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Improvement of fruit juice quality: novel endo-polygalacturonase II from Aspergillus tubingensis FAT 43 for enhanced liquefaction, clarification, and antioxidant potential

Pavlović, Marija; Margetić, Aleksandra; Leonardi, Adrijana; Križaj, Igor; Kojić, Milan; Vujčić, Zoran; Šokarda Slavić, Marinela

(Royal Society of Chemistry, 2024)

TY  - JOUR
AU  - Pavlović, Marija
AU  - Margetić, Aleksandra
AU  - Leonardi, Adrijana
AU  - Križaj, Igor
AU  - Kojić, Milan
AU  - Vujčić, Zoran
AU  - Šokarda Slavić, Marinela
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/863
AB  - This study focuses on the isolation, purification, and characterisation of endo-polygalacturonase II from Aspergillus tubingensis FAT43, particularly emphasising its potential applications in the fruit juice industry. A comprehensive screening test revealed the temporal dynamics of endo-polygalacturonase production during a 96-hour fermentation process. The purification process, involving ammonium sulfate and ethanol precipitation followed by ion-exchange chromatography, resulted in a 3.3-fold purification of PG II with a yield of 16% and a specific activity of 6001.67 U mg−1. Molecular analysis confirmed the identity of PG II, its gene (pgaII), and a high degree of sequence identity with Aspergillus tubingensis in the SWISS-PROT database. The optimal pH for PG II activity was 3.5–4.5, with robust stability across a broad pH spectrum (3–7). The enzyme exhibited optimal temperature activity at 45 °C, with a retention of 90% activity at 50 °C. The calculated activation energy for PG II was 62.1 kJ mol−1, indicating good stability. Inactivation kinetics revealed a half-life of 13.7 h at 40 °C, 5.4 h at 50 °C, and 0.85 h at 60 °C, with an activation energy of denaturation of 32.8 kJ mol−1. Compared to literature-reported PGs, PG II from A. tubingensis FAT43 demonstrated superior thermal stability. Hydrolysis experiments on different pectins revealed the highest specificity for non-methylated substrates (polygalacturonic acid). In fruit juice processing, PG II significantly increased juice yield and clarity, with the highest impact observed in strawberry juice. Antioxidant activity assays indicated enhanced antioxidant potential in enzyme-treated juices, especially strawberry, quince, and apple juices. The study highlights PG II's potential as an industrially valuable enzyme for fruit juice processing, offering improved thermostability and versatility across various fruit types.
PB  - Royal Society of Chemistry
T2  - Food & Function
T1  - Improvement of fruit juice quality: novel endo-polygalacturonase II from Aspergillus tubingensis FAT 43 for enhanced liquefaction, clarification, and antioxidant potential
DO  - 10.1039/D3FO05297D
ER  - 
@article{
author = "Pavlović, Marija and Margetić, Aleksandra and Leonardi, Adrijana and Križaj, Igor and Kojić, Milan and Vujčić, Zoran and Šokarda Slavić, Marinela",
year = "2024",
abstract = "This study focuses on the isolation, purification, and characterisation of endo-polygalacturonase II from Aspergillus tubingensis FAT43, particularly emphasising its potential applications in the fruit juice industry. A comprehensive screening test revealed the temporal dynamics of endo-polygalacturonase production during a 96-hour fermentation process. The purification process, involving ammonium sulfate and ethanol precipitation followed by ion-exchange chromatography, resulted in a 3.3-fold purification of PG II with a yield of 16% and a specific activity of 6001.67 U mg−1. Molecular analysis confirmed the identity of PG II, its gene (pgaII), and a high degree of sequence identity with Aspergillus tubingensis in the SWISS-PROT database. The optimal pH for PG II activity was 3.5–4.5, with robust stability across a broad pH spectrum (3–7). The enzyme exhibited optimal temperature activity at 45 °C, with a retention of 90% activity at 50 °C. The calculated activation energy for PG II was 62.1 kJ mol−1, indicating good stability. Inactivation kinetics revealed a half-life of 13.7 h at 40 °C, 5.4 h at 50 °C, and 0.85 h at 60 °C, with an activation energy of denaturation of 32.8 kJ mol−1. Compared to literature-reported PGs, PG II from A. tubingensis FAT43 demonstrated superior thermal stability. Hydrolysis experiments on different pectins revealed the highest specificity for non-methylated substrates (polygalacturonic acid). In fruit juice processing, PG II significantly increased juice yield and clarity, with the highest impact observed in strawberry juice. Antioxidant activity assays indicated enhanced antioxidant potential in enzyme-treated juices, especially strawberry, quince, and apple juices. The study highlights PG II's potential as an industrially valuable enzyme for fruit juice processing, offering improved thermostability and versatility across various fruit types.",
publisher = "Royal Society of Chemistry",
journal = "Food & Function",
title = "Improvement of fruit juice quality: novel endo-polygalacturonase II from Aspergillus tubingensis FAT 43 for enhanced liquefaction, clarification, and antioxidant potential",
doi = "10.1039/D3FO05297D"
}
Pavlović, M., Margetić, A., Leonardi, A., Križaj, I., Kojić, M., Vujčić, Z.,& Šokarda Slavić, M.. (2024). Improvement of fruit juice quality: novel endo-polygalacturonase II from Aspergillus tubingensis FAT 43 for enhanced liquefaction, clarification, and antioxidant potential. in Food & Function
Royal Society of Chemistry..
https://doi.org/10.1039/D3FO05297D
Pavlović M, Margetić A, Leonardi A, Križaj I, Kojić M, Vujčić Z, Šokarda Slavić M. Improvement of fruit juice quality: novel endo-polygalacturonase II from Aspergillus tubingensis FAT 43 for enhanced liquefaction, clarification, and antioxidant potential. in Food & Function. 2024;.
doi:10.1039/D3FO05297D .
Pavlović, Marija, Margetić, Aleksandra, Leonardi, Adrijana, Križaj, Igor, Kojić, Milan, Vujčić, Zoran, Šokarda Slavić, Marinela, "Improvement of fruit juice quality: novel endo-polygalacturonase II from Aspergillus tubingensis FAT 43 for enhanced liquefaction, clarification, and antioxidant potential" in Food & Function (2024),
https://doi.org/10.1039/D3FO05297D . .
1

NMDA Receptor Antagonist Memantine Ameliorates Experimental Autoimmune Encephalomyelitis in Aged Rats

Bufan, Biljana; Ćuruvija, Ivana; Blagojević, Veljko; Grujić-Milanović, Jelica; Prijić, Ivana; Radosavljević, Tatjana; Samardžić, Janko; Radosavljevic, Milica; Janković, Radmila; Djuretić, Jasmina

(MDPI, 2024)

TY  - JOUR
AU  - Bufan, Biljana
AU  - Ćuruvija, Ivana
AU  - Blagojević, Veljko
AU  - Grujić-Milanović, Jelica
AU  - Prijić, Ivana
AU  - Radosavljević, Tatjana
AU  - Samardžić, Janko
AU  - Radosavljevic, Milica
AU  - Janković, Radmila
AU  - Djuretić, Jasmina
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/867
AB  - Aging is closely related to the main aspects of multiple sclerosis (MS). The average age of the MS population is increasing and the number of elderly MS patients is expected to increase. In addition to neurons, N-methyl-D-aspartate receptors (NMDARs) are also expressed on non-neuronal cells, such as immune cells. The aim of this study was to investigate the role of NMDARs in experimental autoimmune encephalomyelitis (EAE) in young and aged rats. Memantine, a non-competitive NMDAR antagonist, was administered to young and aged Dark Agouti rats from day 7 after immunization. Antagonizing NMDARs had a more favourable effect on clinical disease, reactivation, and apoptosis of CD4+ T cells in the target organ of aged EAE rats. The expression of the fractalkine receptor CX3CR1 was increased in memantine-treated rats, but to a greater extent in aged rats. Additionally, memantine increased Nrf2 and Nrf2-regulated enzymes’ mRNA expression in brain tissue. The concentrations of superoxide anion radicals, malondialdehyde, and advanced oxidation protein products in brain tissue were consistent with previous results. Overall, our results suggest that NMDARs play a more important role in the pathogenesis of EAE in aged than in young rats.
PB  - MDPI
T2  - Biomedicines
T2  - Biomedicines
T1  - NMDA Receptor Antagonist Memantine Ameliorates Experimental Autoimmune Encephalomyelitis in Aged Rats
IS  - 4
SP  - 717
VL  - 12
DO  - 10.3390/biomedicines12040717
ER  - 
@article{
author = "Bufan, Biljana and Ćuruvija, Ivana and Blagojević, Veljko and Grujić-Milanović, Jelica and Prijić, Ivana and Radosavljević, Tatjana and Samardžić, Janko and Radosavljevic, Milica and Janković, Radmila and Djuretić, Jasmina",
year = "2024",
abstract = "Aging is closely related to the main aspects of multiple sclerosis (MS). The average age of the MS population is increasing and the number of elderly MS patients is expected to increase. In addition to neurons, N-methyl-D-aspartate receptors (NMDARs) are also expressed on non-neuronal cells, such as immune cells. The aim of this study was to investigate the role of NMDARs in experimental autoimmune encephalomyelitis (EAE) in young and aged rats. Memantine, a non-competitive NMDAR antagonist, was administered to young and aged Dark Agouti rats from day 7 after immunization. Antagonizing NMDARs had a more favourable effect on clinical disease, reactivation, and apoptosis of CD4+ T cells in the target organ of aged EAE rats. The expression of the fractalkine receptor CX3CR1 was increased in memantine-treated rats, but to a greater extent in aged rats. Additionally, memantine increased Nrf2 and Nrf2-regulated enzymes’ mRNA expression in brain tissue. The concentrations of superoxide anion radicals, malondialdehyde, and advanced oxidation protein products in brain tissue were consistent with previous results. Overall, our results suggest that NMDARs play a more important role in the pathogenesis of EAE in aged than in young rats.",
publisher = "MDPI",
journal = "Biomedicines, Biomedicines",
title = "NMDA Receptor Antagonist Memantine Ameliorates Experimental Autoimmune Encephalomyelitis in Aged Rats",
number = "4",
pages = "717",
volume = "12",
doi = "10.3390/biomedicines12040717"
}
Bufan, B., Ćuruvija, I., Blagojević, V., Grujić-Milanović, J., Prijić, I., Radosavljević, T., Samardžić, J., Radosavljevic, M., Janković, R.,& Djuretić, J.. (2024). NMDA Receptor Antagonist Memantine Ameliorates Experimental Autoimmune Encephalomyelitis in Aged Rats. in Biomedicines
MDPI., 12(4), 717.
https://doi.org/10.3390/biomedicines12040717
Bufan B, Ćuruvija I, Blagojević V, Grujić-Milanović J, Prijić I, Radosavljević T, Samardžić J, Radosavljevic M, Janković R, Djuretić J. NMDA Receptor Antagonist Memantine Ameliorates Experimental Autoimmune Encephalomyelitis in Aged Rats. in Biomedicines. 2024;12(4):717.
doi:10.3390/biomedicines12040717 .
Bufan, Biljana, Ćuruvija, Ivana, Blagojević, Veljko, Grujić-Milanović, Jelica, Prijić, Ivana, Radosavljević, Tatjana, Samardžić, Janko, Radosavljevic, Milica, Janković, Radmila, Djuretić, Jasmina, "NMDA Receptor Antagonist Memantine Ameliorates Experimental Autoimmune Encephalomyelitis in Aged Rats" in Biomedicines, 12, no. 4 (2024):717,
https://doi.org/10.3390/biomedicines12040717 . .

Supplementary Material for: Patiño-Guillén, G.; Pešović, J.; Panić, M.; Savić-Pavićević, D.; Bošković, F.; Keyser, U. F. Single-Molecule RNA Sizing Enables Quantitative Analysis of Alternative Transcription Termination. Nat Commun 2024, 15 (1), 1699. https://doi.org/10.1038/s41467-024-45968-8.

Patiño-Guillén, Gerardo; Pešović, Jovan; Panić, Marko; Savić-Pavićević, Dušanka; Bošković, Filip; Keyser, Ulrich Felix

(Nature, 2024)

TY  - DATA
AU  - Patiño-Guillén, Gerardo
AU  - Pešović, Jovan
AU  - Panić, Marko
AU  - Savić-Pavićević, Dušanka
AU  - Bošković, Filip
AU  - Keyser, Ulrich Felix
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/866
AB  - This PDF file includes: → Supplementary Figures 1 to 29 → Supplementary Tables 1 to 8
PB  - Nature
T2  - Nature Communications
T1  - Supplementary Material for: Patiño-Guillén, G.; Pešović, J.; Panić, M.; Savić-Pavićević, D.; Bošković, F.; Keyser, U. F. Single-Molecule RNA Sizing Enables Quantitative Analysis of Alternative Transcription Termination. Nat Commun 2024, 15 (1), 1699. https://doi.org/10.1038/s41467-024-45968-8.
IS  - 1
VL  - 15
DO  - 10.17863/CAM.104528
ER  - 
@misc{
author = "Patiño-Guillén, Gerardo and Pešović, Jovan and Panić, Marko and Savić-Pavićević, Dušanka and Bošković, Filip and Keyser, Ulrich Felix",
year = "2024",
abstract = "This PDF file includes: → Supplementary Figures 1 to 29 → Supplementary Tables 1 to 8",
publisher = "Nature",
journal = "Nature Communications",
title = "Supplementary Material for: Patiño-Guillén, G.; Pešović, J.; Panić, M.; Savić-Pavićević, D.; Bošković, F.; Keyser, U. F. Single-Molecule RNA Sizing Enables Quantitative Analysis of Alternative Transcription Termination. Nat Commun 2024, 15 (1), 1699. https://doi.org/10.1038/s41467-024-45968-8.",
number = "1",
volume = "15",
doi = "10.17863/CAM.104528"
}
Patiño-Guillén, G., Pešović, J., Panić, M., Savić-Pavićević, D., Bošković, F.,& Keyser, U. F.. (2024). Supplementary Material for: Patiño-Guillén, G.; Pešović, J.; Panić, M.; Savić-Pavićević, D.; Bošković, F.; Keyser, U. F. Single-Molecule RNA Sizing Enables Quantitative Analysis of Alternative Transcription Termination. Nat Commun 2024, 15 (1), 1699. https://doi.org/10.1038/s41467-024-45968-8.. in Nature Communications
Nature., 15(1).
https://doi.org/10.17863/CAM.104528
Patiño-Guillén G, Pešović J, Panić M, Savić-Pavićević D, Bošković F, Keyser UF. Supplementary Material for: Patiño-Guillén, G.; Pešović, J.; Panić, M.; Savić-Pavićević, D.; Bošković, F.; Keyser, U. F. Single-Molecule RNA Sizing Enables Quantitative Analysis of Alternative Transcription Termination. Nat Commun 2024, 15 (1), 1699. https://doi.org/10.1038/s41467-024-45968-8.. in Nature Communications. 2024;15(1).
doi:10.17863/CAM.104528 .
Patiño-Guillén, Gerardo, Pešović, Jovan, Panić, Marko, Savić-Pavićević, Dušanka, Bošković, Filip, Keyser, Ulrich Felix, "Supplementary Material for: Patiño-Guillén, G.; Pešović, J.; Panić, M.; Savić-Pavićević, D.; Bošković, F.; Keyser, U. F. Single-Molecule RNA Sizing Enables Quantitative Analysis of Alternative Transcription Termination. Nat Commun 2024, 15 (1), 1699. https://doi.org/10.1038/s41467-024-45968-8." in Nature Communications, 15, no. 1 (2024),
https://doi.org/10.17863/CAM.104528 . .

Single-molecule RNA sizing enables quantitative analysis of alternative transcription termination

Patiño-Guillén, Gerardo; Pešović, Jovan; Panić, Marko; Savić-Pavićević, Dušanka; Bošković, Filip; Keyser, Ulrich Felix

(Nature, 2024)

TY  - JOUR
AU  - Patiño-Guillén, Gerardo
AU  - Pešović, Jovan
AU  - Panić, Marko
AU  - Savić-Pavićević, Dušanka
AU  - Bošković, Filip
AU  - Keyser, Ulrich Felix
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/865
AB  - Transcription, a critical process in molecular biology, has found many applications in RNA synthesis, including mRNA vaccines and RNA therapeutics. However, current RNA characterization technologies suffer from amplification and enzymatic biases that lead to loss of native information. Here, we introduce a strategy to quantitatively study both transcription and RNA polymerase behaviour by sizing RNA with RNA nanotechnology and nanopores. To begin, we utilize T7 RNA polymerase to transcribe linear DNA lacking termination sequences. Surprisingly, we discover alternative transcription termination in the origin of replication sequence. Next, we employ circular DNA without transcription terminators to perform rolling circle transcription. This allows us to gain valuable insights into the processivity and transcription behaviour of RNA polymerase at the single-molecule level. Our work demonstrates how RNA nanotechnology and nanopores may be used in tandem for the direct and quantitative analysis of RNA transcripts. This methodology provides a promising pathway for accurate RNA structural mapping by enabling the study of full-length RNA transcripts at the single-molecule level.
PB  - Nature
T2  - Nature Communications
T1  - Single-molecule RNA sizing enables quantitative analysis of alternative transcription termination
IS  - 1
SP  - 1699
VL  - 15
DO  - 10.1038/s41467-024-45968-8
ER  - 
@article{
author = "Patiño-Guillén, Gerardo and Pešović, Jovan and Panić, Marko and Savić-Pavićević, Dušanka and Bošković, Filip and Keyser, Ulrich Felix",
year = "2024",
abstract = "Transcription, a critical process in molecular biology, has found many applications in RNA synthesis, including mRNA vaccines and RNA therapeutics. However, current RNA characterization technologies suffer from amplification and enzymatic biases that lead to loss of native information. Here, we introduce a strategy to quantitatively study both transcription and RNA polymerase behaviour by sizing RNA with RNA nanotechnology and nanopores. To begin, we utilize T7 RNA polymerase to transcribe linear DNA lacking termination sequences. Surprisingly, we discover alternative transcription termination in the origin of replication sequence. Next, we employ circular DNA without transcription terminators to perform rolling circle transcription. This allows us to gain valuable insights into the processivity and transcription behaviour of RNA polymerase at the single-molecule level. Our work demonstrates how RNA nanotechnology and nanopores may be used in tandem for the direct and quantitative analysis of RNA transcripts. This methodology provides a promising pathway for accurate RNA structural mapping by enabling the study of full-length RNA transcripts at the single-molecule level.",
publisher = "Nature",
journal = "Nature Communications",
title = "Single-molecule RNA sizing enables quantitative analysis of alternative transcription termination",
number = "1",
pages = "1699",
volume = "15",
doi = "10.1038/s41467-024-45968-8"
}
Patiño-Guillén, G., Pešović, J., Panić, M., Savić-Pavićević, D., Bošković, F.,& Keyser, U. F.. (2024). Single-molecule RNA sizing enables quantitative analysis of alternative transcription termination. in Nature Communications
Nature., 15(1), 1699.
https://doi.org/10.1038/s41467-024-45968-8
Patiño-Guillén G, Pešović J, Panić M, Savić-Pavićević D, Bošković F, Keyser UF. Single-molecule RNA sizing enables quantitative analysis of alternative transcription termination. in Nature Communications. 2024;15(1):1699.
doi:10.1038/s41467-024-45968-8 .
Patiño-Guillén, Gerardo, Pešović, Jovan, Panić, Marko, Savić-Pavićević, Dušanka, Bošković, Filip, Keyser, Ulrich Felix, "Single-molecule RNA sizing enables quantitative analysis of alternative transcription termination" in Nature Communications, 15, no. 1 (2024):1699,
https://doi.org/10.1038/s41467-024-45968-8 . .
28

A novel thermostable YtnP lactonase from Stenotrophomonas maltophilia inhibits Pseudomonas aeruginosa virulence in vitro and in vivo

Ćurčić, Jovana; Dinić, Miroslav; Novović, Katarina; Vasiljević, Zorica; Kojić, Milan; Jovčić, Branko; Malešević, Milka

(2024)

TY  - JOUR
AU  - Ćurčić, Jovana
AU  - Dinić, Miroslav
AU  - Novović, Katarina
AU  - Vasiljević, Zorica
AU  - Kojić, Milan
AU  - Jovčić, Branko
AU  - Malešević, Milka
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/864
AB  - Infections caused by multidrug-resistant pathogens are one of the biggest challenges facing the healthcare system today. Quorum quenching (QQ) enzymes have the potential to be used as innovative enzyme-based antivirulence therapeutics to combat infections caused by multidrug-resistant pathogens. The main objective of this research was to describe the novel YtnP lactonase derived from the clinical isolate Stenotrophomonas maltophilia and to investigate its antivirulence potential against multidrug-resistant Pseudomonas aeruginosa MMA83. YtnP lactonase, the QQ enzyme, belongs to the family of metallo-β-lactamases. The recombinant enzyme has several advantageous biotechnological properties, such as high thermostability, activity in a wide pH range, and no cytotoxic effect. High-performance liquid chromatography analysis revealed the activity of recombinant YtnP lactonase toward a wide range of N-acyl-homoserine lactones (AHLs), quorum sensing signaling molecules, with a higher preference for long-chain AHLs. Recombinant YtnP lactonase was shown to inhibit P. aeruginosa MMA83 biofilm formation, induce biofilm decomposition, and reduce extracellular virulence factors production. Moreover, the lifespan of MMA83-infected Caenorhabditis elegans was prolonged with YtnP lactonase treatment. YtnP lactonase showed synergistic inhibitory activity in combination with gentamicin and acted additively with meropenem against MMA83. The described properties make YtnP lactonase a promising therapeutic candidate for the development of next-generation antivirulence agents.
T2  - International Journal of Biological Macromolecules
T1  - A novel thermostable YtnP lactonase from Stenotrophomonas maltophilia inhibits Pseudomonas aeruginosa virulence in vitro and in vivo
SP  - 130421
DO  - 10.1016/j.ijbiomac.2024.130421
ER  - 
@article{
author = "Ćurčić, Jovana and Dinić, Miroslav and Novović, Katarina and Vasiljević, Zorica and Kojić, Milan and Jovčić, Branko and Malešević, Milka",
year = "2024",
abstract = "Infections caused by multidrug-resistant pathogens are one of the biggest challenges facing the healthcare system today. Quorum quenching (QQ) enzymes have the potential to be used as innovative enzyme-based antivirulence therapeutics to combat infections caused by multidrug-resistant pathogens. The main objective of this research was to describe the novel YtnP lactonase derived from the clinical isolate Stenotrophomonas maltophilia and to investigate its antivirulence potential against multidrug-resistant Pseudomonas aeruginosa MMA83. YtnP lactonase, the QQ enzyme, belongs to the family of metallo-β-lactamases. The recombinant enzyme has several advantageous biotechnological properties, such as high thermostability, activity in a wide pH range, and no cytotoxic effect. High-performance liquid chromatography analysis revealed the activity of recombinant YtnP lactonase toward a wide range of N-acyl-homoserine lactones (AHLs), quorum sensing signaling molecules, with a higher preference for long-chain AHLs. Recombinant YtnP lactonase was shown to inhibit P. aeruginosa MMA83 biofilm formation, induce biofilm decomposition, and reduce extracellular virulence factors production. Moreover, the lifespan of MMA83-infected Caenorhabditis elegans was prolonged with YtnP lactonase treatment. YtnP lactonase showed synergistic inhibitory activity in combination with gentamicin and acted additively with meropenem against MMA83. The described properties make YtnP lactonase a promising therapeutic candidate for the development of next-generation antivirulence agents.",
journal = "International Journal of Biological Macromolecules",
title = "A novel thermostable YtnP lactonase from Stenotrophomonas maltophilia inhibits Pseudomonas aeruginosa virulence in vitro and in vivo",
pages = "130421",
doi = "10.1016/j.ijbiomac.2024.130421"
}
Ćurčić, J., Dinić, M., Novović, K., Vasiljević, Z., Kojić, M., Jovčić, B.,& Malešević, M.. (2024). A novel thermostable YtnP lactonase from Stenotrophomonas maltophilia inhibits Pseudomonas aeruginosa virulence in vitro and in vivo. in International Journal of Biological Macromolecules, 130421.
https://doi.org/10.1016/j.ijbiomac.2024.130421
Ćurčić J, Dinić M, Novović K, Vasiljević Z, Kojić M, Jovčić B, Malešević M. A novel thermostable YtnP lactonase from Stenotrophomonas maltophilia inhibits Pseudomonas aeruginosa virulence in vitro and in vivo. in International Journal of Biological Macromolecules. 2024;:130421.
doi:10.1016/j.ijbiomac.2024.130421 .
Ćurčić, Jovana, Dinić, Miroslav, Novović, Katarina, Vasiljević, Zorica, Kojić, Milan, Jovčić, Branko, Malešević, Milka, "A novel thermostable YtnP lactonase from Stenotrophomonas maltophilia inhibits Pseudomonas aeruginosa virulence in vitro and in vivo" in International Journal of Biological Macromolecules (2024):130421,
https://doi.org/10.1016/j.ijbiomac.2024.130421 . .

rBet v 1a-BanLecwt induce upregulation of IL-10 and IFN-γ gene expression in Caco-2/THP-1 co-culture and secretion of IL-10 and IFN-γ/IL-4 levels in PBMCs of birch pollen allergic donors

Protić-Rosić, Isidora; Lopandić, Zorana; Popović, Dragan; Blagojević, Gordan; Gavrović-Jankulović, Marija

(Elsevier, 2024)

TY  - JOUR
AU  - Protić-Rosić, Isidora
AU  - Lopandić, Zorana
AU  - Popović, Dragan
AU  - Blagojević, Gordan
AU  - Gavrović-Jankulović, Marija
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/861
AB  - Novel allergen immunotherapy (AIT) approaches necessitate the use of more effective and safe therapeutics, which can be accomplished by employing novel adjuvants for improved innate immune cell activation, as well as hypoallergenic allergen forms. In this study, we investigate the immunomodulatory effects of a chimera rBet v 1a-BanLecwt (rBv1a-BLwt; Cwt) composed of the major birch pollen allergen Bet v 1a and banana lectin (BanLecwt; BLwt) and two novel chimeras, rBv1l-BLH84T (rBet v 1l-BanLecH84T; C1) and rBLH84T-Bv1l (rBanLecH84T-Bet v 1l; C2), both composed of BLH84T and hypoallergenic birch pollen allergen Bv1l in the co-culture model Caco-2/THP-1, and PBMCs from donors with birch pollen allergy. The chimeric molecules rBv1l-BLH84T (C1) and rBLH84T-Bv1l (C2) were created in silico and then produced in E. coli using recombinant DNA technology. Real-time PCR analysis of gene expression following compound treatment in the co-culture model revealed that all three chimeras have the potential to induce the anti-inflammatory cytokine IL-10 gene expression in Caco-2 cells and IFN-γ gene expression in THP-1 cells. Sandwich ELISA revealed that Cwt increased IL-10 secretion and IFN-/IL-4 levels in PBMCs from birch pollen allergic donors, whereas C1 and C2 were less effective. The findings suggest that Cwt should be analyzed further due to its potential benefit in AIT.
PB  - Elsevier
T2  - International Immunopharmacology
T1  - rBet v 1a-BanLecwt induce upregulation of IL-10 and IFN-γ gene expression in Caco-2/THP-1 co-culture and secretion of IL-10 and IFN-γ/IL-4 levels in PBMCs of birch pollen allergic donors
SP  - 111607
VL  - 129
DO  - 10.1016/j.intimp.2024.111607
ER  - 
@article{
author = "Protić-Rosić, Isidora and Lopandić, Zorana and Popović, Dragan and Blagojević, Gordan and Gavrović-Jankulović, Marija",
year = "2024",
abstract = "Novel allergen immunotherapy (AIT) approaches necessitate the use of more effective and safe therapeutics, which can be accomplished by employing novel adjuvants for improved innate immune cell activation, as well as hypoallergenic allergen forms. In this study, we investigate the immunomodulatory effects of a chimera rBet v 1a-BanLecwt (rBv1a-BLwt; Cwt) composed of the major birch pollen allergen Bet v 1a and banana lectin (BanLecwt; BLwt) and two novel chimeras, rBv1l-BLH84T (rBet v 1l-BanLecH84T; C1) and rBLH84T-Bv1l (rBanLecH84T-Bet v 1l; C2), both composed of BLH84T and hypoallergenic birch pollen allergen Bv1l in the co-culture model Caco-2/THP-1, and PBMCs from donors with birch pollen allergy. The chimeric molecules rBv1l-BLH84T (C1) and rBLH84T-Bv1l (C2) were created in silico and then produced in E. coli using recombinant DNA technology. Real-time PCR analysis of gene expression following compound treatment in the co-culture model revealed that all three chimeras have the potential to induce the anti-inflammatory cytokine IL-10 gene expression in Caco-2 cells and IFN-γ gene expression in THP-1 cells. Sandwich ELISA revealed that Cwt increased IL-10 secretion and IFN-/IL-4 levels in PBMCs from birch pollen allergic donors, whereas C1 and C2 were less effective. The findings suggest that Cwt should be analyzed further due to its potential benefit in AIT.",
publisher = "Elsevier",
journal = "International Immunopharmacology",
title = "rBet v 1a-BanLecwt induce upregulation of IL-10 and IFN-γ gene expression in Caco-2/THP-1 co-culture and secretion of IL-10 and IFN-γ/IL-4 levels in PBMCs of birch pollen allergic donors",
pages = "111607",
volume = "129",
doi = "10.1016/j.intimp.2024.111607"
}
Protić-Rosić, I., Lopandić, Z., Popović, D., Blagojević, G.,& Gavrović-Jankulović, M.. (2024). rBet v 1a-BanLecwt induce upregulation of IL-10 and IFN-γ gene expression in Caco-2/THP-1 co-culture and secretion of IL-10 and IFN-γ/IL-4 levels in PBMCs of birch pollen allergic donors. in International Immunopharmacology
Elsevier., 129, 111607.
https://doi.org/10.1016/j.intimp.2024.111607
Protić-Rosić I, Lopandić Z, Popović D, Blagojević G, Gavrović-Jankulović M. rBet v 1a-BanLecwt induce upregulation of IL-10 and IFN-γ gene expression in Caco-2/THP-1 co-culture and secretion of IL-10 and IFN-γ/IL-4 levels in PBMCs of birch pollen allergic donors. in International Immunopharmacology. 2024;129:111607.
doi:10.1016/j.intimp.2024.111607 .
Protić-Rosić, Isidora, Lopandić, Zorana, Popović, Dragan, Blagojević, Gordan, Gavrović-Jankulović, Marija, "rBet v 1a-BanLecwt induce upregulation of IL-10 and IFN-γ gene expression in Caco-2/THP-1 co-culture and secretion of IL-10 and IFN-γ/IL-4 levels in PBMCs of birch pollen allergic donors" in International Immunopharmacology, 129 (2024):111607,
https://doi.org/10.1016/j.intimp.2024.111607 . .

Supplementary information for the article: Mladenovic Stokanic, M.; Simovic, A.; Jovanovic, V.; Radomirovic, M.; Udovicki, B.; Krstic Ristivojevic, M.; Djukic, T.; Vasovic, T.; Acimovic, J.; Sabljic, L.; Lukic, I.; Kovacevic, A.; Cujic, D.; Gnjatovic, M.; Smiljanic, K.; Stojadinovic, M.; Radosavljevic, J.; Stanic-Vucinic, D.; Stojanovic, M.; Rajkovic, A.; Cirkovic Velickovic, T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. International Journal of Molecular Sciences 2024, 25 (1), 333. https://doi.org/10.3390/ijms25010333.

Mladenović Stokanić, Maja; Simović, Ana; Jovanović, Vesna; Radomirović, Mirjana; Udovički, Božidar; Krstić Ristivojević, Maja; Djukić, Teodora; Vasović, Tamara; Aćimović, Jelena; Sabljić, Ljiljana; Lukić, Ivana; Kovačević, Ana; Cujic, Danica; Gnjatović, Marija; Smiljanić, Katarina; Stojadinović, Marija; Radosavljević, Jelena; Stanić-Vučinić, Dragana; Stojanović, Marijana; Rajković, Andreja; Ćirkovic Veličković, Tanja

(MDPI, 2024)

TY  - DATA
AU  - Mladenović Stokanić, Maja
AU  - Simović, Ana
AU  - Jovanović, Vesna
AU  - Radomirović, Mirjana
AU  - Udovički, Božidar
AU  - Krstić Ristivojević, Maja
AU  - Djukić, Teodora
AU  - Vasović, Tamara
AU  - Aćimović, Jelena
AU  - Sabljić, Ljiljana
AU  - Lukić, Ivana
AU  - Kovačević, Ana
AU  - Cujic, Danica
AU  - Gnjatović, Marija
AU  - Smiljanić, Katarina
AU  - Stojadinović, Marija
AU  - Radosavljević, Jelena
AU  - Stanić-Vučinić, Dragana
AU  - Stojanović, Marijana
AU  - Rajković, Andreja
AU  - Ćirkovic Veličković, Tanja
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/859
AB  - S1.1. Checking of N protein purity Recombinant N protein purity was checked after imidazole removal and buffer exchange by SDS PAGE (Figure 6.). For comparison, commercial high-purity HSA was also analyzed. S1.2. Identification of N protein Tandem mass spectrometry identification of proteins in an in-gel digested band of N protein (Figure S1, lane 3), confirmed the identity of N protein with high scores and peptide coverage (Fig. S2.). S2. Purification of polyclonal antibodies from mice and rabbit sera For the development of an ELISA test specific for the detection of SARS-CoV-2 N protein, recombinantly produced N protein was used for the immunization of mice and rabbits. Sera obtained from rabbits and mice were then tested for titer and specificity (Figure S3 and Figure 1). To determine the titer of polyclonal sera required to detect N protein in samples, we use wells coated with N protein and serial dilution of sera pools from different animals. After multiple washing steps, we detected the binding of rabbit and mice antibodies using secondary biotinylated antibodies and streptavidin-alkaline phosphatase chimaera or secondary antibodies with previously coupled alkaline phosphatase, where the amount of enzymes’ substrate converted to the product was measured as an increase in absorbance at 405 nm. As shown in Figure S3A, unpurified sera pools from both animals showed very high titers and expected logarithmic decrease of signal with dilution. Based on the obtained data titer for unpurified sera was determined to be X. The same trend was observed for pools purified using AS precipitation and rabbit sera purified using protein A affinity chromatography (Figure S3B and S3C). As shown in Figure S3D, clear bands from antibodies could be observed in both full and purified samples. Western blot analysis showed only one protein band on mass around 40 kDa, a Accession number / Protein Name Score Coverage (%) Unique peptides P0DTC9|NCAP_SARS2 Nucleoprotein OS=Severe acute respiratory syndrome coronavirus 2, 46 kDa 504.9 74.22 183 mass of purified N protein suggesting that the obtained sera is highly specific for N protein (Figure 2). Section S3 Diagnostic validationS3.1. Stabilization of capture antibodies Pre-coated ELISA plates were prepared for usage in clinical practice. To ensure the preservation of the biofunctionality of the surface-bound capture antibodies, the commonly used stabilizing excipient, 3% sucrose with 10% glycerol in MilliQ water was used. The plates were incubated with 300 μL per well of a stabilizing agent for 1 hour at room temperature. After an hour of incubation, the solution was carefully aspirated from each well. The plate was then blotted against clear paper towels to remove any remaining liquid, and the plates were allowed to air dry for 3 hours at RT. Dried plates were wrapped in parafilm and stored at 4 °C for later use. To remove the stabilizing agent coating, wells were washed with slightly acidic distilled water (pH of 6) three times, leaving the plate prepared for subsequent assay steps. Section S4. Characterization of N protein by HRMS S4.1. SDS PAGE and in-gel digestion Characterization of the produced recombinant N protein was done by HRMS after its in-gel digestion. A total of 10 μg of purified protein(s) were loaded in a 0.5 cm wide well and after SDSPAGE gel was stained with Coomassie Brilliant Blue R-250 (CBB). Protein gel bands were washed, reduced with dithiothreitol, and alkylated with iodoacetamide, followed by in-gel trypsin digestion1 (Shevchenko et al. 2006) with some minor modifications. The amount of trypsin was leveled to a trypsin/sample ratio of 1:30 (w/w). The final concentration of MS-grade trypsin (diluted in 25 mM ammonium bicarbonate buffer) was 1 ng/μL. Sample clean-up was performed using zip tips HyperSep C18 (Thermo Fisher Scientific Inc., Bremen, Germany). S5.1 Immunization of rabbits and mice Mice immunization Swiss Webster mice (n=10) were immunized subcutaneously with N protein formulated with Complete Freund`s adjuvant (CFA; 1st dose, 100 μg N protein / dose) or Incomplete Freund`s adjuvant (IFA; 2nd and 3rd doses, 50 μg N protein / dose) in three-week intervals. Mice were housed in small groups of up to six animals and had access to commercial mice food and water ad libitum. N protein solution (500ug/ml in PBS) was sterilized by filtering through 0.22 um filters. Sterile N protein solution was mixed with CFA (Sigma, Cat. No. F5881) at ratio 1:1 (v/v) under aseptic conditions. In total 400 ul of N protein-CFA emulsion (N protein final concentration 250ug/ml) was applied per immunization per mouse. Initial immunization was done by injection of N protein in CFA given subcutaneously (SC) in four sites (thigh pocket, base of tail, and mediastinum) with a 100 ul using 23-25 gauge needle. In total 100 ug of N protein was applied per mouse (25 ug per site). Subsequent immunizations with booster doses were done in the same way, but using IFA (Sigma, Cat. No. F5506) instead of CFA and N protein final concentration was 125 ug/ml. . In total 50 ug of N protein was applied per mouse (12.5 ug per site). Immunizations were done every three weeks. Mice immunization scheme: 1. day 0 – N protein in PBS: CFA = 1:1 (v/v); N protein final concentration was 250 μg/mL; 400 μL per mice (4x100 μL), e.g. 100 μg per mice 2. day 21 - N protein in PBS: IFA = 1:1 (v/v); N protein final concentration was 125 μg/mL; 400 μL per mice (4x100 μL), e.g. 50 μg per mice 3. day 42 - N protein in PBS: IFA = 1:1 (v/v); N protein final concentration was 125 μg/mL; 400 μl per mice (4x100 μL), e.g. 50 μg per mice First bleeding was performed two weeks after the 3rd dose, and then in intervals not shorter than two weeks. The sera obtained after the first bleeding was tested for the production of specific anti-N protein antibodies.
PB  - MDPI
T2  - International Journal of Molecular Sciences
T1  - Supplementary information for the article:       Mladenovic Stokanic, M.; Simovic, A.; Jovanovic, V.; Radomirovic, M.; Udovicki, B.; Krstic Ristivojevic, M.; Djukic, T.; Vasovic, T.; Acimovic, J.; Sabljic, L.; Lukic, I.; Kovacevic, A.; Cujic, D.; Gnjatovic, M.; Smiljanic, K.; Stojadinovic, M.; Radosavljevic, J.; Stanic-Vucinic, D.; Stojanovic, M.; Rajkovic, A.; Cirkovic Velickovic, T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. International Journal of Molecular Sciences 2024, 25 (1), 333. https://doi.org/10.3390/ijms25010333.
IS  - 1
VL  - 25
UR  - https://hdl.handle.net/21.15107/rcub_intor_859
ER  - 
@misc{
author = "Mladenović Stokanić, Maja and Simović, Ana and Jovanović, Vesna and Radomirović, Mirjana and Udovički, Božidar and Krstić Ristivojević, Maja and Djukić, Teodora and Vasović, Tamara and Aćimović, Jelena and Sabljić, Ljiljana and Lukić, Ivana and Kovačević, Ana and Cujic, Danica and Gnjatović, Marija and Smiljanić, Katarina and Stojadinović, Marija and Radosavljević, Jelena and Stanić-Vučinić, Dragana and Stojanović, Marijana and Rajković, Andreja and Ćirkovic Veličković, Tanja",
year = "2024",
abstract = "S1.1. Checking of N protein purity Recombinant N protein purity was checked after imidazole removal and buffer exchange by SDS PAGE (Figure 6.). For comparison, commercial high-purity HSA was also analyzed. S1.2. Identification of N protein Tandem mass spectrometry identification of proteins in an in-gel digested band of N protein (Figure S1, lane 3), confirmed the identity of N protein with high scores and peptide coverage (Fig. S2.). S2. Purification of polyclonal antibodies from mice and rabbit sera For the development of an ELISA test specific for the detection of SARS-CoV-2 N protein, recombinantly produced N protein was used for the immunization of mice and rabbits. Sera obtained from rabbits and mice were then tested for titer and specificity (Figure S3 and Figure 1). To determine the titer of polyclonal sera required to detect N protein in samples, we use wells coated with N protein and serial dilution of sera pools from different animals. After multiple washing steps, we detected the binding of rabbit and mice antibodies using secondary biotinylated antibodies and streptavidin-alkaline phosphatase chimaera or secondary antibodies with previously coupled alkaline phosphatase, where the amount of enzymes’ substrate converted to the product was measured as an increase in absorbance at 405 nm. As shown in Figure S3A, unpurified sera pools from both animals showed very high titers and expected logarithmic decrease of signal with dilution. Based on the obtained data titer for unpurified sera was determined to be X. The same trend was observed for pools purified using AS precipitation and rabbit sera purified using protein A affinity chromatography (Figure S3B and S3C). As shown in Figure S3D, clear bands from antibodies could be observed in both full and purified samples. Western blot analysis showed only one protein band on mass around 40 kDa, a Accession number / Protein Name Score Coverage (%) Unique peptides P0DTC9|NCAP_SARS2 Nucleoprotein OS=Severe acute respiratory syndrome coronavirus 2, 46 kDa 504.9 74.22 183 mass of purified N protein suggesting that the obtained sera is highly specific for N protein (Figure 2). Section S3 Diagnostic validationS3.1. Stabilization of capture antibodies Pre-coated ELISA plates were prepared for usage in clinical practice. To ensure the preservation of the biofunctionality of the surface-bound capture antibodies, the commonly used stabilizing excipient, 3% sucrose with 10% glycerol in MilliQ water was used. The plates were incubated with 300 μL per well of a stabilizing agent for 1 hour at room temperature. After an hour of incubation, the solution was carefully aspirated from each well. The plate was then blotted against clear paper towels to remove any remaining liquid, and the plates were allowed to air dry for 3 hours at RT. Dried plates were wrapped in parafilm and stored at 4 °C for later use. To remove the stabilizing agent coating, wells were washed with slightly acidic distilled water (pH of 6) three times, leaving the plate prepared for subsequent assay steps. Section S4. Characterization of N protein by HRMS S4.1. SDS PAGE and in-gel digestion Characterization of the produced recombinant N protein was done by HRMS after its in-gel digestion. A total of 10 μg of purified protein(s) were loaded in a 0.5 cm wide well and after SDSPAGE gel was stained with Coomassie Brilliant Blue R-250 (CBB). Protein gel bands were washed, reduced with dithiothreitol, and alkylated with iodoacetamide, followed by in-gel trypsin digestion1 (Shevchenko et al. 2006) with some minor modifications. The amount of trypsin was leveled to a trypsin/sample ratio of 1:30 (w/w). The final concentration of MS-grade trypsin (diluted in 25 mM ammonium bicarbonate buffer) was 1 ng/μL. Sample clean-up was performed using zip tips HyperSep C18 (Thermo Fisher Scientific Inc., Bremen, Germany). S5.1 Immunization of rabbits and mice Mice immunization Swiss Webster mice (n=10) were immunized subcutaneously with N protein formulated with Complete Freund`s adjuvant (CFA; 1st dose, 100 μg N protein / dose) or Incomplete Freund`s adjuvant (IFA; 2nd and 3rd doses, 50 μg N protein / dose) in three-week intervals. Mice were housed in small groups of up to six animals and had access to commercial mice food and water ad libitum. N protein solution (500ug/ml in PBS) was sterilized by filtering through 0.22 um filters. Sterile N protein solution was mixed with CFA (Sigma, Cat. No. F5881) at ratio 1:1 (v/v) under aseptic conditions. In total 400 ul of N protein-CFA emulsion (N protein final concentration 250ug/ml) was applied per immunization per mouse. Initial immunization was done by injection of N protein in CFA given subcutaneously (SC) in four sites (thigh pocket, base of tail, and mediastinum) with a 100 ul using 23-25 gauge needle. In total 100 ug of N protein was applied per mouse (25 ug per site). Subsequent immunizations with booster doses were done in the same way, but using IFA (Sigma, Cat. No. F5506) instead of CFA and N protein final concentration was 125 ug/ml. . In total 50 ug of N protein was applied per mouse (12.5 ug per site). Immunizations were done every three weeks. Mice immunization scheme: 1. day 0 – N protein in PBS: CFA = 1:1 (v/v); N protein final concentration was 250 μg/mL; 400 μL per mice (4x100 μL), e.g. 100 μg per mice 2. day 21 - N protein in PBS: IFA = 1:1 (v/v); N protein final concentration was 125 μg/mL; 400 μL per mice (4x100 μL), e.g. 50 μg per mice 3. day 42 - N protein in PBS: IFA = 1:1 (v/v); N protein final concentration was 125 μg/mL; 400 μl per mice (4x100 μL), e.g. 50 μg per mice First bleeding was performed two weeks after the 3rd dose, and then in intervals not shorter than two weeks. The sera obtained after the first bleeding was tested for the production of specific anti-N protein antibodies.",
publisher = "MDPI",
journal = "International Journal of Molecular Sciences",
title = "Supplementary information for the article:       Mladenovic Stokanic, M.; Simovic, A.; Jovanovic, V.; Radomirovic, M.; Udovicki, B.; Krstic Ristivojevic, M.; Djukic, T.; Vasovic, T.; Acimovic, J.; Sabljic, L.; Lukic, I.; Kovacevic, A.; Cujic, D.; Gnjatovic, M.; Smiljanic, K.; Stojadinovic, M.; Radosavljevic, J.; Stanic-Vucinic, D.; Stojanovic, M.; Rajkovic, A.; Cirkovic Velickovic, T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. International Journal of Molecular Sciences 2024, 25 (1), 333. https://doi.org/10.3390/ijms25010333.",
number = "1",
volume = "25",
url = "https://hdl.handle.net/21.15107/rcub_intor_859"
}
Mladenović Stokanić, M., Simović, A., Jovanović, V., Radomirović, M., Udovički, B., Krstić Ristivojević, M., Djukić, T., Vasović, T., Aćimović, J., Sabljić, L., Lukić, I., Kovačević, A., Cujic, D., Gnjatović, M., Smiljanić, K., Stojadinović, M., Radosavljević, J., Stanić-Vučinić, D., Stojanović, M., Rajković, A.,& Ćirkovic Veličković, T.. (2024). Supplementary information for the article:       Mladenovic Stokanic, M.; Simovic, A.; Jovanovic, V.; Radomirovic, M.; Udovicki, B.; Krstic Ristivojevic, M.; Djukic, T.; Vasovic, T.; Acimovic, J.; Sabljic, L.; Lukic, I.; Kovacevic, A.; Cujic, D.; Gnjatovic, M.; Smiljanic, K.; Stojadinovic, M.; Radosavljevic, J.; Stanic-Vucinic, D.; Stojanovic, M.; Rajkovic, A.; Cirkovic Velickovic, T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. International Journal of Molecular Sciences 2024, 25 (1), 333. https://doi.org/10.3390/ijms25010333.. in International Journal of Molecular Sciences
MDPI., 25(1).
https://hdl.handle.net/21.15107/rcub_intor_859
Mladenović Stokanić M, Simović A, Jovanović V, Radomirović M, Udovički B, Krstić Ristivojević M, Djukić T, Vasović T, Aćimović J, Sabljić L, Lukić I, Kovačević A, Cujic D, Gnjatović M, Smiljanić K, Stojadinović M, Radosavljević J, Stanić-Vučinić D, Stojanović M, Rajković A, Ćirkovic Veličković T. Supplementary information for the article:       Mladenovic Stokanic, M.; Simovic, A.; Jovanovic, V.; Radomirovic, M.; Udovicki, B.; Krstic Ristivojevic, M.; Djukic, T.; Vasovic, T.; Acimovic, J.; Sabljic, L.; Lukic, I.; Kovacevic, A.; Cujic, D.; Gnjatovic, M.; Smiljanic, K.; Stojadinovic, M.; Radosavljevic, J.; Stanic-Vucinic, D.; Stojanovic, M.; Rajkovic, A.; Cirkovic Velickovic, T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. International Journal of Molecular Sciences 2024, 25 (1), 333. https://doi.org/10.3390/ijms25010333.. in International Journal of Molecular Sciences. 2024;25(1).
https://hdl.handle.net/21.15107/rcub_intor_859 .
Mladenović Stokanić, Maja, Simović, Ana, Jovanović, Vesna, Radomirović, Mirjana, Udovički, Božidar, Krstić Ristivojević, Maja, Djukić, Teodora, Vasović, Tamara, Aćimović, Jelena, Sabljić, Ljiljana, Lukić, Ivana, Kovačević, Ana, Cujic, Danica, Gnjatović, Marija, Smiljanić, Katarina, Stojadinović, Marija, Radosavljević, Jelena, Stanić-Vučinić, Dragana, Stojanović, Marijana, Rajković, Andreja, Ćirkovic Veličković, Tanja, "Supplementary information for the article:       Mladenovic Stokanic, M.; Simovic, A.; Jovanovic, V.; Radomirovic, M.; Udovicki, B.; Krstic Ristivojevic, M.; Djukic, T.; Vasovic, T.; Acimovic, J.; Sabljic, L.; Lukic, I.; Kovacevic, A.; Cujic, D.; Gnjatovic, M.; Smiljanic, K.; Stojadinovic, M.; Radosavljevic, J.; Stanic-Vucinic, D.; Stojanovic, M.; Rajkovic, A.; Cirkovic Velickovic, T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. International Journal of Molecular Sciences 2024, 25 (1), 333. https://doi.org/10.3390/ijms25010333." in International Journal of Molecular Sciences, 25, no. 1 (2024),
https://hdl.handle.net/21.15107/rcub_intor_859 .

Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species

Mladenović Stokanić, Maja; Simović, Ana; Jovanović, Vesna; Radomirović, Mirjana; Udovički, Božidar; Krstić Ristivojević, Maja; Djukić, Teodora; Vasović, Tamara; Aćimović, Jelena; Sabljić, Ljiljana; Lukić, Ivana; Kovačević, Ana; Cujic, Danica; Gnjatović, Marija; Smiljanić, Katarina; Stojadinović, Marija; Radosavljević, Jelena; Stanić-Vučinić, Dragana; Stojanović, Marijana; Rajković, Andreja; Ćirkovic Veličković, Tanja

(MDPI, 2024)

TY  - JOUR
AU  - Mladenović Stokanić, Maja
AU  - Simović, Ana
AU  - Jovanović, Vesna
AU  - Radomirović, Mirjana
AU  - Udovički, Božidar
AU  - Krstić Ristivojević, Maja
AU  - Djukić, Teodora
AU  - Vasović, Tamara
AU  - Aćimović, Jelena
AU  - Sabljić, Ljiljana
AU  - Lukić, Ivana
AU  - Kovačević, Ana
AU  - Cujic, Danica
AU  - Gnjatović, Marija
AU  - Smiljanić, Katarina
AU  - Stojadinović, Marija
AU  - Radosavljević, Jelena
AU  - Stanić-Vučinić, Dragana
AU  - Stojanović, Marijana
AU  - Rajković, Andreja
AU  - Ćirkovic Veličković, Tanja
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/858
AB  - In this study, a cost-effective sandwich ELISA test, based on polyclonal antibodies, for routine quantification SARS-CoV-2 nucleocapsid (N) protein was developed. The recombinant N protein was produced and used for the production of mice and rabbit antisera. Polyclonal N protein-specific antibodies served as capture and detection antibodies. The prototype ELISA has LOD 0.93 ng/mL and LOQ 5.3 ng/mL, with a linear range of 1.52–48.83 ng/mL. N protein heat pretreatment (56 °C, 1 h) decreased, while pretreatment with 1% Triton X-100 increased analytical ELISA sensitivity. The diagnostic specificity of ELISA was 100% (95% CI, 91.19–100.00%) and sensitivity was 52.94% (95% CI, 35.13–70.22%) compared to rtRT-PCR (Ct < 40). Profoundly higher sensitivity was obtained using patient samples mostly containing Wuhan-similar variants (Wuhan, alpha, and delta), 62.50% (95% CI, 40.59 to 81.20%), in comparison to samples mostly containing Wuhan-distant variants (Omicron) 30.00% (6.67–65.25%). The developed product has relatively high diagnostic sensitivity in relation to its analytical sensitivity due to the usage of polyclonal antibodies from two species, providing a wide repertoire of antibodies against multiple N protein epitopes. Moreover, the fast, simple, and inexpensive production of polyclonal antibodies, as the most expensive assay components, would result in affordable antigen tests.
PB  - MDPI
T2  - International Journal of Molecular Sciences
T1  - Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species
IS  - 1
SP  - 333
VL  - 25
DO  - 10.3390/ijms25010333
ER  - 
@article{
author = "Mladenović Stokanić, Maja and Simović, Ana and Jovanović, Vesna and Radomirović, Mirjana and Udovički, Božidar and Krstić Ristivojević, Maja and Djukić, Teodora and Vasović, Tamara and Aćimović, Jelena and Sabljić, Ljiljana and Lukić, Ivana and Kovačević, Ana and Cujic, Danica and Gnjatović, Marija and Smiljanić, Katarina and Stojadinović, Marija and Radosavljević, Jelena and Stanić-Vučinić, Dragana and Stojanović, Marijana and Rajković, Andreja and Ćirkovic Veličković, Tanja",
year = "2024",
abstract = "In this study, a cost-effective sandwich ELISA test, based on polyclonal antibodies, for routine quantification SARS-CoV-2 nucleocapsid (N) protein was developed. The recombinant N protein was produced and used for the production of mice and rabbit antisera. Polyclonal N protein-specific antibodies served as capture and detection antibodies. The prototype ELISA has LOD 0.93 ng/mL and LOQ 5.3 ng/mL, with a linear range of 1.52–48.83 ng/mL. N protein heat pretreatment (56 °C, 1 h) decreased, while pretreatment with 1% Triton X-100 increased analytical ELISA sensitivity. The diagnostic specificity of ELISA was 100% (95% CI, 91.19–100.00%) and sensitivity was 52.94% (95% CI, 35.13–70.22%) compared to rtRT-PCR (Ct < 40). Profoundly higher sensitivity was obtained using patient samples mostly containing Wuhan-similar variants (Wuhan, alpha, and delta), 62.50% (95% CI, 40.59 to 81.20%), in comparison to samples mostly containing Wuhan-distant variants (Omicron) 30.00% (6.67–65.25%). The developed product has relatively high diagnostic sensitivity in relation to its analytical sensitivity due to the usage of polyclonal antibodies from two species, providing a wide repertoire of antibodies against multiple N protein epitopes. Moreover, the fast, simple, and inexpensive production of polyclonal antibodies, as the most expensive assay components, would result in affordable antigen tests.",
publisher = "MDPI",
journal = "International Journal of Molecular Sciences",
title = "Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species",
number = "1",
pages = "333",
volume = "25",
doi = "10.3390/ijms25010333"
}
Mladenović Stokanić, M., Simović, A., Jovanović, V., Radomirović, M., Udovički, B., Krstić Ristivojević, M., Djukić, T., Vasović, T., Aćimović, J., Sabljić, L., Lukić, I., Kovačević, A., Cujic, D., Gnjatović, M., Smiljanić, K., Stojadinović, M., Radosavljević, J., Stanić-Vučinić, D., Stojanović, M., Rajković, A.,& Ćirkovic Veličković, T.. (2024). Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. in International Journal of Molecular Sciences
MDPI., 25(1), 333.
https://doi.org/10.3390/ijms25010333
Mladenović Stokanić M, Simović A, Jovanović V, Radomirović M, Udovički B, Krstić Ristivojević M, Djukić T, Vasović T, Aćimović J, Sabljić L, Lukić I, Kovačević A, Cujic D, Gnjatović M, Smiljanić K, Stojadinović M, Radosavljević J, Stanić-Vučinić D, Stojanović M, Rajković A, Ćirkovic Veličković T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. in International Journal of Molecular Sciences. 2024;25(1):333.
doi:10.3390/ijms25010333 .
Mladenović Stokanić, Maja, Simović, Ana, Jovanović, Vesna, Radomirović, Mirjana, Udovički, Božidar, Krstić Ristivojević, Maja, Djukić, Teodora, Vasović, Tamara, Aćimović, Jelena, Sabljić, Ljiljana, Lukić, Ivana, Kovačević, Ana, Cujic, Danica, Gnjatović, Marija, Smiljanić, Katarina, Stojadinović, Marija, Radosavljević, Jelena, Stanić-Vučinić, Dragana, Stojanović, Marijana, Rajković, Andreja, Ćirkovic Veličković, Tanja, "Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species" in International Journal of Molecular Sciences, 25, no. 1 (2024):333,
https://doi.org/10.3390/ijms25010333 . .

Seasonal and inter-seasonal RSV activity in the European Region during the COVID-19 pandemic from autumn 2020 to summer 2022

Meslé, Margaux M. I.; Sinnathamby, Mary; Mook, Piers; Pebody, Richard; Protić, Jelena

(Wiley, 2023)

TY  - JOUR
AU  - Meslé, Margaux M. I.
AU  - Sinnathamby, Mary
AU  - Mook, Piers
AU  - Pebody, Richard
AU  - Protić, Jelena
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/846
AB  - Background The emergence of the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) in early 2020 and subsequent implementation of public health and social measures (PHSM) disrupted the epidemiology of respiratory viruses. This work describes the epidemiology of respiratory syncytial virus (RSV) observed during two winter seasons (weeks 40–20) and inter-seasonal periods (weeks 21–39) during the pandemic between October 2020 and September 2022. Methods Using data submitted to The European Surveillance System (TESSy) by countries or territories in the World Health Organization (WHO) European Region between weeks 40/2020 and 39/2022, we aggregated country-specific weekly RSV counts of sentinel, non-sentinel and Severe Acute Respiratory Infection (SARI) surveillance specimens and calculated percentage positivity. Results for both 2020/21 and 2021/22 seasons and inter-seasons were compared with pre-pandemic 2016/17 to 2019/20 seasons and inter-seasons. Results Although more specimens were tested than in pre-COVID-19 pandemic seasons, very few RSV detections were reported during the 2020/21 season in all surveillance systems. During the 2021 inter-season, a gradual increase in detections was observed in all systems. In 2021/22, all systems saw early peaks of RSV infection, and during the 2022 inter-seasonal period, patterns of detections were closer to those seen before the COVID-19 pandemic. Conclusion RSV surveillance continued throughout the COVID-19 pandemic, with an initial reduction in transmission, followed by very high and out-of-season RSV circulation (summer 2021) and then an early start of the 2021/22 season. As of the 2022/23 season, RSV circulation had not yet normalised.
PB  - Wiley
T2  - Influenza and Other Respiratory Viruses
T1  - Seasonal and inter-seasonal RSV activity in the European Region during the COVID-19 pandemic from autumn 2020 to summer 2022
IS  - 11
SP  - e13219
VL  - 17
DO  - 10.1111/irv.13219
ER  - 
@article{
author = "Meslé, Margaux M. I. and Sinnathamby, Mary and Mook, Piers and Pebody, Richard and Protić, Jelena",
year = "2023",
abstract = "Background The emergence of the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) in early 2020 and subsequent implementation of public health and social measures (PHSM) disrupted the epidemiology of respiratory viruses. This work describes the epidemiology of respiratory syncytial virus (RSV) observed during two winter seasons (weeks 40–20) and inter-seasonal periods (weeks 21–39) during the pandemic between October 2020 and September 2022. Methods Using data submitted to The European Surveillance System (TESSy) by countries or territories in the World Health Organization (WHO) European Region between weeks 40/2020 and 39/2022, we aggregated country-specific weekly RSV counts of sentinel, non-sentinel and Severe Acute Respiratory Infection (SARI) surveillance specimens and calculated percentage positivity. Results for both 2020/21 and 2021/22 seasons and inter-seasons were compared with pre-pandemic 2016/17 to 2019/20 seasons and inter-seasons. Results Although more specimens were tested than in pre-COVID-19 pandemic seasons, very few RSV detections were reported during the 2020/21 season in all surveillance systems. During the 2021 inter-season, a gradual increase in detections was observed in all systems. In 2021/22, all systems saw early peaks of RSV infection, and during the 2022 inter-seasonal period, patterns of detections were closer to those seen before the COVID-19 pandemic. Conclusion RSV surveillance continued throughout the COVID-19 pandemic, with an initial reduction in transmission, followed by very high and out-of-season RSV circulation (summer 2021) and then an early start of the 2021/22 season. As of the 2022/23 season, RSV circulation had not yet normalised.",
publisher = "Wiley",
journal = "Influenza and Other Respiratory Viruses",
title = "Seasonal and inter-seasonal RSV activity in the European Region during the COVID-19 pandemic from autumn 2020 to summer 2022",
number = "11",
pages = "e13219",
volume = "17",
doi = "10.1111/irv.13219"
}
Meslé, M. M. I., Sinnathamby, M., Mook, P., Pebody, R.,& Protić, J.. (2023). Seasonal and inter-seasonal RSV activity in the European Region during the COVID-19 pandemic from autumn 2020 to summer 2022. in Influenza and Other Respiratory Viruses
Wiley., 17(11), e13219.
https://doi.org/10.1111/irv.13219
Meslé MMI, Sinnathamby M, Mook P, Pebody R, Protić J. Seasonal and inter-seasonal RSV activity in the European Region during the COVID-19 pandemic from autumn 2020 to summer 2022. in Influenza and Other Respiratory Viruses. 2023;17(11):e13219.
doi:10.1111/irv.13219 .
Meslé, Margaux M. I., Sinnathamby, Mary, Mook, Piers, Pebody, Richard, Protić, Jelena, "Seasonal and inter-seasonal RSV activity in the European Region during the COVID-19 pandemic from autumn 2020 to summer 2022" in Influenza and Other Respiratory Viruses, 17, no. 11 (2023):e13219,
https://doi.org/10.1111/irv.13219 . .
1
3

Antimikrobna antitela u fiziološkim i patološkim stanjima

Minić, Rajna; Đorđević, Brižita; Michaličkova, Danica; Živković, Irena; Živančević Simonović, Snežana; Ilić, Vesna

(Beograd: Srpska akademija nauka i umetnosti, Odeljenje medicinskih nauka SANU, Odbor za imunologiju i alergologiju i Društvo imunologa Srbije, 2023)

TY  - CONF
AU  - Minić, Rajna
AU  - Đorđević, Brižita
AU  - Michaličkova, Danica
AU  - Živković, Irena
AU  - Živančević Simonović, Snežana
AU  - Ilić, Vesna
PY  - 2023
UR  - http://rimi.imi.bg.ac.rs/handle/123456789/1407
UR  - http://intor.torlakinstitut.com/handle/123456789/862
AB  - U ovom izlaganju biće predstavljeni naši dosadašnji rezultati na temu antimikrobnih antitela kod profesionalnih sportista, odraslih osoba koje se ne bave profesionalno sportom; promene nivoa antibakterijskih antitela pri starenju; poređenje specifičnosti serumskih i salivarnih IgA antitela kod mladih odraslih osoba, kao i nivoi ukupnih i antibakterijskih antitela u Covid-19 virusnoj infekciji i u sepsi. Nivoi i titri antimikrobnih antitela različitih klasa i potklasa određivani su ELISA testom iz uzoraka seruma, plazme ili salive. Ispitivano je vezivanje za izolovane antigene mikroorganizama (lipopolisaharid, peptidoglikan i zimozan) i za cele mikroorganizme, različitih vrsta i sojeva. Vezivanje prečišćenog salivarnog IgA za mikroorganizme praćeno je i protočnom citometrijom. Od rezultata izdvajamo da izlaganje fizičkom naporu visokog intenziteta dovodi do promena na nivou antibakterijskih antitela, i to u pravcu sveobuhvatnijeg prepoznavanja lipopolisaharida1. Suplementacija profesionalnih sportista određenim probioticima dovodi do održanja nivoa ukupnih salivarnih i serumskih IgA antitela, kao i IgG antitela specifičnih prema Enterococcus faecalis, koja su u ovoj populaciji pokazala sezonsku varijaciju. Pri starenju dolazi do smanjenja nivoa antipneumokoknih antitela i to zavisno od pola, pa su stariji mukarci naročito pogođeni ovim smanjenjem3. Salivarna IgA antitela razlikuju se po specifičnosti od serumskih IgA antitela i pružaju nespecifičnu zaštitu. Poređenje pacijenata sa sepsom, hospitalizovanih Covid-19 pacijenta i starosnih kontrola pokazalo je brojne razlike u nivoima kako ukupnih, tako i antibakterijskih antitela. Nivoi antibakterijskih antitela kod pacijentata koji nisu preživeli bili su niži nego kod onih koji su preživeli, što potvrđuje značaj ovih anititela za preživljavanje sepse.
PB  - Beograd: Srpska akademija nauka i umetnosti, Odeljenje medicinskih nauka SANU, Odbor za imunologiju i alergologiju i Društvo imunologa Srbije
C3  - Naučni skup Svetski dan imunologije, 27. april 2023. godine Svečana sala SANU - Knjiga sažetaka
T1  - Antimikrobna antitela u fiziološkim i patološkim stanjima
UR  - https://hdl.handle.net/21.15107/rcub_intor_862
ER  - 
@conference{
author = "Minić, Rajna and Đorđević, Brižita and Michaličkova, Danica and Živković, Irena and Živančević Simonović, Snežana and Ilić, Vesna",
year = "2023",
abstract = "U ovom izlaganju biće predstavljeni naši dosadašnji rezultati na temu antimikrobnih antitela kod profesionalnih sportista, odraslih osoba koje se ne bave profesionalno sportom; promene nivoa antibakterijskih antitela pri starenju; poređenje specifičnosti serumskih i salivarnih IgA antitela kod mladih odraslih osoba, kao i nivoi ukupnih i antibakterijskih antitela u Covid-19 virusnoj infekciji i u sepsi. Nivoi i titri antimikrobnih antitela različitih klasa i potklasa određivani su ELISA testom iz uzoraka seruma, plazme ili salive. Ispitivano je vezivanje za izolovane antigene mikroorganizama (lipopolisaharid, peptidoglikan i zimozan) i za cele mikroorganizme, različitih vrsta i sojeva. Vezivanje prečišćenog salivarnog IgA za mikroorganizme praćeno je i protočnom citometrijom. Od rezultata izdvajamo da izlaganje fizičkom naporu visokog intenziteta dovodi do promena na nivou antibakterijskih antitela, i to u pravcu sveobuhvatnijeg prepoznavanja lipopolisaharida1. Suplementacija profesionalnih sportista određenim probioticima dovodi do održanja nivoa ukupnih salivarnih i serumskih IgA antitela, kao i IgG antitela specifičnih prema Enterococcus faecalis, koja su u ovoj populaciji pokazala sezonsku varijaciju. Pri starenju dolazi do smanjenja nivoa antipneumokoknih antitela i to zavisno od pola, pa su stariji mukarci naročito pogođeni ovim smanjenjem3. Salivarna IgA antitela razlikuju se po specifičnosti od serumskih IgA antitela i pružaju nespecifičnu zaštitu. Poređenje pacijenata sa sepsom, hospitalizovanih Covid-19 pacijenta i starosnih kontrola pokazalo je brojne razlike u nivoima kako ukupnih, tako i antibakterijskih antitela. Nivoi antibakterijskih antitela kod pacijentata koji nisu preživeli bili su niži nego kod onih koji su preživeli, što potvrđuje značaj ovih anititela za preživljavanje sepse.",
publisher = "Beograd: Srpska akademija nauka i umetnosti, Odeljenje medicinskih nauka SANU, Odbor za imunologiju i alergologiju i Društvo imunologa Srbije",
journal = "Naučni skup Svetski dan imunologije, 27. april 2023. godine Svečana sala SANU - Knjiga sažetaka",
title = "Antimikrobna antitela u fiziološkim i patološkim stanjima",
url = "https://hdl.handle.net/21.15107/rcub_intor_862"
}
Minić, R., Đorđević, B., Michaličkova, D., Živković, I., Živančević Simonović, S.,& Ilić, V.. (2023). Antimikrobna antitela u fiziološkim i patološkim stanjima. in Naučni skup Svetski dan imunologije, 27. april 2023. godine Svečana sala SANU - Knjiga sažetaka
Beograd: Srpska akademija nauka i umetnosti, Odeljenje medicinskih nauka SANU, Odbor za imunologiju i alergologiju i Društvo imunologa Srbije..
https://hdl.handle.net/21.15107/rcub_intor_862
Minić R, Đorđević B, Michaličkova D, Živković I, Živančević Simonović S, Ilić V. Antimikrobna antitela u fiziološkim i patološkim stanjima. in Naučni skup Svetski dan imunologije, 27. april 2023. godine Svečana sala SANU - Knjiga sažetaka. 2023;.
https://hdl.handle.net/21.15107/rcub_intor_862 .
Minić, Rajna, Đorđević, Brižita, Michaličkova, Danica, Živković, Irena, Živančević Simonović, Snežana, Ilić, Vesna, "Antimikrobna antitela u fiziološkim i patološkim stanjima" in Naučni skup Svetski dan imunologije, 27. april 2023. godine Svečana sala SANU - Knjiga sažetaka (2023),
https://hdl.handle.net/21.15107/rcub_intor_862 .

Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus

Ćirković-Veličković, Tanja; Gnjatović, Marija; Ćujić, Danica; Todorović, Aleksandra; Stanić-Vučinić, Dragana; Đukić, Teodora; Mladenović, Maja; Vasović, Tamara; Stojadinović, Marija; Krstić-Ristivojević, Maja; Jovanović, Vesna; Simović, Ana; Radosavljević, Jelena; Aćimović, Jelena M.; Radomirović, Mirjana Ž.; Stojanović, Marijana

(2023)

TY  - GEN
AU  - Ćirković-Veličković, Tanja
AU  - Gnjatović, Marija
AU  - Ćujić, Danica
AU  - Todorović, Aleksandra
AU  - Stanić-Vučinić, Dragana
AU  - Đukić, Teodora
AU  - Mladenović, Maja
AU  - Vasović, Tamara
AU  - Stojadinović, Marija
AU  - Krstić-Ristivojević, Maja
AU  - Jovanović, Vesna
AU  - Simović, Ana
AU  - Radosavljević, Jelena
AU  - Aćimović, Jelena M.
AU  - Radomirović, Mirjana Ž.
AU  - Stojanović, Marijana
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/860
AB  - Novi korona virus (SARS CoV-2) koji se pojavio u Vuhanu 2019. godine pripada grupi jednolančanih RNK virusa [1]. Predstavlja novi infektivni agens za humanu populaciju i veoma je brzo detektovan u velikom broju zemalja. Uzročnik je respiratornih infekcija koje mogu da budu praćene i veoma teškom kliničkom slikom. Brzo širenje, odsustvo imuniteta na ovaj virus i odsustvo pouzdanih testova za detekciju virusa u trenutku izbijanja pandemije su bolest izazvanu ovim virusom brzo pretvorili u zdravstveni i društveni problem najvišeg prioriteta na globalnom nivou. Iako su najveće biotehnološke kompanije ubrzano počele sa razvojem i masovnom proizvodnjom dijagnostičkih testova i vakcina, njihova dostupnost u trenucima najveće potražnje je i dalje nedovoljna, a cene istih su limitirajući faktor za bolju kontrolu bolesti i širenja pandemije [2]. Razvoj sopstvenih i održiva proizvodnja testova i vakcina za COVID-19 su od velikog društvenog značaja. Važan preduslov za održivu proizvodnju testova je dostupnost rekombinantnih antigena virusa i mogućnost proizvodnje istih na velikoj skali za potrebe proizvodnje domaćih testova. Ovim tehničkim rešenjem se opisuje dobijanje dva ključna antigena novog korona virusa rekombinantnom tehnologijom i njihova primena u serološkom ELISA testu koji proizvodi Institut za primenu nuklearne energije, INEP, kao i za dobijanje reagenasa za detekciju antigena novog korona virusa (specifičnih antitela). U prvoj fazi, optimizovane su sekvence proteina koje su podigle osetljivost postojećih seroloških testova. Inovativnost našeg pristupa se ogleda i u razrađenim eksperimentalnim protokolima za dobijanje rekombinantnih proteina nukleokapsida na velikoj skali, kao i u solubilnoj formi, što olakšava postupak prečišćavanja. Izbor fragmenta nukleokapsida koji se heterologo eksprimira u solubilnoj formi, a specifično detektuje antitela i generiše jak imuni odgovor tokom imunizacije životinja (imunogenost) na osnovu pregleda poznatih epitopskih sekvenci je ključna inovacija ovog tehničkog rešenja. Ovo je prvi primer uspešno primenjenog rekombinatnog proteina proizvedenog u Srbiji u dijagnostičkom testu koji je registrovankod Agencije za lekove i medicinska sredstva Republike Srbije (broj rešenja 515-02-02370-21-002), a koji je primenu našao i na međunarodnom nivou.
T1  - Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus
UR  - https://hdl.handle.net/21.15107/rcub_intor_860
ER  - 
@misc{
author = "Ćirković-Veličković, Tanja and Gnjatović, Marija and Ćujić, Danica and Todorović, Aleksandra and Stanić-Vučinić, Dragana and Đukić, Teodora and Mladenović, Maja and Vasović, Tamara and Stojadinović, Marija and Krstić-Ristivojević, Maja and Jovanović, Vesna and Simović, Ana and Radosavljević, Jelena and Aćimović, Jelena M. and Radomirović, Mirjana Ž. and Stojanović, Marijana",
year = "2023",
abstract = "Novi korona virus (SARS CoV-2) koji se pojavio u Vuhanu 2019. godine pripada grupi jednolančanih RNK virusa [1]. Predstavlja novi infektivni agens za humanu populaciju i veoma je brzo detektovan u velikom broju zemalja. Uzročnik je respiratornih infekcija koje mogu da budu praćene i veoma teškom kliničkom slikom. Brzo širenje, odsustvo imuniteta na ovaj virus i odsustvo pouzdanih testova za detekciju virusa u trenutku izbijanja pandemije su bolest izazvanu ovim virusom brzo pretvorili u zdravstveni i društveni problem najvišeg prioriteta na globalnom nivou. Iako su najveće biotehnološke kompanije ubrzano počele sa razvojem i masovnom proizvodnjom dijagnostičkih testova i vakcina, njihova dostupnost u trenucima najveće potražnje je i dalje nedovoljna, a cene istih su limitirajući faktor za bolju kontrolu bolesti i širenja pandemije [2]. Razvoj sopstvenih i održiva proizvodnja testova i vakcina za COVID-19 su od velikog društvenog značaja. Važan preduslov za održivu proizvodnju testova je dostupnost rekombinantnih antigena virusa i mogućnost proizvodnje istih na velikoj skali za potrebe proizvodnje domaćih testova. Ovim tehničkim rešenjem se opisuje dobijanje dva ključna antigena novog korona virusa rekombinantnom tehnologijom i njihova primena u serološkom ELISA testu koji proizvodi Institut za primenu nuklearne energije, INEP, kao i za dobijanje reagenasa za detekciju antigena novog korona virusa (specifičnih antitela). U prvoj fazi, optimizovane su sekvence proteina koje su podigle osetljivost postojećih seroloških testova. Inovativnost našeg pristupa se ogleda i u razrađenim eksperimentalnim protokolima za dobijanje rekombinantnih proteina nukleokapsida na velikoj skali, kao i u solubilnoj formi, što olakšava postupak prečišćavanja. Izbor fragmenta nukleokapsida koji se heterologo eksprimira u solubilnoj formi, a specifično detektuje antitela i generiše jak imuni odgovor tokom imunizacije životinja (imunogenost) na osnovu pregleda poznatih epitopskih sekvenci je ključna inovacija ovog tehničkog rešenja. Ovo je prvi primer uspešno primenjenog rekombinatnog proteina proizvedenog u Srbiji u dijagnostičkom testu koji je registrovankod Agencije za lekove i medicinska sredstva Republike Srbije (broj rešenja 515-02-02370-21-002), a koji je primenu našao i na međunarodnom nivou.",
title = "Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus",
url = "https://hdl.handle.net/21.15107/rcub_intor_860"
}
Ćirković-Veličković, T., Gnjatović, M., Ćujić, D., Todorović, A., Stanić-Vučinić, D., Đukić, T., Mladenović, M., Vasović, T., Stojadinović, M., Krstić-Ristivojević, M., Jovanović, V., Simović, A., Radosavljević, J., Aćimović, J. M., Radomirović, M. Ž.,& Stojanović, M.. (2023). Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus. .
https://hdl.handle.net/21.15107/rcub_intor_860
Ćirković-Veličković T, Gnjatović M, Ćujić D, Todorović A, Stanić-Vučinić D, Đukić T, Mladenović M, Vasović T, Stojadinović M, Krstić-Ristivojević M, Jovanović V, Simović A, Radosavljević J, Aćimović JM, Radomirović MŽ, Stojanović M. Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus. 2023;.
https://hdl.handle.net/21.15107/rcub_intor_860 .
Ćirković-Veličković, Tanja, Gnjatović, Marija, Ćujić, Danica, Todorović, Aleksandra, Stanić-Vučinić, Dragana, Đukić, Teodora, Mladenović, Maja, Vasović, Tamara, Stojadinović, Marija, Krstić-Ristivojević, Maja, Jovanović, Vesna, Simović, Ana, Radosavljević, Jelena, Aćimović, Jelena M., Radomirović, Mirjana Ž., Stojanović, Marijana, "Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus" (2023),
https://hdl.handle.net/21.15107/rcub_intor_860 .

Multistep Approach Points to Compounds Responsible for the Biological Activity and Safety of Hydrolates from Nine Lamiaceae Medicinal Plants on Human Skin Fibroblasts

Smiljanić, Katarina; Prodić, Ivana; Trifunovic, Sara; Krstić Ristivojević, Maja; Aćimović, Milica; Stanković Jeremić, Jovana; Lončar, Biljana; Tešević, Vele

(MDPI, 2023)

TY  - JOUR
AU  - Smiljanić, Katarina
AU  - Prodić, Ivana
AU  - Trifunovic, Sara
AU  - Krstić Ristivojević, Maja
AU  - Aćimović, Milica
AU  - Stanković Jeremić, Jovana
AU  - Lončar, Biljana
AU  - Tešević, Vele
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/857
AB  - As byproducts of essential oil distillation, hydrolates are used in natural cosmetics/biomedicine due to their beneficial skin effects. However, data on their safety with relevant biological targets, such as human skin cells, are scarce. Therefore, we have tested nine hydrolates from the Lamiaceae family with skin fibroblasts that are responsible for extracellular collagenous matrix builds. Thyme, oregano, and winter savoury hydrolates showed several times higher total phenolics, which correlated strongly with their radical scavenging and antioxidative capacity; there was no correlation between their viability profiles and the reducing sugar levels. No proteins/peptides were detected. All hydrolates appeared safe for prolonged skin exposure except for 10-fold diluted lavender, which showed cytotoxicity (~20%), as well as rosemary and lavandin (~10%) using viability, DNA synthesis, and cell count testing. Clary sage, oregano, lemon balm, and thyme hydrolates (10-fold diluted) increased fibroblast viability and/or proliferation by 10–30% compared with the control, while their viability remained unaffected by Mentha and winter savoury. In line with the STITCH database, increased viability could be attributed to thymol presence in oregano and thyme hydrolates in lemon balm, which is most likely attributable to neral and geranial. The proliferative effect of clary sage could be supported by alpha-terpineol, not linalool. The major volatile organic compounds (VOCs) associated with cytotoxic effects on fibroblasts were borneol, 1,8-cineole, and terpinene-4-ol. Further research with pure compounds is warranted to confirm the roles of VOCs in the observed effects that are relevant to cosmetic and wound healing aspects.
PB  - MDPI
T2  - Antioxidants
T2  - Antioxidants
T1  - Multistep Approach Points to Compounds Responsible for the Biological Activity and Safety of Hydrolates from Nine Lamiaceae Medicinal Plants on Human Skin Fibroblasts
IS  - 11
SP  - 1988
VL  - 12
DO  - 10.3390/antiox12111988
ER  - 
@article{
author = "Smiljanić, Katarina and Prodić, Ivana and Trifunovic, Sara and Krstić Ristivojević, Maja and Aćimović, Milica and Stanković Jeremić, Jovana and Lončar, Biljana and Tešević, Vele",
year = "2023",
abstract = "As byproducts of essential oil distillation, hydrolates are used in natural cosmetics/biomedicine due to their beneficial skin effects. However, data on their safety with relevant biological targets, such as human skin cells, are scarce. Therefore, we have tested nine hydrolates from the Lamiaceae family with skin fibroblasts that are responsible for extracellular collagenous matrix builds. Thyme, oregano, and winter savoury hydrolates showed several times higher total phenolics, which correlated strongly with their radical scavenging and antioxidative capacity; there was no correlation between their viability profiles and the reducing sugar levels. No proteins/peptides were detected. All hydrolates appeared safe for prolonged skin exposure except for 10-fold diluted lavender, which showed cytotoxicity (~20%), as well as rosemary and lavandin (~10%) using viability, DNA synthesis, and cell count testing. Clary sage, oregano, lemon balm, and thyme hydrolates (10-fold diluted) increased fibroblast viability and/or proliferation by 10–30% compared with the control, while their viability remained unaffected by Mentha and winter savoury. In line with the STITCH database, increased viability could be attributed to thymol presence in oregano and thyme hydrolates in lemon balm, which is most likely attributable to neral and geranial. The proliferative effect of clary sage could be supported by alpha-terpineol, not linalool. The major volatile organic compounds (VOCs) associated with cytotoxic effects on fibroblasts were borneol, 1,8-cineole, and terpinene-4-ol. Further research with pure compounds is warranted to confirm the roles of VOCs in the observed effects that are relevant to cosmetic and wound healing aspects.",
publisher = "MDPI",
journal = "Antioxidants, Antioxidants",
title = "Multistep Approach Points to Compounds Responsible for the Biological Activity and Safety of Hydrolates from Nine Lamiaceae Medicinal Plants on Human Skin Fibroblasts",
number = "11",
pages = "1988",
volume = "12",
doi = "10.3390/antiox12111988"
}
Smiljanić, K., Prodić, I., Trifunovic, S., Krstić Ristivojević, M., Aćimović, M., Stanković Jeremić, J., Lončar, B.,& Tešević, V.. (2023). Multistep Approach Points to Compounds Responsible for the Biological Activity and Safety of Hydrolates from Nine Lamiaceae Medicinal Plants on Human Skin Fibroblasts. in Antioxidants
MDPI., 12(11), 1988.
https://doi.org/10.3390/antiox12111988
Smiljanić K, Prodić I, Trifunovic S, Krstić Ristivojević M, Aćimović M, Stanković Jeremić J, Lončar B, Tešević V. Multistep Approach Points to Compounds Responsible for the Biological Activity and Safety of Hydrolates from Nine Lamiaceae Medicinal Plants on Human Skin Fibroblasts. in Antioxidants. 2023;12(11):1988.
doi:10.3390/antiox12111988 .
Smiljanić, Katarina, Prodić, Ivana, Trifunovic, Sara, Krstić Ristivojević, Maja, Aćimović, Milica, Stanković Jeremić, Jovana, Lončar, Biljana, Tešević, Vele, "Multistep Approach Points to Compounds Responsible for the Biological Activity and Safety of Hydrolates from Nine Lamiaceae Medicinal Plants on Human Skin Fibroblasts" in Antioxidants, 12, no. 11 (2023):1988,
https://doi.org/10.3390/antiox12111988 . .
2

Comparative study between virus neutralisation testing and other serological methods detecting anti-SARS-CoV-2 antibodies in Europe, 2021

Mögling, Ramona; Reimerink, Johan; Stanoeva, Kamelia R.; Keramarou, Maria; Guiomar, Raquel; Costa, Inês; Haveri, Anu; Holzer, Barbara; Korukluoğlu, Gülay; Nguyen, Trung; Pakarna, Gatis; Pancer, Katarzyna; Trilar, Katarina Prosenc; Protić, Jelena; Stojanović, Marijana; De Santis, Riccardo; Lista, Florigio; Vremera, Teodora; Leustean, Mihaela; Pistol, Adriana; Zelena, Hana; Reusken, Chantal; Broberg, Eeva K.

(Elsevier, 2023)

TY  - JOUR
AU  - Mögling, Ramona
AU  - Reimerink, Johan
AU  - Stanoeva, Kamelia R.
AU  - Keramarou, Maria
AU  - Guiomar, Raquel
AU  - Costa, Inês
AU  - Haveri, Anu
AU  - Holzer, Barbara
AU  - Korukluoğlu, Gülay
AU  - Nguyen, Trung
AU  - Pakarna, Gatis
AU  - Pancer, Katarzyna
AU  - Trilar, Katarina Prosenc
AU  - Protić, Jelena
AU  - Stojanović, Marijana
AU  - De Santis, Riccardo
AU  - Lista, Florigio
AU  - Vremera, Teodora
AU  - Leustean, Mihaela
AU  - Pistol, Adriana
AU  - Zelena, Hana
AU  - Reusken, Chantal
AU  - Broberg, Eeva K.
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/805
AB  - One consequence of the ongoing coronavirus disease pandemic was the rapid development of both in-house and commercial serological assays detecting anti-SARS-CoV-2 antibodies, in an effort to reliably detect acute and past SARS-CoV-2 infections. It is crucial to evaluate the quality of these serological tests and consequently the sero-epidemiological studies that are performed with the respective tests. Here, we describe the set-up and results of a comparative study, in which a laboratory contracted by the European Centre for Disease Prevention and Control offered a centralised service to EU/EEA Member and pre-accession Member States to test representative serum specimens with known serological results, with the gold standard technique (virus neutralisation tests) to determine the presence of neutralising antibodies. Laboratories from 12 European countries shared 719 serum specimens with the contractor laboratory. We found that in-house serological tests detecting neutralising antibodies showed the highest percent agreement, both positive and negative, with the virus neutralisation test results. Despite extensive differences in virus neutralisation protocols neutralisation titres showed a strong correlation. From the commercial assays, the best positive percent agreement was found for SARS-CoV-2 IgG (sCOVG) (Siemens - Atellica IM Analyzer). Despite lower positive percent agreement of LIAISON SARS-CoV-2 TrimericS IgG kit (Diasorin Inc.), the obtained results showed relatively good correlation with neutralisation titres. The set-up of this study allowed for high comparability between laboratories and enabled laboratories that do not have the capacity or capability to perform VNTs themselves. Given the variety of in-house protocols detecting SARS-CoV-2 specific neutralising antibodies, including the virus strain, it could be of interest to select reference isolates for SARS-CoV-2 diagnostic to be made available for interested EU Member States and pre-accession countries.
PB  - Elsevier
T2  - Journal of Virological Methods
T1  - Comparative study between virus neutralisation testing and other serological methods detecting anti-SARS-CoV-2 antibodies in Europe, 2021
SP  - 114825
VL  - 322
DO  - 10.1016/j.jviromet.2023.114825
ER  - 
@article{
author = "Mögling, Ramona and Reimerink, Johan and Stanoeva, Kamelia R. and Keramarou, Maria and Guiomar, Raquel and Costa, Inês and Haveri, Anu and Holzer, Barbara and Korukluoğlu, Gülay and Nguyen, Trung and Pakarna, Gatis and Pancer, Katarzyna and Trilar, Katarina Prosenc and Protić, Jelena and Stojanović, Marijana and De Santis, Riccardo and Lista, Florigio and Vremera, Teodora and Leustean, Mihaela and Pistol, Adriana and Zelena, Hana and Reusken, Chantal and Broberg, Eeva K.",
year = "2023",
abstract = "One consequence of the ongoing coronavirus disease pandemic was the rapid development of both in-house and commercial serological assays detecting anti-SARS-CoV-2 antibodies, in an effort to reliably detect acute and past SARS-CoV-2 infections. It is crucial to evaluate the quality of these serological tests and consequently the sero-epidemiological studies that are performed with the respective tests. Here, we describe the set-up and results of a comparative study, in which a laboratory contracted by the European Centre for Disease Prevention and Control offered a centralised service to EU/EEA Member and pre-accession Member States to test representative serum specimens with known serological results, with the gold standard technique (virus neutralisation tests) to determine the presence of neutralising antibodies. Laboratories from 12 European countries shared 719 serum specimens with the contractor laboratory. We found that in-house serological tests detecting neutralising antibodies showed the highest percent agreement, both positive and negative, with the virus neutralisation test results. Despite extensive differences in virus neutralisation protocols neutralisation titres showed a strong correlation. From the commercial assays, the best positive percent agreement was found for SARS-CoV-2 IgG (sCOVG) (Siemens - Atellica IM Analyzer). Despite lower positive percent agreement of LIAISON SARS-CoV-2 TrimericS IgG kit (Diasorin Inc.), the obtained results showed relatively good correlation with neutralisation titres. The set-up of this study allowed for high comparability between laboratories and enabled laboratories that do not have the capacity or capability to perform VNTs themselves. Given the variety of in-house protocols detecting SARS-CoV-2 specific neutralising antibodies, including the virus strain, it could be of interest to select reference isolates for SARS-CoV-2 diagnostic to be made available for interested EU Member States and pre-accession countries.",
publisher = "Elsevier",
journal = "Journal of Virological Methods",
title = "Comparative study between virus neutralisation testing and other serological methods detecting anti-SARS-CoV-2 antibodies in Europe, 2021",
pages = "114825",
volume = "322",
doi = "10.1016/j.jviromet.2023.114825"
}
Mögling, R., Reimerink, J., Stanoeva, K. R., Keramarou, M., Guiomar, R., Costa, I., Haveri, A., Holzer, B., Korukluoğlu, G., Nguyen, T., Pakarna, G., Pancer, K., Trilar, K. P., Protić, J., Stojanović, M., De Santis, R., Lista, F., Vremera, T., Leustean, M., Pistol, A., Zelena, H., Reusken, C.,& Broberg, E. K.. (2023). Comparative study between virus neutralisation testing and other serological methods detecting anti-SARS-CoV-2 antibodies in Europe, 2021. in Journal of Virological Methods
Elsevier., 322, 114825.
https://doi.org/10.1016/j.jviromet.2023.114825
Mögling R, Reimerink J, Stanoeva KR, Keramarou M, Guiomar R, Costa I, Haveri A, Holzer B, Korukluoğlu G, Nguyen T, Pakarna G, Pancer K, Trilar KP, Protić J, Stojanović M, De Santis R, Lista F, Vremera T, Leustean M, Pistol A, Zelena H, Reusken C, Broberg EK. Comparative study between virus neutralisation testing and other serological methods detecting anti-SARS-CoV-2 antibodies in Europe, 2021. in Journal of Virological Methods. 2023;322:114825.
doi:10.1016/j.jviromet.2023.114825 .
Mögling, Ramona, Reimerink, Johan, Stanoeva, Kamelia R., Keramarou, Maria, Guiomar, Raquel, Costa, Inês, Haveri, Anu, Holzer, Barbara, Korukluoğlu, Gülay, Nguyen, Trung, Pakarna, Gatis, Pancer, Katarzyna, Trilar, Katarina Prosenc, Protić, Jelena, Stojanović, Marijana, De Santis, Riccardo, Lista, Florigio, Vremera, Teodora, Leustean, Mihaela, Pistol, Adriana, Zelena, Hana, Reusken, Chantal, Broberg, Eeva K., "Comparative study between virus neutralisation testing and other serological methods detecting anti-SARS-CoV-2 antibodies in Europe, 2021" in Journal of Virological Methods, 322 (2023):114825,
https://doi.org/10.1016/j.jviromet.2023.114825 . .
1

Erythrocyte fatty acid aberrations in Amyotrophic Lateral Sclerosis: Correlation with disease duration

Minić, Rajna; Arsić, Aleksandra; Kojadinović, Milica; Palibrk, Aleksa; Đorđević, Brižita; Stević, Zorica

(Društvo medicinskih biohemičara Srbije, 2023)

TY  - JOUR
AU  - Minić, Rajna
AU  - Arsić, Aleksandra
AU  - Kojadinović, Milica
AU  - Palibrk, Aleksa
AU  - Đorđević, Brižita
AU  - Stević, Zorica
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/842
AB  - Background: Recent literature data highlights metabolic changes in amyotrophic lateral sclerosis (ALS). To explore possible early metabolic changes, we aimed to analyse the fatty acids (FA) composition of erythrocytes in newly diagnosed als patients and to see whether fatty acid levels correlate with the ALSFRS-R score or disease duration. Methods: The severity of motor function involvement was assessed by the ALSFRS-R scale at the initial evaluation. The fatty acid profile of erythrocyte membranes was analysed by gas-liquid chromatography. The study comprised 26 clinically diagnosed als patients, with mean ALSFRS-R 38±8. The control group included 26 healthy volunteers. Results: Significantly higher levels of palmitic acid and total saturated FAs were detected in als patients. In als patients, total monounsaturated FA, palmitoleic, vaccenic, and oleic acid were also significantly increased. The levels of eicosapentaenoic acid, docosapentaenoic acid, total polyunsaturated FA (PUFA) and n-6 PUFA were significantly lower in als patients. Additionally, a-linolenic acid, the precursor of the n-3 PUFA family, was not detected in als patients. We found no significant correlation between the ALSFRS-R score and the abundance of individual FAs analysed. A moderate negative correlation was found between disease duration and DHA level, and a positive correlation was detected with MUFA. Conclusion: Experimental evidence presented may contribute to shaping a beneficial nutritional intervention.
PB  - Društvo medicinskih biohemičara Srbije
T2  - Journal of Medical Biochemistry
T2  - Journal of Medical Biochemistry
T1  - Erythrocyte fatty acid aberrations in Amyotrophic Lateral Sclerosis: Correlation with disease duration
EP  - 629
IS  - 4
SP  - 621
VL  - 42
DO  - 10.5937/jomb0-40387
ER  - 
@article{
author = "Minić, Rajna and Arsić, Aleksandra and Kojadinović, Milica and Palibrk, Aleksa and Đorđević, Brižita and Stević, Zorica",
year = "2023",
abstract = "Background: Recent literature data highlights metabolic changes in amyotrophic lateral sclerosis (ALS). To explore possible early metabolic changes, we aimed to analyse the fatty acids (FA) composition of erythrocytes in newly diagnosed als patients and to see whether fatty acid levels correlate with the ALSFRS-R score or disease duration. Methods: The severity of motor function involvement was assessed by the ALSFRS-R scale at the initial evaluation. The fatty acid profile of erythrocyte membranes was analysed by gas-liquid chromatography. The study comprised 26 clinically diagnosed als patients, with mean ALSFRS-R 38±8. The control group included 26 healthy volunteers. Results: Significantly higher levels of palmitic acid and total saturated FAs were detected in als patients. In als patients, total monounsaturated FA, palmitoleic, vaccenic, and oleic acid were also significantly increased. The levels of eicosapentaenoic acid, docosapentaenoic acid, total polyunsaturated FA (PUFA) and n-6 PUFA were significantly lower in als patients. Additionally, a-linolenic acid, the precursor of the n-3 PUFA family, was not detected in als patients. We found no significant correlation between the ALSFRS-R score and the abundance of individual FAs analysed. A moderate negative correlation was found between disease duration and DHA level, and a positive correlation was detected with MUFA. Conclusion: Experimental evidence presented may contribute to shaping a beneficial nutritional intervention.",
publisher = "Društvo medicinskih biohemičara Srbije",
journal = "Journal of Medical Biochemistry, Journal of Medical Biochemistry",
title = "Erythrocyte fatty acid aberrations in Amyotrophic Lateral Sclerosis: Correlation with disease duration",
pages = "629-621",
number = "4",
volume = "42",
doi = "10.5937/jomb0-40387"
}
Minić, R., Arsić, A., Kojadinović, M., Palibrk, A., Đorđević, B.,& Stević, Z.. (2023). Erythrocyte fatty acid aberrations in Amyotrophic Lateral Sclerosis: Correlation with disease duration. in Journal of Medical Biochemistry
Društvo medicinskih biohemičara Srbije., 42(4), 621-629.
https://doi.org/10.5937/jomb0-40387
Minić R, Arsić A, Kojadinović M, Palibrk A, Đorđević B, Stević Z. Erythrocyte fatty acid aberrations in Amyotrophic Lateral Sclerosis: Correlation with disease duration. in Journal of Medical Biochemistry. 2023;42(4):621-629.
doi:10.5937/jomb0-40387 .
Minić, Rajna, Arsić, Aleksandra, Kojadinović, Milica, Palibrk, Aleksa, Đorđević, Brižita, Stević, Zorica, "Erythrocyte fatty acid aberrations in Amyotrophic Lateral Sclerosis: Correlation with disease duration" in Journal of Medical Biochemistry, 42, no. 4 (2023):621-629,
https://doi.org/10.5937/jomb0-40387 . .

Dynamic light scattering analysis of immune complexes in sera of rheumatoid arthritis patients

Đukić, Tamara; Drvenica, Ivana; Kovačić, Marijana; Minić, Rajna; Vučetić, Dušan; Majerič, Dragana; Šefik-Bukilica, Mirjana; Savić, Olivera; Bugarski, Branko; Ilić, Vesna

(Elsevier, 2023)

TY  - JOUR
AU  - Đukić, Tamara
AU  - Drvenica, Ivana
AU  - Kovačić, Marijana
AU  - Minić, Rajna
AU  - Vučetić, Dušan
AU  - Majerič, Dragana
AU  - Šefik-Bukilica, Mirjana
AU  - Savić, Olivera
AU  - Bugarski, Branko
AU  - Ilić, Vesna
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/778
AB  - The size of circulating immune complexes (CICs) in rheumatoid arthritis (RA) could be an emerging criterion in disease diagnosis. This study analyzed size and electrokinetic potential of CICs from RA patients, healthy young adults, and RA patients age-matched controls aiming to establish their unique CIC features. Pooled CIC of 30 RA patients, 30 young adults, and 30 RA group's age-matched controls (middle-aged and oldеr healthy adults), and in vitro IgG aggregates from pooled sera of 300 healthy volunteers were tested using dynamic light scattering (DLS). Size distribution of CIC in healthy young adults exhibited high polydispersity. RA CIC patients and their age-matched control showed distinctly narrower size distributions compared with young adults. In these groups, particles clustered around two well-defined peaks. Particles of peak 1 were 36.1 ± 6.8 nm in RA age-matched control, and 30.8 ± 4.2 nm in RA patients. Particles of peak 2 of the RA age-matched control's CIC was 251.7 ± 41.2 nm, while RA CIC contained larger particles (359.9 ± 50.5 nm). The lower zeta potential of RA CIC, compared to control, indicated a disease-related decrease in colloidal stability. DLS identified RA-specific, but also age-specific distribution of CIC size and opened possibility of becoming a method for CIC size analysis in IC-mediated diseases.
PB  - Elsevier
T2  - Analytical Biochemistry
T2  - Analytical BiochemistryAnalytical Biochemistry
T1  - Dynamic light scattering analysis of immune complexes in sera of rheumatoid arthritis patients
SP  - 115194
VL  - 674
DO  - 10.1016/j.ab.2023.115194
ER  - 
@article{
author = "Đukić, Tamara and Drvenica, Ivana and Kovačić, Marijana and Minić, Rajna and Vučetić, Dušan and Majerič, Dragana and Šefik-Bukilica, Mirjana and Savić, Olivera and Bugarski, Branko and Ilić, Vesna",
year = "2023",
abstract = "The size of circulating immune complexes (CICs) in rheumatoid arthritis (RA) could be an emerging criterion in disease diagnosis. This study analyzed size and electrokinetic potential of CICs from RA patients, healthy young adults, and RA patients age-matched controls aiming to establish their unique CIC features. Pooled CIC of 30 RA patients, 30 young adults, and 30 RA group's age-matched controls (middle-aged and oldеr healthy adults), and in vitro IgG aggregates from pooled sera of 300 healthy volunteers were tested using dynamic light scattering (DLS). Size distribution of CIC in healthy young adults exhibited high polydispersity. RA CIC patients and their age-matched control showed distinctly narrower size distributions compared with young adults. In these groups, particles clustered around two well-defined peaks. Particles of peak 1 were 36.1 ± 6.8 nm in RA age-matched control, and 30.8 ± 4.2 nm in RA patients. Particles of peak 2 of the RA age-matched control's CIC was 251.7 ± 41.2 nm, while RA CIC contained larger particles (359.9 ± 50.5 nm). The lower zeta potential of RA CIC, compared to control, indicated a disease-related decrease in colloidal stability. DLS identified RA-specific, but also age-specific distribution of CIC size and opened possibility of becoming a method for CIC size analysis in IC-mediated diseases.",
publisher = "Elsevier",
journal = "Analytical Biochemistry, Analytical BiochemistryAnalytical Biochemistry",
title = "Dynamic light scattering analysis of immune complexes in sera of rheumatoid arthritis patients",
pages = "115194",
volume = "674",
doi = "10.1016/j.ab.2023.115194"
}
Đukić, T., Drvenica, I., Kovačić, M., Minić, R., Vučetić, D., Majerič, D., Šefik-Bukilica, M., Savić, O., Bugarski, B.,& Ilić, V.. (2023). Dynamic light scattering analysis of immune complexes in sera of rheumatoid arthritis patients. in Analytical Biochemistry
Elsevier., 674, 115194.
https://doi.org/10.1016/j.ab.2023.115194
Đukić T, Drvenica I, Kovačić M, Minić R, Vučetić D, Majerič D, Šefik-Bukilica M, Savić O, Bugarski B, Ilić V. Dynamic light scattering analysis of immune complexes in sera of rheumatoid arthritis patients. in Analytical Biochemistry. 2023;674:115194.
doi:10.1016/j.ab.2023.115194 .
Đukić, Tamara, Drvenica, Ivana, Kovačić, Marijana, Minić, Rajna, Vučetić, Dušan, Majerič, Dragana, Šefik-Bukilica, Mirjana, Savić, Olivera, Bugarski, Branko, Ilić, Vesna, "Dynamic light scattering analysis of immune complexes in sera of rheumatoid arthritis patients" in Analytical Biochemistry, 674 (2023):115194,
https://doi.org/10.1016/j.ab.2023.115194 . .
1

Fatty acid profiling in amyotrophic lateral sclerosis

Minić, Rajna; Stević, Zorica; Arsić, Aleksandra.

(Elsevier, 2023)

TY  - CHAP
AU  - Minić, Rajna
AU  - Stević, Zorica
AU  - Arsić, Aleksandra.
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/656
AB  - Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease, affecting motor neurons, which is characterized by the progressive loss of motor function. Sufficient evidence has been gathered, either in the form of experimental results from animal disease models or from the assessment of affected human subjects, which implicates metabolic changes in the course of ALS. Hypermetabolism, abnormal glucose tolerance and dyslipidaemia were reported to have high incidence in ALS patients. In fact, according to published studies, BMI is significantly and inversely associated with ALS progression. Here, we discuss basic principles of fatty acid metabolism, composition and the existing knowledge of metabolic and fatty acid composition changes in ALS patients. Literature data suggest that ALS patients have unfavorable changes in the fatty acid profile, especially because of the low level of long-chain polyunsaturated fatty acids. Nutritional interventions and supplementation with various foods or food supplements are currently not showing conclusive results, implying there is more to learn about potential ways to modify this illness.
PB  - Elsevier
T2  - Diet and Nutrition in Neurological Disorders
T1  - Fatty acid profiling in amyotrophic lateral sclerosis
EP  - 172
SP  - 155
DO  - 10.1016/B978-0-323-89834-8.00023-4
ER  - 
@inbook{
author = "Minić, Rajna and Stević, Zorica and Arsić, Aleksandra.",
year = "2023",
abstract = "Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease, affecting motor neurons, which is characterized by the progressive loss of motor function. Sufficient evidence has been gathered, either in the form of experimental results from animal disease models or from the assessment of affected human subjects, which implicates metabolic changes in the course of ALS. Hypermetabolism, abnormal glucose tolerance and dyslipidaemia were reported to have high incidence in ALS patients. In fact, according to published studies, BMI is significantly and inversely associated with ALS progression. Here, we discuss basic principles of fatty acid metabolism, composition and the existing knowledge of metabolic and fatty acid composition changes in ALS patients. Literature data suggest that ALS patients have unfavorable changes in the fatty acid profile, especially because of the low level of long-chain polyunsaturated fatty acids. Nutritional interventions and supplementation with various foods or food supplements are currently not showing conclusive results, implying there is more to learn about potential ways to modify this illness.",
publisher = "Elsevier",
journal = "Diet and Nutrition in Neurological Disorders",
booktitle = "Fatty acid profiling in amyotrophic lateral sclerosis",
pages = "172-155",
doi = "10.1016/B978-0-323-89834-8.00023-4"
}
Minić, R., Stević, Z.,& Arsić, Aleksandra.. (2023). Fatty acid profiling in amyotrophic lateral sclerosis. in Diet and Nutrition in Neurological Disorders
Elsevier., 155-172.
https://doi.org/10.1016/B978-0-323-89834-8.00023-4
Minić R, Stević Z, Arsić A. Fatty acid profiling in amyotrophic lateral sclerosis. in Diet and Nutrition in Neurological Disorders. 2023;:155-172.
doi:10.1016/B978-0-323-89834-8.00023-4 .
Minić, Rajna, Stević, Zorica, Arsić, Aleksandra., "Fatty acid profiling in amyotrophic lateral sclerosis" in Diet and Nutrition in Neurological Disorders (2023):155-172,
https://doi.org/10.1016/B978-0-323-89834-8.00023-4 . .

Neurotoksično dejstvo venoma Vipera ammodytes na modelu izolovane dijafragme pacova

Ivanović, Saša; Ćupić, Vitomir; Živković, Irena; Milovanović, Vladimir; Ćupić Miladinović, Dejana; Borozan, Sunčica

(Belgrade : Serbian Society of Toxicology, 2023)

TY  - CONF
AU  - Ivanović, Saša
AU  - Ćupić, Vitomir
AU  - Živković, Irena
AU  - Milovanović, Vladimir
AU  - Ćupić Miladinović, Dejana
AU  - Borozan, Sunčica
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/845
AB  - Poskok (Vipera ammodytes) predstavlja najzastupljeniju zmiju otrovnicu u Srbiji i na Balkanu, kako po brojnosti tako i po arealu rasprostranjenosti. U poređenju sa venomima ostalih otrovnica iz familije Viperidae na Balkanu, toksičnost venoma poskoka je najveća. Neurotoksičnost sirovog venoma poskoka smo ispitivali u in vitro uslovima na osnovu kontrakcija preparata dijafragme izazvanih poljnom električnom stimulacijom i aktivnosti enzima: acetilholinesteraze (AChE) i ukupnih ATP-aza u dijafragmi.Formirano je 5 grupa pacova (n=5): kontrola (dijafragma bez venoma), dijafragma sa venomom, dijafragma sa smešom venom/antivenom („Viekvin“, Torlak, Srbija) u odnosima 1:2, 1:10 i 1:20 (m/m). Kontrakcije dijafragme su opale na 50% kontrolnih kontrakcija: pod uticajem venoma za 62,00±2,31minuta, a sa smešom venom/antivenom 1:2 za 78,50±7,51 minuta, 1:10 za 150,00±17,32 minuta, 1:20 za 307,50±2,89 minuta. Statistička razlika (p0,05) pod dejstvom venoma. Venom inhibira aktivnost ukupnih ATP-aza dijafragme za 51,81% u odnosu na kontrolu. Venom poskoka ne deluje na AChE i nikotinske receptore u neuro-mišićnoj sinapsi dijafragme, već narušava energetski metabolizam mitohondrija.
AB  - Vipera ammodytes is the most common venomous snake in Serbia and the Balkans, both in terms of numbers and area of distribution. Compared to the venoms of other poisonous snakes from the family Viperidae in the Balkans, the toxicity of the V. ammodytes venom is the highest. We investigated the neurotoxicity of crude venom in vitro using contractions of the diaphragm preparation induced by electric field stimulation and enzyme activity: acetylcholinesterase (AChE) and total ATPases in the diaphragm. Five groups of rats were formed (n=5): control (diaphragm without venom), diaphragmwith venom, diaphragm with mixture of venom/antivenom ("Viekvin", Torlak, Serbia) in the ratio 1:2, 1:10 and 1:20 (m/m). Diaphragmatic contractions decreased to 50% of control contractions: under the influence of venom for 62.00±2.31 minutes, and with venom/antivenom mixture 1:2 for 78.50±7.51 minutes, 1:10 for 150.00±17.32 minutes, 1:20 for 307.50±2.89 minutes. A statistical difference (p 0.05) under the influence of the venom. Venom inhibits the total ATPases activity of the diaphragm by 51.81% compared with control.The venom of V. ammodytes does not act on AChE and nicotinic receptors in the neuromuscular synapse of the diaphragm, but interferes with mitochondrial energy metabolism.
PB  - Belgrade : Serbian Society of Toxicology
C3  - 13th International Congress of the Serbian Society of Toxicology and 1st TOXSEE Regional Conference, belgrade, 10 - 12 May, 2023
T1  - Neurotoksično dejstvo venoma Vipera ammodytes na modelu izolovane dijafragme pacova
T1  - Neurotoxic effect of Vipera ammodytes venom on the rat isolated diaphragm model
EP  - 108
SP  - 107
UR  - https://hdl.handle.net/21.15107/rcub_intor_845
ER  - 
@conference{
author = "Ivanović, Saša and Ćupić, Vitomir and Živković, Irena and Milovanović, Vladimir and Ćupić Miladinović, Dejana and Borozan, Sunčica",
year = "2023",
abstract = "Poskok (Vipera ammodytes) predstavlja najzastupljeniju zmiju otrovnicu u Srbiji i na Balkanu, kako po brojnosti tako i po arealu rasprostranjenosti. U poređenju sa venomima ostalih otrovnica iz familije Viperidae na Balkanu, toksičnost venoma poskoka je najveća. Neurotoksičnost sirovog venoma poskoka smo ispitivali u in vitro uslovima na osnovu kontrakcija preparata dijafragme izazvanih poljnom električnom stimulacijom i aktivnosti enzima: acetilholinesteraze (AChE) i ukupnih ATP-aza u dijafragmi.Formirano je 5 grupa pacova (n=5): kontrola (dijafragma bez venoma), dijafragma sa venomom, dijafragma sa smešom venom/antivenom („Viekvin“, Torlak, Srbija) u odnosima 1:2, 1:10 i 1:20 (m/m). Kontrakcije dijafragme su opale na 50% kontrolnih kontrakcija: pod uticajem venoma za 62,00±2,31minuta, a sa smešom venom/antivenom 1:2 za 78,50±7,51 minuta, 1:10 za 150,00±17,32 minuta, 1:20 za 307,50±2,89 minuta. Statistička razlika (p0,05) pod dejstvom venoma. Venom inhibira aktivnost ukupnih ATP-aza dijafragme za 51,81% u odnosu na kontrolu. Venom poskoka ne deluje na AChE i nikotinske receptore u neuro-mišićnoj sinapsi dijafragme, već narušava energetski metabolizam mitohondrija., Vipera ammodytes is the most common venomous snake in Serbia and the Balkans, both in terms of numbers and area of distribution. Compared to the venoms of other poisonous snakes from the family Viperidae in the Balkans, the toxicity of the V. ammodytes venom is the highest. We investigated the neurotoxicity of crude venom in vitro using contractions of the diaphragm preparation induced by electric field stimulation and enzyme activity: acetylcholinesterase (AChE) and total ATPases in the diaphragm. Five groups of rats were formed (n=5): control (diaphragm without venom), diaphragmwith venom, diaphragm with mixture of venom/antivenom ("Viekvin", Torlak, Serbia) in the ratio 1:2, 1:10 and 1:20 (m/m). Diaphragmatic contractions decreased to 50% of control contractions: under the influence of venom for 62.00±2.31 minutes, and with venom/antivenom mixture 1:2 for 78.50±7.51 minutes, 1:10 for 150.00±17.32 minutes, 1:20 for 307.50±2.89 minutes. A statistical difference (p 0.05) under the influence of the venom. Venom inhibits the total ATPases activity of the diaphragm by 51.81% compared with control.The venom of V. ammodytes does not act on AChE and nicotinic receptors in the neuromuscular synapse of the diaphragm, but interferes with mitochondrial energy metabolism.",
publisher = "Belgrade : Serbian Society of Toxicology",
journal = "13th International Congress of the Serbian Society of Toxicology and 1st TOXSEE Regional Conference, belgrade, 10 - 12 May, 2023",
title = "Neurotoksično dejstvo venoma Vipera ammodytes na modelu izolovane dijafragme pacova, Neurotoxic effect of Vipera ammodytes venom on the rat isolated diaphragm model",
pages = "108-107",
url = "https://hdl.handle.net/21.15107/rcub_intor_845"
}
Ivanović, S., Ćupić, V., Živković, I., Milovanović, V., Ćupić Miladinović, D.,& Borozan, S.. (2023). Neurotoksično dejstvo venoma Vipera ammodytes na modelu izolovane dijafragme pacova. in 13th International Congress of the Serbian Society of Toxicology and 1st TOXSEE Regional Conference, belgrade, 10 - 12 May, 2023
Belgrade : Serbian Society of Toxicology., 107-108.
https://hdl.handle.net/21.15107/rcub_intor_845
Ivanović S, Ćupić V, Živković I, Milovanović V, Ćupić Miladinović D, Borozan S. Neurotoksično dejstvo venoma Vipera ammodytes na modelu izolovane dijafragme pacova. in 13th International Congress of the Serbian Society of Toxicology and 1st TOXSEE Regional Conference, belgrade, 10 - 12 May, 2023. 2023;:107-108.
https://hdl.handle.net/21.15107/rcub_intor_845 .
Ivanović, Saša, Ćupić, Vitomir, Živković, Irena, Milovanović, Vladimir, Ćupić Miladinović, Dejana, Borozan, Sunčica, "Neurotoksično dejstvo venoma Vipera ammodytes na modelu izolovane dijafragme pacova" in 13th International Congress of the Serbian Society of Toxicology and 1st TOXSEE Regional Conference, belgrade, 10 - 12 May, 2023 (2023):107-108,
https://hdl.handle.net/21.15107/rcub_intor_845 .

Epidemiological Predictors of Positive SARS-CoV-2 Polymerase Chain Reaction Test in Three Cohorts: Hospitalized Patients, Healthcare Workers, and Military Population, Serbia, 2020

Šuljagić, Vesna; Đurić-Petković, Danijela; Lazić, Srdan; Mladenović, Jovan; Rakonjac, Bojan; Opačić, Dolores; Ljubenović, Nenad; Milojković, Biljana; Radojević, Katarina; Nenezić, Ivana; Rančić, Nemanja

(MDPI, 2023)

TY  - JOUR
AU  - Šuljagić, Vesna
AU  - Đurić-Petković, Danijela
AU  - Lazić, Srdan
AU  - Mladenović, Jovan
AU  - Rakonjac, Bojan
AU  - Opačić, Dolores
AU  - Ljubenović, Nenad
AU  - Milojković, Biljana
AU  - Radojević, Katarina
AU  - Nenezić, Ivana
AU  - Rančić, Nemanja
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/655
AB  - Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its resulting coronavirus disease 2019 (COVID-19) has caused a fast-moving pandemic. Diagnostic testing, aimed to identify patients infected with SARS-CoV-2, plays a key role in controlling the COVID-19 pandemic in different populations. (2) Methods: This retrospective cohort study aimed to investigate predictors associated with positive polymerase chain reaction (PCR) SARS-CoV-2 test results in hospitalized patients, healthcare workers (HCWs), and military personnel (MP) during 2020, before the widespread availability of COVID-19 vaccines. Persons with a positive test result were compared with persons with a negative test result in three cohorts during the study period. (3) Results: A total of 6912 respondents were tested, and 1334 (19.3%) of them had positive PCR SARS-CoV-2 test results. Contact with a known COVID-19 case within 14 days (p < 0.001; OR: 1.48; 95% CI: 1.25–1.76), fever (p < 0.001; OR: 3.66; 95% CI: 3.04–4.41), cough (p < 0.001; OR: 1.91; 95% CI: 1.59–2.30), headache (p = 0.028; OR: 1.24; 95% CI: 1.02–1.50), and myalgia/arthralgia (p < 0.001; OR: 1.99; 95% CI: 1.65–2.42) were independently associated with positive PCR SARS-CoV-2 test results in the cohort of MP. Furthermore, fever (p < 0.001; OR: 2.75; 95% CI: 1.83–4.13), cough (p < 0.001; OR: 2.04; 95% CI: 1.32–3.13), headache (p = 0.008; OR: 1.76; 95% CI: 1.15–2.68), and myalgia/arthralgia (p = 0.039; OR: 1.58; 95% CI: 1.02–2.45) were independently associated with positive PCR SARS-CoV-2 test results in the cohort of HCWs. Moreover, independent predictors of positive PCR SARS-CoV-2 test results in hospitalized patients were contact with a known COVID-19 case within 14 days (p < 0.001; OR: 2.56; 95% CI: 1.71–3.83), fever (p < 0.001; OR: 1.89; 95% CI: 1.38–2.59), pneumonia (p = 0.041; OR: 1.45; 95% CI: 1.01–2.09), and neurological diseases (p = 0.009; OR: 0.375; 95% CI: 0.18–0.78). (4) Conclusions: According to data gathered from cohorts of hospitalized patients, HCWs, and MP, before the widespread availability of COVID-19 vaccines in Serbia, we can conclude that predictors of positive PCR SARS-CoV-2 test results in MP and HCWs were similar. Accurate estimates of COVID-19 in different population groups are important for health authorities.
PB  - MDPI
T2  - International Journal of Environmental Research and Public Health
T1  - Epidemiological Predictors of Positive SARS-CoV-2 Polymerase Chain Reaction Test in Three Cohorts: Hospitalized Patients, Healthcare Workers, and Military Population, Serbia, 2020
IS  - 4
SP  - 3601
VL  - 20
DO  - 10.3390/ijerph20043601
ER  - 
@article{
author = "Šuljagić, Vesna and Đurić-Petković, Danijela and Lazić, Srdan and Mladenović, Jovan and Rakonjac, Bojan and Opačić, Dolores and Ljubenović, Nenad and Milojković, Biljana and Radojević, Katarina and Nenezić, Ivana and Rančić, Nemanja",
year = "2023",
abstract = "Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its resulting coronavirus disease 2019 (COVID-19) has caused a fast-moving pandemic. Diagnostic testing, aimed to identify patients infected with SARS-CoV-2, plays a key role in controlling the COVID-19 pandemic in different populations. (2) Methods: This retrospective cohort study aimed to investigate predictors associated with positive polymerase chain reaction (PCR) SARS-CoV-2 test results in hospitalized patients, healthcare workers (HCWs), and military personnel (MP) during 2020, before the widespread availability of COVID-19 vaccines. Persons with a positive test result were compared with persons with a negative test result in three cohorts during the study period. (3) Results: A total of 6912 respondents were tested, and 1334 (19.3%) of them had positive PCR SARS-CoV-2 test results. Contact with a known COVID-19 case within 14 days (p < 0.001; OR: 1.48; 95% CI: 1.25–1.76), fever (p < 0.001; OR: 3.66; 95% CI: 3.04–4.41), cough (p < 0.001; OR: 1.91; 95% CI: 1.59–2.30), headache (p = 0.028; OR: 1.24; 95% CI: 1.02–1.50), and myalgia/arthralgia (p < 0.001; OR: 1.99; 95% CI: 1.65–2.42) were independently associated with positive PCR SARS-CoV-2 test results in the cohort of MP. Furthermore, fever (p < 0.001; OR: 2.75; 95% CI: 1.83–4.13), cough (p < 0.001; OR: 2.04; 95% CI: 1.32–3.13), headache (p = 0.008; OR: 1.76; 95% CI: 1.15–2.68), and myalgia/arthralgia (p = 0.039; OR: 1.58; 95% CI: 1.02–2.45) were independently associated with positive PCR SARS-CoV-2 test results in the cohort of HCWs. Moreover, independent predictors of positive PCR SARS-CoV-2 test results in hospitalized patients were contact with a known COVID-19 case within 14 days (p < 0.001; OR: 2.56; 95% CI: 1.71–3.83), fever (p < 0.001; OR: 1.89; 95% CI: 1.38–2.59), pneumonia (p = 0.041; OR: 1.45; 95% CI: 1.01–2.09), and neurological diseases (p = 0.009; OR: 0.375; 95% CI: 0.18–0.78). (4) Conclusions: According to data gathered from cohorts of hospitalized patients, HCWs, and MP, before the widespread availability of COVID-19 vaccines in Serbia, we can conclude that predictors of positive PCR SARS-CoV-2 test results in MP and HCWs were similar. Accurate estimates of COVID-19 in different population groups are important for health authorities.",
publisher = "MDPI",
journal = "International Journal of Environmental Research and Public Health",
title = "Epidemiological Predictors of Positive SARS-CoV-2 Polymerase Chain Reaction Test in Three Cohorts: Hospitalized Patients, Healthcare Workers, and Military Population, Serbia, 2020",
number = "4",
pages = "3601",
volume = "20",
doi = "10.3390/ijerph20043601"
}
Šuljagić, V., Đurić-Petković, D., Lazić, S., Mladenović, J., Rakonjac, B., Opačić, D., Ljubenović, N., Milojković, B., Radojević, K., Nenezić, I.,& Rančić, N.. (2023). Epidemiological Predictors of Positive SARS-CoV-2 Polymerase Chain Reaction Test in Three Cohorts: Hospitalized Patients, Healthcare Workers, and Military Population, Serbia, 2020. in International Journal of Environmental Research and Public Health
MDPI., 20(4), 3601.
https://doi.org/10.3390/ijerph20043601
Šuljagić V, Đurić-Petković D, Lazić S, Mladenović J, Rakonjac B, Opačić D, Ljubenović N, Milojković B, Radojević K, Nenezić I, Rančić N. Epidemiological Predictors of Positive SARS-CoV-2 Polymerase Chain Reaction Test in Three Cohorts: Hospitalized Patients, Healthcare Workers, and Military Population, Serbia, 2020. in International Journal of Environmental Research and Public Health. 2023;20(4):3601.
doi:10.3390/ijerph20043601 .
Šuljagić, Vesna, Đurić-Petković, Danijela, Lazić, Srdan, Mladenović, Jovan, Rakonjac, Bojan, Opačić, Dolores, Ljubenović, Nenad, Milojković, Biljana, Radojević, Katarina, Nenezić, Ivana, Rančić, Nemanja, "Epidemiological Predictors of Positive SARS-CoV-2 Polymerase Chain Reaction Test in Three Cohorts: Hospitalized Patients, Healthcare Workers, and Military Population, Serbia, 2020" in International Journal of Environmental Research and Public Health, 20, no. 4 (2023):3601,
https://doi.org/10.3390/ijerph20043601 . .
1
2
1

Thymic changes as a contributing factor in the increased susceptibility of old Albino Oxford rats to EAE development

Petrušić, Marija; Stojić-Vukanić, Zorica; Pilipović, Ivan; Kosec, Duško; Prijić, Ivana; Leposavić, Gordana

(Elsevier, 2023)

TY  - JOUR
AU  - Petrušić, Marija
AU  - Stojić-Vukanić, Zorica
AU  - Pilipović, Ivan
AU  - Kosec, Duško
AU  - Prijić, Ivana
AU  - Leposavić, Gordana
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/633
AB  - The study was aimed to examine putative contribution of thymic involution to ageing-associated increase in susceptibility of Albino Oxford (AO) rats to the development of clinical EAE, and vice versa influence of the disease on the progression of thymic involution. To this end we examined (i) the parameters of thymocyte negative selection efficacy, the thymic generation of CD4+CD25+Foxp3+ T regulatory cells (Tregs) and thymic capacity to instruct/predetermine IL-17-producing T-cell differentiation, and thymopietic efficacy-associated accumulation of “inflammescent” cytotoxic CD28- T cells in the periphery, and (ii) the key underlying mechanisms in young and old non-immunised AO rats and their counterparts immunised for EAE (on the 16th day post-immunisation when the disease in old rats reached the plateau) using flow cytometry analysis and/or RT-qPCR. It was found that thymic involution impairs: (i) the efficacy of negative selection (by affecting thymocyte expression of CD90, negative regulator of selection threshold and the expression of thymic stromal cell integrity factors) and (ii) Treg generation (by diminishing expression of cytokines supporting their differentiation/maturation). Additionally, the results suggest that thymic involution facilitates CD8+ T-cell differentiation into IL-17-producing cells (previously linked to the development of clinical EAE in old AO rats). Furthermore, they confirmed that ageing-related decrease in thymic T-cell output (as indicated by diminished frequency of recent thymic emigrants in peripheral blood) resulted in the accumulation of CD28- T cells in peripheral blood and, upon immunisation, in the target organ. On the other hand, the development of EAE (most likely by increasing circulatory levels of proinflammatory cytokines) contributed to the decline in thymic output of T cells, including Tregs, and thereby to the progression/maintenance of clinical EAE. Thus, in AO rats thymic involution via multi-layered mechanisms may favour the development of clinically manifested autoimmunity, which, in turn, precipitates the thymus atrophy.
PB  - Elsevier
T2  - Experimental Gerontology
T1  - Thymic changes as a contributing factor in the increased susceptibility of old Albino Oxford rats to EAE development
SP  - 112009
VL  - 171
DO  - 10.1016/j.exger.2022.112009
ER  - 
@article{
author = "Petrušić, Marija and Stojić-Vukanić, Zorica and Pilipović, Ivan and Kosec, Duško and Prijić, Ivana and Leposavić, Gordana",
year = "2023",
abstract = "The study was aimed to examine putative contribution of thymic involution to ageing-associated increase in susceptibility of Albino Oxford (AO) rats to the development of clinical EAE, and vice versa influence of the disease on the progression of thymic involution. To this end we examined (i) the parameters of thymocyte negative selection efficacy, the thymic generation of CD4+CD25+Foxp3+ T regulatory cells (Tregs) and thymic capacity to instruct/predetermine IL-17-producing T-cell differentiation, and thymopietic efficacy-associated accumulation of “inflammescent” cytotoxic CD28- T cells in the periphery, and (ii) the key underlying mechanisms in young and old non-immunised AO rats and their counterparts immunised for EAE (on the 16th day post-immunisation when the disease in old rats reached the plateau) using flow cytometry analysis and/or RT-qPCR. It was found that thymic involution impairs: (i) the efficacy of negative selection (by affecting thymocyte expression of CD90, negative regulator of selection threshold and the expression of thymic stromal cell integrity factors) and (ii) Treg generation (by diminishing expression of cytokines supporting their differentiation/maturation). Additionally, the results suggest that thymic involution facilitates CD8+ T-cell differentiation into IL-17-producing cells (previously linked to the development of clinical EAE in old AO rats). Furthermore, they confirmed that ageing-related decrease in thymic T-cell output (as indicated by diminished frequency of recent thymic emigrants in peripheral blood) resulted in the accumulation of CD28- T cells in peripheral blood and, upon immunisation, in the target organ. On the other hand, the development of EAE (most likely by increasing circulatory levels of proinflammatory cytokines) contributed to the decline in thymic output of T cells, including Tregs, and thereby to the progression/maintenance of clinical EAE. Thus, in AO rats thymic involution via multi-layered mechanisms may favour the development of clinically manifested autoimmunity, which, in turn, precipitates the thymus atrophy.",
publisher = "Elsevier",
journal = "Experimental Gerontology",
title = "Thymic changes as a contributing factor in the increased susceptibility of old Albino Oxford rats to EAE development",
pages = "112009",
volume = "171",
doi = "10.1016/j.exger.2022.112009"
}
Petrušić, M., Stojić-Vukanić, Z., Pilipović, I., Kosec, D., Prijić, I.,& Leposavić, G.. (2023). Thymic changes as a contributing factor in the increased susceptibility of old Albino Oxford rats to EAE development. in Experimental Gerontology
Elsevier., 171, 112009.
https://doi.org/10.1016/j.exger.2022.112009
Petrušić M, Stojić-Vukanić Z, Pilipović I, Kosec D, Prijić I, Leposavić G. Thymic changes as a contributing factor in the increased susceptibility of old Albino Oxford rats to EAE development. in Experimental Gerontology. 2023;171:112009.
doi:10.1016/j.exger.2022.112009 .
Petrušić, Marija, Stojić-Vukanić, Zorica, Pilipović, Ivan, Kosec, Duško, Prijić, Ivana, Leposavić, Gordana, "Thymic changes as a contributing factor in the increased susceptibility of old Albino Oxford rats to EAE development" in Experimental Gerontology, 171 (2023):112009,
https://doi.org/10.1016/j.exger.2022.112009 . .
2
2

Short-term effect of Brevibacillus laterosporus supplemented diet on worker honey bee microbiome

Malešević, Milka; Stanisavljević, Nemanja; Rašić, Slađan; Vukotić, Goran; Gardijan, Lazar; Obradović, Mina; Kojić, Milan

(Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade, 2023)

TY  - CONF
AU  - Malešević, Milka
AU  - Stanisavljević, Nemanja
AU  - Rašić, Slađan
AU  - Vukotić, Goran
AU  - Gardijan, Lazar
AU  - Obradović, Mina
AU  - Kojić, Milan
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/823
AB  - Introduction: Brevibacillus laterosporus is a promising microbiological agent that can be used to prevent and control destructive diseases affecting honey bee colonies. In the presentstudy, the short-termeffect of the B. laterosporus BGSP11 bee diet on microbiota and mycobiota was investigated.Methods: The honey bee diet was supplemented with spores of B. laterosporus BGSP11 at a concentration of 1×108 CFU/mL in sucrose solution. Metabarcoding analysis of the bee microbial community profile was performed based on 16S RNA (bacteriobiota) and Internally Transcribes Spacer (ITS) region(mycobiota) obtained using MiSeq Illumina sequencing. The QIIME2 v2021.4 pipeline was used to analyze the obtained amplicon data library.Results: The results show that the BGSP11 bee diet slightly altered the bee microbiota and did not leadto potentially harmful changes in the bacterial microbiota. Moreover, it can potentially induce positivechanges, mainly reflected in the reduction of opportunistic bacteria. On the other hand, the treatmenthad a greater effect on mycobiota. However, the changesin the bee mycobiome caused by the treatmentcannot be considered a priori as beneficial or harmful,since the interaction between the bee and its mycobiome is not sufficiently studied. The observed positive changes in the bee mycobiome are mainlyreflected in the reduction of phytopathogenic fungi that may affect the organoleptic and techno-functional properties of honey.Conclusion: This pilot study suggests that the introduction of BGSP11 in beekeeping practice as a biological agent could be considered due to no harmful effects observed on the microbiota of bees.
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  - Short-term effect of Brevibacillus laterosporus supplemented diet on worker honey bee microbiome
EP  - 112
SP  - 112
UR  - https://hdl.handle.net/21.15107/rcub_intor_823
ER  - 
@conference{
author = "Malešević, Milka and Stanisavljević, Nemanja and Rašić, Slađan and Vukotić, Goran and Gardijan, Lazar and Obradović, Mina and Kojić, Milan",
year = "2023",
abstract = "Introduction: Brevibacillus laterosporus is a promising microbiological agent that can be used to prevent and control destructive diseases affecting honey bee colonies. In the presentstudy, the short-termeffect of the B. laterosporus BGSP11 bee diet on microbiota and mycobiota was investigated.Methods: The honey bee diet was supplemented with spores of B. laterosporus BGSP11 at a concentration of 1×108 CFU/mL in sucrose solution. Metabarcoding analysis of the bee microbial community profile was performed based on 16S RNA (bacteriobiota) and Internally Transcribes Spacer (ITS) region(mycobiota) obtained using MiSeq Illumina sequencing. The QIIME2 v2021.4 pipeline was used to analyze the obtained amplicon data library.Results: The results show that the BGSP11 bee diet slightly altered the bee microbiota and did not leadto potentially harmful changes in the bacterial microbiota. Moreover, it can potentially induce positivechanges, mainly reflected in the reduction of opportunistic bacteria. On the other hand, the treatmenthad a greater effect on mycobiota. However, the changesin the bee mycobiome caused by the treatmentcannot be considered a priori as beneficial or harmful,since the interaction between the bee and its mycobiome is not sufficiently studied. The observed positive changes in the bee mycobiome are mainlyreflected in the reduction of phytopathogenic fungi that may affect the organoleptic and techno-functional properties of honey.Conclusion: This pilot study suggests that the introduction of BGSP11 in beekeeping practice as a biological agent could be considered due to no harmful effects observed on the microbiota of bees.",
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 = "Short-term effect of Brevibacillus laterosporus supplemented diet on worker honey bee microbiome",
pages = "112-112",
url = "https://hdl.handle.net/21.15107/rcub_intor_823"
}
Malešević, M., Stanisavljević, N., Rašić, S., Vukotić, G., Gardijan, L., Obradović, M.,& Kojić, M.. (2023). Short-term effect of Brevibacillus laterosporus supplemented diet on worker honey bee microbiome. 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., 112-112.
https://hdl.handle.net/21.15107/rcub_intor_823
Malešević M, Stanisavljević N, Rašić S, Vukotić G, Gardijan L, Obradović M, Kojić M. Short-term effect of Brevibacillus laterosporus supplemented diet on worker honey bee microbiome. 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;:112-112.
https://hdl.handle.net/21.15107/rcub_intor_823 .
Malešević, Milka, Stanisavljević, Nemanja, Rašić, Slađan, Vukotić, Goran, Gardijan, Lazar, Obradović, Mina, Kojić, Milan, "Short-term effect of Brevibacillus laterosporus supplemented diet on worker honey bee microbiome" 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):112-112,
https://hdl.handle.net/21.15107/rcub_intor_823 .

Drying without dying: revealing the role of late embryogenesis abundant proteins during desiccation in Ramonda serbica

Pantelić, Ana; Prodić, Ivana; Milić, Dejana; Senćanski, Milan; Vidović, Marija

(Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade, 2023)

TY  - CONF
AU  - Pantelić, Ana
AU  - Prodić, Ivana
AU  - Milić, Dejana
AU  - Senćanski, Milan
AU  - Vidović, Marija
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/824
AB  - Introduction: Resurrection plants (such as Ramonda serbica) can survive a long desiccation period andfully resume their metabolism upon watering. The hallmark of desiccation tolerance (DT) is the accumulation of protective, intrinsically disordered proteins(IDPs), called late embryogenesis abundant proteins (LEAPs). Although their high structural plasticity allows them to interact with various partners, nospecific cellular targets of LEAPs have been identified so far.Methods: To identify LEAPsinvolved in DT, differential transcriptome and proteome analyses of hydratedand desiccated R. serbica leaves were performed. The identified LEAPs were structurally characterisedand classified. To evaluate theirstructural propertiesin vitro and their potential functionsin vivo, the representative RsLEA proteins, were produced in Escherichia coli using recombinant DNA technology.Results: Members of the LEA4 protein family represent the majority of desiccation-inducible LEAPs. Even17 proteins belonging to the LEA4 protein family group were induced by desiccation. They show high disorder propensity (82 %), and at the same time, a high tendency to form α-helices (>80%). Although recombinant DNA technology has traditionally been used to overexpress and purify various globularproteins, the production of IDPsis challenging due to their high susceptibility to proteolytic cleavage andaggregation. Nevertheless, the representative LEAPs containing hexa-Histagsimmunoglobulin G-binding protein and a proteolytic TEV site were produced, purified and cleaved by TEV protease.Conclusion: The combination of in silico and in vitro results will be crucial for the identification of endogenous partners of LEAPs, providing further insight into their role in DT.
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  - Drying without dying: revealing the role of late embryogenesis abundant proteins during desiccation in Ramonda serbica
EP  - 110
SP  - 110
UR  - https://hdl.handle.net/21.15107/rcub_intor_824
ER  - 
@conference{
author = "Pantelić, Ana and Prodić, Ivana and Milić, Dejana and Senćanski, Milan and Vidović, Marija",
year = "2023",
abstract = "Introduction: Resurrection plants (such as Ramonda serbica) can survive a long desiccation period andfully resume their metabolism upon watering. The hallmark of desiccation tolerance (DT) is the accumulation of protective, intrinsically disordered proteins(IDPs), called late embryogenesis abundant proteins (LEAPs). Although their high structural plasticity allows them to interact with various partners, nospecific cellular targets of LEAPs have been identified so far.Methods: To identify LEAPsinvolved in DT, differential transcriptome and proteome analyses of hydratedand desiccated R. serbica leaves were performed. The identified LEAPs were structurally characterisedand classified. To evaluate theirstructural propertiesin vitro and their potential functionsin vivo, the representative RsLEA proteins, were produced in Escherichia coli using recombinant DNA technology.Results: Members of the LEA4 protein family represent the majority of desiccation-inducible LEAPs. Even17 proteins belonging to the LEA4 protein family group were induced by desiccation. They show high disorder propensity (82 %), and at the same time, a high tendency to form α-helices (>80%). Although recombinant DNA technology has traditionally been used to overexpress and purify various globularproteins, the production of IDPsis challenging due to their high susceptibility to proteolytic cleavage andaggregation. Nevertheless, the representative LEAPs containing hexa-Histagsimmunoglobulin G-binding protein and a proteolytic TEV site were produced, purified and cleaved by TEV protease.Conclusion: The combination of in silico and in vitro results will be crucial for the identification of endogenous partners of LEAPs, providing further insight into their role in DT.",
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 = "Drying without dying: revealing the role of late embryogenesis abundant proteins during desiccation in Ramonda serbica",
pages = "110-110",
url = "https://hdl.handle.net/21.15107/rcub_intor_824"
}
Pantelić, A., Prodić, I., Milić, D., Senćanski, M.,& Vidović, M.. (2023). Drying without dying: revealing the role of late embryogenesis abundant proteins during desiccation in Ramonda serbica. 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., 110-110.
https://hdl.handle.net/21.15107/rcub_intor_824
Pantelić A, Prodić I, Milić D, Senćanski M, Vidović M. Drying without dying: revealing the role of late embryogenesis abundant proteins during desiccation in Ramonda serbica. 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;:110-110.
https://hdl.handle.net/21.15107/rcub_intor_824 .
Pantelić, Ana, Prodić, Ivana, Milić, Dejana, Senćanski, Milan, Vidović, Marija, "Drying without dying: revealing the role of late embryogenesis abundant proteins during desiccation in Ramonda serbica" 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):110-110,
https://hdl.handle.net/21.15107/rcub_intor_824 .

Influence of amino acid substitution on the antimicrobial activity of bacteriocin lactolisterin BU

Gardijan, Lazar; Kojić, Milan; Jovanović, Goran; Malešević, Milka

(Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade, 2023)

TY  - CONF
AU  - Gardijan, Lazar
AU  - Kojić, Milan
AU  - Jovanović, Goran
AU  - Malešević, Milka
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/804
AB  - Introduction: Lactolisterin BU (LBU) is a potent bacteriocin derived from Lactococcuslactis subsp. lactis
bv. diacetylactis BGBU1-4. It exhibits antimicrobial properties against Gram-positive food spoilage and
foodborne pathogens. This research aimed to explore the impact of amino acid substitution in LBU on
its antimicrobial activity by utilizing in silico prediction of LBU’ssecondary structure and amino acid substitutions.
Methods: The secondary structure of LBU was predicted using Phyre2 software. Five variants of LBU
were selected and chemically synthesized, along with unaltered LBU and BHT-B,serving as controls. Peptides were twofold diluted in distilled water, resulting in final concentrations ranging from 1000 µg/ml
to 0.5 µg/ml. An agarspot test, employing 5 µl of the dilution, was conducted on three indicatorstrains:
Lactococcus lactis BGMN1-596, Listeria monocytogenes ATCC19111, and Staphylococcus aureus
ATCC25923. The presence of inhibition zones was analyzed after overnight incubation at 37°C (S. aureus)
and 30°C (L. lactis and L. monocytogenes).
Results: Phyre2 analysis unveiled the presence of two α-helices in LBU’s structure. The majority of LBU
variants displayed altered antimicrobial activity, with some changes being genusspecific, potentially attributable to variances in cell wall composition. Some variants completely lost their activity, underscoring the significance of native amino acids or their physicochemical properties in the corresponding
positions within LBU’s structure. Furthermore, it was confirmed that chemically synthesized LBU effectively retains its antimicrobial activity.
Conclusion: Changesin amino acid composition give insight on structure-function relationship of LBU.
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  - Influence of amino acid substitution on the antimicrobial activity of bacteriocin lactolisterin BU
EP  - 123
SP  - 123
UR  - https://hdl.handle.net/21.15107/rcub_intor_804
ER  - 
@conference{
author = "Gardijan, Lazar and Kojić, Milan and Jovanović, Goran and Malešević, Milka",
year = "2023",
abstract = "Introduction: Lactolisterin BU (LBU) is a potent bacteriocin derived from Lactococcuslactis subsp. lactis
bv. diacetylactis BGBU1-4. It exhibits antimicrobial properties against Gram-positive food spoilage and
foodborne pathogens. This research aimed to explore the impact of amino acid substitution in LBU on
its antimicrobial activity by utilizing in silico prediction of LBU’ssecondary structure and amino acid substitutions.
Methods: The secondary structure of LBU was predicted using Phyre2 software. Five variants of LBU
were selected and chemically synthesized, along with unaltered LBU and BHT-B,serving as controls. Peptides were twofold diluted in distilled water, resulting in final concentrations ranging from 1000 µg/ml
to 0.5 µg/ml. An agarspot test, employing 5 µl of the dilution, was conducted on three indicatorstrains:
Lactococcus lactis BGMN1-596, Listeria monocytogenes ATCC19111, and Staphylococcus aureus
ATCC25923. The presence of inhibition zones was analyzed after overnight incubation at 37°C (S. aureus)
and 30°C (L. lactis and L. monocytogenes).
Results: Phyre2 analysis unveiled the presence of two α-helices in LBU’s structure. The majority of LBU
variants displayed altered antimicrobial activity, with some changes being genusspecific, potentially attributable to variances in cell wall composition. Some variants completely lost their activity, underscoring the significance of native amino acids or their physicochemical properties in the corresponding
positions within LBU’s structure. Furthermore, it was confirmed that chemically synthesized LBU effectively retains its antimicrobial activity.
Conclusion: Changesin amino acid composition give insight on structure-function relationship of LBU.",
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 = "Influence of amino acid substitution on the antimicrobial activity of bacteriocin lactolisterin BU",
pages = "123-123",
url = "https://hdl.handle.net/21.15107/rcub_intor_804"
}
Gardijan, L., Kojić, M., Jovanović, G.,& Malešević, M.. (2023). Influence of amino acid substitution on the antimicrobial activity of bacteriocin lactolisterin BU. 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., 123-123.
https://hdl.handle.net/21.15107/rcub_intor_804
Gardijan L, Kojić M, Jovanović G, Malešević M. Influence of amino acid substitution on the antimicrobial activity of bacteriocin lactolisterin BU. 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;:123-123.
https://hdl.handle.net/21.15107/rcub_intor_804 .
Gardijan, Lazar, Kojić, Milan, Jovanović, Goran, Malešević, Milka, "Influence of amino acid substitution on the antimicrobial activity of bacteriocin lactolisterin BU" 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):123-123,
https://hdl.handle.net/21.15107/rcub_intor_804 .

Determination of muscle fiber types expressing ANKRD2

Novković, Mirjana; Vasić, Marko; Jasnić, Jovana; Milošević, Emilija; Milovanović, Mina; Savić, Slobodan; Kojić, Snežana

(Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade, 2023)

TY  - CONF
AU  - Novković, Mirjana
AU  - Vasić, Marko
AU  - Jasnić, Jovana
AU  - Milošević, Emilija
AU  - Milovanović, Mina
AU  - Savić, Slobodan
AU  - Kojić, Snežana
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2114
UR  - http://intor.torlakinstitut.com/handle/123456789/806
AB  - Introduction: Ankyrin Repeat Domain 2 (ANKRD2) is expressed in skeletal muscle, where plays a role inmuscle development, differentiation and adaptation to stress. Human skeletal muscle consists of threemajor fiber types: type 1 (slow-twitch, oxidative), type 2A (fast-twitch, oxidative) and type 2X (fast-twitch,glycolytic). ANKRD2 is reported to be primarily expressed in type 1 myofibers. However, recent findingson human single myofibers and our study of chicken muscles have shown that this protein may also beexpressed in type 2A fibers. Hence, our objective was to examine whether ANKRD2 is present in humanfast, type 2A muscle fibers using immunohistochemistry.Methods: Samples of large leg musclessoleus, gastrocnemius, vastusintermedius and vastuslateralis wereobtained from human cadaveric tissue. Serial cryosections were independently stained with anti-ANKRD2and antibodies for different myosin heavy chain isoforms (6H1 for type 2X, BF35 for type 1 and 2A, antiMHCs for type 1 and anti-MHCf for type 2A and 2X fibers). Immunostained tissues were analyzed by fluorescent microscopy.Results: In addition to slow, type 1, ANKRD2 wasfound expressed in fast, type 2A myofibers, which bothhave oxidative metabolism. Further, we did not observe ANDRD2 expression in glycolytic, type 2Xmyiofibers. This pattern of ANKRD2 expression was consistent across all examined muscles.Conclusion: Our resultsimplicate that the regulatory mechanism of ANKRD2 expression in human skeletal muscle is associated with oxidative metabolism, rather than muscle contraction speed.
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  - Determination of muscle fiber types expressing ANKRD2
EP  - 155
SP  - 155
UR  - https://hdl.handle.net/21.15107/rcub_intor_806
ER  - 
@conference{
author = "Novković, Mirjana and Vasić, Marko and Jasnić, Jovana and Milošević, Emilija and Milovanović, Mina and Savić, Slobodan and Kojić, Snežana",
year = "2023",
abstract = "Introduction: Ankyrin Repeat Domain 2 (ANKRD2) is expressed in skeletal muscle, where plays a role inmuscle development, differentiation and adaptation to stress. Human skeletal muscle consists of threemajor fiber types: type 1 (slow-twitch, oxidative), type 2A (fast-twitch, oxidative) and type 2X (fast-twitch,glycolytic). ANKRD2 is reported to be primarily expressed in type 1 myofibers. However, recent findingson human single myofibers and our study of chicken muscles have shown that this protein may also beexpressed in type 2A fibers. Hence, our objective was to examine whether ANKRD2 is present in humanfast, type 2A muscle fibers using immunohistochemistry.Methods: Samples of large leg musclessoleus, gastrocnemius, vastusintermedius and vastuslateralis wereobtained from human cadaveric tissue. Serial cryosections were independently stained with anti-ANKRD2and antibodies for different myosin heavy chain isoforms (6H1 for type 2X, BF35 for type 1 and 2A, antiMHCs for type 1 and anti-MHCf for type 2A and 2X fibers). Immunostained tissues were analyzed by fluorescent microscopy.Results: In addition to slow, type 1, ANKRD2 wasfound expressed in fast, type 2A myofibers, which bothhave oxidative metabolism. Further, we did not observe ANDRD2 expression in glycolytic, type 2Xmyiofibers. This pattern of ANKRD2 expression was consistent across all examined muscles.Conclusion: Our resultsimplicate that the regulatory mechanism of ANKRD2 expression in human skeletal muscle is associated with oxidative metabolism, rather than muscle contraction speed.",
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 = "Determination of muscle fiber types expressing ANKRD2",
pages = "155-155",
url = "https://hdl.handle.net/21.15107/rcub_intor_806"
}
Novković, M., Vasić, M., Jasnić, J., Milošević, E., Milovanović, M., Savić, S.,& Kojić, S.. (2023). Determination of muscle fiber types expressing ANKRD2. 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., 155-155.
https://hdl.handle.net/21.15107/rcub_intor_806
Novković M, Vasić M, Jasnić J, Milošević E, Milovanović M, Savić S, Kojić S. Determination of muscle fiber types expressing ANKRD2. 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;:155-155.
https://hdl.handle.net/21.15107/rcub_intor_806 .
Novković, Mirjana, Vasić, Marko, Jasnić, Jovana, Milošević, Emilija, Milovanović, Mina, Savić, Slobodan, Kojić, Snežana, "Determination of muscle fiber types expressing ANKRD2" 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):155-155,
https://hdl.handle.net/21.15107/rcub_intor_806 .

A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression

Ćurčić, Jovana; Jakovljević, Stefan; Novović, Katarina; Vasiljević, Zorica; Kojić, Milan; Jovčić, Branko; Malešević, Milka

(Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade, 2023)

TY  - CONF
AU  - Ćurčić, Jovana
AU  - Jakovljević, Stefan
AU  - Novović, Katarina
AU  - Vasiljević, Zorica
AU  - Kojić, Milan
AU  - Jovčić, Branko
AU  - Malešević, Milka
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/803
AB  - Introduction: Quorum quenching (QQ) isthe enzymatic degradation of cell-to-cellsignaling molecules.
In this study, the potential of the novel YtnP lactonase, the quorum quenching enzyme derived from S.
maltophilia, to reduce P. aeruginosa quorum sensing and virulence factor gene expression was investigated.
Methods: MMA83 culture (adjusted to 1.5x105 CFU/ml) was treated with recombinant YtnP lactonase
(final concentration 50 μg/ml) at 37°C for 12 hours under aeration. RNA isolation of the treated and untreated MMA83 culture was performed using the RNeasy Mini Kit (Qiagen, Germany) according to the
protocol. Quantitative reverse transcription-polymerase chain reaction (RT-qPCR), was used to analyze
the effect ofYtnP lactonase on the relative mRNA levels of the LasI/LasR, RhiI/RhiR, and PQS signaling network genes of P. aeruginosa MMA83 and virulence factor genes. The rpsL was used as an endogenous
control to normalize obtained data following the 2-ΔΔCt method.
Results: The QS genes belonging to three QS networks – LasI/LasR, RhiI/RhiR, and PQS of P. aeruginosa
MMA83 treated with YtnP lactonase were significantly downregulated. The RT -qPCR results show that
treatment with YtnP-lactonase decreased the relative mRNA levels of genes involved in the production
of elastase (lasB approximately 2-fold), alginate (algK approximately 2.2-fold), pyocyanin (phzM approximately 3.5-fold), pyoverdin (pvdS approximately 2-fold), and rhamnolipid (rhlC approximately 4-fold).
These results suggest that YtnP lactonase exerts an antivirulence effect at the transcription level.
Conclusion: YtnP lactonase, a quorum quenching (QQ) enzyme, has the potential to be used as an innovative enzyme-based antivirulence therapeutic to combat infections caused by P. aeruginosa.
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  - A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression
EP  - 121
SP  - 121
UR  - https://hdl.handle.net/21.15107/rcub_intor_803
ER  - 
@conference{
author = "Ćurčić, Jovana and Jakovljević, Stefan and Novović, Katarina and Vasiljević, Zorica and Kojić, Milan and Jovčić, Branko and Malešević, Milka",
year = "2023",
abstract = "Introduction: Quorum quenching (QQ) isthe enzymatic degradation of cell-to-cellsignaling molecules.
In this study, the potential of the novel YtnP lactonase, the quorum quenching enzyme derived from S.
maltophilia, to reduce P. aeruginosa quorum sensing and virulence factor gene expression was investigated.
Methods: MMA83 culture (adjusted to 1.5x105 CFU/ml) was treated with recombinant YtnP lactonase
(final concentration 50 μg/ml) at 37°C for 12 hours under aeration. RNA isolation of the treated and untreated MMA83 culture was performed using the RNeasy Mini Kit (Qiagen, Germany) according to the
protocol. Quantitative reverse transcription-polymerase chain reaction (RT-qPCR), was used to analyze
the effect ofYtnP lactonase on the relative mRNA levels of the LasI/LasR, RhiI/RhiR, and PQS signaling network genes of P. aeruginosa MMA83 and virulence factor genes. The rpsL was used as an endogenous
control to normalize obtained data following the 2-ΔΔCt method.
Results: The QS genes belonging to three QS networks – LasI/LasR, RhiI/RhiR, and PQS of P. aeruginosa
MMA83 treated with YtnP lactonase were significantly downregulated. The RT -qPCR results show that
treatment with YtnP-lactonase decreased the relative mRNA levels of genes involved in the production
of elastase (lasB approximately 2-fold), alginate (algK approximately 2.2-fold), pyocyanin (phzM approximately 3.5-fold), pyoverdin (pvdS approximately 2-fold), and rhamnolipid (rhlC approximately 4-fold).
These results suggest that YtnP lactonase exerts an antivirulence effect at the transcription level.
Conclusion: YtnP lactonase, a quorum quenching (QQ) enzyme, has the potential to be used as an innovative enzyme-based antivirulence therapeutic to combat infections caused by P. aeruginosa.",
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 = "A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression",
pages = "121-121",
url = "https://hdl.handle.net/21.15107/rcub_intor_803"
}
Ćurčić, J., Jakovljević, S., Novović, K., Vasiljević, Z., Kojić, M., Jovčić, B.,& Malešević, M.. (2023). A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression. 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., 121-121.
https://hdl.handle.net/21.15107/rcub_intor_803
Ćurčić J, Jakovljević S, Novović K, Vasiljević Z, Kojić M, Jovčić B, Malešević M. A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression. 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;:121-121.
https://hdl.handle.net/21.15107/rcub_intor_803 .
Ćurčić, Jovana, Jakovljević, Stefan, Novović, Katarina, Vasiljević, Zorica, Kojić, Milan, Jovčić, Branko, Malešević, Milka, "A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression" 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):121-121,
https://hdl.handle.net/21.15107/rcub_intor_803 .

Antioxidant Properties of Protein-Rich Plant Foods in Gastrointestinal Digestion-Peanuts as Our Antioxidant Friend or Foe in Allergies

Prodić, Ivana; Krstić-Ristivojević, Maja; Smiljanić, Katarina

(MDPI, 2023)

TY  - JOUR
AU  - Prodić, Ivana
AU  - Krstić-Ristivojević, Maja
AU  - Smiljanić, Katarina
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/756
AB  - Thermally processed peanuts are ideal plant models for studying the relationship between allergenicity and antioxidant capacity of protein-rich foods, besides lipids, carbohydrates and phytochemicals. Peanut is highly praised in the human diet; however, it is rich in allergens (>75% of total proteins). One-third of peanut allergens belong to the products of genes responsible for the defence of plants against stress conditions. The proximate composition of major peanut macromolecules and polyphenols is reviewed, focusing on the identity and relative abundance of all peanut proteins derived from recent proteomic studies. The importance of thermal processing, gastrointestinal digestion (performed by INFOGEST protocol) and their influence on allergenicity and antioxidant properties of protein-rich plant food matrices is elaborated. Antioxidant properties of bioactive peptides from nuts were also considered. Moreover, there are no studies dealing simultaneously with the antioxidant and allergenic properties of protein- and polyphenol-rich foods, considering all the molecules that can significantly contribute to the antioxidant capacity during and after gastrointestinal digestion. In summary, proteins and carbohydrates are underappreciated sources of antioxidant power released during the gastrointestinal digestion of protein-rich plant foods, and it is crucial to decipher their antioxidant contribution in addition to polyphenols and vitamins before and after gastrointestinal digestion.
PB  - MDPI
T2  - Antioxidants
T1  - Antioxidant Properties of Protein-Rich Plant Foods in Gastrointestinal Digestion-Peanuts as Our Antioxidant Friend or Foe in Allergies
IS  - 4
SP  - 886
VL  - 12
DO  - 10.3390/antiox12040886
ER  - 
@article{
author = "Prodić, Ivana and Krstić-Ristivojević, Maja and Smiljanić, Katarina",
year = "2023",
abstract = "Thermally processed peanuts are ideal plant models for studying the relationship between allergenicity and antioxidant capacity of protein-rich foods, besides lipids, carbohydrates and phytochemicals. Peanut is highly praised in the human diet; however, it is rich in allergens (>75% of total proteins). One-third of peanut allergens belong to the products of genes responsible for the defence of plants against stress conditions. The proximate composition of major peanut macromolecules and polyphenols is reviewed, focusing on the identity and relative abundance of all peanut proteins derived from recent proteomic studies. The importance of thermal processing, gastrointestinal digestion (performed by INFOGEST protocol) and their influence on allergenicity and antioxidant properties of protein-rich plant food matrices is elaborated. Antioxidant properties of bioactive peptides from nuts were also considered. Moreover, there are no studies dealing simultaneously with the antioxidant and allergenic properties of protein- and polyphenol-rich foods, considering all the molecules that can significantly contribute to the antioxidant capacity during and after gastrointestinal digestion. In summary, proteins and carbohydrates are underappreciated sources of antioxidant power released during the gastrointestinal digestion of protein-rich plant foods, and it is crucial to decipher their antioxidant contribution in addition to polyphenols and vitamins before and after gastrointestinal digestion.",
publisher = "MDPI",
journal = "Antioxidants",
title = "Antioxidant Properties of Protein-Rich Plant Foods in Gastrointestinal Digestion-Peanuts as Our Antioxidant Friend or Foe in Allergies",
number = "4",
pages = "886",
volume = "12",
doi = "10.3390/antiox12040886"
}
Prodić, I., Krstić-Ristivojević, M.,& Smiljanić, K.. (2023). Antioxidant Properties of Protein-Rich Plant Foods in Gastrointestinal Digestion-Peanuts as Our Antioxidant Friend or Foe in Allergies. in Antioxidants
MDPI., 12(4), 886.
https://doi.org/10.3390/antiox12040886
Prodić I, Krstić-Ristivojević M, Smiljanić K. Antioxidant Properties of Protein-Rich Plant Foods in Gastrointestinal Digestion-Peanuts as Our Antioxidant Friend or Foe in Allergies. in Antioxidants. 2023;12(4):886.
doi:10.3390/antiox12040886 .
Prodić, Ivana, Krstić-Ristivojević, Maja, Smiljanić, Katarina, "Antioxidant Properties of Protein-Rich Plant Foods in Gastrointestinal Digestion-Peanuts as Our Antioxidant Friend or Foe in Allergies" in Antioxidants, 12, no. 4 (2023):886,
https://doi.org/10.3390/antiox12040886 . .
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