TY  - CONF
AU  - Đorđević, Miloš
AU  - Grdović, Nevena
AU  - Mihailović, Mirjana
AU  - Arambašić Jovanović, Jelena
AU  - Uskoković, Aleksandra
AU  - Rajić, Jovana
AU  - Đorđević, Marija
AU  - Tolić, Anja
AU  - Mišić, Danijela
AU  - Šiler, Branislav
AU  - Poznanović, Goran
AU  - Vidaković, Melita
AU  - Dinić, Svetlana
PY  - 2019
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5692
AB  - Diabetes is characterized by hyperglycemia resulting from a deficiency in insulin secretion and/or action leading to serve diabetic complications. Despite numerous efforts, recovery, and maintenance of functional beta-cells is still an unresolved task in diabetes therapy. Considering anti-diabetic properties of medicinal herb Centaurium erythraea Rafn (CE), this study aimed to analyze protective effects of the CE extract on Rin-5F beta-cell line exposed to diabetogenic agent streptozotocin (STZ). Cytoprotective concentration of CE extract (0.25 mg/mL) and IC50 dose of STZ (12 mM) were determined using cell viability assay (MTT). The level of insulin mRNA and the concentration of insulin released from beta-cells in a culture medium were analyzed by RT-qPCR and ELISA, respectively. Activity of Akt, ERK and p38 kinases, as well as nuclear levels of islet-enriched Pdx1 and MafA proteins were assessed by Western blot analysis. In comparison to STZ-treated beta-cells, CE extract/STZ co-treatment increased viability of Rin-5F cells for 12%. STZ-treated beta-cells displayed reduction mRNA level of insulin to 63% and reduced insulin secretion to 76% in comparison to control, while application of CE extract improved insulin mRNA level to 77% and insulin secretion to 90% of the control level. Improved viability and functionality of beta-cells could be ascribed to a CE extract-mediated modulation of the activities of pro-survival Akt, ERK and p38 kinases and Pdx1 and MafA factors involved in regulation of beta-cell proliferation and insulin expression/secretion. The results of this study suggest that CE extract promotes proliferative and pro-survival pathways in beta-cells and improves their functional properties.
PB  - Lisboa: Universidade Lusófona
C3  - NutRedOx COST Action CA 16112; 2019 Oct 2-4; Lisboa, Portugal
T1  - Centaurium erithraea extract mediates prosurvival pathways and insulin expression in STZ-treated beta-cells
SP  - 25
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5692
ER  - 

@conference{
author = "Đorđević, Miloš and Grdović, Nevena and Mihailović, Mirjana and Arambašić Jovanović, Jelena and Uskoković, Aleksandra and Rajić, Jovana and Đorđević, Marija and Tolić, Anja and Mišić, Danijela and Šiler, Branislav and Poznanović, Goran and Vidaković, Melita and Dinić, Svetlana",
year = "2019",
abstract = "Diabetes is characterized by hyperglycemia resulting from a deficiency in insulin secretion and/or action leading to serve diabetic complications. Despite numerous efforts, recovery, and maintenance of functional beta-cells is still an unresolved task in diabetes therapy. Considering anti-diabetic properties of medicinal herb Centaurium erythraea Rafn (CE), this study aimed to analyze protective effects of the CE extract on Rin-5F beta-cell line exposed to diabetogenic agent streptozotocin (STZ). Cytoprotective concentration of CE extract (0.25 mg/mL) and IC50 dose of STZ (12 mM) were determined using cell viability assay (MTT). The level of insulin mRNA and the concentration of insulin released from beta-cells in a culture medium were analyzed by RT-qPCR and ELISA, respectively. Activity of Akt, ERK and p38 kinases, as well as nuclear levels of islet-enriched Pdx1 and MafA proteins were assessed by Western blot analysis. In comparison to STZ-treated beta-cells, CE extract/STZ co-treatment increased viability of Rin-5F cells for 12%. STZ-treated beta-cells displayed reduction mRNA level of insulin to 63% and reduced insulin secretion to 76% in comparison to control, while application of CE extract improved insulin mRNA level to 77% and insulin secretion to 90% of the control level. Improved viability and functionality of beta-cells could be ascribed to a CE extract-mediated modulation of the activities of pro-survival Akt, ERK and p38 kinases and Pdx1 and MafA factors involved in regulation of beta-cell proliferation and insulin expression/secretion. The results of this study suggest that CE extract promotes proliferative and pro-survival pathways in beta-cells and improves their functional properties.",
publisher = "Lisboa: Universidade Lusófona",
journal = "NutRedOx COST Action CA 16112; 2019 Oct 2-4; Lisboa, Portugal",
title = "Centaurium erithraea extract mediates prosurvival pathways and insulin expression in STZ-treated beta-cells",
pages = "25",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5692"
}

Đorđević, M., Grdović, N., Mihailović, M., Arambašić Jovanović, J., Uskoković, A., Rajić, J., Đorđević, M., Tolić, A., Mišić, D., Šiler, B., Poznanović, G., Vidaković, M.,& Dinić, S.. (2019). Centaurium erithraea extract mediates prosurvival pathways and insulin expression in STZ-treated beta-cells. in NutRedOx COST Action CA 16112; 2019 Oct 2-4; Lisboa, Portugal
Lisboa: Universidade Lusófona., 25.
https://hdl.handle.net/21.15107/rcub_ibiss_5692

Đorđević M, Grdović N, Mihailović M, Arambašić Jovanović J, Uskoković A, Rajić J, Đorđević M, Tolić A, Mišić D, Šiler B, Poznanović G, Vidaković M, Dinić S. Centaurium erithraea extract mediates prosurvival pathways and insulin expression in STZ-treated beta-cells. in NutRedOx COST Action CA 16112; 2019 Oct 2-4; Lisboa, Portugal. 2019;:25.
https://hdl.handle.net/21.15107/rcub_ibiss_5692 .

Đorđević, Miloš, Grdović, Nevena, Mihailović, Mirjana, Arambašić Jovanović, Jelena, Uskoković, Aleksandra, Rajić, Jovana, Đorđević, Marija, Tolić, Anja, Mišić, Danijela, Šiler, Branislav, Poznanović, Goran, Vidaković, Melita, Dinić, Svetlana, "Centaurium erithraea extract mediates prosurvival pathways and insulin expression in STZ-treated beta-cells" in NutRedOx COST Action CA 16112; 2019 Oct 2-4; Lisboa, Portugal (2019):25,
https://hdl.handle.net/21.15107/rcub_ibiss_5692 .

TY  - CONF
AU  - Rajić, Jovana
AU  - Grdović, Nevena
AU  - Petrović Matić, Sanja
AU  - Dinić, Svetlana
AU  - Uskoković, Aleksandra
AU  - Mihailović, Mirjana
AU  - Arambašić Jovanović, Jelena
AU  - Tolić, Anja
AU  - Đorđević, Marija
AU  - Đorđević, Miloš
AU  - Poznanović, Goran
AU  - Pucar, Ana
AU  - Vidaković, Melita
PY  - 2018
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5800
AB  - Objectives: Procalcitonin (PCT) has recently emerged as an important biomarker for an early and accurate diagnosis of bacte­rial infection, hence suggested as biomarker of periodontal disease caused by oral pathogenic microorganisms. Altered DNA meth­ylation of PCT coding gene - calcitonin-related polypeptide a (CALCA) has been shown during systemic inflammation. The aim of this study was to evaluate the influence oflocal and/or systemic inflammation present during periodontitis and diabetes on DNA methylation status of CALCA and its potential use as epigenetic­based biomarker of these chronic inflammatory conditions. 
Methods: The study included 65 individuals divided in three groups: healthy control (n = 17), periodontitis (n = 27) and diabe­tes/periodontitis group (n = 21). Periodontitis was diagnosed us­ing International Workshop for a Classification of Periodontal Diseases and Conditions criteria (1999) while type 2 diabetes as­sessment was performed according to WHO criteria (2013). DNA methylation profile of CALCA promoter in buccal epithelial cells was analyzed by methylation specific polymerase chain reaction (MSP). 
Results: Decrease in DNA methylation of CALCA promoter was observed in periodontitis and even more pronounced in dia­betes/periodontitis compared to control group, although without statistical significance. Correlation analysis revealed statistically significant relationship between the extent of DNA methylation of the CALCA promoter and glycosylated hemoglobin. Even though it is known that life style affects DNA methylation patterns, there was no difference in DNA methylation of CALCA promoter be­tween smokers/non-smokers and normal/overweight individuals. 
Conclusions: Presented results suggest that local periodontal inflammation contributes to the change, but that only systemic inflammation significantly alters the DNA methylation profile of CALCA in buccal cells. Those results imply that DNA methylation status of CALCA reflects systemic inflammation, but additional studies are needed to estimate usefulness of this epigenetic-based biomarker for periodontal disease.
PB  - Krager Publishers
C3  - Selected Abstracts from the 3rd European Summer School on Nutrigenomics; 2018 Jun 25-29; Jesi, italy
T1  - Evaluation of the DNA Methylation Status of Procalcitonin Gene as a Biomarker of Local and Systemic Inflammation
DO  - 10.1159/000490753
SP  - 12
ER  - 

@conference{
author = "Rajić, Jovana and Grdović, Nevena and Petrović Matić, Sanja and Dinić, Svetlana and Uskoković, Aleksandra and Mihailović, Mirjana and Arambašić Jovanović, Jelena and Tolić, Anja and Đorđević, Marija and Đorđević, Miloš and Poznanović, Goran and Pucar, Ana and Vidaković, Melita",
year = "2018",
abstract = "Objectives: Procalcitonin (PCT) has recently emerged as an important biomarker for an early and accurate diagnosis of bacte­rial infection, hence suggested as biomarker of periodontal disease caused by oral pathogenic microorganisms. Altered DNA meth­ylation of PCT coding gene - calcitonin-related polypeptide a (CALCA) has been shown during systemic inflammation. The aim of this study was to evaluate the influence oflocal and/or systemic inflammation present during periodontitis and diabetes on DNA methylation status of CALCA and its potential use as epigenetic­based biomarker of these chronic inflammatory conditions. 
Methods: The study included 65 individuals divided in three groups: healthy control (n = 17), periodontitis (n = 27) and diabe­tes/periodontitis group (n = 21). Periodontitis was diagnosed us­ing International Workshop for a Classification of Periodontal Diseases and Conditions criteria (1999) while type 2 diabetes as­sessment was performed according to WHO criteria (2013). DNA methylation profile of CALCA promoter in buccal epithelial cells was analyzed by methylation specific polymerase chain reaction (MSP). 
Results: Decrease in DNA methylation of CALCA promoter was observed in periodontitis and even more pronounced in dia­betes/periodontitis compared to control group, although without statistical significance. Correlation analysis revealed statistically significant relationship between the extent of DNA methylation of the CALCA promoter and glycosylated hemoglobin. Even though it is known that life style affects DNA methylation patterns, there was no difference in DNA methylation of CALCA promoter be­tween smokers/non-smokers and normal/overweight individuals. 
Conclusions: Presented results suggest that local periodontal inflammation contributes to the change, but that only systemic inflammation significantly alters the DNA methylation profile of CALCA in buccal cells. Those results imply that DNA methylation status of CALCA reflects systemic inflammation, but additional studies are needed to estimate usefulness of this epigenetic-based biomarker for periodontal disease.",
publisher = "Krager Publishers",
journal = "Selected Abstracts from the 3rd European Summer School on Nutrigenomics; 2018 Jun 25-29; Jesi, italy",
title = "Evaluation of the DNA Methylation Status of Procalcitonin Gene as a Biomarker of Local and Systemic Inflammation",
doi = "10.1159/000490753",
pages = "12"
}

Rajić, J., Grdović, N., Petrović Matić, S., Dinić, S., Uskoković, A., Mihailović, M., Arambašić Jovanović, J., Tolić, A., Đorđević, M., Đorđević, M., Poznanović, G., Pucar, A.,& Vidaković, M.. (2018). Evaluation of the DNA Methylation Status of Procalcitonin Gene as a Biomarker of Local and Systemic Inflammation. in Selected Abstracts from the 3rd European Summer School on Nutrigenomics; 2018 Jun 25-29; Jesi, italy
Krager Publishers., 12.
https://doi.org/10.1159/000490753

Rajić J, Grdović N, Petrović Matić S, Dinić S, Uskoković A, Mihailović M, Arambašić Jovanović J, Tolić A, Đorđević M, Đorđević M, Poznanović G, Pucar A, Vidaković M. Evaluation of the DNA Methylation Status of Procalcitonin Gene as a Biomarker of Local and Systemic Inflammation. in Selected Abstracts from the 3rd European Summer School on Nutrigenomics; 2018 Jun 25-29; Jesi, italy. 2018;:12.
doi:10.1159/000490753 .

Rajić, Jovana, Grdović, Nevena, Petrović Matić, Sanja, Dinić, Svetlana, Uskoković, Aleksandra, Mihailović, Mirjana, Arambašić Jovanović, Jelena, Tolić, Anja, Đorđević, Marija, Đorđević, Miloš, Poznanović, Goran, Pucar, Ana, Vidaković, Melita, "Evaluation of the DNA Methylation Status of Procalcitonin Gene as a Biomarker of Local and Systemic Inflammation" in Selected Abstracts from the 3rd European Summer School on Nutrigenomics; 2018 Jun 25-29; Jesi, italy (2018):12,
https://doi.org/10.1159/000490753 . .

TY  - CONF
AU  - Vidaković, Melita
AU  - Đorđević, Marija
AU  - Rajić, Jovana
AU  - Arambašić Jovanović, Jelena
AU  - Grdović, Nevena
AU  - Tolić, Anja
AU  - Đorđević, Miloš
AU  - Mihailović, Mirjana
AU  - Uskoković, Aleksandra
AU  - Poznanović, Goran
AU  - Dinić, Svetlana
AU  - Jurkowski, Tomasz
PY  - 2017
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5805
AB  - Diabetes is the perfect candidate for cell replacement therapy, since it is caused by either an absolute (type 1 diabetes) or relative (type 2 diabetes) defect of insulin-producing pancreatic beta cells. Our research is focused on applying a novel synthetic epigenetic tool (Epi-CRISPRs) for straightfonward, one-step mouse pancreatic alpha (a-cells) to beta cell transdifferentiation by targeted DNA methylation and suppression of genes essential for maintaining pancreatic cell identity (homeobox Arx gene (Arx)). Up to now, we succeeded to transiently transfect a-cells with Epi-CRISPR constructs and 275 gRNA or mix gRNA. The suppression of Arx in a-cells was confirmed on day 5 and 8 posttransfection. The reduction of glucagon synthesis and beginning of insulin production in transfected a-cell was confirmed and visualised by immunostaining. DNA methylation-mediated suppression of Ao( in a-cells leads to their transdifferentiation to insulin-producing beta cells will be confirmed by bisulfite sequencing (undergoing experiments).
Furthermore, we are also investigating an epithelial-mesenchymal transition (ETM), the mechanism which underlies the progressive decline in organ functioning in diabetes, such as the development of kidney and liver fibrosis. ETM is a process of reprogramming epithelial cells from a fully differentiated epithelial state to a more mesenchymal state. Our aim is to analyse the DNA methylation profile and gene expression of either epithelial (E-cadherin) or mesenchymal markers (a-smooth muscle actin and fibronectin) whose differential methylation and gene suppression could lead to more epithelial-like or mesenchyme-like phenotype. This will be accomplished using an in vitro model system based on epithelial cells treated with TGF-pi and 2. The obtained data should enable ETM reversal and stop fibrosis in diabetes and other pathologies using different compounds that act as DNA methylating/demethylating agents or using Epi-CRISPRs-based targeted DNA methylation/demethylation in future.
We are on the way to develop a clear-cut technology able to provide a perfect delivery system for increase of insulin-producing cells in vitro. This system will allow for targeted gene silencing via increased DNA methylation of gene of interest. In addition, we are able to test if any compound used for treatment in different pathological conditions affects the DNA methylation profile of the examined cells. On the other hand, there is a great need for chemical compounds able to act as DNA hypo or hypermethylated agents
PB  - COST Action CA1611
C3  - NutRedOx COST Action CA16112 & the 6'" NutriOx Atelier 20: Preventing Age-Related Diseases with Redo Active Compounds: a taste of controversory?: 2017 Sep 27-29; Strasbourg, France
T1  - Cellular Reprogramming via Epi-CRISPRs-lnduced Targeted DNA Methylation
SP  - 44
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5805
ER  - 

@conference{
author = "Vidaković, Melita and Đorđević, Marija and Rajić, Jovana and Arambašić Jovanović, Jelena and Grdović, Nevena and Tolić, Anja and Đorđević, Miloš and Mihailović, Mirjana and Uskoković, Aleksandra and Poznanović, Goran and Dinić, Svetlana and Jurkowski, Tomasz",
year = "2017",
abstract = "Diabetes is the perfect candidate for cell replacement therapy, since it is caused by either an absolute (type 1 diabetes) or relative (type 2 diabetes) defect of insulin-producing pancreatic beta cells. Our research is focused on applying a novel synthetic epigenetic tool (Epi-CRISPRs) for straightfonward, one-step mouse pancreatic alpha (a-cells) to beta cell transdifferentiation by targeted DNA methylation and suppression of genes essential for maintaining pancreatic cell identity (homeobox Arx gene (Arx)). Up to now, we succeeded to transiently transfect a-cells with Epi-CRISPR constructs and 275 gRNA or mix gRNA. The suppression of Arx in a-cells was confirmed on day 5 and 8 posttransfection. The reduction of glucagon synthesis and beginning of insulin production in transfected a-cell was confirmed and visualised by immunostaining. DNA methylation-mediated suppression of Ao( in a-cells leads to their transdifferentiation to insulin-producing beta cells will be confirmed by bisulfite sequencing (undergoing experiments).
Furthermore, we are also investigating an epithelial-mesenchymal transition (ETM), the mechanism which underlies the progressive decline in organ functioning in diabetes, such as the development of kidney and liver fibrosis. ETM is a process of reprogramming epithelial cells from a fully differentiated epithelial state to a more mesenchymal state. Our aim is to analyse the DNA methylation profile and gene expression of either epithelial (E-cadherin) or mesenchymal markers (a-smooth muscle actin and fibronectin) whose differential methylation and gene suppression could lead to more epithelial-like or mesenchyme-like phenotype. This will be accomplished using an in vitro model system based on epithelial cells treated with TGF-pi and 2. The obtained data should enable ETM reversal and stop fibrosis in diabetes and other pathologies using different compounds that act as DNA methylating/demethylating agents or using Epi-CRISPRs-based targeted DNA methylation/demethylation in future.
We are on the way to develop a clear-cut technology able to provide a perfect delivery system for increase of insulin-producing cells in vitro. This system will allow for targeted gene silencing via increased DNA methylation of gene of interest. In addition, we are able to test if any compound used for treatment in different pathological conditions affects the DNA methylation profile of the examined cells. On the other hand, there is a great need for chemical compounds able to act as DNA hypo or hypermethylated agents",
publisher = "COST Action CA1611",
journal = "NutRedOx COST Action CA16112 & the 6'" NutriOx Atelier 20: Preventing Age-Related Diseases with Redo Active Compounds: a taste of controversory?: 2017 Sep 27-29; Strasbourg, France",
title = "Cellular Reprogramming via Epi-CRISPRs-lnduced Targeted DNA Methylation",
pages = "44",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5805"
}

Vidaković, M., Đorđević, M., Rajić, J., Arambašić Jovanović, J., Grdović, N., Tolić, A., Đorđević, M., Mihailović, M., Uskoković, A., Poznanović, G., Dinić, S.,& Jurkowski, T.. (2017). Cellular Reprogramming via Epi-CRISPRs-lnduced Targeted DNA Methylation. in NutRedOx COST Action CA16112 & the 6'" NutriOx Atelier 20: Preventing Age-Related Diseases with Redo Active Compounds: a taste of controversory?: 2017 Sep 27-29; Strasbourg, France
COST Action CA1611., 44.
https://hdl.handle.net/21.15107/rcub_ibiss_5805

Vidaković M, Đorđević M, Rajić J, Arambašić Jovanović J, Grdović N, Tolić A, Đorđević M, Mihailović M, Uskoković A, Poznanović G, Dinić S, Jurkowski T. Cellular Reprogramming via Epi-CRISPRs-lnduced Targeted DNA Methylation. in NutRedOx COST Action CA16112 & the 6'" NutriOx Atelier 20: Preventing Age-Related Diseases with Redo Active Compounds: a taste of controversory?: 2017 Sep 27-29; Strasbourg, France. 2017;:44.
https://hdl.handle.net/21.15107/rcub_ibiss_5805 .

Vidaković, Melita, Đorđević, Marija, Rajić, Jovana, Arambašić Jovanović, Jelena, Grdović, Nevena, Tolić, Anja, Đorđević, Miloš, Mihailović, Mirjana, Uskoković, Aleksandra, Poznanović, Goran, Dinić, Svetlana, Jurkowski, Tomasz, "Cellular Reprogramming via Epi-CRISPRs-lnduced Targeted DNA Methylation" in NutRedOx COST Action CA16112 & the 6'" NutriOx Atelier 20: Preventing Age-Related Diseases with Redo Active Compounds: a taste of controversory?: 2017 Sep 27-29; Strasbourg, France (2017):44,
https://hdl.handle.net/21.15107/rcub_ibiss_5805 .