TY  - JOUR
AU  - Spasovski, Vesna
AU  - Romolo, Anna
AU  - Zagorc, Urška
AU  - Arrigler, Vesna
AU  - Kisovec, Matic
AU  - Zavec, Apolonija Bedina
AU  - Arko, Matevž
AU  - Molnár, Adrienn
AU  - Schlosser, Gitta
AU  - Iglič, Aleš
AU  - Kogej, Ksenija
AU  - Kralj-Iglič, Veronika
PY  - 2024
UR  - https://www.dovepress.com/characterization-of-nanohybridosomes-from-lipids-and-spruce-homogenate-peer-reviewed-fulltext-article-IJN
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2332
AB  - Introduction: Lipid nanovesicles associated with bioactive phytochemicals from spruce needle homogenate (here called nano-sized
hybridosomes or nanohybridosomes, NSHs) were considered.
Methods: We formed NSHs by mixing appropriate amounts of lecithin, glycerol and supernatant of isolation of extracellular vesicles
from spruce needle homogenate. We visualized NSHs by light microscopy and cryogenic transmission electron microscopy and
assessed them by flow cytometry, dynamic light scattering, ultraviolet–visual spectroscopy, interferometric light microscopy and liquid
chromatography–mass spectrometry.
Results: We found that the particles consisted of a bilayer membrane and a fluid-like interior. Flow cytometry and
interferometric light microscopy measurements showed that the majority of the particles were nano-sized. Dynamic light
scattering and interferometric light microscopy measurements agreed well on the average hydrodynamic radius of the
particles Rh (between 140 and 180 nm), while the concentrations of the particles were in the range between 1013 and
1014/mL indicating that NSHs present a considerable (more than 25%) of the sample which is much more than the yield of
natural extracellular vesicles (EVs) from spruce needle homogenate (estimated less than 1%). Spruce specific lipids and
proteins were found in hybridosomes.
Discussion: Simple and low-cost preparation method, non-demanding saving process and efficient formation procedure suggest that
large-scale production of NSHs from lipids and spruce needle homogenate is feasible.
Plain Language Summary: Cells shed into their exterior nanoparticles (here referred to as extracellular vesicles – EVs) that
are free to move, reach distant cells and are taken up by them. As they carry bioactive constituents, EVs may have important
impact on the recipient cells. The mechanisms of EV formation and mediation can be employed in designing therapeutic,
prophylactic and diagnostic methods for various medical issues. EVs can be harvested from biological samples; however,
their yield is small,12 and there are potential side effects. Artificial vesicles – liposomes – have high yield; however, in vivo,
they can be degraded before reaching the target and their reproducibility is yet insufficient. In order to combine advantages of
both types of nanoparticles, we have composed nanohybridosomes (NSHs) from soya lecithin, water and supernatant of
isolation of EVs from spruce needle homogenate, visualized them by cryogenic electron microscopy and characterized them
with respect to their size, concentration and protein/nucleic acid content. We have applied a recently developed interferometric light microscopy to determine the hydrodynamic radius and the concentration of EVs. We found that the majority of
composed particles are nano-sized and that they enclose more than 25% of the incoming volume of liquid, which is considerably more than about 1% that can be harvested by isolation of EVs from spruce needle homogenate by (ultra)
centrifugation
PB  - Dove Press Ltd
T2  - International Journal of Nanomedicine
T1  - Characterization of Nanohybridosomes from Lipids and Spruce Homogenate Containing Extracellular Vesicles
EP  - 1721
SP  - 1709
VL  - 19
DO  - 10.2147/IJN.S432836
ER  - 

@article{
author = "Spasovski, Vesna and Romolo, Anna and Zagorc, Urška and Arrigler, Vesna and Kisovec, Matic and Zavec, Apolonija Bedina and Arko, Matevž and Molnár, Adrienn and Schlosser, Gitta and Iglič, Aleš and Kogej, Ksenija and Kralj-Iglič, Veronika",
year = "2024",
abstract = "Introduction: Lipid nanovesicles associated with bioactive phytochemicals from spruce needle homogenate (here called nano-sized
hybridosomes or nanohybridosomes, NSHs) were considered.
Methods: We formed NSHs by mixing appropriate amounts of lecithin, glycerol and supernatant of isolation of extracellular vesicles
from spruce needle homogenate. We visualized NSHs by light microscopy and cryogenic transmission electron microscopy and
assessed them by flow cytometry, dynamic light scattering, ultraviolet–visual spectroscopy, interferometric light microscopy and liquid
chromatography–mass spectrometry.
Results: We found that the particles consisted of a bilayer membrane and a fluid-like interior. Flow cytometry and
interferometric light microscopy measurements showed that the majority of the particles were nano-sized. Dynamic light
scattering and interferometric light microscopy measurements agreed well on the average hydrodynamic radius of the
particles Rh (between 140 and 180 nm), while the concentrations of the particles were in the range between 1013 and
1014/mL indicating that NSHs present a considerable (more than 25%) of the sample which is much more than the yield of
natural extracellular vesicles (EVs) from spruce needle homogenate (estimated less than 1%). Spruce specific lipids and
proteins were found in hybridosomes.
Discussion: Simple and low-cost preparation method, non-demanding saving process and efficient formation procedure suggest that
large-scale production of NSHs from lipids and spruce needle homogenate is feasible.
Plain Language Summary: Cells shed into their exterior nanoparticles (here referred to as extracellular vesicles – EVs) that
are free to move, reach distant cells and are taken up by them. As they carry bioactive constituents, EVs may have important
impact on the recipient cells. The mechanisms of EV formation and mediation can be employed in designing therapeutic,
prophylactic and diagnostic methods for various medical issues. EVs can be harvested from biological samples; however,
their yield is small,12 and there are potential side effects. Artificial vesicles – liposomes – have high yield; however, in vivo,
they can be degraded before reaching the target and their reproducibility is yet insufficient. In order to combine advantages of
both types of nanoparticles, we have composed nanohybridosomes (NSHs) from soya lecithin, water and supernatant of
isolation of EVs from spruce needle homogenate, visualized them by cryogenic electron microscopy and characterized them
with respect to their size, concentration and protein/nucleic acid content. We have applied a recently developed interferometric light microscopy to determine the hydrodynamic radius and the concentration of EVs. We found that the majority of
composed particles are nano-sized and that they enclose more than 25% of the incoming volume of liquid, which is considerably more than about 1% that can be harvested by isolation of EVs from spruce needle homogenate by (ultra)
centrifugation",
publisher = "Dove Press Ltd",
journal = "International Journal of Nanomedicine",
title = "Characterization of Nanohybridosomes from Lipids and Spruce Homogenate Containing Extracellular Vesicles",
pages = "1721-1709",
volume = "19",
doi = "10.2147/IJN.S432836"
}

Spasovski, V., Romolo, A., Zagorc, U., Arrigler, V., Kisovec, M., Zavec, A. B., Arko, M., Molnár, A., Schlosser, G., Iglič, A., Kogej, K.,& Kralj-Iglič, V.. (2024). Characterization of Nanohybridosomes from Lipids and Spruce Homogenate Containing Extracellular Vesicles. in International Journal of Nanomedicine
Dove Press Ltd., 19, 1709-1721.
https://doi.org/10.2147/IJN.S432836

Spasovski V, Romolo A, Zagorc U, Arrigler V, Kisovec M, Zavec AB, Arko M, Molnár A, Schlosser G, Iglič A, Kogej K, Kralj-Iglič V. Characterization of Nanohybridosomes from Lipids and Spruce Homogenate Containing Extracellular Vesicles. in International Journal of Nanomedicine. 2024;19:1709-1721.
doi:10.2147/IJN.S432836 .

Spasovski, Vesna, Romolo, Anna, Zagorc, Urška, Arrigler, Vesna, Kisovec, Matic, Zavec, Apolonija Bedina, Arko, Matevž, Molnár, Adrienn, Schlosser, Gitta, Iglič, Aleš, Kogej, Ksenija, Kralj-Iglič, Veronika, "Characterization of Nanohybridosomes from Lipids and Spruce Homogenate Containing Extracellular Vesicles" in International Journal of Nanomedicine, 19 (2024):1709-1721,
https://doi.org/10.2147/IJN.S432836 . .

TY  - JOUR
AU  - Jelovac, Marina
AU  - Kotur, Nikola
AU  - Ristivojević, Bojan
AU  - Pavlović, Djordje
AU  - Spasovski, Vesna
AU  - Damjanov, Nemanja
AU  - Pavlović, Sonja
AU  - Zukić, Branka
PY  - 2023
UR  - https://www.mdpi.com/1422-0067/24/10/8538
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1859
AB  - Systemic sclerosis (SSc) is a rare connective tissue disorder with highest morbidity and mortality among rheumatologic diseases. Disease progression is highly heterogeneous between patients, implying a strong need for individualization of therapy. Four pharmacogenetic variants, namely TPMT rs1800460, TPMT rs1142345, MTHFR rs1801133 and SLCO1B1 rs4149056 were tested for association with severe disease outcomes in 102 patients with SSc from Serbia treated either with immunosuppressants azathioprine (AZA) and methotrexate (MTX) or with other types of medications. Genotyping was performed using PCR-RFLP and direct Sanger sequencing. R software was used for statistical analysis and development of polygenic risk score (PRS) model. Association was found between MTHFR rs1801133 and higher risk for elevated systolic pressure in all patients except those prescribed with MTX, and higher risk for kidney insufficiency in patients prescribed with other types of drugs. In patients treated with MTX, variant SLCO1B1 rs4149056 was protective against kidney insufficiency. For patients receiving MTX a trend was shown for having a higher PRS rank and elevated systolic pressure. Our results open a door wide for more extensive research on pharmacogenomics markers in patients with SSc. Altogether, pharmacogenomics markers could predict the outcome of patients with SSc and help in prevention of adverse drug reactions.
T2  - International Journal of Molecular Sciences
T2  - International Journal of Molecular Sciences
T1  - Can Pharmacogenetic Variants in TPMT, MTHFR and SLCO1B1 Genes Be Used as Potential Markers of Outcome Prediction in Systemic Sclerosis Patients?
IS  - 10
SP  - 8538
VL  - 24
DO  - 10.3390/ijms24108538
ER  - 

@article{
author = "Jelovac, Marina and Kotur, Nikola and Ristivojević, Bojan and Pavlović, Djordje and Spasovski, Vesna and Damjanov, Nemanja and Pavlović, Sonja and Zukić, Branka",
year = "2023",
abstract = "Systemic sclerosis (SSc) is a rare connective tissue disorder with highest morbidity and mortality among rheumatologic diseases. Disease progression is highly heterogeneous between patients, implying a strong need for individualization of therapy. Four pharmacogenetic variants, namely TPMT rs1800460, TPMT rs1142345, MTHFR rs1801133 and SLCO1B1 rs4149056 were tested for association with severe disease outcomes in 102 patients with SSc from Serbia treated either with immunosuppressants azathioprine (AZA) and methotrexate (MTX) or with other types of medications. Genotyping was performed using PCR-RFLP and direct Sanger sequencing. R software was used for statistical analysis and development of polygenic risk score (PRS) model. Association was found between MTHFR rs1801133 and higher risk for elevated systolic pressure in all patients except those prescribed with MTX, and higher risk for kidney insufficiency in patients prescribed with other types of drugs. In patients treated with MTX, variant SLCO1B1 rs4149056 was protective against kidney insufficiency. For patients receiving MTX a trend was shown for having a higher PRS rank and elevated systolic pressure. Our results open a door wide for more extensive research on pharmacogenomics markers in patients with SSc. Altogether, pharmacogenomics markers could predict the outcome of patients with SSc and help in prevention of adverse drug reactions.",
journal = "International Journal of Molecular Sciences, International Journal of Molecular Sciences",
title = "Can Pharmacogenetic Variants in TPMT, MTHFR and SLCO1B1 Genes Be Used as Potential Markers of Outcome Prediction in Systemic Sclerosis Patients?",
number = "10",
pages = "8538",
volume = "24",
doi = "10.3390/ijms24108538"
}

Jelovac, M., Kotur, N., Ristivojević, B., Pavlović, D., Spasovski, V., Damjanov, N., Pavlović, S.,& Zukić, B.. (2023). Can Pharmacogenetic Variants in TPMT, MTHFR and SLCO1B1 Genes Be Used as Potential Markers of Outcome Prediction in Systemic Sclerosis Patients?. in International Journal of Molecular Sciences, 24(10), 8538.
https://doi.org/10.3390/ijms24108538

Jelovac M, Kotur N, Ristivojević B, Pavlović D, Spasovski V, Damjanov N, Pavlović S, Zukić B. Can Pharmacogenetic Variants in TPMT, MTHFR and SLCO1B1 Genes Be Used as Potential Markers of Outcome Prediction in Systemic Sclerosis Patients?. in International Journal of Molecular Sciences. 2023;24(10):8538.
doi:10.3390/ijms24108538 .

Jelovac, Marina, Kotur, Nikola, Ristivojević, Bojan, Pavlović, Djordje, Spasovski, Vesna, Damjanov, Nemanja, Pavlović, Sonja, Zukić, Branka, "Can Pharmacogenetic Variants in TPMT, MTHFR and SLCO1B1 Genes Be Used as Potential Markers of Outcome Prediction in Systemic Sclerosis Patients?" in International Journal of Molecular Sciences, 24, no. 10 (2023):8538,
https://doi.org/10.3390/ijms24108538 . .

TY  - CONF
AU  - Zala, Jan
AU  - Romolo, Romolo
AU  - Matevž, Arko
AU  - Spasovski, Vesna
AU  - Iglič, Aleš
AU  - Drobne, Damjana
AU  - Kralj-Iglič, Veronika
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2267
AB  - Introduction: Blood plasma is used in regenerative medicine for more than 30 years as it was found to have
impact on angiogenesis and proliferation of endothelial cells. This effect was initially ascribed to growth factors
and cytokines, however, particularly in the last decade researchers are focusing on platelet-derived
extracellular vesicles (PEVs) that are also present in the platelet rich plasma.
Methods: We prepared platelet and extracellular vesicles-rich plasma (PVRP) by centrifugation of the
human blood, and differential (ultra) centrifugation of PVRP to isolate PEVs. We exposed Human Umbilical Vein
Endothelial Cells (HUVEC) to 5% PEVs for 24 hours and assessed inflammation markers (Interleukin(IL)-1beta,
IL-6 and Tumor necrosis factor (TNF)-alpha using ELISA tests), oxidative stress markers (Cholinesterase
(ChE) and glutathione S-transferase (GST) activity by spectrophotometry, as well reactive oxygen species
(ROS) and lipid droplets (LD) by flow cytometry). We observed morphological changes in HUVEC indicating
angiogenesis by using optical microscopy.
Results: We found that after 24 hours caused a decrease of the concentration of IL-6, IL-1beta and TNF-alpha.
Also, ChE and GST activity and ROS count were decreased. LD production, which is triggered in need to protect
the cells from free radicals and oxidative stress damage, was however higher. Treatment of the cells had
impact on cell morphology with progressed formation of the tubes and cell connecting, which is regarded as the
beginning of HUVEC angiogenesis process.
Discussion: Beneficial effect of PVRP in healing and regeneration may include suppression of inflammation
and oxidative stress by PEVs.
PB  - Slovenian Network for Extracellular Vesicles (SiN-EV)
PB  - Austrian Society for Extracellular Vesicles (ASEV)
PB  - Hungarian Society for Extracellular Vesicles (HSEV)
PB  - Slovenian Network for Extracellular Vesicles (SiN-EV)
C3  - Small New World 2.0
T1  - Extracellular vesicles from blood plasma as mediators of anti-inflammatory effects, oxidative stress and angiogenesis in HUVEC
EP  - 118
SP  - 118
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2267
ER  - 

@conference{
author = "Zala, Jan and Romolo, Romolo and Matevž, Arko and Spasovski, Vesna and Iglič, Aleš and Drobne, Damjana and Kralj-Iglič, Veronika",
year = "2023",
abstract = "Introduction: Blood plasma is used in regenerative medicine for more than 30 years as it was found to have
impact on angiogenesis and proliferation of endothelial cells. This effect was initially ascribed to growth factors
and cytokines, however, particularly in the last decade researchers are focusing on platelet-derived
extracellular vesicles (PEVs) that are also present in the platelet rich plasma.
Methods: We prepared platelet and extracellular vesicles-rich plasma (PVRP) by centrifugation of the
human blood, and differential (ultra) centrifugation of PVRP to isolate PEVs. We exposed Human Umbilical Vein
Endothelial Cells (HUVEC) to 5% PEVs for 24 hours and assessed inflammation markers (Interleukin(IL)-1beta,
IL-6 and Tumor necrosis factor (TNF)-alpha using ELISA tests), oxidative stress markers (Cholinesterase
(ChE) and glutathione S-transferase (GST) activity by spectrophotometry, as well reactive oxygen species
(ROS) and lipid droplets (LD) by flow cytometry). We observed morphological changes in HUVEC indicating
angiogenesis by using optical microscopy.
Results: We found that after 24 hours caused a decrease of the concentration of IL-6, IL-1beta and TNF-alpha.
Also, ChE and GST activity and ROS count were decreased. LD production, which is triggered in need to protect
the cells from free radicals and oxidative stress damage, was however higher. Treatment of the cells had
impact on cell morphology with progressed formation of the tubes and cell connecting, which is regarded as the
beginning of HUVEC angiogenesis process.
Discussion: Beneficial effect of PVRP in healing and regeneration may include suppression of inflammation
and oxidative stress by PEVs.",
publisher = "Slovenian Network for Extracellular Vesicles (SiN-EV), Austrian Society for Extracellular Vesicles (ASEV), Hungarian Society for Extracellular Vesicles (HSEV), Slovenian Network for Extracellular Vesicles (SiN-EV)",
journal = "Small New World 2.0",
title = "Extracellular vesicles from blood plasma as mediators of anti-inflammatory effects, oxidative stress and angiogenesis in HUVEC",
pages = "118-118",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2267"
}

Zala, J., Romolo, R., Matevž, A., Spasovski, V., Iglič, A., Drobne, D.,& Kralj-Iglič, V.. (2023). Extracellular vesicles from blood plasma as mediators of anti-inflammatory effects, oxidative stress and angiogenesis in HUVEC. in Small New World 2.0
Slovenian Network for Extracellular Vesicles (SiN-EV)., 118-118.
https://hdl.handle.net/21.15107/rcub_imagine_2267

Zala J, Romolo R, Matevž A, Spasovski V, Iglič A, Drobne D, Kralj-Iglič V. Extracellular vesicles from blood plasma as mediators of anti-inflammatory effects, oxidative stress and angiogenesis in HUVEC. in Small New World 2.0. 2023;:118-118.
https://hdl.handle.net/21.15107/rcub_imagine_2267 .

Zala, Jan, Romolo, Romolo, Matevž, Arko, Spasovski, Vesna, Iglič, Aleš, Drobne, Damjana, Kralj-Iglič, Veronika, "Extracellular vesicles from blood plasma as mediators of anti-inflammatory effects, oxidative stress and angiogenesis in HUVEC" in Small New World 2.0 (2023):118-118,
https://hdl.handle.net/21.15107/rcub_imagine_2267 .

TY  - JOUR
AU  - Jeran, Marko
AU  - Romolo, Anna
AU  - Spasovski, Vesna
AU  - Hočevar, Matej
AU  - Novak, Urban
AU  - Štukelj, Roman
AU  - Šuštar, Vid
AU  - Kisovec, Matic
AU  - Bedina Zavec, Apolonija
AU  - Kogej, Ksenija
AU  - Iglič, Aleš
AU  - Trebše, Polonca
AU  - Kralj-Iglič, Veronika
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2086
AB  - Small cellular particles (SCPs) are being considered for their role in cell-to-cell communication. We harvested and characterized SCPs from spruce needle homogenate. SCPs were isolated by differential ultracentrifugation. They were imaged by scanning electron microscope (SEM) and cryogenic transmission electron microscope (cryo TEM), assessed for their number density and hydrodynamic diameter by interferometric light microscopy (ILM) and flow cytometry (FCM), total phenolic content (TPC) by UV-vis spectroscopy, and terpene content by gas chromatography-mass spectrometry (GC-MS). The supernatant after ultracentrifugation at 50,000× g contained bilayer-enclosed vesicles whereas in the isolate we observed small particles of other types and only a few vesicles. The number density of cell-sized particles (CSPs) (larger than 2 μm) and meso-sized particles (MSPs) (cca 400 nm–2 µm) was about four orders of magnitude lower than the number density of SCPs (sized below 500 nm). The average hydrodynamic diameter of SCPs measured in 10,029 SCPs was 161 ± 133 nm. TCP decreased considerably due to 5-day aging. Volatile terpenoid content was found in the pellet after 300× g. The above results indicate that spruce needle homogenate is a source of vesicles to be explored for potential delivery use.
PB  - MDPI
T2  - International Journal of Molecular Sciences
T1  - Small Cellular Particles from European Spruce Needle Homogenate
IS  - 5
SP  - 4349
VL  - 24
DO  - 10.3390/ijms24054349
ER  - 

@article{
author = "Jeran, Marko and Romolo, Anna and Spasovski, Vesna and Hočevar, Matej and Novak, Urban and Štukelj, Roman and Šuštar, Vid and Kisovec, Matic and Bedina Zavec, Apolonija and Kogej, Ksenija and Iglič, Aleš and Trebše, Polonca and Kralj-Iglič, Veronika",
year = "2023",
abstract = "Small cellular particles (SCPs) are being considered for their role in cell-to-cell communication. We harvested and characterized SCPs from spruce needle homogenate. SCPs were isolated by differential ultracentrifugation. They were imaged by scanning electron microscope (SEM) and cryogenic transmission electron microscope (cryo TEM), assessed for their number density and hydrodynamic diameter by interferometric light microscopy (ILM) and flow cytometry (FCM), total phenolic content (TPC) by UV-vis spectroscopy, and terpene content by gas chromatography-mass spectrometry (GC-MS). The supernatant after ultracentrifugation at 50,000× g contained bilayer-enclosed vesicles whereas in the isolate we observed small particles of other types and only a few vesicles. The number density of cell-sized particles (CSPs) (larger than 2 μm) and meso-sized particles (MSPs) (cca 400 nm–2 µm) was about four orders of magnitude lower than the number density of SCPs (sized below 500 nm). The average hydrodynamic diameter of SCPs measured in 10,029 SCPs was 161 ± 133 nm. TCP decreased considerably due to 5-day aging. Volatile terpenoid content was found in the pellet after 300× g. The above results indicate that spruce needle homogenate is a source of vesicles to be explored for potential delivery use.",
publisher = "MDPI",
journal = "International Journal of Molecular Sciences",
title = "Small Cellular Particles from European Spruce Needle Homogenate",
number = "5",
pages = "4349",
volume = "24",
doi = "10.3390/ijms24054349"
}

Jeran, M., Romolo, A., Spasovski, V., Hočevar, M., Novak, U., Štukelj, R., Šuštar, V., Kisovec, M., Bedina Zavec, A., Kogej, K., Iglič, A., Trebše, P.,& Kralj-Iglič, V.. (2023). Small Cellular Particles from European Spruce Needle Homogenate. in International Journal of Molecular Sciences
MDPI., 24(5), 4349.
https://doi.org/10.3390/ijms24054349

Jeran M, Romolo A, Spasovski V, Hočevar M, Novak U, Štukelj R, Šuštar V, Kisovec M, Bedina Zavec A, Kogej K, Iglič A, Trebše P, Kralj-Iglič V. Small Cellular Particles from European Spruce Needle Homogenate. in International Journal of Molecular Sciences. 2023;24(5):4349.
doi:10.3390/ijms24054349 .

Jeran, Marko, Romolo, Anna, Spasovski, Vesna, Hočevar, Matej, Novak, Urban, Štukelj, Roman, Šuštar, Vid, Kisovec, Matic, Bedina Zavec, Apolonija, Kogej, Ksenija, Iglič, Aleš, Trebše, Polonca, Kralj-Iglič, Veronika, "Small Cellular Particles from European Spruce Needle Homogenate" in International Journal of Molecular Sciences, 24, no. 5 (2023):4349,
https://doi.org/10.3390/ijms24054349 . .

TY  - CONF
AU  - Nikolić, Dragana
AU  - Divac Rankov, Aleksandra
AU  - Samardžić, Jelena
AU  - Pantelić, Ana
AU  - Spasovski, Vesna
AU  - Banović Đeri, Bojana
AU  - Kosanović, Maja
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2266
AB  - Outer membrane vesicles (OMVs), extracellular vesicles (EVs) produced by Gram-negative bacteria, are
increasingly recognised as promising tools in biomedicine due to their innate ability to interact with human
cells and trigger immune responses. The interaction of OMVs of plant growth-promoting bacteria (PGPB) with
plants, as well as with plant-pathogenic microorganisms, is far less explored. Considering the great
importance of PGPBs for the development of sustainable, environmentally friendly solutions in agriculture, the
study of the role of OMVs in PGPB-plant and PGPB-phytopathogen interactions holds valuable application
potential.
To investigate PGPB OMVs, we isolated and characterised OMVs produced by Paraburkholderia phytofirmans
PsJN, a PGPB strain known to enhance plant resistance to various abiotic and biotic stresses. After testing
different methods for isolating and purifying OMVs, a commercially available affinity-based column system
was selected as the most efficient. Outer membrane origin of isolated OMVs was confirmed using an essay for
detection of lipopolysaccharide (LPS).
To examine the interaction of OMVs with plant cells, Arabidopsis thaliana roots were incubated with isolated
P. phytofirmans PsJN vesicles, previously labelled with lipid binding fluorescent dye Vybrant™ DiD. Red signals
were observed, under confocal laser scanning microscope, in root hairs and root surface in DiD-OMV treated
plants, while in control-treated roots the same signals were missing. The results suggest direct contact of
OMVs with root hairs, which are necessary for nutrient acquisition and plant-microbe interactions in
rhizosphere. Our further research is focused on the characterization of OMV-associated RNA and its potential
delivery into host plant cells.
PB  - Serbian Society for Extracellular Vesicles (SrbEVs)
PB  - Austrian Society for Extracellular Vesicles (ASEV)
PB  - Hungarian Society for Extracellular Vesicles (HSEV)
PB  - Slovenian Network for Extracellular Vesicles (SiN-EV)
C3  - Small New World 2.0
T1  - Exploring the interaction of Outer membrane vesicles (OMVs) produced by Paraburkholderia phytofirmans PsJN with Arabidopsis thaliana roots
EP  - 109
SP  - 109
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2266
ER  - 

@conference{
author = "Nikolić, Dragana and Divac Rankov, Aleksandra and Samardžić, Jelena and Pantelić, Ana and Spasovski, Vesna and Banović Đeri, Bojana and Kosanović, Maja",
year = "2023",
abstract = "Outer membrane vesicles (OMVs), extracellular vesicles (EVs) produced by Gram-negative bacteria, are
increasingly recognised as promising tools in biomedicine due to their innate ability to interact with human
cells and trigger immune responses. The interaction of OMVs of plant growth-promoting bacteria (PGPB) with
plants, as well as with plant-pathogenic microorganisms, is far less explored. Considering the great
importance of PGPBs for the development of sustainable, environmentally friendly solutions in agriculture, the
study of the role of OMVs in PGPB-plant and PGPB-phytopathogen interactions holds valuable application
potential.
To investigate PGPB OMVs, we isolated and characterised OMVs produced by Paraburkholderia phytofirmans
PsJN, a PGPB strain known to enhance plant resistance to various abiotic and biotic stresses. After testing
different methods for isolating and purifying OMVs, a commercially available affinity-based column system
was selected as the most efficient. Outer membrane origin of isolated OMVs was confirmed using an essay for
detection of lipopolysaccharide (LPS).
To examine the interaction of OMVs with plant cells, Arabidopsis thaliana roots were incubated with isolated
P. phytofirmans PsJN vesicles, previously labelled with lipid binding fluorescent dye Vybrant™ DiD. Red signals
were observed, under confocal laser scanning microscope, in root hairs and root surface in DiD-OMV treated
plants, while in control-treated roots the same signals were missing. The results suggest direct contact of
OMVs with root hairs, which are necessary for nutrient acquisition and plant-microbe interactions in
rhizosphere. Our further research is focused on the characterization of OMV-associated RNA and its potential
delivery into host plant cells.",
publisher = "Serbian Society for Extracellular Vesicles (SrbEVs), Austrian Society for Extracellular Vesicles (ASEV), Hungarian Society for Extracellular Vesicles (HSEV), Slovenian Network for Extracellular Vesicles (SiN-EV)",
journal = "Small New World 2.0",
title = "Exploring the interaction of Outer membrane vesicles (OMVs) produced by Paraburkholderia phytofirmans PsJN with Arabidopsis thaliana roots",
pages = "109-109",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2266"
}

Nikolić, D., Divac Rankov, A., Samardžić, J., Pantelić, A., Spasovski, V., Banović Đeri, B.,& Kosanović, M.. (2023). Exploring the interaction of Outer membrane vesicles (OMVs) produced by Paraburkholderia phytofirmans PsJN with Arabidopsis thaliana roots. in Small New World 2.0
Serbian Society for Extracellular Vesicles (SrbEVs)., 109-109.
https://hdl.handle.net/21.15107/rcub_imagine_2266

Nikolić D, Divac Rankov A, Samardžić J, Pantelić A, Spasovski V, Banović Đeri B, Kosanović M. Exploring the interaction of Outer membrane vesicles (OMVs) produced by Paraburkholderia phytofirmans PsJN with Arabidopsis thaliana roots. in Small New World 2.0. 2023;:109-109.
https://hdl.handle.net/21.15107/rcub_imagine_2266 .

Nikolić, Dragana, Divac Rankov, Aleksandra, Samardžić, Jelena, Pantelić, Ana, Spasovski, Vesna, Banović Đeri, Bojana, Kosanović, Maja, "Exploring the interaction of Outer membrane vesicles (OMVs) produced by Paraburkholderia phytofirmans PsJN with Arabidopsis thaliana roots" in Small New World 2.0 (2023):109-109,
https://hdl.handle.net/21.15107/rcub_imagine_2266 .

TY  - CONF
AU  - Spasovski, Vesna
AU  - Romolo, Anna
AU  - Kisovec, Matic
AU  - Zagorc, Urška
AU  - Arrigler, Vesna
AU  - Arko, Matevž
AU  - Bedina Zavec, Apolonija
AU  - Iglič, Aleš
AU  - Kogej, Ksenija
AU  - Kralj-Iglič, Veronika
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2265
AB  - Introduction: Being of compatible structure with biomembranes, lipid–based nanoparticles are considered
as convenient platforms for drug delivery systems. In the proposed work we considered formation of lipid
nanovesicles associated with bioactive phytochemicals from spruce needle homogenate (here called
hybridosomes). We formed hybridosomes by mixing appropriate amounts of lecithin, supernatant of isolation
of extracellular particles from spruce needle homogenate and glycerol.
Methods: We visualized hybridosomes by light microscopy and cryogenic transmission electron microscopy
and assessed them by flow cytometry, dynamic light scattering, ultraviolet–visual spectroscopy and
interferometric microscopy.
Results: We found that the particles consisted of a bilayer membrane and a fluid-like interior. Flow cytometry
and interferometric light microscopy measurements showed that the majority of the particles were
nano-sized. Dynamic light scattering and interferometric light microscopy measurements agreed well with the
determined average hydrodynamic radius of the particles Rh (between 140 and 180 nm) while their number
densities were in the range between 10^13 and 10^14/mL indicating that hybridosomes present about 2/3 of the
mixture, excluding solvent and other small molecules.
Discussion: Simple and low-cost preparation method, non-demanding saving process and efficient
formation procedure suggest that large scale production of hybridosomes from lipids and spruce needle
homogenate is feasible.
PB  - Serbian Society for Extracellular Vesicles (SrbEVs)
PB  - Austrian Society for Extracellular Vesicles (ASEV)
PB  - Hungarian Society for Extracellular Vesicles (HSEV)
PB  - Serbian Society for Extracellular Vesicles (SrbEVs)
C3  - Small New World 2.0
T1  - Hybridosomes from spruce needle homogenate
EP  - 85
SP  - 85
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2265
ER  - 

@conference{
author = "Spasovski, Vesna and Romolo, Anna and Kisovec, Matic and Zagorc, Urška and Arrigler, Vesna and Arko, Matevž and Bedina Zavec, Apolonija and Iglič, Aleš and Kogej, Ksenija and Kralj-Iglič, Veronika",
year = "2023",
abstract = "Introduction: Being of compatible structure with biomembranes, lipid–based nanoparticles are considered
as convenient platforms for drug delivery systems. In the proposed work we considered formation of lipid
nanovesicles associated with bioactive phytochemicals from spruce needle homogenate (here called
hybridosomes). We formed hybridosomes by mixing appropriate amounts of lecithin, supernatant of isolation
of extracellular particles from spruce needle homogenate and glycerol.
Methods: We visualized hybridosomes by light microscopy and cryogenic transmission electron microscopy
and assessed them by flow cytometry, dynamic light scattering, ultraviolet–visual spectroscopy and
interferometric microscopy.
Results: We found that the particles consisted of a bilayer membrane and a fluid-like interior. Flow cytometry
and interferometric light microscopy measurements showed that the majority of the particles were
nano-sized. Dynamic light scattering and interferometric light microscopy measurements agreed well with the
determined average hydrodynamic radius of the particles Rh (between 140 and 180 nm) while their number
densities were in the range between 10^13 and 10^14/mL indicating that hybridosomes present about 2/3 of the
mixture, excluding solvent and other small molecules.
Discussion: Simple and low-cost preparation method, non-demanding saving process and efficient
formation procedure suggest that large scale production of hybridosomes from lipids and spruce needle
homogenate is feasible.",
publisher = "Serbian Society for Extracellular Vesicles (SrbEVs), Austrian Society for Extracellular Vesicles (ASEV), Hungarian Society for Extracellular Vesicles (HSEV), Serbian Society for Extracellular Vesicles (SrbEVs)",
journal = "Small New World 2.0",
title = "Hybridosomes from spruce needle homogenate",
pages = "85-85",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2265"
}

Spasovski, V., Romolo, A., Kisovec, M., Zagorc, U., Arrigler, V., Arko, M., Bedina Zavec, A., Iglič, A., Kogej, K.,& Kralj-Iglič, V.. (2023). Hybridosomes from spruce needle homogenate. in Small New World 2.0
Serbian Society for Extracellular Vesicles (SrbEVs)., 85-85.
https://hdl.handle.net/21.15107/rcub_imagine_2265

Spasovski V, Romolo A, Kisovec M, Zagorc U, Arrigler V, Arko M, Bedina Zavec A, Iglič A, Kogej K, Kralj-Iglič V. Hybridosomes from spruce needle homogenate. in Small New World 2.0. 2023;:85-85.
https://hdl.handle.net/21.15107/rcub_imagine_2265 .

Spasovski, Vesna, Romolo, Anna, Kisovec, Matic, Zagorc, Urška, Arrigler, Vesna, Arko, Matevž, Bedina Zavec, Apolonija, Iglič, Aleš, Kogej, Ksenija, Kralj-Iglič, Veronika, "Hybridosomes from spruce needle homogenate" in Small New World 2.0 (2023):85-85,
https://hdl.handle.net/21.15107/rcub_imagine_2265 .

TY  - CONF
AU  - Parezanović, Marina
AU  - Stojiljković, Maja
AU  - Anđelković, Marina
AU  - Stevanović, Nina
AU  - Spasovski, Vesna
AU  - Ugrin, Milena
AU  - Komazec, Jovana
AU  - Tošić, Nataša
AU  - Pavlović, Sonja
AU  - Celic, Dejan
AU  - Vučenović, Jelica
AU  - Skakić, Anita
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2277
AB  - Background/Objectives: Fabry disease (FD) is a rare X-linked
disorder caused by variants in the GLA gene leading to the deficiency
of lysosomal α-galactosidase-A and progressive accumulation
of globotriaosylceramide affecting the heart, nervous system,
and kidneys. FD has overlapping phenotypes and often remains
undiagnosed. Therefore, the precise molecular-genetic diagnosis
and the earliest possible treatment are essential to avoid significant
disease progression.
Methods: We analyzed 95 (34 female and 61 male) hemodialysis
patients with clinical suspicion of FD using Sanger sequencing
of all coding exons (7) and flanking intron regions of the GLA
gene, and measured the relative expression of the GLA gene in
available samples.
Results: The genetic analysis revealed 3 patients with a missense
variant (p.Asp313Tyr), and 10 patients with combinations of
non-coding variants, described as complex intronic haplotypes
(CIHs). CIH1 (c.-10C>T, c.370-81_370-77delCAGCC, c.640-16A>G,
c.1000-22C>T), the most frequent haplotype, was detected in 7
(7.4%) patients. Lyso-Gb3 biomarker levels were within the normal
range in each tested patient. However, RT-qPCR analysis revealed
decreased relative expression of GLA gene in PBMC of 2 female
patients with CIH1 and one female patient carrying only c.-10C>T
variant by 9,1%, 7,4%, 46,3%, respectively, pointing out that further
analyses are needed to confirm/exclude FD in these patients.
Conclusion: Because the effects of CIHs are not yet fully
understood, our work highlights the importance of analyzing
intronic regions of the GLA gene as genetic modifiers and the
need to include expression analysis in the diagnostic algorithm.
PB  - Springer Nature
C3  - European Journal of Human Genetic
T1  - Molecular diagnosis of Fabry disease in patients with chronic renal failure of unknown etiology
EP  - 433
IS  - Supplement S1
SP  - 432
VL  - 31
DO  - 10.1038/s41431-023-01339-3
ER  - 

@conference{
author = "Parezanović, Marina and Stojiljković, Maja and Anđelković, Marina and Stevanović, Nina and Spasovski, Vesna and Ugrin, Milena and Komazec, Jovana and Tošić, Nataša and Pavlović, Sonja and Celic, Dejan and Vučenović, Jelica and Skakić, Anita",
year = "2023",
abstract = "Background/Objectives: Fabry disease (FD) is a rare X-linked
disorder caused by variants in the GLA gene leading to the deficiency
of lysosomal α-galactosidase-A and progressive accumulation
of globotriaosylceramide affecting the heart, nervous system,
and kidneys. FD has overlapping phenotypes and often remains
undiagnosed. Therefore, the precise molecular-genetic diagnosis
and the earliest possible treatment are essential to avoid significant
disease progression.
Methods: We analyzed 95 (34 female and 61 male) hemodialysis
patients with clinical suspicion of FD using Sanger sequencing
of all coding exons (7) and flanking intron regions of the GLA
gene, and measured the relative expression of the GLA gene in
available samples.
Results: The genetic analysis revealed 3 patients with a missense
variant (p.Asp313Tyr), and 10 patients with combinations of
non-coding variants, described as complex intronic haplotypes
(CIHs). CIH1 (c.-10C>T, c.370-81_370-77delCAGCC, c.640-16A>G,
c.1000-22C>T), the most frequent haplotype, was detected in 7
(7.4%) patients. Lyso-Gb3 biomarker levels were within the normal
range in each tested patient. However, RT-qPCR analysis revealed
decreased relative expression of GLA gene in PBMC of 2 female
patients with CIH1 and one female patient carrying only c.-10C>T
variant by 9,1%, 7,4%, 46,3%, respectively, pointing out that further
analyses are needed to confirm/exclude FD in these patients.
Conclusion: Because the effects of CIHs are not yet fully
understood, our work highlights the importance of analyzing
intronic regions of the GLA gene as genetic modifiers and the
need to include expression analysis in the diagnostic algorithm.",
publisher = "Springer Nature",
journal = "European Journal of Human Genetic",
title = "Molecular diagnosis of Fabry disease in patients with chronic renal failure of unknown etiology",
pages = "433-432",
number = "Supplement S1",
volume = "31",
doi = "10.1038/s41431-023-01339-3"
}

Parezanović, M., Stojiljković, M., Anđelković, M., Stevanović, N., Spasovski, V., Ugrin, M., Komazec, J., Tošić, N., Pavlović, S., Celic, D., Vučenović, J.,& Skakić, A.. (2023). Molecular diagnosis of Fabry disease in patients with chronic renal failure of unknown etiology. in European Journal of Human Genetic
Springer Nature., 31(Supplement S1), 432-433.
https://doi.org/10.1038/s41431-023-01339-3

Parezanović M, Stojiljković M, Anđelković M, Stevanović N, Spasovski V, Ugrin M, Komazec J, Tošić N, Pavlović S, Celic D, Vučenović J, Skakić A. Molecular diagnosis of Fabry disease in patients with chronic renal failure of unknown etiology. in European Journal of Human Genetic. 2023;31(Supplement S1):432-433.
doi:10.1038/s41431-023-01339-3 .

Parezanović, Marina, Stojiljković, Maja, Anđelković, Marina, Stevanović, Nina, Spasovski, Vesna, Ugrin, Milena, Komazec, Jovana, Tošić, Nataša, Pavlović, Sonja, Celic, Dejan, Vučenović, Jelica, Skakić, Anita, "Molecular diagnosis of Fabry disease in patients with chronic renal failure of unknown etiology" in European Journal of Human Genetic, 31, no. Supplement S1 (2023):432-433,
https://doi.org/10.1038/s41431-023-01339-3 . .

TY  - CONF
AU  - Spasovski, Vesna
AU  - Romolo, Anna
AU  - Kisovec, Matic
AU  - Zagorc, Urška
AU  - Arrigler, Vesna
AU  - Arko, Matevž
AU  - Bedina Zavec, Apolonija
AU  - Iglič, Aleš
AU  - Kogej, Ksenija
AU  - Kralj-Iglič, Veronika
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2264
AB  - Introduction: Being of compatible structure with biomembranes, lipid–based nanoparticles are considered
as convenient platforms for drug delivery systems. In the proposed work we considered formation of lipid
nanovesicles associated with bioactive phytochemicals from spruce needle homogenate (here called
hybridosomes). We formed hybridosomes by mixing appropriate amounts of lecithin, supernatant of isolation
of extracellular particles from spruce needle homogenate and glycerol.
Methods: We visualized hybridosomes by light microscopy and cryogenic transmission electron microscopy
and assessed them by flow cytometry, dynamic light scattering, ultraviolet–visual spectroscopy and
interferometric microscopy.
Results: We found that the particles consisted of a bilayer membrane and a fluid-like interior. Flow cytometry
and interferometric light microscopy measurements showed that the majority of the particles were
nano-sized. Dynamic light scattering and interferometric light microscopy measurements agreed well with the
determined average hydrodynamic radius of the particles Rh (between 140 and 180 nm) while their number
densities were in the range between 10^13 and 10^14/mL indicating that hybridosomes present about 2/3 of the
mixture, excluding solvent and other small molecules.
Discussion: Simple and low-cost preparation method, non-demanding saving process and efficient
formation procedure suggest that large scale production of hybridosomes from lipids and spruce needle
homogenate is feasible.
PB  - Serbian Society for Extracellular Vesicles (SrbEVs)
PB  - Austrian Society for Extracellular Vesicles (ASEV)
PB  - Hungarian Society for Extracellular Vesicles (HSEV)
PB  - Slovenian Network for Extracellular Vesicles (SiN-EV)
C3  - Small New World 2.0
T1  - Hybridosomes from spruce needle homogenate
EP  - 27
SP  - 27
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2264
ER  - 

@conference{
author = "Spasovski, Vesna and Romolo, Anna and Kisovec, Matic and Zagorc, Urška and Arrigler, Vesna and Arko, Matevž and Bedina Zavec, Apolonija and Iglič, Aleš and Kogej, Ksenija and Kralj-Iglič, Veronika",
year = "2023",
abstract = "Introduction: Being of compatible structure with biomembranes, lipid–based nanoparticles are considered
as convenient platforms for drug delivery systems. In the proposed work we considered formation of lipid
nanovesicles associated with bioactive phytochemicals from spruce needle homogenate (here called
hybridosomes). We formed hybridosomes by mixing appropriate amounts of lecithin, supernatant of isolation
of extracellular particles from spruce needle homogenate and glycerol.
Methods: We visualized hybridosomes by light microscopy and cryogenic transmission electron microscopy
and assessed them by flow cytometry, dynamic light scattering, ultraviolet–visual spectroscopy and
interferometric microscopy.
Results: We found that the particles consisted of a bilayer membrane and a fluid-like interior. Flow cytometry
and interferometric light microscopy measurements showed that the majority of the particles were
nano-sized. Dynamic light scattering and interferometric light microscopy measurements agreed well with the
determined average hydrodynamic radius of the particles Rh (between 140 and 180 nm) while their number
densities were in the range between 10^13 and 10^14/mL indicating that hybridosomes present about 2/3 of the
mixture, excluding solvent and other small molecules.
Discussion: Simple and low-cost preparation method, non-demanding saving process and efficient
formation procedure suggest that large scale production of hybridosomes from lipids and spruce needle
homogenate is feasible.",
publisher = "Serbian Society for Extracellular Vesicles (SrbEVs), Austrian Society for Extracellular Vesicles (ASEV), Hungarian Society for Extracellular Vesicles (HSEV), Slovenian Network for Extracellular Vesicles (SiN-EV)",
journal = "Small New World 2.0",
title = "Hybridosomes from spruce needle homogenate",
pages = "27-27",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2264"
}

Spasovski, V., Romolo, A., Kisovec, M., Zagorc, U., Arrigler, V., Arko, M., Bedina Zavec, A., Iglič, A., Kogej, K.,& Kralj-Iglič, V.. (2023). Hybridosomes from spruce needle homogenate. in Small New World 2.0
Serbian Society for Extracellular Vesicles (SrbEVs)., 27-27.
https://hdl.handle.net/21.15107/rcub_imagine_2264

Spasovski V, Romolo A, Kisovec M, Zagorc U, Arrigler V, Arko M, Bedina Zavec A, Iglič A, Kogej K, Kralj-Iglič V. Hybridosomes from spruce needle homogenate. in Small New World 2.0. 2023;:27-27.
https://hdl.handle.net/21.15107/rcub_imagine_2264 .

Spasovski, Vesna, Romolo, Anna, Kisovec, Matic, Zagorc, Urška, Arrigler, Vesna, Arko, Matevž, Bedina Zavec, Apolonija, Iglič, Aleš, Kogej, Ksenija, Kralj-Iglič, Veronika, "Hybridosomes from spruce needle homogenate" in Small New World 2.0 (2023):27-27,
https://hdl.handle.net/21.15107/rcub_imagine_2264 .

TY  - CONF
AU  - Stevanović, Nina
AU  - Anđelković, Marina
AU  - Skakić, Anita
AU  - Spasovski, Vesna
AU  - Stojiljković, Maja
AU  - Parezanović, Marina
AU  - Ugrin, Milena
AU  - Pavlović, Sonja
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2275
AB  - Background/Objectives: Primary ciliary dyskinesia (PCD) is a
disease caused by impaired ciliary motility and mainly affects the
lungs and reproductive organs. Inheritance is autosomal recessive
and X-linked with more than 40 disease-causing genes, wherefore
PCD patients have diverse clinical manifestations, thus making
diagnosis difficult. The utility of next-generation sequencing (NGS)
technology for diagnostic purposes allows a better understanding
of the PCD genetic background. However, the identification of
specific disease-causing variants is challenging. The objective of
this study was to create a unique guideline that will enable the
standardization of the assessment of novel variants within PCD
associated genes.
Methods: The study included designing a pipeline for the classification
of the rare genetic variants detected using NGS. The pipeline
included in silico (translation, 3D-model, protein-protein interactions,
sequence conservation, posttranslational modifications) and functional
analysis (expressional analysis, Western Blot) of the variants.
Results: The designed pipeline consists of three steps: sequencing,
detection, and identification of genes/variants; classification of
variants according to their effect; and variant characterization using
in silico structural and functional analysis. The pipeline was validated
by the analysis of the variants detected in a disease-causing
gene (DNAI1) and the novel candidate gene (SPAG16).
Conclusion: The application of the pipeline resulted in the
identification of disease-causing variants, as well as pathogenicity
validation, through the analysis on transcriptional, translational,
and posttranslational levels.The application of created pipeline
leads to the confirmation of PCD diagnosis and enables a shift
from candidate to disease-causing gene.
PB  - Springer Nature
C3  - European Journal of Human Genetic
T1  - Unique pipeline for the assessment of novel genetic variants leads to confirmation of PCD diagnosis
EP  - 383
IS  - Supplement S1
SP  - 383
VL  - 31
DO  - 10.1038/s41431-023-01338-4
ER  - 

@conference{
author = "Stevanović, Nina and Anđelković, Marina and Skakić, Anita and Spasovski, Vesna and Stojiljković, Maja and Parezanović, Marina and Ugrin, Milena and Pavlović, Sonja",
year = "2023",
abstract = "Background/Objectives: Primary ciliary dyskinesia (PCD) is a
disease caused by impaired ciliary motility and mainly affects the
lungs and reproductive organs. Inheritance is autosomal recessive
and X-linked with more than 40 disease-causing genes, wherefore
PCD patients have diverse clinical manifestations, thus making
diagnosis difficult. The utility of next-generation sequencing (NGS)
technology for diagnostic purposes allows a better understanding
of the PCD genetic background. However, the identification of
specific disease-causing variants is challenging. The objective of
this study was to create a unique guideline that will enable the
standardization of the assessment of novel variants within PCD
associated genes.
Methods: The study included designing a pipeline for the classification
of the rare genetic variants detected using NGS. The pipeline
included in silico (translation, 3D-model, protein-protein interactions,
sequence conservation, posttranslational modifications) and functional
analysis (expressional analysis, Western Blot) of the variants.
Results: The designed pipeline consists of three steps: sequencing,
detection, and identification of genes/variants; classification of
variants according to their effect; and variant characterization using
in silico structural and functional analysis. The pipeline was validated
by the analysis of the variants detected in a disease-causing
gene (DNAI1) and the novel candidate gene (SPAG16).
Conclusion: The application of the pipeline resulted in the
identification of disease-causing variants, as well as pathogenicity
validation, through the analysis on transcriptional, translational,
and posttranslational levels.The application of created pipeline
leads to the confirmation of PCD diagnosis and enables a shift
from candidate to disease-causing gene.",
publisher = "Springer Nature",
journal = "European Journal of Human Genetic",
title = "Unique pipeline for the assessment of novel genetic variants leads to confirmation of PCD diagnosis",
pages = "383-383",
number = "Supplement S1",
volume = "31",
doi = "10.1038/s41431-023-01338-4"
}

Stevanović, N., Anđelković, M., Skakić, A., Spasovski, V., Stojiljković, M., Parezanović, M., Ugrin, M.,& Pavlović, S.. (2023). Unique pipeline for the assessment of novel genetic variants leads to confirmation of PCD diagnosis. in European Journal of Human Genetic
Springer Nature., 31(Supplement S1), 383-383.
https://doi.org/10.1038/s41431-023-01338-4

Stevanović N, Anđelković M, Skakić A, Spasovski V, Stojiljković M, Parezanović M, Ugrin M, Pavlović S. Unique pipeline for the assessment of novel genetic variants leads to confirmation of PCD diagnosis. in European Journal of Human Genetic. 2023;31(Supplement S1):383-383.
doi:10.1038/s41431-023-01338-4 .

Stevanović, Nina, Anđelković, Marina, Skakić, Anita, Spasovski, Vesna, Stojiljković, Maja, Parezanović, Marina, Ugrin, Milena, Pavlović, Sonja, "Unique pipeline for the assessment of novel genetic variants leads to confirmation of PCD diagnosis" in European Journal of Human Genetic, 31, no. Supplement S1 (2023):383-383,
https://doi.org/10.1038/s41431-023-01338-4 . .

TY  - DATA
AU  - Spasovski, Vesna
AU  - Srzentić Dražilov, Sanja
AU  - Nikčević, Gordana
AU  - Baščarević, Zoran
AU  - Stojiljković, Maja
AU  - Pavlović, Sonja
AU  - Spasovski, Duško
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2089
AB  - Background: Perthes disease is a juvenile form of osteonecrosis of the femoral head that affects children under the age of 15. One hundred years after its discovery, some light has been shed on its etiology and the biological factors relevant to its etiology and disease severity. Methods: The aim of this study was to summarize the literature findings on the biological factors relevant to the pathogenesis of Perthes disease, their diagnostic and clinical significance, and their therapeutic potential. A special focus on candidate genes as susceptibility factors and factors relevant to clinical severity was made, where studies reporting clinical or preclinical results were considered as the inclusion criteria. PubMed databases were searched by two independent researchers. Sixty-eight articles were included in this review. Results on the factors relevant to vascular involvement and inflammatory molecules indicated as factors that contribute to impaired bone remodeling have been summarized. Moreover, several candidate genes relevant to an active phase of the disease have been suggested as possible biological therapeutic targets. Conclusions: Delineation of molecular biomarkers that underlie the pathophysiological process of Perthes disease can allow for the provision of earlier and more accurate diagnoses of the disease and more precise follow-ups and treatment in the early phases of the disease.
PB  - MDPI
T2  - Diagnostics
T1  - Supplementary material for: Spasovski, V.; Srzentić Dražilov, S.; Nikčević, G.; Baščarević, Z.; Stojiljković, M.; Pavlović, S.; Spasovski, D. Molecular Biomarkers in Perthes Disease: A Review. Diagnostics 2023, 13 (3), 471. https://doi.org/10.3390/diagnostics13030471.
IS  - 3
SP  - 471
VL  - 13
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2089
ER  - 

@misc{
author = "Spasovski, Vesna and Srzentić Dražilov, Sanja and Nikčević, Gordana and Baščarević, Zoran and Stojiljković, Maja and Pavlović, Sonja and Spasovski, Duško",
year = "2023",
abstract = "Background: Perthes disease is a juvenile form of osteonecrosis of the femoral head that affects children under the age of 15. One hundred years after its discovery, some light has been shed on its etiology and the biological factors relevant to its etiology and disease severity. Methods: The aim of this study was to summarize the literature findings on the biological factors relevant to the pathogenesis of Perthes disease, their diagnostic and clinical significance, and their therapeutic potential. A special focus on candidate genes as susceptibility factors and factors relevant to clinical severity was made, where studies reporting clinical or preclinical results were considered as the inclusion criteria. PubMed databases were searched by two independent researchers. Sixty-eight articles were included in this review. Results on the factors relevant to vascular involvement and inflammatory molecules indicated as factors that contribute to impaired bone remodeling have been summarized. Moreover, several candidate genes relevant to an active phase of the disease have been suggested as possible biological therapeutic targets. Conclusions: Delineation of molecular biomarkers that underlie the pathophysiological process of Perthes disease can allow for the provision of earlier and more accurate diagnoses of the disease and more precise follow-ups and treatment in the early phases of the disease.",
publisher = "MDPI",
journal = "Diagnostics",
title = "Supplementary material for: Spasovski, V.; Srzentić Dražilov, S.; Nikčević, G.; Baščarević, Z.; Stojiljković, M.; Pavlović, S.; Spasovski, D. Molecular Biomarkers in Perthes Disease: A Review. Diagnostics 2023, 13 (3), 471. https://doi.org/10.3390/diagnostics13030471.",
number = "3",
pages = "471",
volume = "13",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2089"
}

Spasovski, V., Srzentić Dražilov, S., Nikčević, G., Baščarević, Z., Stojiljković, M., Pavlović, S.,& Spasovski, D.. (2023). Supplementary material for: Spasovski, V.; Srzentić Dražilov, S.; Nikčević, G.; Baščarević, Z.; Stojiljković, M.; Pavlović, S.; Spasovski, D. Molecular Biomarkers in Perthes Disease: A Review. Diagnostics 2023, 13 (3), 471. https://doi.org/10.3390/diagnostics13030471.. in Diagnostics
MDPI., 13(3), 471.
https://hdl.handle.net/21.15107/rcub_imagine_2089

Spasovski V, Srzentić Dražilov S, Nikčević G, Baščarević Z, Stojiljković M, Pavlović S, Spasovski D. Supplementary material for: Spasovski, V.; Srzentić Dražilov, S.; Nikčević, G.; Baščarević, Z.; Stojiljković, M.; Pavlović, S.; Spasovski, D. Molecular Biomarkers in Perthes Disease: A Review. Diagnostics 2023, 13 (3), 471. https://doi.org/10.3390/diagnostics13030471.. in Diagnostics. 2023;13(3):471.
https://hdl.handle.net/21.15107/rcub_imagine_2089 .

Spasovski, Vesna, Srzentić Dražilov, Sanja, Nikčević, Gordana, Baščarević, Zoran, Stojiljković, Maja, Pavlović, Sonja, Spasovski, Duško, "Supplementary material for: Spasovski, V.; Srzentić Dražilov, S.; Nikčević, G.; Baščarević, Z.; Stojiljković, M.; Pavlović, S.; Spasovski, D. Molecular Biomarkers in Perthes Disease: A Review. Diagnostics 2023, 13 (3), 471. https://doi.org/10.3390/diagnostics13030471." in Diagnostics, 13, no. 3 (2023):471,
https://hdl.handle.net/21.15107/rcub_imagine_2089 .

TY  - CONF
AU  - Ugrin, Milena
AU  - Komazec, Jovana
AU  - Klaassen, Kristel
AU  - Skakić, Anita
AU  - Anđelković, Marina
AU  - Spasovski, Vesna
AU  - Stevanović, Nina
AU  - Parezanović, Marina
AU  - Stanković, Sara
AU  - Stojiljković, Maja
AU  - Pavlović, Sonja
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2139
AB  - Introduction: Thalassemia syndromes are heterogeneous group of hereditary anemias characterized
by defects in the synthesis of hemoglobin (Hb) polypeptide chains. These disorders comprise thalassemias and thalassemic hemoglobin variants which are predominantly caused by mutationsin a- and
b-globin genes (HBA and HBB genes). Clinical manifestations of thalassemia syndromes range from
asymptomatic thalassemia minor to severe anemia in thalassemia major cases. The aim of thisstudy was
to update our previous findings on frequency of thalassemia mutations which result from a 13-year-old
systematic survey in Serbia.
Methods: Two hundred and fourteen patients from 149 unrelated families presented with hematological parameters indicative of thalassemia syndromes were studied. Detection of α- and β-globin gene
mutations was performed using PCR and direct sequencing.
Results: Two Hb variants and twelve different β-thalassemia mutations, including two mutations previously not reported in Serbian population, were detected. Hb variant Lepore Boston-Washington wasthe
most common cause of thalassemia, with frequency of 24.3%, followed by HBB:c.316-106C>G mutation
detected in 18.1% of families. The third most frequent cause of β-thalassemia were HBB:c.118C>T and
HBB:c.93-21G>A mutations with 16.6% incidence each. Together, these four variants account for over
75% of all mutated β-globin alleles. In addition, five families affected with α-thalassemia were detected.
Conclusion: Despite the increase in cohort size by 50% between this and our previous studies, the frequency of mutations affecting HBB gene remained unchanged. Results presented in this study will update Serbian national mutation database and contribute to better understanding of geographic history
of South European and Balkan populations.
PB  - 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  - Molecular basis of thalassemia syndromes in Serbia: an update
EP  - 86
SP  - 86
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2139
ER  - 

@conference{
author = "Ugrin, Milena and Komazec, Jovana and Klaassen, Kristel and Skakić, Anita and Anđelković, Marina and Spasovski, Vesna and Stevanović, Nina and Parezanović, Marina and Stanković, Sara and Stojiljković, Maja and Pavlović, Sonja",
year = "2023",
abstract = "Introduction: Thalassemia syndromes are heterogeneous group of hereditary anemias characterized
by defects in the synthesis of hemoglobin (Hb) polypeptide chains. These disorders comprise thalassemias and thalassemic hemoglobin variants which are predominantly caused by mutationsin a- and
b-globin genes (HBA and HBB genes). Clinical manifestations of thalassemia syndromes range from
asymptomatic thalassemia minor to severe anemia in thalassemia major cases. The aim of thisstudy was
to update our previous findings on frequency of thalassemia mutations which result from a 13-year-old
systematic survey in Serbia.
Methods: Two hundred and fourteen patients from 149 unrelated families presented with hematological parameters indicative of thalassemia syndromes were studied. Detection of α- and β-globin gene
mutations was performed using PCR and direct sequencing.
Results: Two Hb variants and twelve different β-thalassemia mutations, including two mutations previously not reported in Serbian population, were detected. Hb variant Lepore Boston-Washington wasthe
most common cause of thalassemia, with frequency of 24.3%, followed by HBB:c.316-106C>G mutation
detected in 18.1% of families. The third most frequent cause of β-thalassemia were HBB:c.118C>T and
HBB:c.93-21G>A mutations with 16.6% incidence each. Together, these four variants account for over
75% of all mutated β-globin alleles. In addition, five families affected with α-thalassemia were detected.
Conclusion: Despite the increase in cohort size by 50% between this and our previous studies, the frequency of mutations affecting HBB gene remained unchanged. Results presented in this study will update Serbian national mutation database and contribute to better understanding of geographic history
of South European and Balkan populations.",
publisher = "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 = "Molecular basis of thalassemia syndromes in Serbia: an update",
pages = "86-86",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2139"
}

Ugrin, M., Komazec, J., Klaassen, K., Skakić, A., Anđelković, M., Spasovski, V., Stevanović, N., Parezanović, M., Stanković, S., Stojiljković, M.,& Pavlović, S.. (2023). Molecular basis of thalassemia syndromes in Serbia: an update. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 86-86.
https://hdl.handle.net/21.15107/rcub_imagine_2139

Ugrin M, Komazec J, Klaassen K, Skakić A, Anđelković M, Spasovski V, Stevanović N, Parezanović M, Stanković S, Stojiljković M, Pavlović S. Molecular basis of thalassemia syndromes in Serbia: an update. 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;:86-86.
https://hdl.handle.net/21.15107/rcub_imagine_2139 .

Ugrin, Milena, Komazec, Jovana, Klaassen, Kristel, Skakić, Anita, Anđelković, Marina, Spasovski, Vesna, Stevanović, Nina, Parezanović, Marina, Stanković, Sara, Stojiljković, Maja, Pavlović, Sonja, "Molecular basis of thalassemia syndromes in Serbia: an update" 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):86-86,
https://hdl.handle.net/21.15107/rcub_imagine_2139 .

TY  - CONF
AU  - Nešić, Sofija
AU  - Divac Rankov, Aleksandra
AU  - Spasovski, Vesna
AU  - Kosanović, Maja
AU  - Nikolić, Dragana
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2157
AB  - Extracellular vesicles (EVs) are recognized as important mediators of intercellular
communication in both eukaryotes and prokaryotes. These lipid membrane –
coated spherical nanoparticles carry proteins, nucleic acids and other cellular
products, and facilitate exchange of these biomolecules among cells within an
organism, but also between cells of different organisms, belonging to different
species and even kingdoms. Outer membrane vesicles (OMVs), EVs produced
by Gram-negative bacteria, are a significant mediator of microbial communication,
involved in biofilm formation, virulence, and modulation of host immunity.
OMVs of both pathogenic and plant beneficial bacteria have been shown
to elicit plant immune responses. Investigations on the modes of OMV-plant
cells interactions are still in their infancy, but gain rising attention. Aiming to
monitor the interaction between OMVs of Paraburkholderia phytofirmans PsJN,
a plant growth promoting bacteria, and Arabidopsis thaliana roots, we isolated
OMVs from bacterial culture in mineral medium, using an ion-exchange chromatography
system. Isolated OMVs were labeled with lipid binding fluorescent
dye Vybrant™ DiD and unbound dye was removed by washing vesicles on ultrafiltration
columns. The same dye concentration in phosphate buffer saline,
equivalently washed, was used as a control. A. thaliana roots, grown on Murashige
and Skoog medium, were incubated with DiD-OMVs or control dye/buffer
mixture, washed and observed under confocal laser scanning microscope. Red
signals were observed in root hairs and epidermis in DiD-OMV treated plants,
while in control-treated roots the same signals were missing. The results indicate
direct contact of bacterial vesicles with epidermis and root hairs, which are
indispensable for nutrient acquisition and plant-microbe interactions in rhizosphere.
Further investigation will address the questions of the nature of OMVplant
cell interaction, including potential delivery of OMVs cargo into host plant
cells. Considering that OMVs are increasingly recognized as promising tools in
biomedicine, exploring their potential for agronomical applications would be
highly appreciated.
PB  - Belgrade : Faculty of Biology
C3  - ICGEB WORKSHOP: Trends in microbial solutions for sustainable agriculture
T1  - Outer membrane vesicles of plant beneficial bacterial strain
Paraburkholderia phytofirmans PsJN make a contact with
Arabidopsis thaliana roots
EP  - 89
SP  - 89
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2157
ER  - 

@conference{
author = "Nešić, Sofija and Divac Rankov, Aleksandra and Spasovski, Vesna and Kosanović, Maja and Nikolić, Dragana",
year = "2023",
abstract = "Extracellular vesicles (EVs) are recognized as important mediators of intercellular
communication in both eukaryotes and prokaryotes. These lipid membrane –
coated spherical nanoparticles carry proteins, nucleic acids and other cellular
products, and facilitate exchange of these biomolecules among cells within an
organism, but also between cells of different organisms, belonging to different
species and even kingdoms. Outer membrane vesicles (OMVs), EVs produced
by Gram-negative bacteria, are a significant mediator of microbial communication,
involved in biofilm formation, virulence, and modulation of host immunity.
OMVs of both pathogenic and plant beneficial bacteria have been shown
to elicit plant immune responses. Investigations on the modes of OMV-plant
cells interactions are still in their infancy, but gain rising attention. Aiming to
monitor the interaction between OMVs of Paraburkholderia phytofirmans PsJN,
a plant growth promoting bacteria, and Arabidopsis thaliana roots, we isolated
OMVs from bacterial culture in mineral medium, using an ion-exchange chromatography
system. Isolated OMVs were labeled with lipid binding fluorescent
dye Vybrant™ DiD and unbound dye was removed by washing vesicles on ultrafiltration
columns. The same dye concentration in phosphate buffer saline,
equivalently washed, was used as a control. A. thaliana roots, grown on Murashige
and Skoog medium, were incubated with DiD-OMVs or control dye/buffer
mixture, washed and observed under confocal laser scanning microscope. Red
signals were observed in root hairs and epidermis in DiD-OMV treated plants,
while in control-treated roots the same signals were missing. The results indicate
direct contact of bacterial vesicles with epidermis and root hairs, which are
indispensable for nutrient acquisition and plant-microbe interactions in rhizosphere.
Further investigation will address the questions of the nature of OMVplant
cell interaction, including potential delivery of OMVs cargo into host plant
cells. Considering that OMVs are increasingly recognized as promising tools in
biomedicine, exploring their potential for agronomical applications would be
highly appreciated.",
publisher = "Belgrade : Faculty of Biology",
journal = "ICGEB WORKSHOP: Trends in microbial solutions for sustainable agriculture",
title = "Outer membrane vesicles of plant beneficial bacterial strain
Paraburkholderia phytofirmans PsJN make a contact with
Arabidopsis thaliana roots",
pages = "89-89",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2157"
}

Nešić, S., Divac Rankov, A., Spasovski, V., Kosanović, M.,& Nikolić, D.. (2023). Outer membrane vesicles of plant beneficial bacterial strain
Paraburkholderia phytofirmans PsJN make a contact with
Arabidopsis thaliana roots. in ICGEB WORKSHOP: Trends in microbial solutions for sustainable agriculture
Belgrade : Faculty of Biology., 89-89.
https://hdl.handle.net/21.15107/rcub_imagine_2157

Nešić S, Divac Rankov A, Spasovski V, Kosanović M, Nikolić D. Outer membrane vesicles of plant beneficial bacterial strain
Paraburkholderia phytofirmans PsJN make a contact with
Arabidopsis thaliana roots. in ICGEB WORKSHOP: Trends in microbial solutions for sustainable agriculture. 2023;:89-89.
https://hdl.handle.net/21.15107/rcub_imagine_2157 .

Nešić, Sofija, Divac Rankov, Aleksandra, Spasovski, Vesna, Kosanović, Maja, Nikolić, Dragana, "Outer membrane vesicles of plant beneficial bacterial strain
Paraburkholderia phytofirmans PsJN make a contact with
Arabidopsis thaliana roots" in ICGEB WORKSHOP: Trends in microbial solutions for sustainable agriculture (2023):89-89,
https://hdl.handle.net/21.15107/rcub_imagine_2157 .

TY  - JOUR
AU  - Spasovski, Vesna
AU  - Srzentić Dražilov, Sanja
AU  - Nikčević, Gordana
AU  - Baščarević, Zoran
AU  - Stojiljković, Maja
AU  - Pavlović, Sonja
AU  - Spasovski, Duško
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2088
AB  - Background: Perthes disease is a juvenile form of osteonecrosis of the femoral head that affects children under the age of 15. One hundred years after its discovery, some light has been shed on its etiology and the biological factors relevant to its etiology and disease severity. Methods: The aim of this study was to summarize the literature findings on the biological factors relevant to the pathogenesis of Perthes disease, their diagnostic and clinical significance, and their therapeutic potential. A special focus on candidate genes as susceptibility factors and factors relevant to clinical severity was made, where studies reporting clinical or preclinical results were considered as the inclusion criteria. PubMed databases were searched by two independent researchers. Sixty-eight articles were included in this review. Results on the factors relevant to vascular involvement and inflammatory molecules indicated as factors that contribute to impaired bone remodeling have been summarized. Moreover, several candidate genes relevant to an active phase of the disease have been suggested as possible biological therapeutic targets. Conclusions: Delineation of molecular biomarkers that underlie the pathophysiological process of Perthes disease can allow for the provision of earlier and more accurate diagnoses of the disease and more precise follow-ups and treatment in the early phases of the disease.
PB  - MDPI
T2  - Diagnostics
T1  - Molecular Biomarkers in Perthes Disease: A Review
IS  - 3
SP  - 471
VL  - 13
DO  - 10.3390/diagnostics13030471
ER  - 

@article{
author = "Spasovski, Vesna and Srzentić Dražilov, Sanja and Nikčević, Gordana and Baščarević, Zoran and Stojiljković, Maja and Pavlović, Sonja and Spasovski, Duško",
year = "2023",
abstract = "Background: Perthes disease is a juvenile form of osteonecrosis of the femoral head that affects children under the age of 15. One hundred years after its discovery, some light has been shed on its etiology and the biological factors relevant to its etiology and disease severity. Methods: The aim of this study was to summarize the literature findings on the biological factors relevant to the pathogenesis of Perthes disease, their diagnostic and clinical significance, and their therapeutic potential. A special focus on candidate genes as susceptibility factors and factors relevant to clinical severity was made, where studies reporting clinical or preclinical results were considered as the inclusion criteria. PubMed databases were searched by two independent researchers. Sixty-eight articles were included in this review. Results on the factors relevant to vascular involvement and inflammatory molecules indicated as factors that contribute to impaired bone remodeling have been summarized. Moreover, several candidate genes relevant to an active phase of the disease have been suggested as possible biological therapeutic targets. Conclusions: Delineation of molecular biomarkers that underlie the pathophysiological process of Perthes disease can allow for the provision of earlier and more accurate diagnoses of the disease and more precise follow-ups and treatment in the early phases of the disease.",
publisher = "MDPI",
journal = "Diagnostics",
title = "Molecular Biomarkers in Perthes Disease: A Review",
number = "3",
pages = "471",
volume = "13",
doi = "10.3390/diagnostics13030471"
}

Spasovski, V., Srzentić Dražilov, S., Nikčević, G., Baščarević, Z., Stojiljković, M., Pavlović, S.,& Spasovski, D.. (2023). Molecular Biomarkers in Perthes Disease: A Review. in Diagnostics
MDPI., 13(3), 471.
https://doi.org/10.3390/diagnostics13030471

Spasovski V, Srzentić Dražilov S, Nikčević G, Baščarević Z, Stojiljković M, Pavlović S, Spasovski D. Molecular Biomarkers in Perthes Disease: A Review. in Diagnostics. 2023;13(3):471.
doi:10.3390/diagnostics13030471 .

Spasovski, Vesna, Srzentić Dražilov, Sanja, Nikčević, Gordana, Baščarević, Zoran, Stojiljković, Maja, Pavlović, Sonja, Spasovski, Duško, "Molecular Biomarkers in Perthes Disease: A Review" in Diagnostics, 13, no. 3 (2023):471,
https://doi.org/10.3390/diagnostics13030471 . .

TY  - CONF
AU  - Skakić, Anita
AU  - Stevanović, Nina
AU  - Spasovski, Vesna
AU  - Parezanović, Marina
AU  - Ugrin, Milena
AU  - Komazec, Jovana
AU  - Klaassen, Kristel
AU  - Stanković, Sara
AU  - Pavlović, Sonja
AU  - Stojiljković, Maja
AU  - Anđelković, Marina
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1903
AB  - Primary ciliary dyskinesia (PCD) is a rare motor ciliopathy, which predominantly affects the lungs and
reproductive organs. PCD has a heterogeneous genetic basis, and it is necessary to analyze more than
40 causative genes in order to establish a precise diagnosis, which is essential for optimal treatment and
adequate genetic counseling. Five patients suspected of PCD were analyzed using next-generation
sequencing (NGS). The pathogenicity of the genetic variants was tested by in silico, qRT-PCR and
Western blot methods. Two newly discovered variants p.N450Lfs*6 and p.D562N in the DNAI1 gene
were detected in one patient suspected of PCD. The results of in silico prediction showed that the
p.N450Lfs*6 variant affects the structure of the 3D model of the protein, abolishes ligand binding sites
and post-translational modifications, thereby disrupting protein-protein interactions (PPI). The
p.D562N variant has no effect on the 3D structure of the protein, but affects the ligand binding site and
is located in the WD-40 domain, which most likely disrupts PPI. The results of the qRT-PCR method
showed a decreased expression level of DNAI1 mRNA by about 50% in the patient compared to the
control group, while Western blot analysis showed the presence of two protein products (699 ak and
455 ak). By analyzing the obtained results, it was concluded that the changes p.N450Lfs*6 and
p.D562N affect the length and quantity of the DNAI1 protein, leading to the loss of protein function
and are responsible for the occurrence of primary ciliary dyskinesia in the analyzed patient.
PB  - Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo
C3  - Genetics & Applications
T1  - Functional characterization of novel variants in the dnai1 gene in a patient with primary ciliary dyskinesia
IS  - 2 (Special edition)
SP  - 110
VL  - 7
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1903
ER  - 

@conference{
author = "Skakić, Anita and Stevanović, Nina and Spasovski, Vesna and Parezanović, Marina and Ugrin, Milena and Komazec, Jovana and Klaassen, Kristel and Stanković, Sara and Pavlović, Sonja and Stojiljković, Maja and Anđelković, Marina",
year = "2023",
abstract = "Primary ciliary dyskinesia (PCD) is a rare motor ciliopathy, which predominantly affects the lungs and
reproductive organs. PCD has a heterogeneous genetic basis, and it is necessary to analyze more than
40 causative genes in order to establish a precise diagnosis, which is essential for optimal treatment and
adequate genetic counseling. Five patients suspected of PCD were analyzed using next-generation
sequencing (NGS). The pathogenicity of the genetic variants was tested by in silico, qRT-PCR and
Western blot methods. Two newly discovered variants p.N450Lfs*6 and p.D562N in the DNAI1 gene
were detected in one patient suspected of PCD. The results of in silico prediction showed that the
p.N450Lfs*6 variant affects the structure of the 3D model of the protein, abolishes ligand binding sites
and post-translational modifications, thereby disrupting protein-protein interactions (PPI). The
p.D562N variant has no effect on the 3D structure of the protein, but affects the ligand binding site and
is located in the WD-40 domain, which most likely disrupts PPI. The results of the qRT-PCR method
showed a decreased expression level of DNAI1 mRNA by about 50% in the patient compared to the
control group, while Western blot analysis showed the presence of two protein products (699 ak and
455 ak). By analyzing the obtained results, it was concluded that the changes p.N450Lfs*6 and
p.D562N affect the length and quantity of the DNAI1 protein, leading to the loss of protein function
and are responsible for the occurrence of primary ciliary dyskinesia in the analyzed patient.",
publisher = "Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo",
journal = "Genetics & Applications",
title = "Functional characterization of novel variants in the dnai1 gene in a patient with primary ciliary dyskinesia",
number = "2 (Special edition)",
pages = "110",
volume = "7",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1903"
}

Skakić, A., Stevanović, N., Spasovski, V., Parezanović, M., Ugrin, M., Komazec, J., Klaassen, K., Stanković, S., Pavlović, S., Stojiljković, M.,& Anđelković, M.. (2023). Functional characterization of novel variants in the dnai1 gene in a patient with primary ciliary dyskinesia. in Genetics & Applications
Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo., 7(2 (Special edition)), 110.
https://hdl.handle.net/21.15107/rcub_imagine_1903

Skakić A, Stevanović N, Spasovski V, Parezanović M, Ugrin M, Komazec J, Klaassen K, Stanković S, Pavlović S, Stojiljković M, Anđelković M. Functional characterization of novel variants in the dnai1 gene in a patient with primary ciliary dyskinesia. in Genetics & Applications. 2023;7(2 (Special edition)):110.
https://hdl.handle.net/21.15107/rcub_imagine_1903 .

Skakić, Anita, Stevanović, Nina, Spasovski, Vesna, Parezanović, Marina, Ugrin, Milena, Komazec, Jovana, Klaassen, Kristel, Stanković, Sara, Pavlović, Sonja, Stojiljković, Maja, Anđelković, Marina, "Functional characterization of novel variants in the dnai1 gene in a patient with primary ciliary dyskinesia" in Genetics & Applications, 7, no. 2 (Special edition) (2023):110,
https://hdl.handle.net/21.15107/rcub_imagine_1903 .

TY  - CONF
AU  - Parezanović, Marina
AU  - Anđelković, Marina
AU  - Stevanović, Nina
AU  - Klaassen, Kristel
AU  - Spasovski, Vesna
AU  - Ugrin, Milena
AU  - Komazec, Jovana
AU  - Stanković, Sara
AU  - Jocić, Nikola
AU  - Pavlović, Sonja
AU  - Stojiljković, Maja
AU  - Skakić, Anita
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2121
AB  - Introduction: Glycogen storage disease type Ib (GSD-Ib) is an autosomal recessive disorder characterized by fasting hypoglycemia and the accumulation of glycogen in the liver, kidneys and intestinal mucosa. Recent studies revealed that chronic endoplasmic reticulum (ER) stress and increased apoptosis
play a role in the progression of disease manifestations. Although dietary control is commonly utilized
to manage hypoglycemia, there is still a lack of effective pharmacological therapy. Therefore, the establishment of proper model system is essential for testing novel treatment approaches.
Methods: To create GSD-Ib in vitro model system, CRISPR/Cas9-knockout (KO) method was used to introduce a deletion in SLC37A4 gene in the FlpInHEK293 cells. Characterization of CRISPR/Cas9-KO model
system was performed using Sanger sequencing, RT-qPCR and Western Blot. Additionally, the expression analysis of ER stress and apoptotic markers was performed.
Results: Sanger sequencing confirmed the presence of c.14_100del in SLC37A4 gene. The expression
level of SLC37A4 was decreased to 26.8% in the SLC37A4-/- cell line compared to the SLC37A4 wild-type
along with Western blot analysis, which confirmed reduced target protein level in SLC37A4-/- clones. Furthermore, ER stress (ATF4, DDIT3, HSPA5, XBP1s) and apoptotic (BCL2, BAX, CASP3, CASP7) markers expression levels were significantly altered in SLC37A4-/- clones compared to wild-type, which proved that
we created a suitable GSD-Ib in vitro model system.
Conclusion: Utilizing CRISPR/Cas9 technology, we established cellular GSD-Ib modelsystem that mirrors
increased ER stress and apoptosis. This model system could be used to facilitate a comprehensive understanding of disease mechanisms and enable testing of potential treatment effectiveness.
PB  - 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  - Application of CRISPR/cas9 technology for in vitro disease modelling in glycogen storage disease type IB
EP  - 65
SP  - 65
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2121
ER  - 

@conference{
author = "Parezanović, Marina and Anđelković, Marina and Stevanović, Nina and Klaassen, Kristel and Spasovski, Vesna and Ugrin, Milena and Komazec, Jovana and Stanković, Sara and Jocić, Nikola and Pavlović, Sonja and Stojiljković, Maja and Skakić, Anita",
year = "2023",
abstract = "Introduction: Glycogen storage disease type Ib (GSD-Ib) is an autosomal recessive disorder characterized by fasting hypoglycemia and the accumulation of glycogen in the liver, kidneys and intestinal mucosa. Recent studies revealed that chronic endoplasmic reticulum (ER) stress and increased apoptosis
play a role in the progression of disease manifestations. Although dietary control is commonly utilized
to manage hypoglycemia, there is still a lack of effective pharmacological therapy. Therefore, the establishment of proper model system is essential for testing novel treatment approaches.
Methods: To create GSD-Ib in vitro model system, CRISPR/Cas9-knockout (KO) method was used to introduce a deletion in SLC37A4 gene in the FlpInHEK293 cells. Characterization of CRISPR/Cas9-KO model
system was performed using Sanger sequencing, RT-qPCR and Western Blot. Additionally, the expression analysis of ER stress and apoptotic markers was performed.
Results: Sanger sequencing confirmed the presence of c.14_100del in SLC37A4 gene. The expression
level of SLC37A4 was decreased to 26.8% in the SLC37A4-/- cell line compared to the SLC37A4 wild-type
along with Western blot analysis, which confirmed reduced target protein level in SLC37A4-/- clones. Furthermore, ER stress (ATF4, DDIT3, HSPA5, XBP1s) and apoptotic (BCL2, BAX, CASP3, CASP7) markers expression levels were significantly altered in SLC37A4-/- clones compared to wild-type, which proved that
we created a suitable GSD-Ib in vitro model system.
Conclusion: Utilizing CRISPR/Cas9 technology, we established cellular GSD-Ib modelsystem that mirrors
increased ER stress and apoptosis. This model system could be used to facilitate a comprehensive understanding of disease mechanisms and enable testing of potential treatment effectiveness.",
publisher = "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 = "Application of CRISPR/cas9 technology for in vitro disease modelling in glycogen storage disease type IB",
pages = "65-65",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2121"
}

Parezanović, M., Anđelković, M., Stevanović, N., Klaassen, K., Spasovski, V., Ugrin, M., Komazec, J., Stanković, S., Jocić, N., Pavlović, S., Stojiljković, M.,& Skakić, A.. (2023). Application of CRISPR/cas9 technology for in vitro disease modelling in glycogen storage disease type IB. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 65-65.
https://hdl.handle.net/21.15107/rcub_imagine_2121

Parezanović M, Anđelković M, Stevanović N, Klaassen K, Spasovski V, Ugrin M, Komazec J, Stanković S, Jocić N, Pavlović S, Stojiljković M, Skakić A. Application of CRISPR/cas9 technology for in vitro disease modelling in glycogen storage disease type IB. 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;:65-65.
https://hdl.handle.net/21.15107/rcub_imagine_2121 .

Parezanović, Marina, Anđelković, Marina, Stevanović, Nina, Klaassen, Kristel, Spasovski, Vesna, Ugrin, Milena, Komazec, Jovana, Stanković, Sara, Jocić, Nikola, Pavlović, Sonja, Stojiljković, Maja, Skakić, Anita, "Application of CRISPR/cas9 technology for in vitro disease modelling in glycogen storage disease type IB" 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):65-65,
https://hdl.handle.net/21.15107/rcub_imagine_2121 .

TY  - JOUR
AU  - Troha, Kaja
AU  - Vozel, Domen
AU  - Arko, Matevž
AU  - Bedina Zavec, Apolonija
AU  - Dolinar, Drago
AU  - Hočevar, Matej
AU  - Jan, Zala
AU  - Kisovec, Matic
AU  - Kocjančič, Boštjan
AU  - Pađen, Ljubiša
AU  - Pajnič, Manca
AU  - Penič, Samo
AU  - Romolo, Anna
AU  - Repar, Neža
AU  - Spasovski, Vesna
AU  - Steiner, Nejc
AU  - Šuštar, Vid
AU  - Iglič, Aleš
AU  - Drobne, Damjana
AU  - Kogej, Ksenija
AU  - Battelino, Saba
AU  - Kralj-Iglič, Veronika
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2087
AB  - The preparation of autologous platelet and extracellular vesicle-rich plasma (PVRP) has been explored in many medical fields with the aim to benefit from its healing potential. In parallel, efforts are being invested to understand the function and dynamics of PVRP that is complex in its composition and interactions. Some clinical evidence reveals beneficial effects of PVRP, while some report that there were no effects. To optimize the preparation methods, functions and mechanisms of PVRP, its constituents should be better understood. With the intention to promote further studies of autologous therapeutic PVRP, we performed a review on some topics regarding PVRP composition, harvesting, assessment and preservation, and also on clinical experience following PVRP application in humans and animals. Besides the acknowledged actions of platelets, leukocytes and different molecules, we focus on extracellular vesicles that were found abundant in PVRP.
PB  - MDPI
T2  - International Journal of Molecular Sciences
T1  - Autologous Platelet and Extracellular Vesicle-Rich Plasma as Therapeutic Fluid: A Review
IS  - 4
SP  - 3420
VL  - 24
DO  - 10.3390/ijms24043420
ER  - 

@article{
author = "Troha, Kaja and Vozel, Domen and Arko, Matevž and Bedina Zavec, Apolonija and Dolinar, Drago and Hočevar, Matej and Jan, Zala and Kisovec, Matic and Kocjančič, Boštjan and Pađen, Ljubiša and Pajnič, Manca and Penič, Samo and Romolo, Anna and Repar, Neža and Spasovski, Vesna and Steiner, Nejc and Šuštar, Vid and Iglič, Aleš and Drobne, Damjana and Kogej, Ksenija and Battelino, Saba and Kralj-Iglič, Veronika",
year = "2023",
abstract = "The preparation of autologous platelet and extracellular vesicle-rich plasma (PVRP) has been explored in many medical fields with the aim to benefit from its healing potential. In parallel, efforts are being invested to understand the function and dynamics of PVRP that is complex in its composition and interactions. Some clinical evidence reveals beneficial effects of PVRP, while some report that there were no effects. To optimize the preparation methods, functions and mechanisms of PVRP, its constituents should be better understood. With the intention to promote further studies of autologous therapeutic PVRP, we performed a review on some topics regarding PVRP composition, harvesting, assessment and preservation, and also on clinical experience following PVRP application in humans and animals. Besides the acknowledged actions of platelets, leukocytes and different molecules, we focus on extracellular vesicles that were found abundant in PVRP.",
publisher = "MDPI",
journal = "International Journal of Molecular Sciences",
title = "Autologous Platelet and Extracellular Vesicle-Rich Plasma as Therapeutic Fluid: A Review",
number = "4",
pages = "3420",
volume = "24",
doi = "10.3390/ijms24043420"
}

Troha, K., Vozel, D., Arko, M., Bedina Zavec, A., Dolinar, D., Hočevar, M., Jan, Z., Kisovec, M., Kocjančič, B., Pađen, L., Pajnič, M., Penič, S., Romolo, A., Repar, N., Spasovski, V., Steiner, N., Šuštar, V., Iglič, A., Drobne, D., Kogej, K., Battelino, S.,& Kralj-Iglič, V.. (2023). Autologous Platelet and Extracellular Vesicle-Rich Plasma as Therapeutic Fluid: A Review. in International Journal of Molecular Sciences
MDPI., 24(4), 3420.
https://doi.org/10.3390/ijms24043420

Troha K, Vozel D, Arko M, Bedina Zavec A, Dolinar D, Hočevar M, Jan Z, Kisovec M, Kocjančič B, Pađen L, Pajnič M, Penič S, Romolo A, Repar N, Spasovski V, Steiner N, Šuštar V, Iglič A, Drobne D, Kogej K, Battelino S, Kralj-Iglič V. Autologous Platelet and Extracellular Vesicle-Rich Plasma as Therapeutic Fluid: A Review. in International Journal of Molecular Sciences. 2023;24(4):3420.
doi:10.3390/ijms24043420 .

Troha, Kaja, Vozel, Domen, Arko, Matevž, Bedina Zavec, Apolonija, Dolinar, Drago, Hočevar, Matej, Jan, Zala, Kisovec, Matic, Kocjančič, Boštjan, Pađen, Ljubiša, Pajnič, Manca, Penič, Samo, Romolo, Anna, Repar, Neža, Spasovski, Vesna, Steiner, Nejc, Šuštar, Vid, Iglič, Aleš, Drobne, Damjana, Kogej, Ksenija, Battelino, Saba, Kralj-Iglič, Veronika, "Autologous Platelet and Extracellular Vesicle-Rich Plasma as Therapeutic Fluid: A Review" in International Journal of Molecular Sciences, 24, no. 4 (2023):3420,
https://doi.org/10.3390/ijms24043420 . .

TY  - CONF
AU  - Stojiljković, Maja
AU  - Klaassen, Kristel
AU  - Skakić, Anita
AU  - Anđelković, Marina
AU  - Komazec, Jovana
AU  - Ugrin, Milena
AU  - Spasovski, Vesna
AU  - Pavlović, Sonja
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2111
AB  - Introduction: All inborn metabolic diseases are rare, having a prevalence less than 1:2000. Vast majority of them are monogenic and finding pathogenic genetic variantsis needed to setthe correct diagnosis, enable adequate treatment and provide genetic counseling to members of affected family. Thisstudy is an overview of genomic studies of rare metabolic diseases in Serbia. Methods: Since 2005, more than 300 patients suspected to have a rare metabolic or neurometabolic disease have been analyzed using sanger sequencing, clinical-exome sequencing, whole-exome sequencing or whole-genome sequencing in order to find disease-causing or disease-modifying variants. Novel variants were characterized using in silico modelling or in in vitro eukaryotic assays (standard or CRISPR/Cas9 developed). Results: Disease-causing variants were found in more than 60 different genes associated with a metabolic or neurometabolic disease. The most frequent disease was phenylketonuria (109 patients), followed by glycogen storage disease Ib (30 patients), while majority of diseases is seen only in a single patient. More than 40 new genetic variants were comprehensively characterized in silico or in vitro. For the first time, candidate modifiers (SHANK gene family) were identified in a group of phenylketonuria patients with an unusual phenotype. Conclusion: In the genomics era, next-generation sequencing significantly shortens time to diagnosis and allowsstudying genetic modifiers of monogenic diseases and genotype-phenotype correlation. Furthermore, characterization of novel genetic targets boosts development of precision medicine
PB  - 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  - Rare metabolic diseases in the genomics era
EP  - 36
SP  - 36
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2111
ER  - 

@conference{
author = "Stojiljković, Maja and Klaassen, Kristel and Skakić, Anita and Anđelković, Marina and Komazec, Jovana and Ugrin, Milena and Spasovski, Vesna and Pavlović, Sonja",
year = "2023",
abstract = "Introduction: All inborn metabolic diseases are rare, having a prevalence less than 1:2000. Vast majority of them are monogenic and finding pathogenic genetic variantsis needed to setthe correct diagnosis, enable adequate treatment and provide genetic counseling to members of affected family. Thisstudy is an overview of genomic studies of rare metabolic diseases in Serbia. Methods: Since 2005, more than 300 patients suspected to have a rare metabolic or neurometabolic disease have been analyzed using sanger sequencing, clinical-exome sequencing, whole-exome sequencing or whole-genome sequencing in order to find disease-causing or disease-modifying variants. Novel variants were characterized using in silico modelling or in in vitro eukaryotic assays (standard or CRISPR/Cas9 developed). Results: Disease-causing variants were found in more than 60 different genes associated with a metabolic or neurometabolic disease. The most frequent disease was phenylketonuria (109 patients), followed by glycogen storage disease Ib (30 patients), while majority of diseases is seen only in a single patient. More than 40 new genetic variants were comprehensively characterized in silico or in vitro. For the first time, candidate modifiers (SHANK gene family) were identified in a group of phenylketonuria patients with an unusual phenotype. Conclusion: In the genomics era, next-generation sequencing significantly shortens time to diagnosis and allowsstudying genetic modifiers of monogenic diseases and genotype-phenotype correlation. Furthermore, characterization of novel genetic targets boosts development of precision medicine",
publisher = "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 = "Rare metabolic diseases in the genomics era",
pages = "36-36",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2111"
}

Stojiljković, M., Klaassen, K., Skakić, A., Anđelković, M., Komazec, J., Ugrin, M., Spasovski, V.,& Pavlović, S.. (2023). Rare metabolic diseases in the genomics era. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 36-36.
https://hdl.handle.net/21.15107/rcub_imagine_2111

Stojiljković M, Klaassen K, Skakić A, Anđelković M, Komazec J, Ugrin M, Spasovski V, Pavlović S. Rare metabolic diseases in the genomics era. 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;:36-36.
https://hdl.handle.net/21.15107/rcub_imagine_2111 .

Stojiljković, Maja, Klaassen, Kristel, Skakić, Anita, Anđelković, Marina, Komazec, Jovana, Ugrin, Milena, Spasovski, Vesna, Pavlović, Sonja, "Rare metabolic diseases in the genomics era" 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):36-36,
https://hdl.handle.net/21.15107/rcub_imagine_2111 .

TY  - CONF
AU  - Parezanović, Marina
AU  - Anđelković, Marina
AU  - Stevanović, Nina
AU  - Spasovski, Vesna
AU  - Ugrin, Milena
AU  - Komazec, Jovana
AU  - Klaassen, Kristel
AU  - Stanković, Sara
AU  - Pavlović, Sonja
AU  - Stojiljković, Maja
AU  - Skakić, Anita
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1901
AB  - Fabry disease (FD) is a rare X-linked disorder caused by variants in the GLA gene leading to the
deficiency of lysosomal α-galactosidase-A and progressive accumulation of globotriaosylceramide
affecting the heart, nervous system, and kidneys. FD has overlapping phenotypes and often remains
undiagnosed. Therefore, a precise molecular-genetic diagnosis and the earliest possible treatment are
essential to avoid significant disease progression. The study aimed to determine the strategy for
establishing routine molecular genetic diagnostics of FD in Serbia to provide an early application of
appropriate therapy and genetic advice to families with a high risk for the birth of a child with FD. We
analyzed 95 (34 female and 61 male) hemodialysis patients with clinical suspicion of FD using Sanger
sequencing of all coding exons (7) and flanking intron regions of the GLA gene and measured the
relative expression of the GLA gene in available samples. The genetic analysis revealed 3 patients with
a missense variant (p.Asp313Tyr), and 10 patients with combinations of non-coding variants, described
as complex intronic haplotypes (CIHs). CIH1 (c.-10C>T, c.370-81_370-77delCAGCC, c.640-16A>G,
c.1000-22C>T), the most frequent haplotype, was detected in 7 (7.4%) patients. Lyso-Gb3 biomarker
levels were within the normal range in each tested patient. However, RT-qPCR analysis revealed
decreased relative expression of the GLA gene in PBMC of 2 female patients with CIH1 and one
female patient carrying only c.-10C>T variant by 9,1%, 7,4%, 46,3%, respectively, pointing out that
further analyses are needed to confirm/exclude FD in these patients. Because the effects of CIHs are not
yet fully understood, our work highlights the importance of analyzing intronic regions of the GLA gene
as genetic modifiers and the need to include expression analysis in the diagnostic algorithm.
PB  - Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo
C3  - Genetics & Applications
T1  - High-risk population screening for fabry disease in patients with chronic renal failure of unknown etiology
IS  - 2 (Special edition)
IS  - 107
VL  - 7
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1901
ER  - 

@conference{
author = "Parezanović, Marina and Anđelković, Marina and Stevanović, Nina and Spasovski, Vesna and Ugrin, Milena and Komazec, Jovana and Klaassen, Kristel and Stanković, Sara and Pavlović, Sonja and Stojiljković, Maja and Skakić, Anita",
year = "2023",
abstract = "Fabry disease (FD) is a rare X-linked disorder caused by variants in the GLA gene leading to the
deficiency of lysosomal α-galactosidase-A and progressive accumulation of globotriaosylceramide
affecting the heart, nervous system, and kidneys. FD has overlapping phenotypes and often remains
undiagnosed. Therefore, a precise molecular-genetic diagnosis and the earliest possible treatment are
essential to avoid significant disease progression. The study aimed to determine the strategy for
establishing routine molecular genetic diagnostics of FD in Serbia to provide an early application of
appropriate therapy and genetic advice to families with a high risk for the birth of a child with FD. We
analyzed 95 (34 female and 61 male) hemodialysis patients with clinical suspicion of FD using Sanger
sequencing of all coding exons (7) and flanking intron regions of the GLA gene and measured the
relative expression of the GLA gene in available samples. The genetic analysis revealed 3 patients with
a missense variant (p.Asp313Tyr), and 10 patients with combinations of non-coding variants, described
as complex intronic haplotypes (CIHs). CIH1 (c.-10C>T, c.370-81_370-77delCAGCC, c.640-16A>G,
c.1000-22C>T), the most frequent haplotype, was detected in 7 (7.4%) patients. Lyso-Gb3 biomarker
levels were within the normal range in each tested patient. However, RT-qPCR analysis revealed
decreased relative expression of the GLA gene in PBMC of 2 female patients with CIH1 and one
female patient carrying only c.-10C>T variant by 9,1%, 7,4%, 46,3%, respectively, pointing out that
further analyses are needed to confirm/exclude FD in these patients. Because the effects of CIHs are not
yet fully understood, our work highlights the importance of analyzing intronic regions of the GLA gene
as genetic modifiers and the need to include expression analysis in the diagnostic algorithm.",
publisher = "Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo",
journal = "Genetics & Applications",
title = "High-risk population screening for fabry disease in patients with chronic renal failure of unknown etiology",
number = "2 (Special edition), 107",
volume = "7",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1901"
}

Parezanović, M., Anđelković, M., Stevanović, N., Spasovski, V., Ugrin, M., Komazec, J., Klaassen, K., Stanković, S., Pavlović, S., Stojiljković, M.,& Skakić, A.. (2023). High-risk population screening for fabry disease in patients with chronic renal failure of unknown etiology. in Genetics & Applications
Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo., 7(2 (Special edition)).
https://hdl.handle.net/21.15107/rcub_imagine_1901

Parezanović M, Anđelković M, Stevanović N, Spasovski V, Ugrin M, Komazec J, Klaassen K, Stanković S, Pavlović S, Stojiljković M, Skakić A. High-risk population screening for fabry disease in patients with chronic renal failure of unknown etiology. in Genetics & Applications. 2023;7(2 (Special edition)).
https://hdl.handle.net/21.15107/rcub_imagine_1901 .

Parezanović, Marina, Anđelković, Marina, Stevanović, Nina, Spasovski, Vesna, Ugrin, Milena, Komazec, Jovana, Klaassen, Kristel, Stanković, Sara, Pavlović, Sonja, Stojiljković, Maja, Skakić, Anita, "High-risk population screening for fabry disease in patients with chronic renal failure of unknown etiology" in Genetics & Applications, 7, no. 2 (Special edition) (2023),
https://hdl.handle.net/21.15107/rcub_imagine_1901 .

TY  - CONF
AU  - Klaassen, Kristel
AU  - Šinžar, Ksenija
AU  - Stanković, Sara
AU  - Đorđević Milošević, Maja
AU  - Kecman, Božica
AU  - Anđelković, Marina
AU  - Skakić, Anita
AU  - Spasovski, Vesna
AU  - Ugrin, Milena
AU  - Komazec, Jovana
AU  - Parezanović, Marina
AU  - Stevanović, Nina
AU  - Pavlović, Sonja
AU  - Stojiljković, Maja
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2142
AB  - Introduction: Phenylketonuria (PKU) is the most frequent inborn disorder of amino acid metabolism
caused by variants in human phenylalanine hydroxylase gene (PAH).
Methods: In thisstudy (an update for the time period of 10 years, with patientsfrom our previousstudies included) a total of 109 PKU patients from Serbia were analyzed. They were classified into three phenotypic categories in accordance with pre-treatment plasma phenylalanine level: classic PKU, mild PKU
and mild hyperphenylalaninemia. For genetic analyses, we combined Sanger sequencing, MLPA and
next generation sequencing to identify disease-causing variantsin PAH gene, which were further classified using ACMG classification. Additionally, we used in silico and/or eukaryotic expression studiesto assess the effect of novel genetic variants identified in our patients.
Results: Disease-causing variants were identified in 217 of 218 alleles, reaching detection rate of 99.5%.
We detected a total of 32 different variants, of which 29 previously described and three novel ones:
p.Gln226Lys, p.Pro244His and p.Pro416Leu. In silico and/or eukaryotic expression studies confirmed pathogenic effect of all novel genetic variants. The most frequent variant was p.Leu48Ser (31.2%), followed
by p.Arg408Trp (13.8%), p.Ile306Val (9.2%). p.Glu390Gly (5%), p.Pro281Leu (4.6%), and p.Arg261Gln
(3.2%). All detected disease-causing variants were classified as pathogenic using ACMG classification.
Conclusion: Our study brings the updated spectrum of molecular genetic data, variant classification
and detailed phenotypic characteristicsfor PKU patientsfrom Serbia. Therefore, ourstudy contributesto
better understanding of molecular landscape of PKU in Europe and to general knowledge on genotype–
phenotype correlation in PKU.
PB  - 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  - Molecular basis of phenylketonuria in Serbia: an update
EP  - 93
SP  - 93
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2142
ER  - 

@conference{
author = "Klaassen, Kristel and Šinžar, Ksenija and Stanković, Sara and Đorđević Milošević, Maja and Kecman, Božica and Anđelković, Marina and Skakić, Anita and Spasovski, Vesna and Ugrin, Milena and Komazec, Jovana and Parezanović, Marina and Stevanović, Nina and Pavlović, Sonja and Stojiljković, Maja",
year = "2023",
abstract = "Introduction: Phenylketonuria (PKU) is the most frequent inborn disorder of amino acid metabolism
caused by variants in human phenylalanine hydroxylase gene (PAH).
Methods: In thisstudy (an update for the time period of 10 years, with patientsfrom our previousstudies included) a total of 109 PKU patients from Serbia were analyzed. They were classified into three phenotypic categories in accordance with pre-treatment plasma phenylalanine level: classic PKU, mild PKU
and mild hyperphenylalaninemia. For genetic analyses, we combined Sanger sequencing, MLPA and
next generation sequencing to identify disease-causing variantsin PAH gene, which were further classified using ACMG classification. Additionally, we used in silico and/or eukaryotic expression studiesto assess the effect of novel genetic variants identified in our patients.
Results: Disease-causing variants were identified in 217 of 218 alleles, reaching detection rate of 99.5%.
We detected a total of 32 different variants, of which 29 previously described and three novel ones:
p.Gln226Lys, p.Pro244His and p.Pro416Leu. In silico and/or eukaryotic expression studies confirmed pathogenic effect of all novel genetic variants. The most frequent variant was p.Leu48Ser (31.2%), followed
by p.Arg408Trp (13.8%), p.Ile306Val (9.2%). p.Glu390Gly (5%), p.Pro281Leu (4.6%), and p.Arg261Gln
(3.2%). All detected disease-causing variants were classified as pathogenic using ACMG classification.
Conclusion: Our study brings the updated spectrum of molecular genetic data, variant classification
and detailed phenotypic characteristicsfor PKU patientsfrom Serbia. Therefore, ourstudy contributesto
better understanding of molecular landscape of PKU in Europe and to general knowledge on genotype–
phenotype correlation in PKU.",
publisher = "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 = "Molecular basis of phenylketonuria in Serbia: an update",
pages = "93-93",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2142"
}

Klaassen, K., Šinžar, K., Stanković, S., Đorđević Milošević, M., Kecman, B., Anđelković, M., Skakić, A., Spasovski, V., Ugrin, M., Komazec, J., Parezanović, M., Stevanović, N., Pavlović, S.,& Stojiljković, M.. (2023). Molecular basis of phenylketonuria in Serbia: an update. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 93-93.
https://hdl.handle.net/21.15107/rcub_imagine_2142

Klaassen K, Šinžar K, Stanković S, Đorđević Milošević M, Kecman B, Anđelković M, Skakić A, Spasovski V, Ugrin M, Komazec J, Parezanović M, Stevanović N, Pavlović S, Stojiljković M. Molecular basis of phenylketonuria in Serbia: an update. 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;:93-93.
https://hdl.handle.net/21.15107/rcub_imagine_2142 .

Klaassen, Kristel, Šinžar, Ksenija, Stanković, Sara, Đorđević Milošević, Maja, Kecman, Božica, Anđelković, Marina, Skakić, Anita, Spasovski, Vesna, Ugrin, Milena, Komazec, Jovana, Parezanović, Marina, Stevanović, Nina, Pavlović, Sonja, Stojiljković, Maja, "Molecular basis of phenylketonuria in Serbia: an update" 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):93-93,
https://hdl.handle.net/21.15107/rcub_imagine_2142 .

TY  - CONF
AU  - Anđelković, Marina
AU  - Skakić, Anita
AU  - Stevanović, Nina
AU  - Parezanović, Marina
AU  - Komazec, Jovana
AU  - Klaassen, Kristel
AU  - Spasovski, Vesna
AU  - Stojiljković, Maja
AU  - Pavlović, Sonja
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1900
AB  - Rare lung diseases (RLDs) are a group of diseases that individually affect one in 2,000 people, with an
estimate that about 80% of RLDs have a genetic origin. Despite the variations among RLDs in clinical
characteristics and manifestations, most of these diseases similarly damage the lungs, making diagnosis
difficult. The utility of NGS technology in RLDs for diagnostic purposes allows a better understanding
of the genetic background, however, the identification and classification of disease-causing variants are
challenging. Further, numerous VUS (variants of uncertain significance) that cannot be precisely
defined and classified are produced. The main goal of this study was to create a unique guideline that
will enable the standardization of the assessment of novel genetic variants in RLDs causative genes.
The designed pipeline consists of three main steps: (1) sequencing, detection, and identification of
genes/variants, (2) classification of variants, and (3) characterization of variants using in silico
structural and functional analysis. The pipeline validation was performed through the analysis of
variants detected in a disease-causing and candidate genes of one of the RLDSs, and detected VUS
variants have gained diagnostic significance. The application of this pipeline resulted in the
identification and classification of novel variants, through analysis at the transcriptional, translational,
and posttranslational levels, and led to accurate diagnosis.
PB  - Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo
C3  - Genetics & Applications
T1  - Improving the diagnostics of rare lung disorders using a uniquely designed pipeline for analysis of ngs data
IS  - 2 (Special edition)
SP  - 104
VL  - 7
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1900
ER  - 

@conference{
author = "Anđelković, Marina and Skakić, Anita and Stevanović, Nina and Parezanović, Marina and Komazec, Jovana and Klaassen, Kristel and Spasovski, Vesna and Stojiljković, Maja and Pavlović, Sonja",
year = "2023",
abstract = "Rare lung diseases (RLDs) are a group of diseases that individually affect one in 2,000 people, with an
estimate that about 80% of RLDs have a genetic origin. Despite the variations among RLDs in clinical
characteristics and manifestations, most of these diseases similarly damage the lungs, making diagnosis
difficult. The utility of NGS technology in RLDs for diagnostic purposes allows a better understanding
of the genetic background, however, the identification and classification of disease-causing variants are
challenging. Further, numerous VUS (variants of uncertain significance) that cannot be precisely
defined and classified are produced. The main goal of this study was to create a unique guideline that
will enable the standardization of the assessment of novel genetic variants in RLDs causative genes.
The designed pipeline consists of three main steps: (1) sequencing, detection, and identification of
genes/variants, (2) classification of variants, and (3) characterization of variants using in silico
structural and functional analysis. The pipeline validation was performed through the analysis of
variants detected in a disease-causing and candidate genes of one of the RLDSs, and detected VUS
variants have gained diagnostic significance. The application of this pipeline resulted in the
identification and classification of novel variants, through analysis at the transcriptional, translational,
and posttranslational levels, and led to accurate diagnosis.",
publisher = "Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo",
journal = "Genetics & Applications",
title = "Improving the diagnostics of rare lung disorders using a uniquely designed pipeline for analysis of ngs data",
number = "2 (Special edition)",
pages = "104",
volume = "7",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1900"
}

Anđelković, M., Skakić, A., Stevanović, N., Parezanović, M., Komazec, J., Klaassen, K., Spasovski, V., Stojiljković, M.,& Pavlović, S.. (2023). Improving the diagnostics of rare lung disorders using a uniquely designed pipeline for analysis of ngs data. in Genetics & Applications
Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo., 7(2 (Special edition)), 104.
https://hdl.handle.net/21.15107/rcub_imagine_1900

Anđelković M, Skakić A, Stevanović N, Parezanović M, Komazec J, Klaassen K, Spasovski V, Stojiljković M, Pavlović S. Improving the diagnostics of rare lung disorders using a uniquely designed pipeline for analysis of ngs data. in Genetics & Applications. 2023;7(2 (Special edition)):104.
https://hdl.handle.net/21.15107/rcub_imagine_1900 .

Anđelković, Marina, Skakić, Anita, Stevanović, Nina, Parezanović, Marina, Komazec, Jovana, Klaassen, Kristel, Spasovski, Vesna, Stojiljković, Maja, Pavlović, Sonja, "Improving the diagnostics of rare lung disorders using a uniquely designed pipeline for analysis of ngs data" in Genetics & Applications, 7, no. 2 (Special edition) (2023):104,
https://hdl.handle.net/21.15107/rcub_imagine_1900 .

TY  - CONF
AU  - Jelovac, Marina
AU  - Kotur, Nikola
AU  - Ristivojević, Bojan
AU  - Pavlović, Đorđe
AU  - Spasovski, Vesna
AU  - Damjanov, Nemanja
AU  - Pavlović, Sonja
AU  - Zukić, Branka
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2119
AB  - Introduction: Systemic sclerosis(SSc) is a rare autoimmune disorder that affects connective tissues and
hasthe highest morbidity and mortality among rheumatologic diseases. Clinical presentations as well as
disease progression are highly heterogeneous between patients, implying a strong need for individualization of therapy.
Methods: Four pharmacogenetic variants, namely TPMT rs1800460, TPMT rs1142345, MTHFR rs1801133
and SLCO1B1 rs4149056 were tested for association with severe disease outcomes in 102 patients with
SSc from Serbia treated either with immunosuppressants azathioprine (AZA) and methotrexate (MTX) or
with other types of medications. Genotyping was performed using PCR-RFLP and direct Sanger sequencing. R software was used for statistical analysis and development of polygenic risk score (PRS)
model.
Results: Association wasfound between MTHFR rs1801133 and higher risk for elevated systolic pressure
in all patients except those prescribed with MTX, and higher risk for kidney insufficiency in patients prescribed with other types of drugs. In patients treated with MTX, variant SLCO1B1 rs4149056 was protective against kidney insufficiency. For patients receiving MTX a trend was shown for having a higher PRS
rank and elevated systolic pressure.
Conclusion: Our results open a door wide for more extensive research on pharmacogenomics markers
in patients with SSc. Altogether, pharmacogenomics markers could predict the outcome of patients with
SSc and help in prevention of adverse drug reactions
PB  - 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  - Can pharmacogenetic variants in TPMT, MTHFR and SLCO1B1 genes be used as potential markers of outcome prediction in systemic sclerosis patients?
EP  - 64
SP  - 64
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2119
ER  - 

@conference{
author = "Jelovac, Marina and Kotur, Nikola and Ristivojević, Bojan and Pavlović, Đorđe and Spasovski, Vesna and Damjanov, Nemanja and Pavlović, Sonja and Zukić, Branka",
year = "2023",
abstract = "Introduction: Systemic sclerosis(SSc) is a rare autoimmune disorder that affects connective tissues and
hasthe highest morbidity and mortality among rheumatologic diseases. Clinical presentations as well as
disease progression are highly heterogeneous between patients, implying a strong need for individualization of therapy.
Methods: Four pharmacogenetic variants, namely TPMT rs1800460, TPMT rs1142345, MTHFR rs1801133
and SLCO1B1 rs4149056 were tested for association with severe disease outcomes in 102 patients with
SSc from Serbia treated either with immunosuppressants azathioprine (AZA) and methotrexate (MTX) or
with other types of medications. Genotyping was performed using PCR-RFLP and direct Sanger sequencing. R software was used for statistical analysis and development of polygenic risk score (PRS)
model.
Results: Association wasfound between MTHFR rs1801133 and higher risk for elevated systolic pressure
in all patients except those prescribed with MTX, and higher risk for kidney insufficiency in patients prescribed with other types of drugs. In patients treated with MTX, variant SLCO1B1 rs4149056 was protective against kidney insufficiency. For patients receiving MTX a trend was shown for having a higher PRS
rank and elevated systolic pressure.
Conclusion: Our results open a door wide for more extensive research on pharmacogenomics markers
in patients with SSc. Altogether, pharmacogenomics markers could predict the outcome of patients with
SSc and help in prevention of adverse drug reactions",
publisher = "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 = "Can pharmacogenetic variants in TPMT, MTHFR and SLCO1B1 genes be used as potential markers of outcome prediction in systemic sclerosis patients?",
pages = "64-64",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2119"
}

Jelovac, M., Kotur, N., Ristivojević, B., Pavlović, Đ., Spasovski, V., Damjanov, N., Pavlović, S.,& Zukić, B.. (2023). Can pharmacogenetic variants in TPMT, MTHFR and SLCO1B1 genes be used as potential markers of outcome prediction in systemic sclerosis patients?. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 64-64.
https://hdl.handle.net/21.15107/rcub_imagine_2119

Jelovac M, Kotur N, Ristivojević B, Pavlović Đ, Spasovski V, Damjanov N, Pavlović S, Zukić B. Can pharmacogenetic variants in TPMT, MTHFR and SLCO1B1 genes be used as potential markers of outcome prediction in systemic sclerosis patients?. 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;:64-64.
https://hdl.handle.net/21.15107/rcub_imagine_2119 .

Jelovac, Marina, Kotur, Nikola, Ristivojević, Bojan, Pavlović, Đorđe, Spasovski, Vesna, Damjanov, Nemanja, Pavlović, Sonja, Zukić, Branka, "Can pharmacogenetic variants in TPMT, MTHFR and SLCO1B1 genes be used as potential markers of outcome prediction in systemic sclerosis patients?" 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):64-64,
https://hdl.handle.net/21.15107/rcub_imagine_2119 .

TY  - JOUR
AU  - Spasovski, Vesna
AU  - Anđelković, Marina
AU  - Parezanović, Marina
AU  - Komazec, Jovana
AU  - Ugrin, Milena
AU  - Klaassen, Kristel
AU  - Stojiljković, Maja
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2085
AB  - Systemic sclerosis (SSc) is a complex autoimmune inflammatory disorder with multiple organ involvement. Skin changes present the hallmark of SSc and coincide with poor prognosis. Interstitial lung diseases (ILD) are the most widely reported complications in SSc patients and the primary cause of death. It has been proposed that the processes of autophagy and apoptosis could play a significant role in the pathogenesis and clinical course of different autoimmune diseases, and accordingly in SSc. In this manuscript, we review the current knowledge of autophagy and apoptosis processes in the skin and lungs of patients with SSc. Profiling of markers involved in these processes in skin cells can be useful to recognize the stage of fibrosis and can be used in the clinical stratification of patients. Furthermore, the knowledge of the molecular mechanisms underlying these processes enables the repurposing of already known drugs and the development of new biological therapeutics that aim to reverse fibrosis by promoting apoptosis and regulate autophagy in personalized treatment approach. In SSc-ILD patients, the molecular signature of the lung tissues of each patient could be a distinctive criterion in order to establish the correct lung pattern, which directly impacts the course and prognosis of the disease. In this case, resolving the role of tissue-specific markers, which could be detected in the circulation using sensitive molecular methods, would be an important step toward development of non-invasive diagnostic procedures that enable early and precise diagnosis and preventing the high mortality of this rare disease.
PB  - MDPI
T2  - International Journal of Molecular Sciences
T1  - The Role of Autophagy and Apoptosis in Affected Skin and Lungs in Patients with Systemic Sclerosis
IS  - 13
SP  - 11212
VL  - 24
DO  - 10.3390/ijms241311212
ER  - 

@article{
author = "Spasovski, Vesna and Anđelković, Marina and Parezanović, Marina and Komazec, Jovana and Ugrin, Milena and Klaassen, Kristel and Stojiljković, Maja",
year = "2023",
abstract = "Systemic sclerosis (SSc) is a complex autoimmune inflammatory disorder with multiple organ involvement. Skin changes present the hallmark of SSc and coincide with poor prognosis. Interstitial lung diseases (ILD) are the most widely reported complications in SSc patients and the primary cause of death. It has been proposed that the processes of autophagy and apoptosis could play a significant role in the pathogenesis and clinical course of different autoimmune diseases, and accordingly in SSc. In this manuscript, we review the current knowledge of autophagy and apoptosis processes in the skin and lungs of patients with SSc. Profiling of markers involved in these processes in skin cells can be useful to recognize the stage of fibrosis and can be used in the clinical stratification of patients. Furthermore, the knowledge of the molecular mechanisms underlying these processes enables the repurposing of already known drugs and the development of new biological therapeutics that aim to reverse fibrosis by promoting apoptosis and regulate autophagy in personalized treatment approach. In SSc-ILD patients, the molecular signature of the lung tissues of each patient could be a distinctive criterion in order to establish the correct lung pattern, which directly impacts the course and prognosis of the disease. In this case, resolving the role of tissue-specific markers, which could be detected in the circulation using sensitive molecular methods, would be an important step toward development of non-invasive diagnostic procedures that enable early and precise diagnosis and preventing the high mortality of this rare disease.",
publisher = "MDPI",
journal = "International Journal of Molecular Sciences",
title = "The Role of Autophagy and Apoptosis in Affected Skin and Lungs in Patients with Systemic Sclerosis",
number = "13",
pages = "11212",
volume = "24",
doi = "10.3390/ijms241311212"
}

Spasovski, V., Anđelković, M., Parezanović, M., Komazec, J., Ugrin, M., Klaassen, K.,& Stojiljković, M.. (2023). The Role of Autophagy and Apoptosis in Affected Skin and Lungs in Patients with Systemic Sclerosis. in International Journal of Molecular Sciences
MDPI., 24(13), 11212.
https://doi.org/10.3390/ijms241311212

Spasovski V, Anđelković M, Parezanović M, Komazec J, Ugrin M, Klaassen K, Stojiljković M. The Role of Autophagy and Apoptosis in Affected Skin and Lungs in Patients with Systemic Sclerosis. in International Journal of Molecular Sciences. 2023;24(13):11212.
doi:10.3390/ijms241311212 .

Spasovski, Vesna, Anđelković, Marina, Parezanović, Marina, Komazec, Jovana, Ugrin, Milena, Klaassen, Kristel, Stojiljković, Maja, "The Role of Autophagy and Apoptosis in Affected Skin and Lungs in Patients with Systemic Sclerosis" in International Journal of Molecular Sciences, 24, no. 13 (2023):11212,
https://doi.org/10.3390/ijms241311212 . .

TY  - CONF
AU  - Stevanović, Nina
AU  - Skakić, Anita
AU  - Parezanović, Marina
AU  - Spasovski, Vesna
AU  - Ugrin, Milena
AU  - Komazec, Jovana
AU  - Klaassen, Kristel
AU  - Stanković, Sara
AU  - Pavlović, Sonja
AU  - Stojiljković, Maja
AU  - Anđelković, Marina
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2140
AB  - Introduction: Primary ciliary dyskinesia is a rare and heterogeneous disorder primarily affecting the respiratory organs, with impaired mucociliary clearance being a common characteristic. Recently, the importance of the miR34/449 family in ciliogenesisin animal models has been described. Thisstudy aimed
to establish a modelsystem to study respiratory diseases and assessfor the first time the role of the miR34 family on the mucociliary process in humans.
Methods: We cultured the primary Normal Human Bronchial Epithelial (NHBE) cells in the air-liquid interface system, enabling the differentiation of multiciliated cells(MCCs) and goblet cells(GCs). During the
differentiation process, transient overexpression of miR-34a/b/c members was conducted. The model
system and treatments were validated through confocal microscopy (β-tubulin, MUC5B, MUC5AC antibodies) and qRT-PCR of miRNAs,specifically ciliogenesis markers(NOTCH1, MCIDAS, GEMC1, CCNO, RFX3),
and differentiated cell markers (FOXJ1 and TFF3).
Results: Expression levels of ciliogenesis and differentiated cells markers and detection of cilia and
mucins at confocal microscopy confirmed the successful establishment of cellular modelsystem. During
the initial differentiation stage, an overexpression of miR34a/b/c changed the expression profile of ciliogenesis and differentiated cell markers.
Conclusion: The established model system provides a valuable platform for exploring innovative treatment approaches for lung diseases. These findings suggest that overexpression of miR34a/b/c has impact on mucociliary process by reducing the duration required for the process of ciliogenesis.
Furthermore, the expression levels of differentiated cell markerssuggest increased number of MCCs and
decreased number of GCs, indicating the role of miR34a/b/c in enhancing mucociliary clearance.
PB  - 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  - The role of MIR-34 family members on the mucociliary process in the cellular respiratory model system
EP  - 90
SP  - 90
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2140
ER  - 

@conference{
author = "Stevanović, Nina and Skakić, Anita and Parezanović, Marina and Spasovski, Vesna and Ugrin, Milena and Komazec, Jovana and Klaassen, Kristel and Stanković, Sara and Pavlović, Sonja and Stojiljković, Maja and Anđelković, Marina",
year = "2023",
abstract = "Introduction: Primary ciliary dyskinesia is a rare and heterogeneous disorder primarily affecting the respiratory organs, with impaired mucociliary clearance being a common characteristic. Recently, the importance of the miR34/449 family in ciliogenesisin animal models has been described. Thisstudy aimed
to establish a modelsystem to study respiratory diseases and assessfor the first time the role of the miR34 family on the mucociliary process in humans.
Methods: We cultured the primary Normal Human Bronchial Epithelial (NHBE) cells in the air-liquid interface system, enabling the differentiation of multiciliated cells(MCCs) and goblet cells(GCs). During the
differentiation process, transient overexpression of miR-34a/b/c members was conducted. The model
system and treatments were validated through confocal microscopy (β-tubulin, MUC5B, MUC5AC antibodies) and qRT-PCR of miRNAs,specifically ciliogenesis markers(NOTCH1, MCIDAS, GEMC1, CCNO, RFX3),
and differentiated cell markers (FOXJ1 and TFF3).
Results: Expression levels of ciliogenesis and differentiated cells markers and detection of cilia and
mucins at confocal microscopy confirmed the successful establishment of cellular modelsystem. During
the initial differentiation stage, an overexpression of miR34a/b/c changed the expression profile of ciliogenesis and differentiated cell markers.
Conclusion: The established model system provides a valuable platform for exploring innovative treatment approaches for lung diseases. These findings suggest that overexpression of miR34a/b/c has impact on mucociliary process by reducing the duration required for the process of ciliogenesis.
Furthermore, the expression levels of differentiated cell markerssuggest increased number of MCCs and
decreased number of GCs, indicating the role of miR34a/b/c in enhancing mucociliary clearance.",
publisher = "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 = "The role of MIR-34 family members on the mucociliary process in the cellular respiratory model system",
pages = "90-90",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2140"
}

Stevanović, N., Skakić, A., Parezanović, M., Spasovski, V., Ugrin, M., Komazec, J., Klaassen, K., Stanković, S., Pavlović, S., Stojiljković, M.,& Anđelković, M.. (2023). The role of MIR-34 family members on the mucociliary process in the cellular respiratory model system. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 90-90.
https://hdl.handle.net/21.15107/rcub_imagine_2140

Stevanović N, Skakić A, Parezanović M, Spasovski V, Ugrin M, Komazec J, Klaassen K, Stanković S, Pavlović S, Stojiljković M, Anđelković M. The role of MIR-34 family members on the mucociliary process in the cellular respiratory model system. 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;:90-90.
https://hdl.handle.net/21.15107/rcub_imagine_2140 .

Stevanović, Nina, Skakić, Anita, Parezanović, Marina, Spasovski, Vesna, Ugrin, Milena, Komazec, Jovana, Klaassen, Kristel, Stanković, Sara, Pavlović, Sonja, Stojiljković, Maja, Anđelković, Marina, "The role of MIR-34 family members on the mucociliary process in the cellular respiratory model system" 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):90-90,
https://hdl.handle.net/21.15107/rcub_imagine_2140 .

TY  - CONF
AU  - Jocić, Nikola
AU  - Parezanović, Marina
AU  - Anđelković, Marina
AU  - Stevanović, Nina
AU  - Ugrin, Milena
AU  - Spasovski, Vesna
AU  - Klaassen, Kristel
AU  - Stanković, Sara
AU  - Komazec, Jovana
AU  - Pavlović, Sonja
AU  - Stojiljković, Maja
AU  - Skakić, Anita
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2141
AB  - Introduction: Glycogen storage disease type Ib (GSD-Ib) is characterized by a deficiency of glucose-6-
phosphate translocase (G6PT) encoded by the SLC37A4 gene, affecting glucose homeostasis and disrupting autophagy. Recent findings suggest that G6PT may also play a role in autophagy and
glycogen-selective autophagy (glycophagy) activation independent of its transport function. To investigate this hypothesis, two groups of GSD-Ib patients carrying variants with different effects on G6PT
transport activity and stability (p.Asn27Lys and p.Leu348Valfs*53), were compared to the control group
of subjects.
Methods: The relative expression levels of SLC37A4 gene, autophagy (mTOR, ULK1, PRKAG1), and glycophagy markers(GABARAPL1, GAA, STBD1) were assessed in mononuclear cells of GSD Ib patients(four
carrying p.Asn27Lys and four carrying p.Leu348Valfs*53 variant) compared to control group using RTqPCR. Statistical analysis was performed using one-way ANOVA followed by a post-hoc t-test.
Results: The p.Asn27Lys group exhibited 1.5-2.5 times higher expression of SLC37A4 and autophagy
markers, while the p.Leu348Valfs*53 group showed downregulation by approximately 50% compared to
the control group. Glycophagy markers were increased twofold in both patient groups, except for GAA,
which had similar expression levels as the control group.
Conclusion: Individuals carrying the p.Asn27Lys variant display the presence of SLC37A4 transcript in
their cells, which correlates with autophagy activation. Conversely, in patients with the p.Leu348Valfs*53
variant SLC37A4 is downregulated, indicating compromised autophagy activation. These findings support the role of G6PT in autophagy activation, independent of itstransport activity. Furthermore, the elevated expression of glycophagy markers observed in both patient groups can be attributed to the
accumulated glycogen.
PB  - 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  - Investigation of the role of the glucose-6-phosphate translocase in the activation of autophagy and glycogen-selective autophagy in glycogen storage disease type IB patients
EP  - 92
SP  - 92
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2141
ER  - 

@conference{
author = "Jocić, Nikola and Parezanović, Marina and Anđelković, Marina and Stevanović, Nina and Ugrin, Milena and Spasovski, Vesna and Klaassen, Kristel and Stanković, Sara and Komazec, Jovana and Pavlović, Sonja and Stojiljković, Maja and Skakić, Anita",
year = "2023",
abstract = "Introduction: Glycogen storage disease type Ib (GSD-Ib) is characterized by a deficiency of glucose-6-
phosphate translocase (G6PT) encoded by the SLC37A4 gene, affecting glucose homeostasis and disrupting autophagy. Recent findings suggest that G6PT may also play a role in autophagy and
glycogen-selective autophagy (glycophagy) activation independent of its transport function. To investigate this hypothesis, two groups of GSD-Ib patients carrying variants with different effects on G6PT
transport activity and stability (p.Asn27Lys and p.Leu348Valfs*53), were compared to the control group
of subjects.
Methods: The relative expression levels of SLC37A4 gene, autophagy (mTOR, ULK1, PRKAG1), and glycophagy markers(GABARAPL1, GAA, STBD1) were assessed in mononuclear cells of GSD Ib patients(four
carrying p.Asn27Lys and four carrying p.Leu348Valfs*53 variant) compared to control group using RTqPCR. Statistical analysis was performed using one-way ANOVA followed by a post-hoc t-test.
Results: The p.Asn27Lys group exhibited 1.5-2.5 times higher expression of SLC37A4 and autophagy
markers, while the p.Leu348Valfs*53 group showed downregulation by approximately 50% compared to
the control group. Glycophagy markers were increased twofold in both patient groups, except for GAA,
which had similar expression levels as the control group.
Conclusion: Individuals carrying the p.Asn27Lys variant display the presence of SLC37A4 transcript in
their cells, which correlates with autophagy activation. Conversely, in patients with the p.Leu348Valfs*53
variant SLC37A4 is downregulated, indicating compromised autophagy activation. These findings support the role of G6PT in autophagy activation, independent of itstransport activity. Furthermore, the elevated expression of glycophagy markers observed in both patient groups can be attributed to the
accumulated glycogen.",
publisher = "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 = "Investigation of the role of the glucose-6-phosphate translocase in the activation of autophagy and glycogen-selective autophagy in glycogen storage disease type IB patients",
pages = "92-92",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2141"
}

Jocić, N., Parezanović, M., Anđelković, M., Stevanović, N., Ugrin, M., Spasovski, V., Klaassen, K., Stanković, S., Komazec, J., Pavlović, S., Stojiljković, M.,& Skakić, A.. (2023). Investigation of the role of the glucose-6-phosphate translocase in the activation of autophagy and glycogen-selective autophagy in glycogen storage disease type IB patients. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 92-92.
https://hdl.handle.net/21.15107/rcub_imagine_2141

Jocić N, Parezanović M, Anđelković M, Stevanović N, Ugrin M, Spasovski V, Klaassen K, Stanković S, Komazec J, Pavlović S, Stojiljković M, Skakić A. Investigation of the role of the glucose-6-phosphate translocase in the activation of autophagy and glycogen-selective autophagy in glycogen storage disease type IB patients. 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;:92-92.
https://hdl.handle.net/21.15107/rcub_imagine_2141 .

Jocić, Nikola, Parezanović, Marina, Anđelković, Marina, Stevanović, Nina, Ugrin, Milena, Spasovski, Vesna, Klaassen, Kristel, Stanković, Sara, Komazec, Jovana, Pavlović, Sonja, Stojiljković, Maja, Skakić, Anita, "Investigation of the role of the glucose-6-phosphate translocase in the activation of autophagy and glycogen-selective autophagy in glycogen storage disease type IB patients" 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):92-92,
https://hdl.handle.net/21.15107/rcub_imagine_2141 .

TY  - CONF
AU  - Stojiljković, Maja
AU  - Ugrin, Milena
AU  - Klaassen, Kristel
AU  - Skakić, Anita
AU  - Anđelković, Marina
AU  - Komazec, Jovana
AU  - Spasovski, Vesna
AU  - Pavlović, Sonja
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2175
AB  - Introduction: Rare diseases are heterogeneous group of diseases, with one common characteristics, a
prevalence less than 1 in 2000 people. Vast majority of them are monogenic and finding pathogenic genetic
variants is needed to set the correct diagnosis, enable adequate treatment and provide genetic counselling to
members of affected family. This study is an overview of genomic studies of rare diseases in Serbia.
Methods: More than 1200 patients suspected to have a rare disease have been analyzed using sanger
sequencing, clinical-exome sequencing, whole-exome sequencing or whole-genome sequencing in order to
find disease-causing or disease-modifying variants. Novel variants were characterized using in silico modelling
or in in vitro eukaryotic assays (standard or CRISPR/Cas9 developed).
Results: Disease-causing variants were found in more than 150 different genes associated with a rare
disease. The most frequent were thalassemia syndromes (214 patients), followed by phenylketonuria (109
patients), congenital adrenal hyperplasia (>90 patients) and glycogen storage disease Ib (30 patients), while
majority of diseases is seen only in a single patient. More than 40 new genetic variants were comprehensively
characterized in silico or in vitro. For the first time, candidate modifiers (SHANK gene family) were identified
in a group of phenylketonuria patients with an unusual phenotype.
Conclusion: In the genomics era, next-generation sequencing significantly shortens time to diagnosis
and allows studying genetic modifiers of monogenic diseases and genotype-phenotype correlation. Furthermore,
characterization of novel genetic targets boosts development of precision medicine.
PB  - Macedonian Academy of Sciences and Arts
C3  - International Journal of Medical Genetics
T1  - THE IMPACT OF NEXT-GENERATION SEQUENCING ON DIAGNOSIS AND TREATMENT OF RARE DISEASES
EP  - 38
IS  - Supplement
SP  - 38
VL  - 26
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2175
ER  - 

@conference{
author = "Stojiljković, Maja and Ugrin, Milena and Klaassen, Kristel and Skakić, Anita and Anđelković, Marina and Komazec, Jovana and Spasovski, Vesna and Pavlović, Sonja",
year = "2023",
abstract = "Introduction: Rare diseases are heterogeneous group of diseases, with one common characteristics, a
prevalence less than 1 in 2000 people. Vast majority of them are monogenic and finding pathogenic genetic
variants is needed to set the correct diagnosis, enable adequate treatment and provide genetic counselling to
members of affected family. This study is an overview of genomic studies of rare diseases in Serbia.
Methods: More than 1200 patients suspected to have a rare disease have been analyzed using sanger
sequencing, clinical-exome sequencing, whole-exome sequencing or whole-genome sequencing in order to
find disease-causing or disease-modifying variants. Novel variants were characterized using in silico modelling
or in in vitro eukaryotic assays (standard or CRISPR/Cas9 developed).
Results: Disease-causing variants were found in more than 150 different genes associated with a rare
disease. The most frequent were thalassemia syndromes (214 patients), followed by phenylketonuria (109
patients), congenital adrenal hyperplasia (>90 patients) and glycogen storage disease Ib (30 patients), while
majority of diseases is seen only in a single patient. More than 40 new genetic variants were comprehensively
characterized in silico or in vitro. For the first time, candidate modifiers (SHANK gene family) were identified
in a group of phenylketonuria patients with an unusual phenotype.
Conclusion: In the genomics era, next-generation sequencing significantly shortens time to diagnosis
and allows studying genetic modifiers of monogenic diseases and genotype-phenotype correlation. Furthermore,
characterization of novel genetic targets boosts development of precision medicine.",
publisher = "Macedonian Academy of Sciences and Arts",
journal = "International Journal of Medical Genetics",
title = "THE IMPACT OF NEXT-GENERATION SEQUENCING ON DIAGNOSIS AND TREATMENT OF RARE DISEASES",
pages = "38-38",
number = "Supplement",
volume = "26",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2175"
}

Stojiljković, M., Ugrin, M., Klaassen, K., Skakić, A., Anđelković, M., Komazec, J., Spasovski, V.,& Pavlović, S.. (2023). THE IMPACT OF NEXT-GENERATION SEQUENCING ON DIAGNOSIS AND TREATMENT OF RARE DISEASES. in International Journal of Medical Genetics
Macedonian Academy of Sciences and Arts., 26(Supplement), 38-38.
https://hdl.handle.net/21.15107/rcub_imagine_2175

Stojiljković M, Ugrin M, Klaassen K, Skakić A, Anđelković M, Komazec J, Spasovski V, Pavlović S. THE IMPACT OF NEXT-GENERATION SEQUENCING ON DIAGNOSIS AND TREATMENT OF RARE DISEASES. in International Journal of Medical Genetics. 2023;26(Supplement):38-38.
https://hdl.handle.net/21.15107/rcub_imagine_2175 .

Stojiljković, Maja, Ugrin, Milena, Klaassen, Kristel, Skakić, Anita, Anđelković, Marina, Komazec, Jovana, Spasovski, Vesna, Pavlović, Sonja, "THE IMPACT OF NEXT-GENERATION SEQUENCING ON DIAGNOSIS AND TREATMENT OF RARE DISEASES" in International Journal of Medical Genetics, 26, no. Supplement (2023):38-38,
https://hdl.handle.net/21.15107/rcub_imagine_2175 .