro-liferation-related AKT/mTOR signaling pathway and the relationship with inflammatory factors in chronic myelogenous leukemia (CML). The activity of the antioxidant enzymes superoxide dis-mutase, glutathione peroxidase, and catalase is reduced in erythrocytes while levels of the oxidative stress markers malondialdehyde and protein carbonyl are elevated in the plasma of patients with CML. In addition, nitrogen species (nitrotyrosine, iNOS, eNOS) and inflammation markers (IL-6, NFkB, and S100 protein) were increased in granulocytes of CML while anti-inflammatory levels of IL-10 were decreased in plasma. CML granulocytes exhibited greater resistance to cytotoxic H2O2 activity compared to healthy subjects. Moreover, phosphorylation of the apoptotic p53 protein was reduced while the activity of the AKT/mTOR signaling pathway was increased, which was further enhanced by oxidative stress (H2O2) in granulocytes and erythroleukemic K562 cells. IL-6 caused oxidative stress and DNA damage that was mitigated using antioxidant or inhibition of inflammatory NFkB transcription factor in K562 cells. We demonstrated the presence of oxidative and ni-trosative stress in CML, with the former mediated by AKT/mTOR signaling and stimulated by in-flammation.
PB  - MDPI
T2  - Biomolecules
T1  - Inflammation Promotes Oxidative and Nitrosative Stress in Chronic Myelogenous Leukemia
IS  - 2
SP  - 247
VL  - 12
DO  - 10.3390/biom12020247
ER  - 

@article{
author = "Đikić, Dragoslava and Bogdanović, Andrija and Marković, Dragana and Mitrović-Ajtić, Olivera and Subotički, Tijana and Diklić, Miloš and Vukotić, Milica and Dragojević, Teodora and Živković, Emilija and Santibanez, Juan F. and Čokić, Vladan",
year = "2022",
abstract = "Chronic inflammation is characterized by the production of reactive oxygen species (ROS), reactive nitrogen species, and inflammatory cytokines in myeloproliferative neoplasms (MPNs). In addition to these parameters, the aim of this study was to analyze the influence of ROS on the pro-liferation-related AKT/mTOR signaling pathway and the relationship with inflammatory factors in chronic myelogenous leukemia (CML). The activity of the antioxidant enzymes superoxide dis-mutase, glutathione peroxidase, and catalase is reduced in erythrocytes while levels of the oxidative stress markers malondialdehyde and protein carbonyl are elevated in the plasma of patients with CML. In addition, nitrogen species (nitrotyrosine, iNOS, eNOS) and inflammation markers (IL-6, NFkB, and S100 protein) were increased in granulocytes of CML while anti-inflammatory levels of IL-10 were decreased in plasma. CML granulocytes exhibited greater resistance to cytotoxic H2O2 activity compared to healthy subjects. Moreover, phosphorylation of the apoptotic p53 protein was reduced while the activity of the AKT/mTOR signaling pathway was increased, which was further enhanced by oxidative stress (H2O2) in granulocytes and erythroleukemic K562 cells. IL-6 caused oxidative stress and DNA damage that was mitigated using antioxidant or inhibition of inflammatory NFkB transcription factor in K562 cells. We demonstrated the presence of oxidative and ni-trosative stress in CML, with the former mediated by AKT/mTOR signaling and stimulated by in-flammation.",
publisher = "MDPI",
journal = "Biomolecules",
title = "Inflammation Promotes Oxidative and Nitrosative Stress in Chronic Myelogenous Leukemia",
number = "2",
pages = "247",
volume = "12",
doi = "10.3390/biom12020247"
}

Đikić, D., Bogdanović, A., Marković, D., Mitrović-Ajtić, O., Subotički, T., Diklić, M., Vukotić, M., Dragojević, T., Živković, E., Santibanez, J. F.,& Čokić, V.. (2022). Inflammation Promotes Oxidative and Nitrosative Stress in Chronic Myelogenous Leukemia. in Biomolecules
MDPI., 12(2), 247.
https://doi.org/10.3390/biom12020247

Đikić D, Bogdanović A, Marković D, Mitrović-Ajtić O, Subotički T, Diklić M, Vukotić M, Dragojević T, Živković E, Santibanez JF, Čokić V. Inflammation Promotes Oxidative and Nitrosative Stress in Chronic Myelogenous Leukemia. in Biomolecules. 2022;12(2):247.
doi:10.3390/biom12020247 .

Đikić, Dragoslava, Bogdanović, Andrija, Marković, Dragana, Mitrović-Ajtić, Olivera, Subotički, Tijana, Diklić, Miloš, Vukotić, Milica, Dragojević, Teodora, Živković, Emilija, Santibanez, Juan F., Čokić, Vladan, "Inflammation Promotes Oxidative and Nitrosative Stress in Chronic Myelogenous Leukemia" in Biomolecules, 12, no. 2 (2022):247,
https://doi.org/10.3390/biom12020247 . .

TY  - JOUR
AU  - Vukotić, Milica
AU  - Kapor, Sunčica
AU  - Dragojević, Teodora
AU  - Đikić, Dragoslava
AU  - Mitrović-Ajtić, Olivera
AU  - Diklić, Miloš
AU  - Subotički, Tijana
AU  - Živković, Emilija
AU  - Beleslin-Čokić, Bojana
AU  - Vojvodić, Aleksandar
AU  - Santibanez, Juan F.
AU  - Gotić, Mirjana
AU  - Čokić, Vladan
PY  - 2022
UR  - http://rimi.imi.bg.ac.rs/handle/123456789/1226
AB  - Although bone marrow-derived mesenchymal stromal cells (BM-MSCs) have been identified as a major cellular source of fibrosis, the exact molecular mechanism and signaling pathways involved have not been identified thus far. Here, we show that BM-MSCs contribute to fibrosis in myeloproliferative neoplasms (MPNs) by differentiating into αSMA-positive myofibroblasts. These cells display a dysregulated extracellular matrix with increased FN1 production and secretion of profibrotic MMP9 compared to healthy donor cells. Fibrogenic TGFβ and inflammatory JAK2/STAT3 and NFκB signaling pathway activity is increased in BM-MSCs of MPN patients. Moreover, coculture with mononuclear cells from MPN patients was sufficient to induce fibrosis in healthy BM-MSCs. Inhibition of JAK1/2, SMAD3 or NFκB significantly reduced the fibrotic phenotype of MPN BM-MSCs and was able to prevent the development of fibrosis induced by coculture of healthy BM-MSCs and MPN mononuclear cells with overly active JAK/STAT signaling, underlining their involvement in fibrosis. Combined treatment with JAK1/2 and SMAD3 inhibitors showed synergistic and the most favorable effects on αSMA and FN1 expression in BM-MSCs. These results support the combined inhibition of TGFβ and inflammatory signaling to extenuate fibrosis in MPN.
PB  - Korean Society of Medical Biochemistry and Molecular Biology [Associate Organisation]
PB  - Springer Nature [Commercial Publisher]
T2  - Experimental & Molecular Medicine
T1  - Inhibition of proinflammatory signaling impairs fibrosis of bone marrow mesenchymal stromal cells in myeloproliferative neoplasms
EP  - 284
IS  - 3
SP  - 273
VL  - 54
DO  - 10.1038/s12276-022-00742-y
ER  - 

@article{
author = "Vukotić, Milica and Kapor, Sunčica and Dragojević, Teodora and Đikić, Dragoslava and Mitrović-Ajtić, Olivera and Diklić, Miloš and Subotički, Tijana and Živković, Emilija and Beleslin-Čokić, Bojana and Vojvodić, Aleksandar and Santibanez, Juan F. and Gotić, Mirjana and Čokić, Vladan",
year = "2022",
abstract = "Although bone marrow-derived mesenchymal stromal cells (BM-MSCs) have been identified as a major cellular source of fibrosis, the exact molecular mechanism and signaling pathways involved have not been identified thus far. Here, we show that BM-MSCs contribute to fibrosis in myeloproliferative neoplasms (MPNs) by differentiating into αSMA-positive myofibroblasts. These cells display a dysregulated extracellular matrix with increased FN1 production and secretion of profibrotic MMP9 compared to healthy donor cells. Fibrogenic TGFβ and inflammatory JAK2/STAT3 and NFκB signaling pathway activity is increased in BM-MSCs of MPN patients. Moreover, coculture with mononuclear cells from MPN patients was sufficient to induce fibrosis in healthy BM-MSCs. Inhibition of JAK1/2, SMAD3 or NFκB significantly reduced the fibrotic phenotype of MPN BM-MSCs and was able to prevent the development of fibrosis induced by coculture of healthy BM-MSCs and MPN mononuclear cells with overly active JAK/STAT signaling, underlining their involvement in fibrosis. Combined treatment with JAK1/2 and SMAD3 inhibitors showed synergistic and the most favorable effects on αSMA and FN1 expression in BM-MSCs. These results support the combined inhibition of TGFβ and inflammatory signaling to extenuate fibrosis in MPN.",
publisher = "Korean Society of Medical Biochemistry and Molecular Biology [Associate Organisation], Springer Nature [Commercial Publisher]",
journal = "Experimental & Molecular Medicine",
title = "Inhibition of proinflammatory signaling impairs fibrosis of bone marrow mesenchymal stromal cells in myeloproliferative neoplasms",
pages = "284-273",
number = "3",
volume = "54",
doi = "10.1038/s12276-022-00742-y"
}

Vukotić, M., Kapor, S., Dragojević, T., Đikić, D., Mitrović-Ajtić, O., Diklić, M., Subotički, T., Živković, E., Beleslin-Čokić, B., Vojvodić, A., Santibanez, J. F., Gotić, M.,& Čokić, V.. (2022). Inhibition of proinflammatory signaling impairs fibrosis of bone marrow mesenchymal stromal cells in myeloproliferative neoplasms. in Experimental & Molecular Medicine
Korean Society of Medical Biochemistry and Molecular Biology [Associate Organisation]., 54(3), 273-284.
https://doi.org/10.1038/s12276-022-00742-y

Vukotić M, Kapor S, Dragojević T, Đikić D, Mitrović-Ajtić O, Diklić M, Subotički T, Živković E, Beleslin-Čokić B, Vojvodić A, Santibanez JF, Gotić M, Čokić V. Inhibition of proinflammatory signaling impairs fibrosis of bone marrow mesenchymal stromal cells in myeloproliferative neoplasms. in Experimental & Molecular Medicine. 2022;54(3):273-284.
doi:10.1038/s12276-022-00742-y .

Vukotić, Milica, Kapor, Sunčica, Dragojević, Teodora, Đikić, Dragoslava, Mitrović-Ajtić, Olivera, Diklić, Miloš, Subotički, Tijana, Živković, Emilija, Beleslin-Čokić, Bojana, Vojvodić, Aleksandar, Santibanez, Juan F., Gotić, Mirjana, Čokić, Vladan, "Inhibition of proinflammatory signaling impairs fibrosis of bone marrow mesenchymal stromal cells in myeloproliferative neoplasms" in Experimental & Molecular Medicine, 54, no. 3 (2022):273-284,
https://doi.org/10.1038/s12276-022-00742-y . .

TY  - JOUR
AU  - Mitrović-Ajtić, Olivera
AU  - Subotički, Tijana
AU  - Diklić, Miloš
AU  - Đikić, Dragoslava
AU  - Vukotić, Milica
AU  - Dragojević, Teodora
AU  - Živković, Emilija
AU  - Antić, Darko
AU  - Čokić, Vladan
PY  - 2022
UR  - http://rimi.imi.bg.ac.rs/handle/123456789/1244
AB  - The calcium-binding proteins S100A4, S100A8, and S100A9 are upregulated in chronic lymphocytic leukemia (CLL), while the S100A9 promotes NF-κB activity during disease progression. The S100-protein family has been involved in several malignancies as mediators of inflammation and proliferation. The hypothesis of our study is that S100A proteins are mediators in signaling pathways associated with inflammation-induced proliferation, such as NF-κB, PI3K/AKT, and JAK/STAT. The mononuclear cells (MNCs) of CLL were treated with proinflammatory IL-6, anti-inflammatory IL-10 cytokines, inhibitors of JAK1/2, NF-κB, and PI3K signaling pathways, to evaluate S100A4, S100A8, S100A9, and S100A12 expression as well as NF-κB activation by qRT-PCR, immunocytochemistry, and immunoblotting. The quantity of S100A4, S100A8, and S100A9 positive cells (p < 0.05) and their protein expression (p < 0.01) were significantly decreased in MNCs of CLL patients compared to healthy controls. The S100A levels were generally increased in CD19+ cells compared to MNCs of CLL. The S100A4 gene expression was significantly stimulated (p < 0.05) by the inhibition of the PI3K/AKT signaling pathway in MNCs. IL-6 stimulated S100A4 and S100A8 protein expression, prevented by the NF-κB and JAK1/2 inhibitors. In contrast, IL-10 reduced S100A8, S100A9, and S100A12 protein expressions in MNCs of CLL. Moreover, IL-10 inhibited activation of NF-κB signaling (4-fold, p < 0.05). In conclusion, inflammation stimulated the S100A protein expression mediated via the proliferation-related signaling and balanced by the cytokines in CLL.
PB  - MDPI
T2  - International Journal of Molecular Sciences
T1  - Regulation of S100As Expression by Inflammatory Cytokines in Chronic Lymphocytic Leukemia
IS  - 13
SP  - 6952
VL  - 23
DO  - 10.3390/ijms23136952
ER  - 

@article{
author = "Mitrović-Ajtić, Olivera and Subotički, Tijana and Diklić, Miloš and Đikić, Dragoslava and Vukotić, Milica and Dragojević, Teodora and Živković, Emilija and Antić, Darko and Čokić, Vladan",
year = "2022",
abstract = "The calcium-binding proteins S100A4, S100A8, and S100A9 are upregulated in chronic lymphocytic leukemia (CLL), while the S100A9 promotes NF-κB activity during disease progression. The S100-protein family has been involved in several malignancies as mediators of inflammation and proliferation. The hypothesis of our study is that S100A proteins are mediators in signaling pathways associated with inflammation-induced proliferation, such as NF-κB, PI3K/AKT, and JAK/STAT. The mononuclear cells (MNCs) of CLL were treated with proinflammatory IL-6, anti-inflammatory IL-10 cytokines, inhibitors of JAK1/2, NF-κB, and PI3K signaling pathways, to evaluate S100A4, S100A8, S100A9, and S100A12 expression as well as NF-κB activation by qRT-PCR, immunocytochemistry, and immunoblotting. The quantity of S100A4, S100A8, and S100A9 positive cells (p < 0.05) and their protein expression (p < 0.01) were significantly decreased in MNCs of CLL patients compared to healthy controls. The S100A levels were generally increased in CD19+ cells compared to MNCs of CLL. The S100A4 gene expression was significantly stimulated (p < 0.05) by the inhibition of the PI3K/AKT signaling pathway in MNCs. IL-6 stimulated S100A4 and S100A8 protein expression, prevented by the NF-κB and JAK1/2 inhibitors. In contrast, IL-10 reduced S100A8, S100A9, and S100A12 protein expressions in MNCs of CLL. Moreover, IL-10 inhibited activation of NF-κB signaling (4-fold, p < 0.05). In conclusion, inflammation stimulated the S100A protein expression mediated via the proliferation-related signaling and balanced by the cytokines in CLL.",
publisher = "MDPI",
journal = "International Journal of Molecular Sciences",
title = "Regulation of S100As Expression by Inflammatory Cytokines in Chronic Lymphocytic Leukemia",
number = "13",
pages = "6952",
volume = "23",
doi = "10.3390/ijms23136952"
}

Mitrović-Ajtić, O., Subotički, T., Diklić, M., Đikić, D., Vukotić, M., Dragojević, T., Živković, E., Antić, D.,& Čokić, V.. (2022). Regulation of S100As Expression by Inflammatory Cytokines in Chronic Lymphocytic Leukemia. in International Journal of Molecular Sciences
MDPI., 23(13), 6952.
https://doi.org/10.3390/ijms23136952

Mitrović-Ajtić O, Subotički T, Diklić M, Đikić D, Vukotić M, Dragojević T, Živković E, Antić D, Čokić V. Regulation of S100As Expression by Inflammatory Cytokines in Chronic Lymphocytic Leukemia. in International Journal of Molecular Sciences. 2022;23(13):6952.
doi:10.3390/ijms23136952 .

Mitrović-Ajtić, Olivera, Subotički, Tijana, Diklić, Miloš, Đikić, Dragoslava, Vukotić, Milica, Dragojević, Teodora, Živković, Emilija, Antić, Darko, Čokić, Vladan, "Regulation of S100As Expression by Inflammatory Cytokines in Chronic Lymphocytic Leukemia" in International Journal of Molecular Sciences, 23, no. 13 (2022):6952,
https://doi.org/10.3390/ijms23136952 . .

TY  - JOUR
AU  - Mitrović-Ajtić, Olivera
AU  - Stanisavljević, Dejana
AU  - Miljatović, Sanja
AU  - Dragojević, Teodora
AU  - Živković, Emilija
AU  - Šabanović, Miloš
AU  - Čokić, Vladan
PY  - 2022
UR  - http://rimi.imi.bg.ac.rs/handle/123456789/1258
AB  - The COVID-19 pandemic has had a strong impact on people’s quality of life (QoL), which is affected by social and economic changes as well as by mental and physical health. The aim of this study was to determine QoL in post-COVID-19 patients who had required hospitalization, and to identify relevant sociodemographic data. We used questionnaires which considered demographic and socioeconomic data, health and vaccination status, the pandemic situation, and EQ-5D scoring. The interactions of all data and the scores of EQ-5D were analyzed. Multivariate logistic regression analysis was applied to the five dimensions of EQ-5D. In this single-hospital-cohort study, the average times elapsed since initial diagnosis and hospital admission were 2.5 (76.3 ± 18.1 days) and 5 months (155.4 ± 33.9 days), respectively. Post-COVID-19 females were 3–5 times more likely to be affected in terms of anxiety/depression, and in negative impact upon their usual activities, at 5 months after diagnosis. At the same time, reductions in mobility were 3–4 times more likely in elderly post-COVID-19 patients, whose levels of pain and discomfort increased. Single patients, those with low incomes, and those with severe clinical outcomes were 2–4 times more likely to experience a reduction in their usual activities, while the presence of co-morbidities and lower levels of education were associated with increased pain and discomfort. Aging-induced pain/discomfort and anxiety/depression were significantly exacerbated in elderly patients with widespread vaccination. Our study revealed effects of demographic and socioeconomic factors upon lower QoL in post-COVID-19 patients in four dimensions of EQ-5D: mobility, usual activity, pain/discomfort, and anxiety/depression, 5 months after first diagnosis and hospitalization.
PB  - MDPI
T2  - Healthcare
T1  - Quality of Life in Post-COVID-19 Patients after Hospitalization
IS  - 9
SP  - 1666
VL  - 10
DO  - 10.3390/healthcare10091666
ER  - 

@article{
author = "Mitrović-Ajtić, Olivera and Stanisavljević, Dejana and Miljatović, Sanja and Dragojević, Teodora and Živković, Emilija and Šabanović, Miloš and Čokić, Vladan",
year = "2022",
abstract = "The COVID-19 pandemic has had a strong impact on people’s quality of life (QoL), which is affected by social and economic changes as well as by mental and physical health. The aim of this study was to determine QoL in post-COVID-19 patients who had required hospitalization, and to identify relevant sociodemographic data. We used questionnaires which considered demographic and socioeconomic data, health and vaccination status, the pandemic situation, and EQ-5D scoring. The interactions of all data and the scores of EQ-5D were analyzed. Multivariate logistic regression analysis was applied to the five dimensions of EQ-5D. In this single-hospital-cohort study, the average times elapsed since initial diagnosis and hospital admission were 2.5 (76.3 ± 18.1 days) and 5 months (155.4 ± 33.9 days), respectively. Post-COVID-19 females were 3–5 times more likely to be affected in terms of anxiety/depression, and in negative impact upon their usual activities, at 5 months after diagnosis. At the same time, reductions in mobility were 3–4 times more likely in elderly post-COVID-19 patients, whose levels of pain and discomfort increased. Single patients, those with low incomes, and those with severe clinical outcomes were 2–4 times more likely to experience a reduction in their usual activities, while the presence of co-morbidities and lower levels of education were associated with increased pain and discomfort. Aging-induced pain/discomfort and anxiety/depression were significantly exacerbated in elderly patients with widespread vaccination. Our study revealed effects of demographic and socioeconomic factors upon lower QoL in post-COVID-19 patients in four dimensions of EQ-5D: mobility, usual activity, pain/discomfort, and anxiety/depression, 5 months after first diagnosis and hospitalization.",
publisher = "MDPI",
journal = "Healthcare",
title = "Quality of Life in Post-COVID-19 Patients after Hospitalization",
number = "9",
pages = "1666",
volume = "10",
doi = "10.3390/healthcare10091666"
}

Mitrović-Ajtić, O., Stanisavljević, D., Miljatović, S., Dragojević, T., Živković, E., Šabanović, M.,& Čokić, V.. (2022). Quality of Life in Post-COVID-19 Patients after Hospitalization. in Healthcare
MDPI., 10(9), 1666.
https://doi.org/10.3390/healthcare10091666

Mitrović-Ajtić O, Stanisavljević D, Miljatović S, Dragojević T, Živković E, Šabanović M, Čokić V. Quality of Life in Post-COVID-19 Patients after Hospitalization. in Healthcare. 2022;10(9):1666.
doi:10.3390/healthcare10091666 .

Mitrović-Ajtić, Olivera, Stanisavljević, Dejana, Miljatović, Sanja, Dragojević, Teodora, Živković, Emilija, Šabanović, Miloš, Čokić, Vladan, "Quality of Life in Post-COVID-19 Patients after Hospitalization" in Healthcare, 10, no. 9 (2022):1666,
https://doi.org/10.3390/healthcare10091666 . .

TY  - CONF
AU  - Miljatović, Sanja
AU  - Dragojević, Teodora
AU  - Đikić, Dragoslava
AU  - Živković, Emilija
AU  - Stevanović, Goran
AU  - Čokić, Vladan
PY  - 2022
UR  - http://rimi.imi.bg.ac.rs/handle/123456789/1457
AB  - Background. A vascular system inflammation is a risk of venous thromboembolism
and can result in widespread microangiopathy with microvascular thrombosis.
Methods. We performed laboratory clinical follow-up of patients with COVID-19 to
compare gender differences. To study the sex difference in COVID-19 outcome we
will measure estradiol and androgens: dihydrotestosterone (DHT) and sex hormone
binding globulin (SHBP) in plasma of 63 COVID-19 patients, analyzed by ELISA. Their
levels will be correlated to the adhesion molecules: soluble intercellular adhesion
molecule 1 (sICAM-1), soluble vascular adhesion molecule 1 (sVCAM-1), sE-selectin,
and sP-selectin as biomarkers for inflammation and thrombosis.
Results. DHT was increased (1.9 fold) in male COVID-19 patients compared to healthy
male volunteers. SHBP was significantly increased in COVID-19 patients compared
to healthy volunteers (p<0.05) as well as female vs. male COVID-19 patients (p<0.001,
2.5 fold). sVCAM-1 and sICAM-1 were increased in female COVID-19 patients compared
to male COVID-19 patients and female volunteers, respectively (p<0.05). The
sP-selection was significantly (p<0.01) increased in male vs. female COVID-19 patients.
SHBP was in negative correlation with sP-selectin (p<0.05). DHT was in positive
correlation with sVCAM-1 (p<0.05). Ferritin had 3-fold higher levels in male than
female COVID-19 patients (p<0.001).
Conclusions. Upregulation of androgen hormones and thrombotic biomarkers in
COVID-19 patients demonstrate sex dependence.
PB  - Belgrade: MEDAPP Association
C3  - The First World Conference Fighting COVID-19 Pandemic - Health Challenges, 26-28 March 2022, Belgrade, Serbia -Abstract book, 2022
T1  - Androgen Dependence in Thrombosis of Patients With COVID-19
EP  - 391
SP  - 391
UR  - https://hdl.handle.net/21.15107/rcub_rimi_1457
ER  - 

@conference{
author = "Miljatović, Sanja and Dragojević, Teodora and Đikić, Dragoslava and Živković, Emilija and Stevanović, Goran and Čokić, Vladan",
year = "2022",
abstract = "Background. A vascular system inflammation is a risk of venous thromboembolism
and can result in widespread microangiopathy with microvascular thrombosis.
Methods. We performed laboratory clinical follow-up of patients with COVID-19 to
compare gender differences. To study the sex difference in COVID-19 outcome we
will measure estradiol and androgens: dihydrotestosterone (DHT) and sex hormone
binding globulin (SHBP) in plasma of 63 COVID-19 patients, analyzed by ELISA. Their
levels will be correlated to the adhesion molecules: soluble intercellular adhesion
molecule 1 (sICAM-1), soluble vascular adhesion molecule 1 (sVCAM-1), sE-selectin,
and sP-selectin as biomarkers for inflammation and thrombosis.
Results. DHT was increased (1.9 fold) in male COVID-19 patients compared to healthy
male volunteers. SHBP was significantly increased in COVID-19 patients compared
to healthy volunteers (p<0.05) as well as female vs. male COVID-19 patients (p<0.001,
2.5 fold). sVCAM-1 and sICAM-1 were increased in female COVID-19 patients compared
to male COVID-19 patients and female volunteers, respectively (p<0.05). The
sP-selection was significantly (p<0.01) increased in male vs. female COVID-19 patients.
SHBP was in negative correlation with sP-selectin (p<0.05). DHT was in positive
correlation with sVCAM-1 (p<0.05). Ferritin had 3-fold higher levels in male than
female COVID-19 patients (p<0.001).
Conclusions. Upregulation of androgen hormones and thrombotic biomarkers in
COVID-19 patients demonstrate sex dependence.",
publisher = "Belgrade: MEDAPP Association",
journal = "The First World Conference Fighting COVID-19 Pandemic - Health Challenges, 26-28 March 2022, Belgrade, Serbia -Abstract book, 2022",
title = "Androgen Dependence in Thrombosis of Patients With COVID-19",
pages = "391-391",
url = "https://hdl.handle.net/21.15107/rcub_rimi_1457"
}

Miljatović, S., Dragojević, T., Đikić, D., Živković, E., Stevanović, G.,& Čokić, V.. (2022). Androgen Dependence in Thrombosis of Patients With COVID-19. in The First World Conference Fighting COVID-19 Pandemic - Health Challenges, 26-28 March 2022, Belgrade, Serbia -Abstract book, 2022
Belgrade: MEDAPP Association., 391-391.
https://hdl.handle.net/21.15107/rcub_rimi_1457

Miljatović S, Dragojević T, Đikić D, Živković E, Stevanović G, Čokić V. Androgen Dependence in Thrombosis of Patients With COVID-19. in The First World Conference Fighting COVID-19 Pandemic - Health Challenges, 26-28 March 2022, Belgrade, Serbia -Abstract book, 2022. 2022;:391-391.
https://hdl.handle.net/21.15107/rcub_rimi_1457 .

Miljatović, Sanja, Dragojević, Teodora, Đikić, Dragoslava, Živković, Emilija, Stevanović, Goran, Čokić, Vladan, "Androgen Dependence in Thrombosis of Patients With COVID-19" in The First World Conference Fighting COVID-19 Pandemic - Health Challenges, 26-28 March 2022, Belgrade, Serbia -Abstract book, 2022 (2022):391-391,
https://hdl.handle.net/21.15107/rcub_rimi_1457 .

TY  - JOUR
AU  - Subotički, Tijana
AU  - Mitrović-Ajtić, Olivera
AU  - Živković, Emilija
AU  - Diklić, Miloš
AU  - Đikić, Dragoslava
AU  - Tošić, Milica
AU  - Beleslin-Čokić, Bojana
AU  - Dragojević, Teodora
AU  - Gotić, Mirjana
AU  - Santibanez, Juan F.
AU  - Čokić, Vladan
PY  - 2021
UR  - http://rimi.imi.bg.ac.rs/handle/123456789/1136
AB  - Background: Chronic inflammation has been recognized in neoplastic disorders, including myeloproliferative neoplasm (MPN), as an important regulator of angiogenesis. Aims: We investigated the influence of vascular endothelial growth factor (VEGF) and pro-inflammatory interleukin-6 (IL-6) on the expression of angiogenic factors, as well as inflammation-related signaling in mononuclear cells (MNC) of patients with MPN and JAK2V617F positive human erythroleukemic (HEL) cells. Results: We found that IL-6 did not change the expression of angiogenic factors in the MNC of patients with MPN and HEL cells. However, IL-6 and the JAK1/2 inhibitor Ruxolitinib significantly increased angiogenic factors—endothelial nitric oxide synthase (eNOS), VEGF, and hypoxia-inducible factor-1 alpha (HIF-1α)—in patients with polycythemia vera (PV). Furthermore, VEGF significantly increased the expression of HIF-1α and eNOS genes, the latter inversely regulated by PI3K and mTOR signaling in the MNC of primary myelofibrosis (PMF). VEGF and inhibitors of inflammatory JAK1/2, PI3K, and mTOR signaling reduced the eNOS protein expression in HEL cells. VEGF also decreased the expression of eNOS and HIF-1α proteins in the MNC of PMF. In contrast, VEGF increased eNOS and HIF-1α protein expression in the MNC of patients with PV, which was mediated by the inflammatory signaling. VEGF increased the level of IL-6 immunopositive MNC of MPN. In summary, VEGF conversely regulated gene and protein expression of angiogenic factors in the MNC of PMF, while VEGF increased angiogenic factor expression in PV mediated by the inflammation-related signaling. Conclusion: The angiogenic VEGF induction of IL-6 supports chronic inflammation that, through positive feedback, further promotes angiogenesis with concomitant JAK1/2 inhibition.
PB  - MDPI
T2  - International Journal of Molecular Sciences
T1  - VEGF Regulation of Angiogenic Factors via Inflammatory Signaling in Myeloproliferative Neoplasms
IS  - 13
SP  - 6671
VL  - 22
DO  - 10.3390/ijms22136671
ER  - 

@article{
author = "Subotički, Tijana and Mitrović-Ajtić, Olivera and Živković, Emilija and Diklić, Miloš and Đikić, Dragoslava and Tošić, Milica and Beleslin-Čokić, Bojana and Dragojević, Teodora and Gotić, Mirjana and Santibanez, Juan F. and Čokić, Vladan",
year = "2021",
abstract = "Background: Chronic inflammation has been recognized in neoplastic disorders, including myeloproliferative neoplasm (MPN), as an important regulator of angiogenesis. Aims: We investigated the influence of vascular endothelial growth factor (VEGF) and pro-inflammatory interleukin-6 (IL-6) on the expression of angiogenic factors, as well as inflammation-related signaling in mononuclear cells (MNC) of patients with MPN and JAK2V617F positive human erythroleukemic (HEL) cells. Results: We found that IL-6 did not change the expression of angiogenic factors in the MNC of patients with MPN and HEL cells. However, IL-6 and the JAK1/2 inhibitor Ruxolitinib significantly increased angiogenic factors—endothelial nitric oxide synthase (eNOS), VEGF, and hypoxia-inducible factor-1 alpha (HIF-1α)—in patients with polycythemia vera (PV). Furthermore, VEGF significantly increased the expression of HIF-1α and eNOS genes, the latter inversely regulated by PI3K and mTOR signaling in the MNC of primary myelofibrosis (PMF). VEGF and inhibitors of inflammatory JAK1/2, PI3K, and mTOR signaling reduced the eNOS protein expression in HEL cells. VEGF also decreased the expression of eNOS and HIF-1α proteins in the MNC of PMF. In contrast, VEGF increased eNOS and HIF-1α protein expression in the MNC of patients with PV, which was mediated by the inflammatory signaling. VEGF increased the level of IL-6 immunopositive MNC of MPN. In summary, VEGF conversely regulated gene and protein expression of angiogenic factors in the MNC of PMF, while VEGF increased angiogenic factor expression in PV mediated by the inflammation-related signaling. Conclusion: The angiogenic VEGF induction of IL-6 supports chronic inflammation that, through positive feedback, further promotes angiogenesis with concomitant JAK1/2 inhibition.",
publisher = "MDPI",
journal = "International Journal of Molecular Sciences",
title = "VEGF Regulation of Angiogenic Factors via Inflammatory Signaling in Myeloproliferative Neoplasms",
number = "13",
pages = "6671",
volume = "22",
doi = "10.3390/ijms22136671"
}

Subotički, T., Mitrović-Ajtić, O., Živković, E., Diklić, M., Đikić, D., Tošić, M., Beleslin-Čokić, B., Dragojević, T., Gotić, M., Santibanez, J. F.,& Čokić, V.. (2021). VEGF Regulation of Angiogenic Factors via Inflammatory Signaling in Myeloproliferative Neoplasms. in International Journal of Molecular Sciences
MDPI., 22(13), 6671.
https://doi.org/10.3390/ijms22136671

Subotički T, Mitrović-Ajtić O, Živković E, Diklić M, Đikić D, Tošić M, Beleslin-Čokić B, Dragojević T, Gotić M, Santibanez JF, Čokić V. VEGF Regulation of Angiogenic Factors via Inflammatory Signaling in Myeloproliferative Neoplasms. in International Journal of Molecular Sciences. 2021;22(13):6671.
doi:10.3390/ijms22136671 .

Subotički, Tijana, Mitrović-Ajtić, Olivera, Živković, Emilija, Diklić, Miloš, Đikić, Dragoslava, Tošić, Milica, Beleslin-Čokić, Bojana, Dragojević, Teodora, Gotić, Mirjana, Santibanez, Juan F., Čokić, Vladan, "VEGF Regulation of Angiogenic Factors via Inflammatory Signaling in Myeloproliferative Neoplasms" in International Journal of Molecular Sciences, 22, no. 13 (2021):6671,
https://doi.org/10.3390/ijms22136671 . .