TY  - CONF
AU  - Kovačević, Sanja
AU  - Matić, Gordana
AU  - Elaković, Ivana
PY  - 2018
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3986
AB  - Introduction 
Increased fructose consumption, mainly through sweetened beverages, coincides with growing rate of obesity, women being more prone than men. Chronic low-grade inflammation has been implicated in the pathogenesis of obesity-related disorders including metabolic syndrome and insulin resistance.
The aim 
We investigated whether fructose overconsumption causes inflammation in the visceral adipose tissue (VAT) and hypothalamus of female rats contributing to development of obesity and insulin resistance.
Methods 
Using qPCR and Western blot, we examined the effects of 9-week fructose-enriched diet on inflammatory status, insulin and leptin signaling in the VAT and hypothalamus, as well as on the expression of orexigenic and anorexigenic neuropeptides in the hypothalamus.
Results 
Fructose-fed rats had increased nuclear accumulation of nuclear factor κB (NF-κB) and elevated expression of pro-inflammatory cytokines (IL-1β, IL6, and TNFα), as well as increased protein level of macrophage-specific marker F4/80 in the VAT. In the same tissue, fructose overconsumption reduced protein content and stimulatory phosphorylation of Akt kinase, while increasing inhibitory phosphorylation of insulin receptor substrate-1 (IRS-1). There were no changes in VAT mass, nor in inflammatory markers, insulin and leptin signaling (leptin receptor and SOCS3 expression) and appetite regulation (NPY, AgRP, POMC and CART) in the hypothalamus.
Conclusions 
The results suggest that fructose overconsumption causes alterations in pro-inflammatory markers and reduces insulin signaling in the VAT of female rats. These alterations could be one of the first consequences of fructose overconsumption, since they were detected in the absence of obesity, and hypothalamic inflammation and insulin and leptin resistance.
PB  - Belgrade: Institute for Biological Research “Siniša Stanković”
C3  - Program & Book of Abstracts : IUBMB Advanced School Nutrition, Metabolism and Aging
T1  - Effects Of Fructose-enriched Diet On Inflammation And Insulin Signaling In The Hypothalamus And Visceral Adipose Tissue Of Female Rats
SP  - 34
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_3986
ER  - 

@conference{
editor = "Đorđević, Ana",
author = "Kovačević, Sanja and Matić, Gordana and Elaković, Ivana",
year = "2018",
abstract = "Introduction 
Increased fructose consumption, mainly through sweetened beverages, coincides with growing rate of obesity, women being more prone than men. Chronic low-grade inflammation has been implicated in the pathogenesis of obesity-related disorders including metabolic syndrome and insulin resistance.
The aim 
We investigated whether fructose overconsumption causes inflammation in the visceral adipose tissue (VAT) and hypothalamus of female rats contributing to development of obesity and insulin resistance.
Methods 
Using qPCR and Western blot, we examined the effects of 9-week fructose-enriched diet on inflammatory status, insulin and leptin signaling in the VAT and hypothalamus, as well as on the expression of orexigenic and anorexigenic neuropeptides in the hypothalamus.
Results 
Fructose-fed rats had increased nuclear accumulation of nuclear factor κB (NF-κB) and elevated expression of pro-inflammatory cytokines (IL-1β, IL6, and TNFα), as well as increased protein level of macrophage-specific marker F4/80 in the VAT. In the same tissue, fructose overconsumption reduced protein content and stimulatory phosphorylation of Akt kinase, while increasing inhibitory phosphorylation of insulin receptor substrate-1 (IRS-1). There were no changes in VAT mass, nor in inflammatory markers, insulin and leptin signaling (leptin receptor and SOCS3 expression) and appetite regulation (NPY, AgRP, POMC and CART) in the hypothalamus.
Conclusions 
The results suggest that fructose overconsumption causes alterations in pro-inflammatory markers and reduces insulin signaling in the VAT of female rats. These alterations could be one of the first consequences of fructose overconsumption, since they were detected in the absence of obesity, and hypothalamic inflammation and insulin and leptin resistance.",
publisher = "Belgrade: Institute for Biological Research “Siniša Stanković”",
journal = "Program & Book of Abstracts : IUBMB Advanced School Nutrition, Metabolism and Aging",
title = "Effects Of Fructose-enriched Diet On Inflammation And Insulin Signaling In The Hypothalamus And Visceral Adipose Tissue Of Female Rats",
pages = "34",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_3986"
}

Đorđević, A., Kovačević, S., Matić, G.,& Elaković, I.. (2018). Effects Of Fructose-enriched Diet On Inflammation And Insulin Signaling In The Hypothalamus And Visceral Adipose Tissue Of Female Rats. in Program & Book of Abstracts : IUBMB Advanced School Nutrition, Metabolism and Aging
Belgrade: Institute for Biological Research “Siniša Stanković”., 34.
https://hdl.handle.net/21.15107/rcub_ibiss_3986

Đorđević A, Kovačević S, Matić G, Elaković I. Effects Of Fructose-enriched Diet On Inflammation And Insulin Signaling In The Hypothalamus And Visceral Adipose Tissue Of Female Rats. in Program & Book of Abstracts : IUBMB Advanced School Nutrition, Metabolism and Aging. 2018;:34.
https://hdl.handle.net/21.15107/rcub_ibiss_3986 .

Đorđević, Ana, Kovačević, Sanja, Matić, Gordana, Elaković, Ivana, "Effects Of Fructose-enriched Diet On Inflammation And Insulin Signaling In The Hypothalamus And Visceral Adipose Tissue Of Female Rats" in Program & Book of Abstracts : IUBMB Advanced School Nutrition, Metabolism and Aging (2018):34,
https://hdl.handle.net/21.15107/rcub_ibiss_3986 .

TY  - CONF
AU  - Rajić, Jovana
AU  - Grdović, Nevena
AU  - Inic-Kanada, Aleksandra
AU  - Stein, Elisabeth
AU  - Schuerer, Nadine
AU  - Uskoković, Aleksandra
AU  - Ghasemian, Ehsan
AU  - Dinić, Svetlana
AU  - Mihailović, Mirjana
AU  - Arambašić Jovanović, Jelena
AU  - Tolić, Anja
AU  - Đorđević, Marija
AU  - Đorđević, Miloš
AU  - Poznanović, Goran
AU  - Barisani-Asenbauer, Talin
AU  - Vidaković, Melita
PY  - 2017
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2869
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2866
AB  - Introduction: Trachoma is the most common cause of infectious blindness worldwide, initiated by repeated infection of the conjunctiva with Chlamydia trachomatis (Ct). The resulting chronic inflammation and formation of fibrotic tissue eventually lead to corneal damage. Based on the facts that epithelial to mesenchymal transition (EMT) plays an important role in the development of fibrosis and that EMT is epigenetically regulated process, the aims of this study were to reveal the capacity of Ct to induce EMT in vitro and to unveil potential underlying epigenetic mechanisms. Methods: Human conjunctival epithelial (HCjE) cells were infected with 107 IFU of Ct for 72 h. EMT-inducing signaling pathways, as well as mRNA and protein expression of EMT markers (E-cadherin, fibronectin and α-SMA) were evaluated by RT-qPCR, Immunoblotting and Immunocytochemistry. DNA methylation patterns of selected regions of E-cadherin, fibronectin and α-SMA genes were examined by Methylation-Specific PCR, High Resolution Melting analysis and Bisulfite Sequencing. Results: Infection with Ct was accompanied with the activation of EMT-inducing signaling pathways, downregulation of epithelial marker E-cadherin and upregulation of mesenchymal markers fibronectin and α-SMA. While DNA methylation status of E-cadherin gene promoter correlated with its expression, methylation status of fibronectin and α-SMA genes couldn’t be related to their expression levels. Conclusion: Ct infection of HCjE cells triggers EMT that goes along with changes in the methylation profile of the E-cadherin promoter. Sequence of events described herein could contribute to scarring process in trachoma and open up possibilities for development of new therapeutic strategies in trachoma treatment.
PB  - Belgrade: University of Belgrade, Faculty of Biology
C3  - 1st Congress of Molecular Biologists of Serbia
T1  - Chlamydia trachomatis infection and development of epithelial mesenchymal transition in conjunctiva : possible epigenetic mechanisms unveiled
SP  - 70
EP  - 70
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_2869
ER  - 

@conference{
editor = "Brajušković, Goran, Đorđević, Ana",
author = "Rajić, Jovana and Grdović, Nevena and Inic-Kanada, Aleksandra and Stein, Elisabeth and Schuerer, Nadine and Uskoković, Aleksandra and Ghasemian, Ehsan and Dinić, Svetlana and Mihailović, Mirjana and Arambašić Jovanović, Jelena and Tolić, Anja and Đorđević, Marija and Đorđević, Miloš and Poznanović, Goran and Barisani-Asenbauer, Talin and Vidaković, Melita",
year = "2017",
abstract = "Introduction: Trachoma is the most common cause of infectious blindness worldwide, initiated by repeated infection of the conjunctiva with Chlamydia trachomatis (Ct). The resulting chronic inflammation and formation of fibrotic tissue eventually lead to corneal damage. Based on the facts that epithelial to mesenchymal transition (EMT) plays an important role in the development of fibrosis and that EMT is epigenetically regulated process, the aims of this study were to reveal the capacity of Ct to induce EMT in vitro and to unveil potential underlying epigenetic mechanisms. Methods: Human conjunctival epithelial (HCjE) cells were infected with 107 IFU of Ct for 72 h. EMT-inducing signaling pathways, as well as mRNA and protein expression of EMT markers (E-cadherin, fibronectin and α-SMA) were evaluated by RT-qPCR, Immunoblotting and Immunocytochemistry. DNA methylation patterns of selected regions of E-cadherin, fibronectin and α-SMA genes were examined by Methylation-Specific PCR, High Resolution Melting analysis and Bisulfite Sequencing. Results: Infection with Ct was accompanied with the activation of EMT-inducing signaling pathways, downregulation of epithelial marker E-cadherin and upregulation of mesenchymal markers fibronectin and α-SMA. While DNA methylation status of E-cadherin gene promoter correlated with its expression, methylation status of fibronectin and α-SMA genes couldn’t be related to their expression levels. Conclusion: Ct infection of HCjE cells triggers EMT that goes along with changes in the methylation profile of the E-cadherin promoter. Sequence of events described herein could contribute to scarring process in trachoma and open up possibilities for development of new therapeutic strategies in trachoma treatment.",
publisher = "Belgrade: University of Belgrade, Faculty of Biology",
journal = "1st Congress of Molecular Biologists of Serbia",
title = "Chlamydia trachomatis infection and development of epithelial mesenchymal transition in conjunctiva : possible epigenetic mechanisms unveiled",
pages = "70-70",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_2869"
}

Brajušković, G., Đorđević, A., Rajić, J., Grdović, N., Inic-Kanada, A., Stein, E., Schuerer, N., Uskoković, A., Ghasemian, E., Dinić, S., Mihailović, M., Arambašić Jovanović, J., Tolić, A., Đorđević, M., Đorđević, M., Poznanović, G., Barisani-Asenbauer, T.,& Vidaković, M.. (2017). Chlamydia trachomatis infection and development of epithelial mesenchymal transition in conjunctiva : possible epigenetic mechanisms unveiled. in 1st Congress of Molecular Biologists of Serbia
Belgrade: University of Belgrade, Faculty of Biology., 70-70.
https://hdl.handle.net/21.15107/rcub_ibiss_2869

Brajušković G, Đorđević A, Rajić J, Grdović N, Inic-Kanada A, Stein E, Schuerer N, Uskoković A, Ghasemian E, Dinić S, Mihailović M, Arambašić Jovanović J, Tolić A, Đorđević M, Đorđević M, Poznanović G, Barisani-Asenbauer T, Vidaković M. Chlamydia trachomatis infection and development of epithelial mesenchymal transition in conjunctiva : possible epigenetic mechanisms unveiled. in 1st Congress of Molecular Biologists of Serbia. 2017;:70-70.
https://hdl.handle.net/21.15107/rcub_ibiss_2869 .

Brajušković, Goran, Đorđević, Ana, Rajić, Jovana, Grdović, Nevena, Inic-Kanada, Aleksandra, Stein, Elisabeth, Schuerer, Nadine, Uskoković, Aleksandra, Ghasemian, Ehsan, Dinić, Svetlana, Mihailović, Mirjana, Arambašić Jovanović, Jelena, Tolić, Anja, Đorđević, Marija, Đorđević, Miloš, Poznanović, Goran, Barisani-Asenbauer, Talin, Vidaković, Melita, "Chlamydia trachomatis infection and development of epithelial mesenchymal transition in conjunctiva : possible epigenetic mechanisms unveiled" in 1st Congress of Molecular Biologists of Serbia (2017):70-70,
https://hdl.handle.net/21.15107/rcub_ibiss_2869 .

TY  - CONF
AU  - Đorđević, Marija
AU  - Arambašić Jovanović, Jelena
AU  - Tolić, Anja
AU  - Đorđević, Miloš
AU  - Mihailović, Mirjana
AU  - Grdović, Nevena
AU  - Uskoković, Aleksandra
AU  - Rajić, Jovana
AU  - Dinić, Svetlana
AU  - Jurkowski, Tomasz P.
AU  - Vidaković, Melita
PY  - 2017
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2868
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2865
AB  - Introduction: Since diabetes is characterized by impaired ability of pancreatic betacells to respond and/or produce insulin, new approaches for renewal and replacement of deficient beta-cells are indispensable. The aim of this study is direct pancreatic alpha- to beta-cells transdifferentiation by using a new synthetic epigenetic tool, Epi-CRISPR system. Using Epi-CRISPR system we aim to introduce targeted DNA methylation and subsequent repression of genes responsible for maintaining alpha-cell identity. Methods: AlphaTC1-6 cells (α-cells) were transiently transfected with dCas9-Dnmt3a- Dnmt3L constructs and one or four different vectors containing guide RNA components for specific targeting the promoter region of aristaless-related homeobox gene (Arx). The success of α-cells transdifferentiation into insulinproducing cells was evaluated by measuring Arx and insulin mRNA level, amount of secreted insulin and by immunostaining of insulin/glucagon in the cells. Results: We observed Arx transcriptional repression in α-cell transfected with Epi- CRISPR construct that targets the Arx gene promoter inducing subsequent methylation. At fifth day post-transfection the expression of Arx was decreased in α- cells followed by consequent increase in insulin (mRNA and protein level). At the same time, the glucagon levels remained unchanged. At twelfth day posttransfection the transfected cells start to lose glucagon while still secreting insulin. Conclusion: This study is near to confirm Epi-CRISPR system functionality and to verify the concept of cell transdifferentiation through silencing of genes responsible for maintaining cell phenotype. The obtained results will be valuable for later Epi- CRISPRs use in mouse in vivo models of diabetes and eventually as a future therapy for diabetes attenuation in humans.
PB  - Belgrade: University of Belgrade, Faculty of Biology, Belgrade
C3  - 1st Congress of Molecular Biologists of Serbia
T1  - Transdifferentiation of pancreatic alpha to beta cells using Epi-CRISPR directed DNA methylation
SP  - 73
EP  - 73
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_2868
ER  - 

@conference{
editor = "Brajušković, Goran, Đorđević, Ana",
author = "Đorđević, Marija and Arambašić Jovanović, Jelena and Tolić, Anja and Đorđević, Miloš and Mihailović, Mirjana and Grdović, Nevena and Uskoković, Aleksandra and Rajić, Jovana and Dinić, Svetlana and Jurkowski, Tomasz P. and Vidaković, Melita",
year = "2017",
abstract = "Introduction: Since diabetes is characterized by impaired ability of pancreatic betacells to respond and/or produce insulin, new approaches for renewal and replacement of deficient beta-cells are indispensable. The aim of this study is direct pancreatic alpha- to beta-cells transdifferentiation by using a new synthetic epigenetic tool, Epi-CRISPR system. Using Epi-CRISPR system we aim to introduce targeted DNA methylation and subsequent repression of genes responsible for maintaining alpha-cell identity. Methods: AlphaTC1-6 cells (α-cells) were transiently transfected with dCas9-Dnmt3a- Dnmt3L constructs and one or four different vectors containing guide RNA components for specific targeting the promoter region of aristaless-related homeobox gene (Arx). The success of α-cells transdifferentiation into insulinproducing cells was evaluated by measuring Arx and insulin mRNA level, amount of secreted insulin and by immunostaining of insulin/glucagon in the cells. Results: We observed Arx transcriptional repression in α-cell transfected with Epi- CRISPR construct that targets the Arx gene promoter inducing subsequent methylation. At fifth day post-transfection the expression of Arx was decreased in α- cells followed by consequent increase in insulin (mRNA and protein level). At the same time, the glucagon levels remained unchanged. At twelfth day posttransfection the transfected cells start to lose glucagon while still secreting insulin. Conclusion: This study is near to confirm Epi-CRISPR system functionality and to verify the concept of cell transdifferentiation through silencing of genes responsible for maintaining cell phenotype. The obtained results will be valuable for later Epi- CRISPRs use in mouse in vivo models of diabetes and eventually as a future therapy for diabetes attenuation in humans.",
publisher = "Belgrade: University of Belgrade, Faculty of Biology, Belgrade",
journal = "1st Congress of Molecular Biologists of Serbia",
title = "Transdifferentiation of pancreatic alpha to beta cells using Epi-CRISPR directed DNA methylation",
pages = "73-73",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_2868"
}

Brajušković, G., Đorđević, A., Đorđević, M., Arambašić Jovanović, J., Tolić, A., Đorđević, M., Mihailović, M., Grdović, N., Uskoković, A., Rajić, J., Dinić, S., Jurkowski, T. P.,& Vidaković, M.. (2017). Transdifferentiation of pancreatic alpha to beta cells using Epi-CRISPR directed DNA methylation. in 1st Congress of Molecular Biologists of Serbia
Belgrade: University of Belgrade, Faculty of Biology, Belgrade., 73-73.
https://hdl.handle.net/21.15107/rcub_ibiss_2868

Brajušković G, Đorđević A, Đorđević M, Arambašić Jovanović J, Tolić A, Đorđević M, Mihailović M, Grdović N, Uskoković A, Rajić J, Dinić S, Jurkowski TP, Vidaković M. Transdifferentiation of pancreatic alpha to beta cells using Epi-CRISPR directed DNA methylation. in 1st Congress of Molecular Biologists of Serbia. 2017;:73-73.
https://hdl.handle.net/21.15107/rcub_ibiss_2868 .

Brajušković, Goran, Đorđević, Ana, Đorđević, Marija, Arambašić Jovanović, Jelena, Tolić, Anja, Đorđević, Miloš, Mihailović, Mirjana, Grdović, Nevena, Uskoković, Aleksandra, Rajić, Jovana, Dinić, Svetlana, Jurkowski, Tomasz P., Vidaković, Melita, "Transdifferentiation of pancreatic alpha to beta cells using Epi-CRISPR directed DNA methylation" in 1st Congress of Molecular Biologists of Serbia (2017):73-73,
https://hdl.handle.net/21.15107/rcub_ibiss_2868 .