TY  - JOUR
AU  - Virijević, Katarina
AU  - Živanović, Marko
AU  - Pavić, Jelena
AU  - Dragačević, Luka
AU  - Ljujić, Biljana
AU  - Miletić Kovačević, Marina
AU  - Papić, Miloš
AU  - Živanović, Suzana
AU  - Milenković, Strahinja
AU  - Radojević, Ivana
AU  - Filipović, Nenad
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/892
AB  - Recent advances in regenerative medicine provide encouraging strategies to produce artificial skin substitutes. Gelatin scaffolds are successfully used as wound-dressing materials due to their superior properties, such as biocompatibility and the ability to mimic the extracellular matrix of the surrounding environment. In this study, five gelatin combination solutions were prepared and successfully electrospun using an electrospinning technique. After careful screening, the optimal concentration of the most promising combination was selected for further investigation. The obtained scaffolds were crosslinked with 25% glutaraldehyde vapor and characterized by scanning electron microscopy, energy-dispersive X-ray spectroscopy, and Fourier-transform infrared spectroscopy. The incorporation of antibiotic agents such as ciprofloxacin hydrochloride and gentamicin sulfate into gelatin membranes improved the already existing antibacterial properties of antibiotic-free gelatin scaffolds against Pseudomonas aeruginosa and Staphylococcus aureus. Also, the outcomes from the in vivo model study revealed that skin regeneration was significantly accelerated with gelatin/ciprofloxacin scaffold treatment. Moreover, the gelatin nanofibers were found to strongly promote the neoangiogenic process in the in vivo chick embryo chorioallantoic membrane assay. Finally, the combination of gelatin’s extracellular matrix and antibacterial agents in the scaffold suggests its potential for effective wound-healing treatments, emphasizing the importance of gelatin scaffolds in tissue engineering.
PB  - MDPI
T2  - Pharmaceuticals
T1  - Electrospun Gelatin Scaffolds with Incorporated Antibiotics for Skin Wound Healing
IS  - 7
SP  - 851
VL  - 17
DO  - 10.3390/ph17070851
ER  - 

@article{
author = "Virijević, Katarina and Živanović, Marko and Pavić, Jelena and Dragačević, Luka and Ljujić, Biljana and Miletić Kovačević, Marina and Papić, Miloš and Živanović, Suzana and Milenković, Strahinja and Radojević, Ivana and Filipović, Nenad",
year = "2024",
abstract = "Recent advances in regenerative medicine provide encouraging strategies to produce artificial skin substitutes. Gelatin scaffolds are successfully used as wound-dressing materials due to their superior properties, such as biocompatibility and the ability to mimic the extracellular matrix of the surrounding environment. In this study, five gelatin combination solutions were prepared and successfully electrospun using an electrospinning technique. After careful screening, the optimal concentration of the most promising combination was selected for further investigation. The obtained scaffolds were crosslinked with 25% glutaraldehyde vapor and characterized by scanning electron microscopy, energy-dispersive X-ray spectroscopy, and Fourier-transform infrared spectroscopy. The incorporation of antibiotic agents such as ciprofloxacin hydrochloride and gentamicin sulfate into gelatin membranes improved the already existing antibacterial properties of antibiotic-free gelatin scaffolds against Pseudomonas aeruginosa and Staphylococcus aureus. Also, the outcomes from the in vivo model study revealed that skin regeneration was significantly accelerated with gelatin/ciprofloxacin scaffold treatment. Moreover, the gelatin nanofibers were found to strongly promote the neoangiogenic process in the in vivo chick embryo chorioallantoic membrane assay. Finally, the combination of gelatin’s extracellular matrix and antibacterial agents in the scaffold suggests its potential for effective wound-healing treatments, emphasizing the importance of gelatin scaffolds in tissue engineering.",
publisher = "MDPI",
journal = "Pharmaceuticals",
title = "Electrospun Gelatin Scaffolds with Incorporated Antibiotics for Skin Wound Healing",
number = "7",
pages = "851",
volume = "17",
doi = "10.3390/ph17070851"
}

Virijević, K., Živanović, M., Pavić, J., Dragačević, L., Ljujić, B., Miletić Kovačević, M., Papić, M., Živanović, S., Milenković, S., Radojević, I.,& Filipović, N.. (2024). Electrospun Gelatin Scaffolds with Incorporated Antibiotics for Skin Wound Healing. in Pharmaceuticals
MDPI., 17(7), 851.
https://doi.org/10.3390/ph17070851

Virijević K, Živanović M, Pavić J, Dragačević L, Ljujić B, Miletić Kovačević M, Papić M, Živanović S, Milenković S, Radojević I, Filipović N. Electrospun Gelatin Scaffolds with Incorporated Antibiotics for Skin Wound Healing. in Pharmaceuticals. 2024;17(7):851.
doi:10.3390/ph17070851 .

Virijević, Katarina, Živanović, Marko, Pavić, Jelena, Dragačević, Luka, Ljujić, Biljana, Miletić Kovačević, Marina, Papić, Miloš, Živanović, Suzana, Milenković, Strahinja, Radojević, Ivana, Filipović, Nenad, "Electrospun Gelatin Scaffolds with Incorporated Antibiotics for Skin Wound Healing" in Pharmaceuticals, 17, no. 7 (2024):851,
https://doi.org/10.3390/ph17070851 . .

TY  - DATA
AU  - Filipić, Brankica
AU  - Kojić, Milan
AU  - Vasiljević, Zorica
AU  - Sovtić, Aleksandar
AU  - Dimkić, Ivica
AU  - Wood, Emily
AU  - Esposito, Alfonso
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/988
AB  - Supplementary Figure S1: Pan-genome analyses of assembled genomes and number of core and shell genes. Supplementary Figure S2. Average nucleotide identity between genome sequences of E. coli isolates. Supplementary Figure S3. Alignment of the plasmid homologous to pMB2910_1 in the four strains, in order from top: 5381a, 5381b, 5681 and 5848. Tables: 5381b_VFDB_Jan_23-9085838786, 5381aVFDB_Jan_23-3173772185, 58848_VFDB_Jan_23-7074876386, 5681_VFDB_Jan_23-2749062099
PB  - MDPI
T2  - Microorganisms
T1  - Supplementary information for the article:  Filipic, B.; Kojic, M.; Vasiljevic, Z.; Sovtic, A.; Dimkic, I.; Wood, E.; Esposito, A. A Longitudinal Study of Escherichia Coli Clinical Isolates from the Tracheal Aspirates of a Paediatric Patient—Strain Type Similar to Pandemic ST131. Microorganisms 2024, 12 (10), 1990. https://doi.org/10.3390/microorganisms12101990.
DO  - 10.3390/microorganisms12101990
ER  - 

@misc{
author = "Filipić, Brankica and Kojić, Milan and Vasiljević, Zorica and Sovtić, Aleksandar and Dimkić, Ivica and Wood, Emily and Esposito, Alfonso",
year = "2024",
abstract = "Supplementary Figure S1: Pan-genome analyses of assembled genomes and number of core and shell genes. Supplementary Figure S2. Average nucleotide identity between genome sequences of E. coli isolates. Supplementary Figure S3. Alignment of the plasmid homologous to pMB2910_1 in the four strains, in order from top: 5381a, 5381b, 5681 and 5848. Tables: 5381b_VFDB_Jan_23-9085838786, 5381aVFDB_Jan_23-3173772185, 58848_VFDB_Jan_23-7074876386, 5681_VFDB_Jan_23-2749062099",
publisher = "MDPI",
journal = "Microorganisms",
title = "Supplementary information for the article:  Filipic, B.; Kojic, M.; Vasiljevic, Z.; Sovtic, A.; Dimkic, I.; Wood, E.; Esposito, A. A Longitudinal Study of Escherichia Coli Clinical Isolates from the Tracheal Aspirates of a Paediatric Patient—Strain Type Similar to Pandemic ST131. Microorganisms 2024, 12 (10), 1990. https://doi.org/10.3390/microorganisms12101990.",
doi = "10.3390/microorganisms12101990"
}

Filipić, B., Kojić, M., Vasiljević, Z., Sovtić, A., Dimkić, I., Wood, E.,& Esposito, A.. (2024). Supplementary information for the article:  Filipic, B.; Kojic, M.; Vasiljevic, Z.; Sovtic, A.; Dimkic, I.; Wood, E.; Esposito, A. A Longitudinal Study of Escherichia Coli Clinical Isolates from the Tracheal Aspirates of a Paediatric Patient—Strain Type Similar to Pandemic ST131. Microorganisms 2024, 12 (10), 1990. https://doi.org/10.3390/microorganisms12101990.. in Microorganisms
MDPI..
https://doi.org/10.3390/microorganisms12101990

Filipić B, Kojić M, Vasiljević Z, Sovtić A, Dimkić I, Wood E, Esposito A. Supplementary information for the article:  Filipic, B.; Kojic, M.; Vasiljevic, Z.; Sovtic, A.; Dimkic, I.; Wood, E.; Esposito, A. A Longitudinal Study of Escherichia Coli Clinical Isolates from the Tracheal Aspirates of a Paediatric Patient—Strain Type Similar to Pandemic ST131. Microorganisms 2024, 12 (10), 1990. https://doi.org/10.3390/microorganisms12101990.. in Microorganisms. 2024;.
doi:10.3390/microorganisms12101990 .

Filipić, Brankica, Kojić, Milan, Vasiljević, Zorica, Sovtić, Aleksandar, Dimkić, Ivica, Wood, Emily, Esposito, Alfonso, "Supplementary information for the article:  Filipic, B.; Kojic, M.; Vasiljevic, Z.; Sovtic, A.; Dimkic, I.; Wood, E.; Esposito, A. A Longitudinal Study of Escherichia Coli Clinical Isolates from the Tracheal Aspirates of a Paediatric Patient—Strain Type Similar to Pandemic ST131. Microorganisms 2024, 12 (10), 1990. https://doi.org/10.3390/microorganisms12101990." in Microorganisms (2024),
https://doi.org/10.3390/microorganisms12101990 . .

TY  - JOUR
AU  - Lukić, Ivana
AU  - Dragačević, Luka
AU  - Panić, Marko
AU  - Stamenković, Marina
AU  - Kojić, Milan
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/990
AB  - In the post-COVID-19 era, there has been a notable surge in the development of mRNA vaccines. These vaccines are not only targeting various pathogens beyond SARS-CoV-2 but also hold promise in treating cancer and genetic disorders. These type of vaccines are revolutionizing vaccinology through their inherent possibility for rapid pandemic response, high efficacy, minimal side effects, and cost-effectiveness. Achieving these benefits hinges on seamlessly integrating mRNA production steps, from plasmid DNA (pDNA) design and antigen cloning, in vitro transcription to lipid nanoparticle formulation. A critical initial step in mRNA vaccine production is pDNA cloning vector design. The vector should be carefully constructed considering a copy number of plasmid, vector backbone with a promoter, the origin of replication, multiple cloning sites, polyadenylation signal, and markers for selection. However, despite careful design, challenges like poly-A tail deletion may arise, prompting the exploration of stable large-size and low-copy vectors, as well as linear and bacteriophage vectors. Additionally, for large-scale production and regulatory compliance, vector systems must be scalable and well-documented. This overview aims to elaborate upon the intricacies in pDNA cloning vector design. The focus is on achieving accurate insert sequence, especially those encoding the complex antigens and gene expression, highlighting the critical role of pDNA design in ensuring vaccine effectiveness. Although commercial vectors and automated synthesis facilitate gene construction, challenges still exist. This emphasizes that a multifaceted approach, combining molecular biology techniques, computational tools, and collaboration with experts in microbiology, molecular biology, and vaccine development, is required for successful mRNA vaccine development.
PB  - Serbian Society for Microbiology
T2  - Microbiology/Mikrobiologija
T1  - mRNA vaccine manufacturing – challenges in plasmid DNA cloning vector design
EP  - 9
IS  - 1
SP  - 3
VL  - 45
UR  - https://hdl.handle.net/21.15107/rcub_intor_990
ER  - 

@article{
author = "Lukić, Ivana and Dragačević, Luka and Panić, Marko and Stamenković, Marina and Kojić, Milan",
year = "2024",
abstract = "In the post-COVID-19 era, there has been a notable surge in the development of mRNA vaccines. These vaccines are not only targeting various pathogens beyond SARS-CoV-2 but also hold promise in treating cancer and genetic disorders. These type of vaccines are revolutionizing vaccinology through their inherent possibility for rapid pandemic response, high efficacy, minimal side effects, and cost-effectiveness. Achieving these benefits hinges on seamlessly integrating mRNA production steps, from plasmid DNA (pDNA) design and antigen cloning, in vitro transcription to lipid nanoparticle formulation. A critical initial step in mRNA vaccine production is pDNA cloning vector design. The vector should be carefully constructed considering a copy number of plasmid, vector backbone with a promoter, the origin of replication, multiple cloning sites, polyadenylation signal, and markers for selection. However, despite careful design, challenges like poly-A tail deletion may arise, prompting the exploration of stable large-size and low-copy vectors, as well as linear and bacteriophage vectors. Additionally, for large-scale production and regulatory compliance, vector systems must be scalable and well-documented. This overview aims to elaborate upon the intricacies in pDNA cloning vector design. The focus is on achieving accurate insert sequence, especially those encoding the complex antigens and gene expression, highlighting the critical role of pDNA design in ensuring vaccine effectiveness. Although commercial vectors and automated synthesis facilitate gene construction, challenges still exist. This emphasizes that a multifaceted approach, combining molecular biology techniques, computational tools, and collaboration with experts in microbiology, molecular biology, and vaccine development, is required for successful mRNA vaccine development.",
publisher = "Serbian Society for Microbiology",
journal = "Microbiology/Mikrobiologija",
title = "mRNA vaccine manufacturing – challenges in plasmid DNA cloning vector design",
pages = "9-3",
number = "1",
volume = "45",
url = "https://hdl.handle.net/21.15107/rcub_intor_990"
}

Lukić, I., Dragačević, L., Panić, M., Stamenković, M.,& Kojić, M.. (2024). mRNA vaccine manufacturing – challenges in plasmid DNA cloning vector design. in Microbiology/Mikrobiologija
Serbian Society for Microbiology., 45(1), 3-9.
https://hdl.handle.net/21.15107/rcub_intor_990

Lukić I, Dragačević L, Panić M, Stamenković M, Kojić M. mRNA vaccine manufacturing – challenges in plasmid DNA cloning vector design. in Microbiology/Mikrobiologija. 2024;45(1):3-9.
https://hdl.handle.net/21.15107/rcub_intor_990 .

Lukić, Ivana, Dragačević, Luka, Panić, Marko, Stamenković, Marina, Kojić, Milan, "mRNA vaccine manufacturing – challenges in plasmid DNA cloning vector design" in Microbiology/Mikrobiologija, 45, no. 1 (2024):3-9,
https://hdl.handle.net/21.15107/rcub_intor_990 .

TY  - CHAP
AU  - Lukić, Ivana
AU  - Minić, Rajna
AU  - Dragačević, Luka
AU  - Blagojević, Veljko
AU  - Stamenković, Marina
AU  - Petković, Katarina
AU  - Radojević, Katarina
AU  - Kojić, Milan
AU  - Panić, Marko
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/989
AB  - Razvoj vakcina ima bogatu istoriju koja je obeležena revolucionarnim iskoracima i razvojem tehnologija koje su značajno uticale na javno zdravlje. Ovaj rad prikazuje istorijski razvoj vakcina, sa posebnim fokusom na nastanak i razvoj iRNK vakcina. Počevši od ranih inovacija u oblasti vakcina, rad pruža sveobuhvatan pregled kako je iRNK tehnologija unela revoluciju ovu oblast. Razmatra se jedinstveni aspekt imunološkog odgovora koji izazivaju iRNK vakcine, ističući njihovu sposobnost da proizvedu snažan i ciljan imunitet protiv patogena. Takođe, rad analizira proces proizvodnje iRNK vakcina, objašnjavajući korake od dizajna plazmida preko sinteze iRNK do enkapsulacije u lipidne nanopartikule. Diskusija uključuje analizu izazova tokom faza razvoja i kako su ti izazovi prevaziđeni. Praćenjem evolucije različitih tehnologija vakcina i naglašavanjem napretka koji su donele iRNK vakcine, ovaj rad naglašava njihov transformativni potencijal u prevenciji infektivnih i tretmanu malignih bolesti.
AB  - The development of vaccines has a rich history marked by groundbreaking innovations and evolving technologies that have significantly improved public health. This paper explores the historical progression of vaccines, with a particular focus on the emergence and development of mRNA vaccines. Starting from early vaccine innovations, the study provides a comprehensive overview of how mRNA technology has revolutionized the field. It delves into the unique aspects of the immune response elicited by mRNA vaccines, highlighting their ability to produce robust and targeted immunity against pathogens. Additionally, the paper examines the production process of mRNA vaccines, from plasmid design and the synthesis of mRNA strands to their encapsulation in lipid nanoparticles. The discussion includes an analysis of the challenges faced during the development phases and how these have been addressed. By tracing the evolution of vaccine technology and emphasizing the advancements brought by mRNA vaccines, this paper underscores their transformative potential in preventing infectious diseases and treating various conditions. The findings suggest that mRNA vaccines represent a pivotal shift in vaccine development, offering new avenues for future research and application in both infectious diseases and personalized medicine.
PB  - IMGGI
T2  - Trendovi u molekularnoj biologiji / Trends in Molecular Biology
T1  - Razvoj iRNK vakcina: od eksperimentalnih početaka do Nobelove nagrade
T1  - mRNA Vaccines Development: From Experimental Beginnings to the Nobel Prize
EP  - 19
IS  - 4
SP  - 8
UR  - https://hdl.handle.net/21.15107/rcub_intor_989
ER  - 

@inbook{
author = "Lukić, Ivana and Minić, Rajna and Dragačević, Luka and Blagojević, Veljko and Stamenković, Marina and Petković, Katarina and Radojević, Katarina and Kojić, Milan and Panić, Marko",
year = "2024",
abstract = "Razvoj vakcina ima bogatu istoriju koja je obeležena revolucionarnim iskoracima i razvojem tehnologija koje su značajno uticale na javno zdravlje. Ovaj rad prikazuje istorijski razvoj vakcina, sa posebnim fokusom na nastanak i razvoj iRNK vakcina. Počevši od ranih inovacija u oblasti vakcina, rad pruža sveobuhvatan pregled kako je iRNK tehnologija unela revoluciju ovu oblast. Razmatra se jedinstveni aspekt imunološkog odgovora koji izazivaju iRNK vakcine, ističući njihovu sposobnost da proizvedu snažan i ciljan imunitet protiv patogena. Takođe, rad analizira proces proizvodnje iRNK vakcina, objašnjavajući korake od dizajna plazmida preko sinteze iRNK do enkapsulacije u lipidne nanopartikule. Diskusija uključuje analizu izazova tokom faza razvoja i kako su ti izazovi prevaziđeni. Praćenjem evolucije različitih tehnologija vakcina i naglašavanjem napretka koji su donele iRNK vakcine, ovaj rad naglašava njihov transformativni potencijal u prevenciji infektivnih i tretmanu malignih bolesti., The development of vaccines has a rich history marked by groundbreaking innovations and evolving technologies that have significantly improved public health. This paper explores the historical progression of vaccines, with a particular focus on the emergence and development of mRNA vaccines. Starting from early vaccine innovations, the study provides a comprehensive overview of how mRNA technology has revolutionized the field. It delves into the unique aspects of the immune response elicited by mRNA vaccines, highlighting their ability to produce robust and targeted immunity against pathogens. Additionally, the paper examines the production process of mRNA vaccines, from plasmid design and the synthesis of mRNA strands to their encapsulation in lipid nanoparticles. The discussion includes an analysis of the challenges faced during the development phases and how these have been addressed. By tracing the evolution of vaccine technology and emphasizing the advancements brought by mRNA vaccines, this paper underscores their transformative potential in preventing infectious diseases and treating various conditions. The findings suggest that mRNA vaccines represent a pivotal shift in vaccine development, offering new avenues for future research and application in both infectious diseases and personalized medicine.",
publisher = "IMGGI",
journal = "Trendovi u molekularnoj biologiji / Trends in Molecular Biology",
booktitle = "Razvoj iRNK vakcina: od eksperimentalnih početaka do Nobelove nagrade, mRNA Vaccines Development: From Experimental Beginnings to the Nobel Prize",
pages = "19-8",
number = "4",
url = "https://hdl.handle.net/21.15107/rcub_intor_989"
}

Lukić, I., Minić, R., Dragačević, L., Blagojević, V., Stamenković, M., Petković, K., Radojević, K., Kojić, M.,& Panić, M.. (2024). Razvoj iRNK vakcina: od eksperimentalnih početaka do Nobelove nagrade. in Trendovi u molekularnoj biologiji / Trends in Molecular Biology
IMGGI.(4), 8-19.
https://hdl.handle.net/21.15107/rcub_intor_989

Lukić I, Minić R, Dragačević L, Blagojević V, Stamenković M, Petković K, Radojević K, Kojić M, Panić M. Razvoj iRNK vakcina: od eksperimentalnih početaka do Nobelove nagrade. in Trendovi u molekularnoj biologiji / Trends in Molecular Biology. 2024;(4):8-19.
https://hdl.handle.net/21.15107/rcub_intor_989 .

Lukić, Ivana, Minić, Rajna, Dragačević, Luka, Blagojević, Veljko, Stamenković, Marina, Petković, Katarina, Radojević, Katarina, Kojić, Milan, Panić, Marko, "Razvoj iRNK vakcina: od eksperimentalnih početaka do Nobelove nagrade" in Trendovi u molekularnoj biologiji / Trends in Molecular Biology, no. 4 (2024):8-19,
https://hdl.handle.net/21.15107/rcub_intor_989 .

TY  - JOUR
AU  - Filipić, Brankica
AU  - Kojić, Milan
AU  - Vasiljević, Zorica
AU  - Sovtić, Aleksandar
AU  - Dimkić, Ivica
AU  - Wood, Emily
AU  - Esposito, Alfonso
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/987
AB  - Escherichia coli is a Gram-negative bacterium and part of the intestinal microbiota. However, it can cause various diarrhoeal illnesses, i.e., traveller’s diarrhoea, dysentery, and extraintestinal infections when the bacteria are translocated from the intestine to other organs, such as urinary tract infections, abdominal and pelvic infections, pneumonia, bacteraemia, and meningitis. It is also an important pathogen in intensive care units where cross-infection may cause intrahospital spread with serious consequences. Within this study, four E. coli isolates from the tracheal aspirates of a tracheotomised paediatric patient on chronic respiratory support were analysed and compared for antibiotic resistance and virulence potential. Genomes of all four isolates (5381a, 5381b, 5681, 5848) were sequenced using Oxford Nanopore Technology. According to PFGE analysis, the clones of isolates 5681 and 5848 were highly similar, and differ from 5381a and 5381b which were isolated first chronologically. All four E. coli isolates belonged to an unknown sequence type, related to the E. coli ST131, a pandemic clone that is evolving rapidly with increasing levels of antimicrobial resistance. All four E. coli isolates in this study exhibited a multidrug-resistant phenotype as, according to MIC data, they were resistant to ceftriaxone, ciprofloxacin, doxycycline, minocycline, and tetracycline. In addition, principal component analyses revealed that isolates 5681 and 5848, which were recovered later than 5381a and 5381b (two weeks and three weeks, respectively) possessed more complex antibiotic resistance genes and virulence profiles, which is concerning considering the short time period during which the strains were isolated.
PB  - MDPI
T2  - Microorganisms
T1  - A Longitudinal Study of Escherichia coli Clinical Isolates from the Tracheal Aspirates of a Paediatric Patient—Strain Type Similar to Pandemic ST131
IS  - 10
SP  - 1990
VL  - 12
DO  - 10.3390/microorganisms12101990
ER  - 

@article{
author = "Filipić, Brankica and Kojić, Milan and Vasiljević, Zorica and Sovtić, Aleksandar and Dimkić, Ivica and Wood, Emily and Esposito, Alfonso",
year = "2024",
abstract = "Escherichia coli is a Gram-negative bacterium and part of the intestinal microbiota. However, it can cause various diarrhoeal illnesses, i.e., traveller’s diarrhoea, dysentery, and extraintestinal infections when the bacteria are translocated from the intestine to other organs, such as urinary tract infections, abdominal and pelvic infections, pneumonia, bacteraemia, and meningitis. It is also an important pathogen in intensive care units where cross-infection may cause intrahospital spread with serious consequences. Within this study, four E. coli isolates from the tracheal aspirates of a tracheotomised paediatric patient on chronic respiratory support were analysed and compared for antibiotic resistance and virulence potential. Genomes of all four isolates (5381a, 5381b, 5681, 5848) were sequenced using Oxford Nanopore Technology. According to PFGE analysis, the clones of isolates 5681 and 5848 were highly similar, and differ from 5381a and 5381b which were isolated first chronologically. All four E. coli isolates belonged to an unknown sequence type, related to the E. coli ST131, a pandemic clone that is evolving rapidly with increasing levels of antimicrobial resistance. All four E. coli isolates in this study exhibited a multidrug-resistant phenotype as, according to MIC data, they were resistant to ceftriaxone, ciprofloxacin, doxycycline, minocycline, and tetracycline. In addition, principal component analyses revealed that isolates 5681 and 5848, which were recovered later than 5381a and 5381b (two weeks and three weeks, respectively) possessed more complex antibiotic resistance genes and virulence profiles, which is concerning considering the short time period during which the strains were isolated.",
publisher = "MDPI",
journal = "Microorganisms",
title = "A Longitudinal Study of Escherichia coli Clinical Isolates from the Tracheal Aspirates of a Paediatric Patient—Strain Type Similar to Pandemic ST131",
number = "10",
pages = "1990",
volume = "12",
doi = "10.3390/microorganisms12101990"
}

Filipić, B., Kojić, M., Vasiljević, Z., Sovtić, A., Dimkić, I., Wood, E.,& Esposito, A.. (2024). A Longitudinal Study of Escherichia coli Clinical Isolates from the Tracheal Aspirates of a Paediatric Patient—Strain Type Similar to Pandemic ST131. in Microorganisms
MDPI., 12(10), 1990.
https://doi.org/10.3390/microorganisms12101990

Filipić B, Kojić M, Vasiljević Z, Sovtić A, Dimkić I, Wood E, Esposito A. A Longitudinal Study of Escherichia coli Clinical Isolates from the Tracheal Aspirates of a Paediatric Patient—Strain Type Similar to Pandemic ST131. in Microorganisms. 2024;12(10):1990.
doi:10.3390/microorganisms12101990 .

Filipić, Brankica, Kojić, Milan, Vasiljević, Zorica, Sovtić, Aleksandar, Dimkić, Ivica, Wood, Emily, Esposito, Alfonso, "A Longitudinal Study of Escherichia coli Clinical Isolates from the Tracheal Aspirates of a Paediatric Patient—Strain Type Similar to Pandemic ST131" in Microorganisms, 12, no. 10 (2024):1990,
https://doi.org/10.3390/microorganisms12101990 . .

TY  - JOUR
AU  - Mirković, Nemanja
AU  - Radulović, Zorica
AU  - Jovcic, Branko
AU  - Stanisavljević, Nemanja
AU  - Kojić, Milan
AU  - Oz, Fatih
AU  - Proestos, Charalampos
AU  - Heinz, Volker
AU  - Tomašević, Igor
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/977
AB  - The presence of pathogens in food has increased awareness of food safety, but it also causes large economic losses. Fresh meat and meat products contain a sufficient quantity of proteins, lipids, water, and a favorable pH that stimulates the growth of various microorganisms, including pathogens.The aim of this study was to investigate the efficacy of Brevibacillus laterosporus BGSP7 (CFS-BGSP7) and BGSP9 (CFS-BGSP9) cell-free supernatants in the control of Listeria monocytogenes and Staphylococcus aureus in raw meat and meat products.Raw meat and meat products were sliced and then aseptically treated by immersion for 2 minutes into solutions containing: i) CFS-BGSP7; ii) CFS-BGSP9; iii) no treatment. The samples were then artificially contaminated with: Group I – L. monocytogenes (~4 log cfu g-1); Group II – S. aureus LMM322 (~4 log cfu g-1). Each sample was individually aseptically vacuum-packed and stored at 4°C for 8 weeks. The number of surviving bacteria in the samples were analyzed immediately after contamination with L. monocytogenes and S. aureus and at regular time-intervals: after 1, 3, 5 and 8 weeks of storage at 4°C.Meat samples treated with CFS-BGSP7 and CFS-SP9 showed a significant decrease in the cell counts of L. monocytogenes and S. aureus. When meat samples treated with CFS-BGSP7 and CFS-BGSP9 are compared, the results show a more intense reduction rate of both L. monocytogenes and S. aureus in all samples treated with CFS-BGSP7.
PB  - Slovak University of Agriculture in Nitra
T2  - Journal of microbiology, biotechnology and food sciences
T1  - Control of Listeria monocytogenes and Staphylococcus aureus in meat and meat products by cell-free supernatant of Brevibacillus laterosporus BGSP7 and BGSP9
SP  - e11182
DO  - 10.55251/jmbfs.11182
ER  - 

@article{
author = "Mirković, Nemanja and Radulović, Zorica and Jovcic, Branko and Stanisavljević, Nemanja and Kojić, Milan and Oz, Fatih and Proestos, Charalampos and Heinz, Volker and Tomašević, Igor",
year = "2024",
abstract = "The presence of pathogens in food has increased awareness of food safety, but it also causes large economic losses. Fresh meat and meat products contain a sufficient quantity of proteins, lipids, water, and a favorable pH that stimulates the growth of various microorganisms, including pathogens.The aim of this study was to investigate the efficacy of Brevibacillus laterosporus BGSP7 (CFS-BGSP7) and BGSP9 (CFS-BGSP9) cell-free supernatants in the control of Listeria monocytogenes and Staphylococcus aureus in raw meat and meat products.Raw meat and meat products were sliced and then aseptically treated by immersion for 2 minutes into solutions containing: i) CFS-BGSP7; ii) CFS-BGSP9; iii) no treatment. The samples were then artificially contaminated with: Group I – L. monocytogenes (~4 log cfu g-1); Group II – S. aureus LMM322 (~4 log cfu g-1). Each sample was individually aseptically vacuum-packed and stored at 4°C for 8 weeks. The number of surviving bacteria in the samples were analyzed immediately after contamination with L. monocytogenes and S. aureus and at regular time-intervals: after 1, 3, 5 and 8 weeks of storage at 4°C.Meat samples treated with CFS-BGSP7 and CFS-SP9 showed a significant decrease in the cell counts of L. monocytogenes and S. aureus. When meat samples treated with CFS-BGSP7 and CFS-BGSP9 are compared, the results show a more intense reduction rate of both L. monocytogenes and S. aureus in all samples treated with CFS-BGSP7.",
publisher = "Slovak University of Agriculture in Nitra",
journal = "Journal of microbiology, biotechnology and food sciences",
title = "Control of Listeria monocytogenes and Staphylococcus aureus in meat and meat products by cell-free supernatant of Brevibacillus laterosporus BGSP7 and BGSP9",
pages = "e11182",
doi = "10.55251/jmbfs.11182"
}

Mirković, N., Radulović, Z., Jovcic, B., Stanisavljević, N., Kojić, M., Oz, F., Proestos, C., Heinz, V.,& Tomašević, I.. (2024). Control of Listeria monocytogenes and Staphylococcus aureus in meat and meat products by cell-free supernatant of Brevibacillus laterosporus BGSP7 and BGSP9. in Journal of microbiology, biotechnology and food sciences
Slovak University of Agriculture in Nitra., e11182.
https://doi.org/10.55251/jmbfs.11182

Mirković N, Radulović Z, Jovcic B, Stanisavljević N, Kojić M, Oz F, Proestos C, Heinz V, Tomašević I. Control of Listeria monocytogenes and Staphylococcus aureus in meat and meat products by cell-free supernatant of Brevibacillus laterosporus BGSP7 and BGSP9. in Journal of microbiology, biotechnology and food sciences. 2024;:e11182.
doi:10.55251/jmbfs.11182 .

Mirković, Nemanja, Radulović, Zorica, Jovcic, Branko, Stanisavljević, Nemanja, Kojić, Milan, Oz, Fatih, Proestos, Charalampos, Heinz, Volker, Tomašević, Igor, "Control of Listeria monocytogenes and Staphylococcus aureus in meat and meat products by cell-free supernatant of Brevibacillus laterosporus BGSP7 and BGSP9" in Journal of microbiology, biotechnology and food sciences (2024):e11182,
https://doi.org/10.55251/jmbfs.11182 . .

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 . .

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.
PB  - Elsevier
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.",
publisher = "Elsevier",
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
Elsevier., 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 . .

TY  - CONF
AU  - Tsibulskaya, Darya
AU  - Blagojević, Veljko
AU  - Terzić-Vidojević, Amarela
AU  - Lukić, Ivana
AU  - Vasić, Marko
AU  - Dragačević, Luka
AU  - Kojić, Milan
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/875
AB  - Autoaggregation, the ability to self-aggregate, is widespread among both Gram-positive and Gram-negative bacteria. The functional role of aggregation is not fully understood, but it is believed to be involved in the adaptation of bacteria to environmental conditions (PMID: 31294207). One interesting class of compounds responsible for the aggregation of lactic acid bacteria is aggregation factors—surface high-molecular-weight proteins rich in threonine and lysine (PMID: 30027759). Recently, our research group discovered a new strain of Streptococcus thermophilus in the mountainous regions of Serbia, exhibiting an aggregation phenotype. Aggregation phenotype was confirmed visually and using microscopy. Complete genome of Agg+ strain was sequenced using NGS and a gene encoding a potential aggregation factor, which was named aggS was identified. The predicted threonine (12.5%) and lysine (10.5%) rich protein contains 2367 amino acids, with an average molecular weight of 255986.63 Da. AggS also contains two cysteine residues, whereas previously well-described aggregation factors of this type did not contain any cysteine residues. The predicted protein includes an N-terminal YSIRK-like signal sequence and an LPXTG cell wall anchor domain. It has 6 Mucin binding domain repeats alternating with 6 Mub B2-like domain repeats. Additionally, we found a region resembling an ice-binding domain. Given that these bacteria endure prolonged periods of low temperatures, it can be speculated that this surface membrane protein also helps the bacteria withstand freezing. The fact that the alignment using BLASTp revealed AggS to be most closely related to an uncharacterised protein from the genome of Lactococcus garvieae, along with the discovery of a transposase gene sequence upstream of the gene, suggests that the aggregation factor was likely acquired through horizontal gene transfer. We plan to clone it into a shuttle vector and investigate the aggregation phenotype using a heterologous expression system in Lactococcus lactis, as well as explore its other functions.
PB  - Serbian Society for Microbiology
C3  - XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april
T1  - Description of a new potential aggregation factor from the Streptococcus thermophilus genome
EP  - 110
SP  - 110
UR  - https://hdl.handle.net/21.15107/rcub_intor_875
ER  - 

@conference{
author = "Tsibulskaya, Darya and Blagojević, Veljko and Terzić-Vidojević, Amarela and Lukić, Ivana and Vasić, Marko and Dragačević, Luka and Kojić, Milan",
year = "2024",
abstract = "Autoaggregation, the ability to self-aggregate, is widespread among both Gram-positive and Gram-negative bacteria. The functional role of aggregation is not fully understood, but it is believed to be involved in the adaptation of bacteria to environmental conditions (PMID: 31294207). One interesting class of compounds responsible for the aggregation of lactic acid bacteria is aggregation factors—surface high-molecular-weight proteins rich in threonine and lysine (PMID: 30027759). Recently, our research group discovered a new strain of Streptococcus thermophilus in the mountainous regions of Serbia, exhibiting an aggregation phenotype. Aggregation phenotype was confirmed visually and using microscopy. Complete genome of Agg+ strain was sequenced using NGS and a gene encoding a potential aggregation factor, which was named aggS was identified. The predicted threonine (12.5%) and lysine (10.5%) rich protein contains 2367 amino acids, with an average molecular weight of 255986.63 Da. AggS also contains two cysteine residues, whereas previously well-described aggregation factors of this type did not contain any cysteine residues. The predicted protein includes an N-terminal YSIRK-like signal sequence and an LPXTG cell wall anchor domain. It has 6 Mucin binding domain repeats alternating with 6 Mub B2-like domain repeats. Additionally, we found a region resembling an ice-binding domain. Given that these bacteria endure prolonged periods of low temperatures, it can be speculated that this surface membrane protein also helps the bacteria withstand freezing. The fact that the alignment using BLASTp revealed AggS to be most closely related to an uncharacterised protein from the genome of Lactococcus garvieae, along with the discovery of a transposase gene sequence upstream of the gene, suggests that the aggregation factor was likely acquired through horizontal gene transfer. We plan to clone it into a shuttle vector and investigate the aggregation phenotype using a heterologous expression system in Lactococcus lactis, as well as explore its other functions.",
publisher = "Serbian Society for Microbiology",
journal = "XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april",
title = "Description of a new potential aggregation factor from the Streptococcus thermophilus genome",
pages = "110-110",
url = "https://hdl.handle.net/21.15107/rcub_intor_875"
}

Tsibulskaya, D., Blagojević, V., Terzić-Vidojević, A., Lukić, I., Vasić, M., Dragačević, L.,& Kojić, M.. (2024). Description of a new potential aggregation factor from the Streptococcus thermophilus genome. in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april
Serbian Society for Microbiology., 110-110.
https://hdl.handle.net/21.15107/rcub_intor_875

Tsibulskaya D, Blagojević V, Terzić-Vidojević A, Lukić I, Vasić M, Dragačević L, Kojić M. Description of a new potential aggregation factor from the Streptococcus thermophilus genome. in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april. 2024;:110-110.
https://hdl.handle.net/21.15107/rcub_intor_875 .

Tsibulskaya, Darya, Blagojević, Veljko, Terzić-Vidojević, Amarela, Lukić, Ivana, Vasić, Marko, Dragačević, Luka, Kojić, Milan, "Description of a new potential aggregation factor from the Streptococcus thermophilus genome" in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april (2024):110-110,
https://hdl.handle.net/21.15107/rcub_intor_875 .

TY  - CONF
AU  - Panić, Marko
AU  - Prijić, Ivana
AU  - Simić, Mihajlo
AU  - Lukić, Ivana
AU  - Petrušić, Marija
AU  - Živković, Irena
AU  - Kojić, Milan
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/876
AB  - Tetanus toxin, a highly potent neurotoxin produced by Clostridium tetani, is the primary agent responsible for causing tetanus. This serious, potentially fatal disease can be effectively prevented through vaccination. Thanks to successful vaccination campaigns, tetanus has become exceedingly rare in both developed and most developing countries. However, the widespread presence of C. tetani spores in the environment means that tetanus cannot be completely eradicated, underscoring the ongoing need for vaccination. Traditionally, tetanus vaccines are produced by cultivating C. tetani, extracting a crude form of the tetanus toxin, and then chemically inactivating it for use in immunization. This method has proven clinically effective and is in widespread use. A challenge with this approach, however, is that the vaccine contains hundreds of various C. tetani proteins, with the active component making up only a variable and small fraction of the overall vaccine mass. To improve the current tetanus vaccine, there is potential in the recombinant production of a genetically inactivated tetanus vaccine. Prior studies have demonstrated the feasibility of engineering the full-length tetanus toxin in E. coli, and our current work builds on this foundation. We have successfully cloned the complete tetanus toxin open reading frame into the pMAL expression vector. This step was followed by the creation of a genetically inactivated protein, achieved through standard site-directed mutagenesis which altered 8 critical amino acid residues. These mutations have been confirmed via sequencing, ensuring that the toxin is genetically inactivated and thus does not require chemical inactivation for vaccine production. Our present focus is on optimizing the expression of this protein in E. coli. Following this, we intend to conduct thorough assessments of the biochemical and immunological properties of the recombinant tetanus toxin. This research represents a promising avenue towards enhancing the efficacy and specificity of tetanus vaccines, potentially improving global health outcomes.
PB  - Serbian Society for Microbiology
C3  - XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april
T1  - Exploring E. coli-based expression of genetically inactivated tetanus toxin for vaccine development
EP  - 113
SP  - 113
UR  - https://hdl.handle.net/21.15107/rcub_intor_876
ER  - 

@conference{
author = "Panić, Marko and Prijić, Ivana and Simić, Mihajlo and Lukić, Ivana and Petrušić, Marija and Živković, Irena and Kojić, Milan",
year = "2024",
abstract = "Tetanus toxin, a highly potent neurotoxin produced by Clostridium tetani, is the primary agent responsible for causing tetanus. This serious, potentially fatal disease can be effectively prevented through vaccination. Thanks to successful vaccination campaigns, tetanus has become exceedingly rare in both developed and most developing countries. However, the widespread presence of C. tetani spores in the environment means that tetanus cannot be completely eradicated, underscoring the ongoing need for vaccination. Traditionally, tetanus vaccines are produced by cultivating C. tetani, extracting a crude form of the tetanus toxin, and then chemically inactivating it for use in immunization. This method has proven clinically effective and is in widespread use. A challenge with this approach, however, is that the vaccine contains hundreds of various C. tetani proteins, with the active component making up only a variable and small fraction of the overall vaccine mass. To improve the current tetanus vaccine, there is potential in the recombinant production of a genetically inactivated tetanus vaccine. Prior studies have demonstrated the feasibility of engineering the full-length tetanus toxin in E. coli, and our current work builds on this foundation. We have successfully cloned the complete tetanus toxin open reading frame into the pMAL expression vector. This step was followed by the creation of a genetically inactivated protein, achieved through standard site-directed mutagenesis which altered 8 critical amino acid residues. These mutations have been confirmed via sequencing, ensuring that the toxin is genetically inactivated and thus does not require chemical inactivation for vaccine production. Our present focus is on optimizing the expression of this protein in E. coli. Following this, we intend to conduct thorough assessments of the biochemical and immunological properties of the recombinant tetanus toxin. This research represents a promising avenue towards enhancing the efficacy and specificity of tetanus vaccines, potentially improving global health outcomes.",
publisher = "Serbian Society for Microbiology",
journal = "XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april",
title = "Exploring E. coli-based expression of genetically inactivated tetanus toxin for vaccine development",
pages = "113-113",
url = "https://hdl.handle.net/21.15107/rcub_intor_876"
}

Panić, M., Prijić, I., Simić, M., Lukić, I., Petrušić, M., Živković, I.,& Kojić, M.. (2024). Exploring E. coli-based expression of genetically inactivated tetanus toxin for vaccine development. in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april
Serbian Society for Microbiology., 113-113.
https://hdl.handle.net/21.15107/rcub_intor_876

Panić M, Prijić I, Simić M, Lukić I, Petrušić M, Živković I, Kojić M. Exploring E. coli-based expression of genetically inactivated tetanus toxin for vaccine development. in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april. 2024;:113-113.
https://hdl.handle.net/21.15107/rcub_intor_876 .

Panić, Marko, Prijić, Ivana, Simić, Mihajlo, Lukić, Ivana, Petrušić, Marija, Živković, Irena, Kojić, Milan, "Exploring E. coli-based expression of genetically inactivated tetanus toxin for vaccine development" in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april (2024):113-113,
https://hdl.handle.net/21.15107/rcub_intor_876 .

TY  - CONF
AU  - Lukić, Ivana
AU  - Dragačević, Luka
AU  - Panić, Marko
AU  - Stamenković, Marina
AU  - Kojić, Milan
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/878
AB  - In the post-COVID-19 era, there has been a significant increase in the development of mRNA vaccines not only against various diseases besides SARS-CoV-2, but also to treat cancer and genetic disorders. These vaccines, revolutionizing vaccinology, offer rapid pandemic response, high efficacy, minimal side effects, and cost-effectiveness. Achieving these benefits hinges on seamlessly integrating mRNA production steps, from plasmid DNA cloning to lipid nanoparticle formulation. This overview aims to comprehend or circumvent pitfalls in plasmid DNA cloning, a critical initial step in mRNA vaccine production. The focus is on achieving accurate insert sequence and gene expression, and it highlights the critical role of plasmid DNA design in ensuring vaccine effectiveness. Our research project entitled “Role of macroautophagy in lipid nanoparticle mRNA delivery and adjuvanticity” recognized the significance of this aspect. During our research, we designed a plasmid DNA cloning vector to incorporate the GFP-SARS-CoV-2 Spike gene. The vector was carefully constructed with several key features, including a high-copy plasmid, pUC18/pUC19 vector backbone with a robust T7 promoter, origin of replication, multiple cloning sites, polyadenylation signal, and ampicillin resistance for bacterial selection. Despite careful design, challenges like poly-A tail deletion may arise, prompting the exploration of stable large-size and low-copy vectors, as well as linear and bacteriophage vectors. But, for largescale production and regulatory compliance, vector systems must be scalable and well-documented. Commercial vectors and automated synthesis facilitate gene construction, with artificial intelligence ensuring sequence accuracy. Precision is crucial for complex antigens, as seen in tuberculosis mRNA vaccine development. Addressing these challenges demands a combining of molecular biology techniques, computational tools, and collaboration with experts in microbiology, molecular biology, and vaccine development. The design’s scalability and documentation are vital for large-scale production and regulatory compliance, emphasizing the multifaceted approach required for successful mRNA vaccine development.
PB  - Serbian Society for Microbiology
C3  - XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april
T1  - mRNA vaccine manufacturing – challenges in plasmid DNA cloning vector design
EP  - 157
SP  - 157
UR  - https://hdl.handle.net/21.15107/rcub_intor_878
ER  - 

@conference{
author = "Lukić, Ivana and Dragačević, Luka and Panić, Marko and Stamenković, Marina and Kojić, Milan",
year = "2024",
abstract = "In the post-COVID-19 era, there has been a significant increase in the development of mRNA vaccines not only against various diseases besides SARS-CoV-2, but also to treat cancer and genetic disorders. These vaccines, revolutionizing vaccinology, offer rapid pandemic response, high efficacy, minimal side effects, and cost-effectiveness. Achieving these benefits hinges on seamlessly integrating mRNA production steps, from plasmid DNA cloning to lipid nanoparticle formulation. This overview aims to comprehend or circumvent pitfalls in plasmid DNA cloning, a critical initial step in mRNA vaccine production. The focus is on achieving accurate insert sequence and gene expression, and it highlights the critical role of plasmid DNA design in ensuring vaccine effectiveness. Our research project entitled “Role of macroautophagy in lipid nanoparticle mRNA delivery and adjuvanticity” recognized the significance of this aspect. During our research, we designed a plasmid DNA cloning vector to incorporate the GFP-SARS-CoV-2 Spike gene. The vector was carefully constructed with several key features, including a high-copy plasmid, pUC18/pUC19 vector backbone with a robust T7 promoter, origin of replication, multiple cloning sites, polyadenylation signal, and ampicillin resistance for bacterial selection. Despite careful design, challenges like poly-A tail deletion may arise, prompting the exploration of stable large-size and low-copy vectors, as well as linear and bacteriophage vectors. But, for largescale production and regulatory compliance, vector systems must be scalable and well-documented. Commercial vectors and automated synthesis facilitate gene construction, with artificial intelligence ensuring sequence accuracy. Precision is crucial for complex antigens, as seen in tuberculosis mRNA vaccine development. Addressing these challenges demands a combining of molecular biology techniques, computational tools, and collaboration with experts in microbiology, molecular biology, and vaccine development. The design’s scalability and documentation are vital for large-scale production and regulatory compliance, emphasizing the multifaceted approach required for successful mRNA vaccine development.",
publisher = "Serbian Society for Microbiology",
journal = "XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april",
title = "mRNA vaccine manufacturing – challenges in plasmid DNA cloning vector design",
pages = "157-157",
url = "https://hdl.handle.net/21.15107/rcub_intor_878"
}

Lukić, I., Dragačević, L., Panić, M., Stamenković, M.,& Kojić, M.. (2024). mRNA vaccine manufacturing – challenges in plasmid DNA cloning vector design. in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april
Serbian Society for Microbiology., 157-157.
https://hdl.handle.net/21.15107/rcub_intor_878

Lukić I, Dragačević L, Panić M, Stamenković M, Kojić M. mRNA vaccine manufacturing – challenges in plasmid DNA cloning vector design. in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april. 2024;:157-157.
https://hdl.handle.net/21.15107/rcub_intor_878 .

Lukić, Ivana, Dragačević, Luka, Panić, Marko, Stamenković, Marina, Kojić, Milan, "mRNA vaccine manufacturing – challenges in plasmid DNA cloning vector design" in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april (2024):157-157,
https://hdl.handle.net/21.15107/rcub_intor_878 .

TY  - CONF
AU  - Panić, Marko
AU  - Prijić, Ivana
AU  - Simić, Mihajlo
AU  - Ćuruvija, Ivana
AU  - Lukić, Ivana
AU  - Drgačević, Luka
AU  - Kojić, Milan
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/880
AB  - Diphtheria and tetanus, once formidable causes of morbidity and mortality worldwide, have seen their threats markedly diminished through the advent and widespread use of vaccines. This review article delves into the historical journey of diphtheria and tetanus vaccines, evaluates their current status in global immunization programs, and explores future perspectives in their evolution and implementation. The inception of diphtheria and tetanus vaccines marked a pivotal shift in infectious disease control. The development of diphtheria toxoid by Emil von Behring in the late 19th century and the subsequent creation of tetanus toxoid in the early 20th century set the stage for large-scale immunization efforts. These efforts were bolstered in the mid-20th century with the integration of these toxoids into combination vaccines, notably the DTP (diphtheria-tetanus-pertussis) vaccine, facilitating broader immunization coverage and enhanced public health outcomes. Currently, the inclusion of diphtheria and tetanus vaccines in national immunization schedules has led to a significant decline in the incidence of these diseases globally. However, challenges remain, including disparities in vaccine coverage and the emergence of non-toxigenic strains causing diphtheria. The review highlights the WHO’s strategies towards achieving higher immunization coverage and the importance of maintaining high vaccination rates to prevent resurgence. Looking forward, the review discusses the ongoing research and development aimed at improving vaccine formulations, reducing adverse reactions, and enhancing the efficacy and durability of protection. Innovations such as nanoparticle vaccines and DNA vaccines are explored as potential avenues for future advancements. Additionally, the review addresses the critical role of global health governance in addressing vaccine hesitancy, improving access in low-resource settings, and coordinating responses to outbreaks. In conclusion, while the battle against diphtheria and tetanus has seen significant victories, continuous efforts in vaccine innovation, policy implementation, and global cooperation are essential to sustain these gains and achieve the ultimate goal of global eradication.
PB  - Serbian Society for Microbiology
C3  - XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april
T1  - Diphtheria and tetanus vaccines: a historical overview, present achievements, and future directions
EP  - 169
SP  - 169
UR  - https://hdl.handle.net/21.15107/rcub_intor_880
ER  - 

@conference{
author = "Panić, Marko and Prijić, Ivana and Simić, Mihajlo and Ćuruvija, Ivana and Lukić, Ivana and Drgačević, Luka and Kojić, Milan",
year = "2024",
abstract = "Diphtheria and tetanus, once formidable causes of morbidity and mortality worldwide, have seen their threats markedly diminished through the advent and widespread use of vaccines. This review article delves into the historical journey of diphtheria and tetanus vaccines, evaluates their current status in global immunization programs, and explores future perspectives in their evolution and implementation. The inception of diphtheria and tetanus vaccines marked a pivotal shift in infectious disease control. The development of diphtheria toxoid by Emil von Behring in the late 19th century and the subsequent creation of tetanus toxoid in the early 20th century set the stage for large-scale immunization efforts. These efforts were bolstered in the mid-20th century with the integration of these toxoids into combination vaccines, notably the DTP (diphtheria-tetanus-pertussis) vaccine, facilitating broader immunization coverage and enhanced public health outcomes. Currently, the inclusion of diphtheria and tetanus vaccines in national immunization schedules has led to a significant decline in the incidence of these diseases globally. However, challenges remain, including disparities in vaccine coverage and the emergence of non-toxigenic strains causing diphtheria. The review highlights the WHO’s strategies towards achieving higher immunization coverage and the importance of maintaining high vaccination rates to prevent resurgence. Looking forward, the review discusses the ongoing research and development aimed at improving vaccine formulations, reducing adverse reactions, and enhancing the efficacy and durability of protection. Innovations such as nanoparticle vaccines and DNA vaccines are explored as potential avenues for future advancements. Additionally, the review addresses the critical role of global health governance in addressing vaccine hesitancy, improving access in low-resource settings, and coordinating responses to outbreaks. In conclusion, while the battle against diphtheria and tetanus has seen significant victories, continuous efforts in vaccine innovation, policy implementation, and global cooperation are essential to sustain these gains and achieve the ultimate goal of global eradication.",
publisher = "Serbian Society for Microbiology",
journal = "XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april",
title = "Diphtheria and tetanus vaccines: a historical overview, present achievements, and future directions",
pages = "169-169",
url = "https://hdl.handle.net/21.15107/rcub_intor_880"
}

Panić, M., Prijić, I., Simić, M., Ćuruvija, I., Lukić, I., Drgačević, L.,& Kojić, M.. (2024). Diphtheria and tetanus vaccines: a historical overview, present achievements, and future directions. in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april
Serbian Society for Microbiology., 169-169.
https://hdl.handle.net/21.15107/rcub_intor_880

Panić M, Prijić I, Simić M, Ćuruvija I, Lukić I, Drgačević L, Kojić M. Diphtheria and tetanus vaccines: a historical overview, present achievements, and future directions. in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april. 2024;:169-169.
https://hdl.handle.net/21.15107/rcub_intor_880 .

Panić, Marko, Prijić, Ivana, Simić, Mihajlo, Ćuruvija, Ivana, Lukić, Ivana, Drgačević, Luka, Kojić, Milan, "Diphtheria and tetanus vaccines: a historical overview, present achievements, and future directions" in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april (2024):169-169,
https://hdl.handle.net/21.15107/rcub_intor_880 .

TY  - CONF
AU  - Vukotić, Goran
AU  - Obradović, Mina
AU  - Plačkić, Nikola
AU  - Kljajević, Nemanja
AU  - Pavić, Aleksandar
AU  - Kekić, Dušan
AU  - Gajić, Ina
AU  - Kojić, Milan
AU  - Stanisavljević, Nemanja
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/885
AB  - Antimicrobial resistance (AMR) arises whenbacteria and other microbes stop respondingto medications. AMR is now recognized as oneof serious global health threats, repeatedlyappearing in the World Health Organization’s(WHO) lists of urgent global health challenges,including the 2024 list. It is taking a fatal toll– nearly 5 million deaths globally per year areassociated with AMR, encompassing 1.27 milliondirectly attributed to AMR. The COVID-19pandemic paved the way for aggravation ofbacterial AMR – primarily due to enhancementin unspecific and unjustified prescription anduse of broad-spectrum antibiotics, resulting inwhat is now recognized as „silent pandemic ofAMR“. Bacteriophages (phages) are natural andspecific predators of bacteria - viruses that caninfect, replicate inside and lyse arguably anybacteria. Their therapeutic potential is beinghastily evaluated through different approaches:in silico, in vitro, ex vivo and in vivo – in laboratoryanimals as well as in human case and clinicalstudies. Although the results are promising,bacteria rapidly develop resistance againstphages, which why the isolation and researchof new phages is needed. Our work is concentratedon three bacterial species for which criticalpriority by WHO has been declared – carbapenem-resistant Acinetobacter baumannii,Pseudomonas aeruginosa and Klebsiella pneumoniae.Twenty distinct pathogenic strains ofA. baumannii, 6 K. pneumoniae and 6 P. aeruginosawere used as targets for bacteriophageisolation, and total of 14, 22 and 8 potentiallydistinct phages were collected, respectively. Allstrains were nosocomial isolates obtained fromvarious tissues, including from terminally ill patients.Six phages were characterized in detail.In particular, phage vB_AbaM_ISTD was appliedagainst A. baumannii in zebrafish embryomodel of systemic infection, and demonstratedpowerful therapeutic potential, eradicating theinfection. Interestingly, its DNA was characterizedwith highly modified thymidine (amassing1228 Da), making it the largest non-canonicaldeoxynucleoside reported so far.
PB  - Serbian Society for Microbiology
C3  - XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health
T1  - Bacteriophages of multidrug-resistant nosocomial pathogens – Belgrade experience
EP  - 121
SP  - 121
UR  - https://hdl.handle.net/21.15107/rcub_intor_885
ER  - 

@conference{
author = "Vukotić, Goran and Obradović, Mina and Plačkić, Nikola and Kljajević, Nemanja and Pavić, Aleksandar and Kekić, Dušan and Gajić, Ina and Kojić, Milan and Stanisavljević, Nemanja",
year = "2024",
abstract = "Antimicrobial resistance (AMR) arises whenbacteria and other microbes stop respondingto medications. AMR is now recognized as oneof serious global health threats, repeatedlyappearing in the World Health Organization’s(WHO) lists of urgent global health challenges,including the 2024 list. It is taking a fatal toll– nearly 5 million deaths globally per year areassociated with AMR, encompassing 1.27 milliondirectly attributed to AMR. The COVID-19pandemic paved the way for aggravation ofbacterial AMR – primarily due to enhancementin unspecific and unjustified prescription anduse of broad-spectrum antibiotics, resulting inwhat is now recognized as „silent pandemic ofAMR“. Bacteriophages (phages) are natural andspecific predators of bacteria - viruses that caninfect, replicate inside and lyse arguably anybacteria. Their therapeutic potential is beinghastily evaluated through different approaches:in silico, in vitro, ex vivo and in vivo – in laboratoryanimals as well as in human case and clinicalstudies. Although the results are promising,bacteria rapidly develop resistance againstphages, which why the isolation and researchof new phages is needed. Our work is concentratedon three bacterial species for which criticalpriority by WHO has been declared – carbapenem-resistant Acinetobacter baumannii,Pseudomonas aeruginosa and Klebsiella pneumoniae.Twenty distinct pathogenic strains ofA. baumannii, 6 K. pneumoniae and 6 P. aeruginosawere used as targets for bacteriophageisolation, and total of 14, 22 and 8 potentiallydistinct phages were collected, respectively. Allstrains were nosocomial isolates obtained fromvarious tissues, including from terminally ill patients.Six phages were characterized in detail.In particular, phage vB_AbaM_ISTD was appliedagainst A. baumannii in zebrafish embryomodel of systemic infection, and demonstratedpowerful therapeutic potential, eradicating theinfection. Interestingly, its DNA was characterizedwith highly modified thymidine (amassing1228 Da), making it the largest non-canonicaldeoxynucleoside reported so far.",
publisher = "Serbian Society for Microbiology",
journal = "XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health",
title = "Bacteriophages of multidrug-resistant nosocomial pathogens – Belgrade experience",
pages = "121-121",
url = "https://hdl.handle.net/21.15107/rcub_intor_885"
}

Vukotić, G., Obradović, M., Plačkić, N., Kljajević, N., Pavić, A., Kekić, D., Gajić, I., Kojić, M.,& Stanisavljević, N.. (2024). Bacteriophages of multidrug-resistant nosocomial pathogens – Belgrade experience. in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health
Serbian Society for Microbiology., 121-121.
https://hdl.handle.net/21.15107/rcub_intor_885

Vukotić G, Obradović M, Plačkić N, Kljajević N, Pavić A, Kekić D, Gajić I, Kojić M, Stanisavljević N. Bacteriophages of multidrug-resistant nosocomial pathogens – Belgrade experience. in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health. 2024;:121-121.
https://hdl.handle.net/21.15107/rcub_intor_885 .

Vukotić, Goran, Obradović, Mina, Plačkić, Nikola, Kljajević, Nemanja, Pavić, Aleksandar, Kekić, Dušan, Gajić, Ina, Kojić, Milan, Stanisavljević, Nemanja, "Bacteriophages of multidrug-resistant nosocomial pathogens – Belgrade experience" in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health (2024):121-121,
https://hdl.handle.net/21.15107/rcub_intor_885 .

TY  - JOUR
AU  - Pavlović, Marija
AU  - Slavić, Marinela Šokarda
AU  - Kojić, Milan
AU  - Margetić, Aleksandra
AU  - Ristović, Marina
AU  - Drulović, Nenad
AU  - Vujčić, Zoran
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/888
AB  - Microbial pectinolytic enzymes are important in various industries, particularly food processing. This study focuses on uncovering insights into a novel pectin lyase, BvPelB, from Bacillus velezensis 16B, with the aim of enhancing fruit juice processing. The study examines the structural and functional characteristics of pectinolytic enzyme, underscoring the critical nature of substrate specificity and enzymatic reaction mechanisms. BvPelB was successfully expressed and purified, exhibiting robust activity under alkaline conditions and thermal stability. Structural analysis revealed similarities with other pectin lyases, despite limited sequence identity. Biochemical characterization showed BvPelB's preference for highly methylated pectins and its endo-acting mode of cleavage. Treatment with BvPelB significantly increased juice yield and clarity without generating excessive methanol, making it a promising candidate for fruit juice processing. Overall, this study provides valuable insights into the enzymatic properties of BvPelB and its potential industrial applications in improving fruit juice processing efficiency and quality.
PB  - Elsevier
T2  - Food Chemistry
T1  - Unveiling novel insights into Bacillus velezensis 16B pectin lyase for improved fruit juice processing
SP  - 140030
DO  - 10.1016/j.foodchem.2024.140030
ER  - 

@article{
author = "Pavlović, Marija and Slavić, Marinela Šokarda and Kojić, Milan and Margetić, Aleksandra and Ristović, Marina and Drulović, Nenad and Vujčić, Zoran",
year = "2024",
abstract = "Microbial pectinolytic enzymes are important in various industries, particularly food processing. This study focuses on uncovering insights into a novel pectin lyase, BvPelB, from Bacillus velezensis 16B, with the aim of enhancing fruit juice processing. The study examines the structural and functional characteristics of pectinolytic enzyme, underscoring the critical nature of substrate specificity and enzymatic reaction mechanisms. BvPelB was successfully expressed and purified, exhibiting robust activity under alkaline conditions and thermal stability. Structural analysis revealed similarities with other pectin lyases, despite limited sequence identity. Biochemical characterization showed BvPelB's preference for highly methylated pectins and its endo-acting mode of cleavage. Treatment with BvPelB significantly increased juice yield and clarity without generating excessive methanol, making it a promising candidate for fruit juice processing. Overall, this study provides valuable insights into the enzymatic properties of BvPelB and its potential industrial applications in improving fruit juice processing efficiency and quality.",
publisher = "Elsevier",
journal = "Food Chemistry",
title = "Unveiling novel insights into Bacillus velezensis 16B pectin lyase for improved fruit juice processing",
pages = "140030",
doi = "10.1016/j.foodchem.2024.140030"
}

Pavlović, M., Slavić, M. Š., Kojić, M., Margetić, A., Ristović, M., Drulović, N.,& Vujčić, Z.. (2024). Unveiling novel insights into Bacillus velezensis 16B pectin lyase for improved fruit juice processing. in Food Chemistry
Elsevier., 140030.
https://doi.org/10.1016/j.foodchem.2024.140030

Pavlović M, Slavić MŠ, Kojić M, Margetić A, Ristović M, Drulović N, Vujčić Z. Unveiling novel insights into Bacillus velezensis 16B pectin lyase for improved fruit juice processing. in Food Chemistry. 2024;:140030.
doi:10.1016/j.foodchem.2024.140030 .

Pavlović, Marija, Slavić, Marinela Šokarda, Kojić, Milan, Margetić, Aleksandra, Ristović, Marina, Drulović, Nenad, Vujčić, Zoran, "Unveiling novel insights into Bacillus velezensis 16B pectin lyase for improved fruit juice processing" in Food Chemistry (2024):140030,
https://doi.org/10.1016/j.foodchem.2024.140030 . .

TY  - CONF
AU  - Pavlović, Marija
AU  - Kojić, Milan
AU  - Ristović, Marina
AU  - Stojanović, Sanja
AU  - Margetić, Aleksandra
AU  - Vujčić, Zoran
AU  - Šokarda Slavić, Marinela
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/936
AB  - This research deals with the characterization of a thermostable endo-1,4-beta-xylanase enzyme from Anoxybacillus vranjensis ST4, a thermophilic bacterium isolated from Vranjska Banja hot spring, Serbia. The enzyme shows a high degree of identity with the same type of enzyme from other species of the genera Anoxybacillus (97%), Geobacillus (74%) and Paenibacillus (65%). The gene for endo-1,4-beta-xylanase from the thermophilic strain ST4 was cloned into the pQE_Ek expression vector and successfully expressed and purified from the Escherichia coli M15[pREP4]. The study encompasses recombinant production, purification, and the comprehensive characterization of the enzymatic properties of endo-1,4-beta-xylanase. This is the first successful overexpression, purification and characterization of a recombinant thermostable endo-1,4-beta-xylanase enzyme from Anoxybacillus. With a monomeric structure of 38.7 kDa, the enzyme demonstrates peak activity at 70°C and pH 6.5. Notably, it exhibits remarkable stability across a wide pH range and at high temperatures, rendering it suitable for diverse industrial applications. Investigation into the enzyme’s kinetic parameters, substrate specificity, and its ability to degrade xylan into high-energy value products further enhances understanding of its biotechnological potential. These findings underscore the significance of thermophilic bacteria and their thermostable enzymes in various industrial processes.
PB  - Serbian Society for Microbiology
C3  - XIII Congress of microbiologists of Serbia, Book of abstracts, April 4-6, 2024, Belgrade, Serbia
T1  - Cloning, overexpression and characterization of a thermostable endo-1,4-beta-xylanase from Anoxybacillus vranjensis ST4
EP  - 31
SP  - 31
UR  - https://hdl.handle.net/21.15107/rcub_intor_936
ER  - 

@conference{
author = "Pavlović, Marija and Kojić, Milan and Ristović, Marina and Stojanović, Sanja and Margetić, Aleksandra and Vujčić, Zoran and Šokarda Slavić, Marinela",
year = "2024",
abstract = "This research deals with the characterization of a thermostable endo-1,4-beta-xylanase enzyme from Anoxybacillus vranjensis ST4, a thermophilic bacterium isolated from Vranjska Banja hot spring, Serbia. The enzyme shows a high degree of identity with the same type of enzyme from other species of the genera Anoxybacillus (97%), Geobacillus (74%) and Paenibacillus (65%). The gene for endo-1,4-beta-xylanase from the thermophilic strain ST4 was cloned into the pQE_Ek expression vector and successfully expressed and purified from the Escherichia coli M15[pREP4]. The study encompasses recombinant production, purification, and the comprehensive characterization of the enzymatic properties of endo-1,4-beta-xylanase. This is the first successful overexpression, purification and characterization of a recombinant thermostable endo-1,4-beta-xylanase enzyme from Anoxybacillus. With a monomeric structure of 38.7 kDa, the enzyme demonstrates peak activity at 70°C and pH 6.5. Notably, it exhibits remarkable stability across a wide pH range and at high temperatures, rendering it suitable for diverse industrial applications. Investigation into the enzyme’s kinetic parameters, substrate specificity, and its ability to degrade xylan into high-energy value products further enhances understanding of its biotechnological potential. These findings underscore the significance of thermophilic bacteria and their thermostable enzymes in various industrial processes.",
publisher = "Serbian Society for Microbiology",
journal = "XIII Congress of microbiologists of Serbia, Book of abstracts, April 4-6, 2024, Belgrade, Serbia",
title = "Cloning, overexpression and characterization of a thermostable endo-1,4-beta-xylanase from Anoxybacillus vranjensis ST4",
pages = "31-31",
url = "https://hdl.handle.net/21.15107/rcub_intor_936"
}

Pavlović, M., Kojić, M., Ristović, M., Stojanović, S., Margetić, A., Vujčić, Z.,& Šokarda Slavić, M.. (2024). Cloning, overexpression and characterization of a thermostable endo-1,4-beta-xylanase from Anoxybacillus vranjensis ST4. in XIII Congress of microbiologists of Serbia, Book of abstracts, April 4-6, 2024, Belgrade, Serbia
Serbian Society for Microbiology., 31-31.
https://hdl.handle.net/21.15107/rcub_intor_936

Pavlović M, Kojić M, Ristović M, Stojanović S, Margetić A, Vujčić Z, Šokarda Slavić M. Cloning, overexpression and characterization of a thermostable endo-1,4-beta-xylanase from Anoxybacillus vranjensis ST4. in XIII Congress of microbiologists of Serbia, Book of abstracts, April 4-6, 2024, Belgrade, Serbia. 2024;:31-31.
https://hdl.handle.net/21.15107/rcub_intor_936 .

Pavlović, Marija, Kojić, Milan, Ristović, Marina, Stojanović, Sanja, Margetić, Aleksandra, Vujčić, Zoran, Šokarda Slavić, Marinela, "Cloning, overexpression and characterization of a thermostable endo-1,4-beta-xylanase from Anoxybacillus vranjensis ST4" in XIII Congress of microbiologists of Serbia, Book of abstracts, April 4-6, 2024, Belgrade, Serbia (2024):31-31,
https://hdl.handle.net/21.15107/rcub_intor_936 .

TY  - JOUR
AU  - Babić, Tamara
AU  - Ugrin, Milena
AU  - Jeremić, Sanja
AU  - Kojić, Milan
AU  - Dinić, Jelena
AU  - Banović Đeri, Bojana
AU  - Zodiakis, Jerome
AU  - Nikolić, Aleksandra
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/958
AB  - ackground: The pervasive role of alternative promoters in context-specific isoform expression and
the importance of promoter choice over its level of transcriptional activity have been recently implied
based on pan-cancer in silico studies. We aimed to explore this phenomenon at the cellular level on the
example of a major tumor suppressor SMAD4 in search of molecular mechanisms in colorectal cancer
that could be exploited for novel biomarkers or therapeutic approaches.
Methods: Multi-omics technologies, in silico tools and in vitro functional assays were applied to analyze
the transcripts expression and the alternative promoters’ function of the SMAD4 gene in colon cell lines
HCEC-1CT, HCT116, DLD-1, SW480 and SW620.
Results: High expression of the transcript SMAD4-213 emerged as a hallmark of colon cancer cells,
while in silico tools point to its possible additional role and potential for sponging miRNAs. Based on the
observed dysregulation of SMAD4-209 and SMAD4-213 in malignant vs. non-malignant colon cells, we
propose that their expression ratio might be a solid biomarker candidate for colorectal cancer detection.
Conclusions: A differential pattern of the respective promoters’ activity was observed that corresponds
to the expression of transcripts, confirming the role of alternative promoters in context-specific isoform
expression. The investigated SMAD4 promoters and transcripts harbor translational potential that should
be further investigated.
PB  - Ivyspring International Publisher
T2  - Journal of Cancer
T1  - Dysregulation of transcripts SMAD4-209 and SMAD4-213 and their respective promoters in colon cancer cell lines
EP  - 5131
IS  - 15
SP  - 5118
VL  - 15
DO  - 10.7150/jca.98911
ER  - 

@article{
author = "Babić, Tamara and Ugrin, Milena and Jeremić, Sanja and Kojić, Milan and Dinić, Jelena and Banović Đeri, Bojana and Zodiakis, Jerome and Nikolić, Aleksandra",
year = "2024",
abstract = "ackground: The pervasive role of alternative promoters in context-specific isoform expression and
the importance of promoter choice over its level of transcriptional activity have been recently implied
based on pan-cancer in silico studies. We aimed to explore this phenomenon at the cellular level on the
example of a major tumor suppressor SMAD4 in search of molecular mechanisms in colorectal cancer
that could be exploited for novel biomarkers or therapeutic approaches.
Methods: Multi-omics technologies, in silico tools and in vitro functional assays were applied to analyze
the transcripts expression and the alternative promoters’ function of the SMAD4 gene in colon cell lines
HCEC-1CT, HCT116, DLD-1, SW480 and SW620.
Results: High expression of the transcript SMAD4-213 emerged as a hallmark of colon cancer cells,
while in silico tools point to its possible additional role and potential for sponging miRNAs. Based on the
observed dysregulation of SMAD4-209 and SMAD4-213 in malignant vs. non-malignant colon cells, we
propose that their expression ratio might be a solid biomarker candidate for colorectal cancer detection.
Conclusions: A differential pattern of the respective promoters’ activity was observed that corresponds
to the expression of transcripts, confirming the role of alternative promoters in context-specific isoform
expression. The investigated SMAD4 promoters and transcripts harbor translational potential that should
be further investigated.",
publisher = "Ivyspring International Publisher",
journal = "Journal of Cancer",
title = "Dysregulation of transcripts SMAD4-209 and SMAD4-213 and their respective promoters in colon cancer cell lines",
pages = "5131-5118",
number = "15",
volume = "15",
doi = "10.7150/jca.98911"
}

Babić, T., Ugrin, M., Jeremić, S., Kojić, M., Dinić, J., Banović Đeri, B., Zodiakis, J.,& Nikolić, A.. (2024). Dysregulation of transcripts SMAD4-209 and SMAD4-213 and their respective promoters in colon cancer cell lines. in Journal of Cancer
Ivyspring International Publisher., 15(15), 5118-5131.
https://doi.org/10.7150/jca.98911

Babić T, Ugrin M, Jeremić S, Kojić M, Dinić J, Banović Đeri B, Zodiakis J, Nikolić A. Dysregulation of transcripts SMAD4-209 and SMAD4-213 and their respective promoters in colon cancer cell lines. in Journal of Cancer. 2024;15(15):5118-5131.
doi:10.7150/jca.98911 .

Babić, Tamara, Ugrin, Milena, Jeremić, Sanja, Kojić, Milan, Dinić, Jelena, Banović Đeri, Bojana, Zodiakis, Jerome, Nikolić, Aleksandra, "Dysregulation of transcripts SMAD4-209 and SMAD4-213 and their respective promoters in colon cancer cell lines" in Journal of Cancer, 15, no. 15 (2024):5118-5131,
https://doi.org/10.7150/jca.98911 . .

TY  - JOUR
AU  - Popović, Nikola
AU  - Veljović, Katarina
AU  - Radojević, Dušan
AU  - Brdarić, Emilija
AU  - Stevanović, Dušan
AU  - Živković, Milica
AU  - Kojić, Milan
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/975
AB  - This study aimed to investigate the probiogenomic features of artisanal bacteriocin-producing Enterococcus faecium BGPAS1-3 and the use of the improved pMALc5HisEk expression vector for overexpressing class II bacteriocins and the application of purified bacteriocin 31 in a milk model as a preservative against L. monocytogenes. The BGPAS1-3 strain was isolated from traditional fresh soft cheese manufactured in households on a small scale in rural locations surrounding Pale Mountain City in Bosnia and Herzegovina. The whole-genome sequencing approach and bioinformatics analyses revealed that the strain BGPAS1-3 was non-pathogenic to humans. The presence of bacteriocin operons suggested the ability of the isolate to suppress the growth of pathogens. Coding regions for three maturated bacteriocins (bacteriocin 31, bacteriocin 32, and enterocin P) produced by BGPAS1-3 were amplified and expressed in Escherichia coli ER2523 using the pMALc5HisEk system. All three bacteriocins were successfully overexpressed and purified after enterokinase cleavage but showed different antimicrobial activity. Bacteriocin 31 showed significantly stronger antimicrobial activity compared with bacteriocin 32. It was the only one that proved to be suitable for use as a food preservative against L. monocytogenes in a milk model.
PB  - MDPI
T2  - Foods
T1  - Insight into the Probiogenomic Potential of Enterococcus faecium BGPAS1-3 and Application of a Potent Thermostable Bacteriocin
IS  - 16
SP  - 2637
VL  - 13
DO  - 10.3390/foods13162637
ER  - 

@article{
author = "Popović, Nikola and Veljović, Katarina and Radojević, Dušan and Brdarić, Emilija and Stevanović, Dušan and Živković, Milica and Kojić, Milan",
year = "2024",
abstract = "This study aimed to investigate the probiogenomic features of artisanal bacteriocin-producing Enterococcus faecium BGPAS1-3 and the use of the improved pMALc5HisEk expression vector for overexpressing class II bacteriocins and the application of purified bacteriocin 31 in a milk model as a preservative against L. monocytogenes. The BGPAS1-3 strain was isolated from traditional fresh soft cheese manufactured in households on a small scale in rural locations surrounding Pale Mountain City in Bosnia and Herzegovina. The whole-genome sequencing approach and bioinformatics analyses revealed that the strain BGPAS1-3 was non-pathogenic to humans. The presence of bacteriocin operons suggested the ability of the isolate to suppress the growth of pathogens. Coding regions for three maturated bacteriocins (bacteriocin 31, bacteriocin 32, and enterocin P) produced by BGPAS1-3 were amplified and expressed in Escherichia coli ER2523 using the pMALc5HisEk system. All three bacteriocins were successfully overexpressed and purified after enterokinase cleavage but showed different antimicrobial activity. Bacteriocin 31 showed significantly stronger antimicrobial activity compared with bacteriocin 32. It was the only one that proved to be suitable for use as a food preservative against L. monocytogenes in a milk model.",
publisher = "MDPI",
journal = "Foods",
title = "Insight into the Probiogenomic Potential of Enterococcus faecium BGPAS1-3 and Application of a Potent Thermostable Bacteriocin",
number = "16",
pages = "2637",
volume = "13",
doi = "10.3390/foods13162637"
}

Popović, N., Veljović, K., Radojević, D., Brdarić, E., Stevanović, D., Živković, M.,& Kojić, M.. (2024). Insight into the Probiogenomic Potential of Enterococcus faecium BGPAS1-3 and Application of a Potent Thermostable Bacteriocin. in Foods
MDPI., 13(16), 2637.
https://doi.org/10.3390/foods13162637

Popović N, Veljović K, Radojević D, Brdarić E, Stevanović D, Živković M, Kojić M. Insight into the Probiogenomic Potential of Enterococcus faecium BGPAS1-3 and Application of a Potent Thermostable Bacteriocin. in Foods. 2024;13(16):2637.
doi:10.3390/foods13162637 .

Popović, Nikola, Veljović, Katarina, Radojević, Dušan, Brdarić, Emilija, Stevanović, Dušan, Živković, Milica, Kojić, Milan, "Insight into the Probiogenomic Potential of Enterococcus faecium BGPAS1-3 and Application of a Potent Thermostable Bacteriocin" in Foods, 13, no. 16 (2024):2637,
https://doi.org/10.3390/foods13162637 . .

TY  - CONF
AU  - Bufan, Biljana
AU  - Marčetić, Mirjana
AU  - Đuretić, Jasmina
AU  - Ćuruvija, Ivana
AU  - Blagojević, Veljko
AU  - Božić, Dragana
AU  - Milutinović, Violeta
AU  - Sopta, Jelena
AU  - Kotur-Stevuljić, Jelena
AU  - Arsenović-Ranin, Nevena
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/979
AB  - Purpose: Traditional herbal plants attract atention as potential therapeutics in the treatment of arthritis. Teucrium
montanum L. (TM) is a widely distributed plant in almost all Mediterranean countries. The therapeutic potential of TM
extract is insufficiently examined, but its composition implies potential anti-inflammatory effect. We tested amelorating
effects of TM extract on rheumatoid arthritis (RA), in rat collagen-induced arthritis (CIA) model.
Methods: Phytochemical analysis of TM methanol extract was performed using LC-DAD-ESI-MS. Female Dark Agouti
rats were immunized with bovine type II collagen (CII) in incomplete Freund`s adjuvant to induce CIA. CIA rats were
treated with TM extract daily per os (100 or 200 mg/kg) from day when the first signs of disease started. Clinical
evaluation of CIA was performed by scoring and histological analysis. Phenotypic and functional characteristics of
splenocytes and draining lymph nodes (dLNs) cells were analyzed by flow cytometry. Cytokine levels in supernatants
from hind paw tissue culture, and IgG CII-specific antibodies level in blood were determined by ELISA.
Results: TM extract contained phenylethanoid glycosides, mainly verbascoside, phenolic acids, flavonoid and iridoid
glycosides. Applied TM extract doses significantly reduced arthritic score and severity of ankle joint inflammation in CIA
rats. This was accompanied by the more pro-oxidant profile in serum, and shifted pro-inflammatory (TNF-α and IL-6)
and anti-inflammatory (IL-10) cytokines balance toward later in paws of treated rats. This was in conjunction with less
inflammatory phenotype of dLN and spleen CD11b+ cells (monocytes/macrophages) judged by their lower expression of
CD86, MHCII, and TLR-4, and lower production of TNF-α and IL-1β. The frequency of TCR+ cells, and IL-17- and
IFN-γ- producing CD4+ cells were lower in dLNs and spleen, while frequency of regulatory CD4+CD25+FoxP3+ cells
was higher. The lower frequency of CD45RA (B cells) in dLNs and spleen was accompanied with lower levels of serum
anti-CII antibodies in treated rats.
Conclusion: These findings suggest, for the first time, that TM extract effectively ameliorated clinical presentation of RA
in model of rat CIA, and that this therapeutic effect may be associated with its immunoregulatory/anti-inflammatory
action.
PB  - Wiley
C3  - 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland
T1  - Teucrium montanum extract ameliorates collagen-induced arthritis in rats
EP  - 887
SP  - 887
UR  - https://hdl.handle.net/21.15107/rcub_intor_979
ER  - 

@conference{
author = "Bufan, Biljana and Marčetić, Mirjana and Đuretić, Jasmina and Ćuruvija, Ivana and Blagojević, Veljko and Božić, Dragana and Milutinović, Violeta and Sopta, Jelena and Kotur-Stevuljić, Jelena and Arsenović-Ranin, Nevena",
year = "2024",
abstract = "Purpose: Traditional herbal plants attract atention as potential therapeutics in the treatment of arthritis. Teucrium
montanum L. (TM) is a widely distributed plant in almost all Mediterranean countries. The therapeutic potential of TM
extract is insufficiently examined, but its composition implies potential anti-inflammatory effect. We tested amelorating
effects of TM extract on rheumatoid arthritis (RA), in rat collagen-induced arthritis (CIA) model.
Methods: Phytochemical analysis of TM methanol extract was performed using LC-DAD-ESI-MS. Female Dark Agouti
rats were immunized with bovine type II collagen (CII) in incomplete Freund`s adjuvant to induce CIA. CIA rats were
treated with TM extract daily per os (100 or 200 mg/kg) from day when the first signs of disease started. Clinical
evaluation of CIA was performed by scoring and histological analysis. Phenotypic and functional characteristics of
splenocytes and draining lymph nodes (dLNs) cells were analyzed by flow cytometry. Cytokine levels in supernatants
from hind paw tissue culture, and IgG CII-specific antibodies level in blood were determined by ELISA.
Results: TM extract contained phenylethanoid glycosides, mainly verbascoside, phenolic acids, flavonoid and iridoid
glycosides. Applied TM extract doses significantly reduced arthritic score and severity of ankle joint inflammation in CIA
rats. This was accompanied by the more pro-oxidant profile in serum, and shifted pro-inflammatory (TNF-α and IL-6)
and anti-inflammatory (IL-10) cytokines balance toward later in paws of treated rats. This was in conjunction with less
inflammatory phenotype of dLN and spleen CD11b+ cells (monocytes/macrophages) judged by their lower expression of
CD86, MHCII, and TLR-4, and lower production of TNF-α and IL-1β. The frequency of TCR+ cells, and IL-17- and
IFN-γ- producing CD4+ cells were lower in dLNs and spleen, while frequency of regulatory CD4+CD25+FoxP3+ cells
was higher. The lower frequency of CD45RA (B cells) in dLNs and spleen was accompanied with lower levels of serum
anti-CII antibodies in treated rats.
Conclusion: These findings suggest, for the first time, that TM extract effectively ameliorated clinical presentation of RA
in model of rat CIA, and that this therapeutic effect may be associated with its immunoregulatory/anti-inflammatory
action.",
publisher = "Wiley",
journal = "7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland",
title = "Teucrium montanum extract ameliorates collagen-induced arthritis in rats",
pages = "887-887",
url = "https://hdl.handle.net/21.15107/rcub_intor_979"
}

Bufan, B., Marčetić, M., Đuretić, J., Ćuruvija, I., Blagojević, V., Božić, D., Milutinović, V., Sopta, J., Kotur-Stevuljić, J.,& Arsenović-Ranin, N.. (2024). Teucrium montanum extract ameliorates collagen-induced arthritis in rats. in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland
Wiley., 887-887.
https://hdl.handle.net/21.15107/rcub_intor_979

Bufan B, Marčetić M, Đuretić J, Ćuruvija I, Blagojević V, Božić D, Milutinović V, Sopta J, Kotur-Stevuljić J, Arsenović-Ranin N. Teucrium montanum extract ameliorates collagen-induced arthritis in rats. in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland. 2024;:887-887.
https://hdl.handle.net/21.15107/rcub_intor_979 .

Bufan, Biljana, Marčetić, Mirjana, Đuretić, Jasmina, Ćuruvija, Ivana, Blagojević, Veljko, Božić, Dragana, Milutinović, Violeta, Sopta, Jelena, Kotur-Stevuljić, Jelena, Arsenović-Ranin, Nevena, "Teucrium montanum extract ameliorates collagen-induced arthritis in rats" in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland (2024):887-887,
https://hdl.handle.net/21.15107/rcub_intor_979 .

TY  - CONF
AU  - Blagojević, Veljko
AU  - Ćuruvija, Ivana
AU  - Anđelović, Ivana
AU  - Vasić, Marko
AU  - Miljković, Radmila
AU  - Prijić, Ivana
AU  - Bufan, Biljana
AU  - Đuretić, Jasmina
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/978
AB  - Memantine is an uncompetitive NMDA antagonist, blocking the ionic channels of NMDA receptors, influencing the Ca2+
metabolism. This drug is clinically used for treating Alzheimer’s disease, and there have been studies on animal models
covering a variety of disease models and tissues, including the heart, liver and brain. Results presented here are from a
peculiar observation made in the following study: female Dark Agouti rats of two ages (young – 3 months, and old – 24
months) were fed a memantine solution (60 mg/kg body weight/day for 7 consecutive days) through oral gavage from the
first day after experimental autoimmune encephalomyelitis was induced, and were sacrificed on day 8 (inductive phase
of disease). We confirmed the effects of systemic memantine application through histological analysis – livers of young
control group rats showed signs of steatosis, which was ameliorated in the treated group; rats from both old and young
control groups showed signs of liver mononuclear infiltration, which was absent in all rats from treated groups, regardless
of age. However, the curious unexpected result was of a technical nature – peritoneal macrophages isolated from the rats
showed substantially greater viability even 11 days after the experiment. Memantine had a stronger effect on peritoneal
cells of young rats – the peritoneal cell yield was two times lower in the memantine treated group, the percentage of
CD45RA+ peritoneal B-cells was substantially higher than in the control group, and 11 days post-experiment there were
3 times more live macrophages (CD11b+CD163+
). Macrophages are usually short living cells upon extraction from
tissues, and these results may point in a direction of an intrinsic mechanism for longevity of macrophages and potentially
other cells expressing the NMDAR. Since memantine is known to block extravasation of blood mononuclear cells into
peripheral tissues, it is possible that the dynamic of peritoneal cells replenishment has been disturbed. The implications
of our observations are unclear and further research will be need to establish the full scope of possibilities it unlocks,
however we feel confident that memantine could be the proverbial shamrock for macrophages, bringing them long and
happy lives.
PB  - Wiley
C3  - 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland
T1  - Long live the macrophages – memantine as cellular youth elixir?
EP  - 830
SP  - 830
UR  - https://hdl.handle.net/21.15107/rcub_intor_978
ER  - 

@conference{
author = "Blagojević, Veljko and Ćuruvija, Ivana and Anđelović, Ivana and Vasić, Marko and Miljković, Radmila and Prijić, Ivana and Bufan, Biljana and Đuretić, Jasmina",
year = "2024",
abstract = "Memantine is an uncompetitive NMDA antagonist, blocking the ionic channels of NMDA receptors, influencing the Ca2+
metabolism. This drug is clinically used for treating Alzheimer’s disease, and there have been studies on animal models
covering a variety of disease models and tissues, including the heart, liver and brain. Results presented here are from a
peculiar observation made in the following study: female Dark Agouti rats of two ages (young – 3 months, and old – 24
months) were fed a memantine solution (60 mg/kg body weight/day for 7 consecutive days) through oral gavage from the
first day after experimental autoimmune encephalomyelitis was induced, and were sacrificed on day 8 (inductive phase
of disease). We confirmed the effects of systemic memantine application through histological analysis – livers of young
control group rats showed signs of steatosis, which was ameliorated in the treated group; rats from both old and young
control groups showed signs of liver mononuclear infiltration, which was absent in all rats from treated groups, regardless
of age. However, the curious unexpected result was of a technical nature – peritoneal macrophages isolated from the rats
showed substantially greater viability even 11 days after the experiment. Memantine had a stronger effect on peritoneal
cells of young rats – the peritoneal cell yield was two times lower in the memantine treated group, the percentage of
CD45RA+ peritoneal B-cells was substantially higher than in the control group, and 11 days post-experiment there were
3 times more live macrophages (CD11b+CD163+
). Macrophages are usually short living cells upon extraction from
tissues, and these results may point in a direction of an intrinsic mechanism for longevity of macrophages and potentially
other cells expressing the NMDAR. Since memantine is known to block extravasation of blood mononuclear cells into
peripheral tissues, it is possible that the dynamic of peritoneal cells replenishment has been disturbed. The implications
of our observations are unclear and further research will be need to establish the full scope of possibilities it unlocks,
however we feel confident that memantine could be the proverbial shamrock for macrophages, bringing them long and
happy lives.",
publisher = "Wiley",
journal = "7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland",
title = "Long live the macrophages – memantine as cellular youth elixir?",
pages = "830-830",
url = "https://hdl.handle.net/21.15107/rcub_intor_978"
}

Blagojević, V., Ćuruvija, I., Anđelović, I., Vasić, M., Miljković, R., Prijić, I., Bufan, B.,& Đuretić, J.. (2024). Long live the macrophages – memantine as cellular youth elixir?. in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland
Wiley., 830-830.
https://hdl.handle.net/21.15107/rcub_intor_978

Blagojević V, Ćuruvija I, Anđelović I, Vasić M, Miljković R, Prijić I, Bufan B, Đuretić J. Long live the macrophages – memantine as cellular youth elixir?. in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland. 2024;:830-830.
https://hdl.handle.net/21.15107/rcub_intor_978 .

Blagojević, Veljko, Ćuruvija, Ivana, Anđelović, Ivana, Vasić, Marko, Miljković, Radmila, Prijić, Ivana, Bufan, Biljana, Đuretić, Jasmina, "Long live the macrophages – memantine as cellular youth elixir?" in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland (2024):830-830,
https://hdl.handle.net/21.15107/rcub_intor_978 .

TY  - CONF
AU  - Petrušić, Marija
AU  - Blagojević, Veljko
AU  - Anđelović, Ivana
AU  - Vasić, Marko
AU  - Dragačević, Luka
AU  - Živković, Irena
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/984
AB  - Purpose: The senescence of the immune system bears the burden of poor response to vaccination. The Influenza virus is
a common etiological factor that humans encounter every year. The elderly are at greater risk of undesirable outcomes,
even when vaccinated, since the vaccine response rate is 45-65%. To address the poor immunological response to
vaccination in the elderly, we sought a non-invasive and comfortable “adjuvant” in the form of probiotic
Lacticaseibacillus rhamnosus that could boost the immune response and affect the vaccination outcomes.
Methods: Young (3) and old (24 months) Dark Agouti female rats were set into four experimental groups (n=6). One
from each group of young and old rats received L.rhamnosus supplemented to the water (⁓1x109 CFU per day) 7 days
before the vaccination and during the next 3 weeks after it (L.rhamnosus+vaccinated). The control groups drank tap water
ad libitum (control+vaccinated). All 4 groups were vaccinated with the seasonal Influenza vaccine (inactivated,
fragmented virus). Three weeks after the vaccination, the experiment was ended and the thymuses and blood were
retrieved for flow cytometry, qPCR, and ELISA analysis.
Results: Our results indicated that supplementation with L.rhamnosus induced a significant increase (p<0.05) in the
frequency of Foxp3+CD25+CD4+TCRαβ+ cells in the blood of young, but not old vaccinated rats. Also, their thymuses
were significantly (p<0.05) larger compared to the control vaccinated group of the same age. Following this was the
significantly lower (p<0.05) frequency of memory CD90-CD45RC-CD4+ T cells in the blood of the young
L.rhamnosus+vaccinated group and CD28nullCD8+ T cells (p<0.05) in both young and old L.rhamnosus+vaccinated
groups compared to the control group. Interestingly, the concentration of proinflammatory IFNγ was significantly lower
(p<0.01) in sera retrieved from both young and old vaccinated rats that received L.rhamnosus supplementation.
Conclusion: The immunomodulatory effect of L.rhamnosus is complex and affects both primary and secondary lymphoid
systems. Our preliminary results suggest that L.rhamnosus supplementation: can be beneficial to vaccination outcomes
for both young and old but is guided by different mechanisms; it could potentially affect slower thymus involution and
take part in postponing the immunosenescence. Future experiments should provide us with additional answers.
PB  - Wiley
C3  - 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland
T1  - Can Lacticaseibacillus rhamnosus modulate the immune response to inactivated Influenza vaccine in young and old rats?
EP  - 1649
SP  - 1649
UR  - https://hdl.handle.net/21.15107/rcub_intor_984
ER  - 

@conference{
author = "Petrušić, Marija and Blagojević, Veljko and Anđelović, Ivana and Vasić, Marko and Dragačević, Luka and Živković, Irena",
year = "2024",
abstract = "Purpose: The senescence of the immune system bears the burden of poor response to vaccination. The Influenza virus is
a common etiological factor that humans encounter every year. The elderly are at greater risk of undesirable outcomes,
even when vaccinated, since the vaccine response rate is 45-65%. To address the poor immunological response to
vaccination in the elderly, we sought a non-invasive and comfortable “adjuvant” in the form of probiotic
Lacticaseibacillus rhamnosus that could boost the immune response and affect the vaccination outcomes.
Methods: Young (3) and old (24 months) Dark Agouti female rats were set into four experimental groups (n=6). One
from each group of young and old rats received L.rhamnosus supplemented to the water (⁓1x109 CFU per day) 7 days
before the vaccination and during the next 3 weeks after it (L.rhamnosus+vaccinated). The control groups drank tap water
ad libitum (control+vaccinated). All 4 groups were vaccinated with the seasonal Influenza vaccine (inactivated,
fragmented virus). Three weeks after the vaccination, the experiment was ended and the thymuses and blood were
retrieved for flow cytometry, qPCR, and ELISA analysis.
Results: Our results indicated that supplementation with L.rhamnosus induced a significant increase (p<0.05) in the
frequency of Foxp3+CD25+CD4+TCRαβ+ cells in the blood of young, but not old vaccinated rats. Also, their thymuses
were significantly (p<0.05) larger compared to the control vaccinated group of the same age. Following this was the
significantly lower (p<0.05) frequency of memory CD90-CD45RC-CD4+ T cells in the blood of the young
L.rhamnosus+vaccinated group and CD28nullCD8+ T cells (p<0.05) in both young and old L.rhamnosus+vaccinated
groups compared to the control group. Interestingly, the concentration of proinflammatory IFNγ was significantly lower
(p<0.01) in sera retrieved from both young and old vaccinated rats that received L.rhamnosus supplementation.
Conclusion: The immunomodulatory effect of L.rhamnosus is complex and affects both primary and secondary lymphoid
systems. Our preliminary results suggest that L.rhamnosus supplementation: can be beneficial to vaccination outcomes
for both young and old but is guided by different mechanisms; it could potentially affect slower thymus involution and
take part in postponing the immunosenescence. Future experiments should provide us with additional answers.",
publisher = "Wiley",
journal = "7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland",
title = "Can Lacticaseibacillus rhamnosus modulate the immune response to inactivated Influenza vaccine in young and old rats?",
pages = "1649-1649",
url = "https://hdl.handle.net/21.15107/rcub_intor_984"
}

Petrušić, M., Blagojević, V., Anđelović, I., Vasić, M., Dragačević, L.,& Živković, I.. (2024). Can Lacticaseibacillus rhamnosus modulate the immune response to inactivated Influenza vaccine in young and old rats?. in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland
Wiley., 1649-1649.
https://hdl.handle.net/21.15107/rcub_intor_984

Petrušić M, Blagojević V, Anđelović I, Vasić M, Dragačević L, Živković I. Can Lacticaseibacillus rhamnosus modulate the immune response to inactivated Influenza vaccine in young and old rats?. in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland. 2024;:1649-1649.
https://hdl.handle.net/21.15107/rcub_intor_984 .

Petrušić, Marija, Blagojević, Veljko, Anđelović, Ivana, Vasić, Marko, Dragačević, Luka, Živković, Irena, "Can Lacticaseibacillus rhamnosus modulate the immune response to inactivated Influenza vaccine in young and old rats?" in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland (2024):1649-1649,
https://hdl.handle.net/21.15107/rcub_intor_984 .

TY  - CONF
AU  - Bufan, Biljana
AU  - Arsenović-Ranin, Nevena
AU  - Živković, Irena
AU  - Blagojević, Veljko
AU  - Kotur-Stevuljević, Jelena
AU  - Leposavić, Gordana
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/983
AB  - Purpose: Although nutritional gaps are prevalent in several micronutrients reported to support immune function, the
significance of their deficits/supplementation for efficacy of vaccines, particularly those with low efficacy, as it is seasonal
influenza vaccine, has not been fully investigated, yet. The present study examined influence of supplementation
combining vitamins C and D, oligoelements zinc, selenium and manganese, and N-acetyl-cysteine on germinal centre
(GC) and serum IgG responses to seasonal quadrivalent influenza vaccine (QIV) in mice.
Methods: Study encompas female BALB/c mice that were given QIV in two doses (28 days
apart). The supplementation started five days before the first injection. Phenotypic and
functional characteristics of cells from secondary lymphoid organs (SLOs) (draining lymph
nodes and spleens) were analyzed by flow cytometry, whereas serum IgG response to QIV and
levels of cytokines from SLO cell cultures were determined by ELISA. Redox status
parameters were evaluated in spleens by spectrophotometric methods.
Results: The supplementation increased the magnitude of serum IgG response 28 days post the first QIV dose, through
stimulation of GC reaction in SLOs, as indicated by increase in the frequency of GC B cells and follicular CD4+ T helper
(Th) cells, and Th cell IL-21 production. Additionally, 14 days following the second QIV dose the supplementation
supported more favorable (viz. more effective in the context of virus infection clearance) IgG2a response through favoring
Th1 response. This could be ascribed not only to its indirect action related to antioxidant properties (confirmed by redox
status analyses), but also to direct action on Th cell differentiation, as indicating by the ratio in production levels of Th1
(INF-γ) /Th2 (IL-4) signature cytokine ratio upon QIV restimulation in SLO cell cultures.
Conclusion: Thus, the study forms solid base for further studies aimed at repurposing use this safe and inexpensive
micronutrient preparation as an adjuvant for virus influenza vaccine and possible some other virus vaccines
PB  - Wiley
C3  - 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland
T1  - Micronutrient supplementation supports immune response to seasonal influenza vaccine in mice
EP  - 1645
SP  - 1645
UR  - https://hdl.handle.net/21.15107/rcub_intor_983
ER  - 

@conference{
author = "Bufan, Biljana and Arsenović-Ranin, Nevena and Živković, Irena and Blagojević, Veljko and Kotur-Stevuljević, Jelena and Leposavić, Gordana",
year = "2024",
abstract = "Purpose: Although nutritional gaps are prevalent in several micronutrients reported to support immune function, the
significance of their deficits/supplementation for efficacy of vaccines, particularly those with low efficacy, as it is seasonal
influenza vaccine, has not been fully investigated, yet. The present study examined influence of supplementation
combining vitamins C and D, oligoelements zinc, selenium and manganese, and N-acetyl-cysteine on germinal centre
(GC) and serum IgG responses to seasonal quadrivalent influenza vaccine (QIV) in mice.
Methods: Study encompas female BALB/c mice that were given QIV in two doses (28 days
apart). The supplementation started five days before the first injection. Phenotypic and
functional characteristics of cells from secondary lymphoid organs (SLOs) (draining lymph
nodes and spleens) were analyzed by flow cytometry, whereas serum IgG response to QIV and
levels of cytokines from SLO cell cultures were determined by ELISA. Redox status
parameters were evaluated in spleens by spectrophotometric methods.
Results: The supplementation increased the magnitude of serum IgG response 28 days post the first QIV dose, through
stimulation of GC reaction in SLOs, as indicated by increase in the frequency of GC B cells and follicular CD4+ T helper
(Th) cells, and Th cell IL-21 production. Additionally, 14 days following the second QIV dose the supplementation
supported more favorable (viz. more effective in the context of virus infection clearance) IgG2a response through favoring
Th1 response. This could be ascribed not only to its indirect action related to antioxidant properties (confirmed by redox
status analyses), but also to direct action on Th cell differentiation, as indicating by the ratio in production levels of Th1
(INF-γ) /Th2 (IL-4) signature cytokine ratio upon QIV restimulation in SLO cell cultures.
Conclusion: Thus, the study forms solid base for further studies aimed at repurposing use this safe and inexpensive
micronutrient preparation as an adjuvant for virus influenza vaccine and possible some other virus vaccines",
publisher = "Wiley",
journal = "7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland",
title = "Micronutrient supplementation supports immune response to seasonal influenza vaccine in mice",
pages = "1645-1645",
url = "https://hdl.handle.net/21.15107/rcub_intor_983"
}

Bufan, B., Arsenović-Ranin, N., Živković, I., Blagojević, V., Kotur-Stevuljević, J.,& Leposavić, G.. (2024). Micronutrient supplementation supports immune response to seasonal influenza vaccine in mice. in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland
Wiley., 1645-1645.
https://hdl.handle.net/21.15107/rcub_intor_983

Bufan B, Arsenović-Ranin N, Živković I, Blagojević V, Kotur-Stevuljević J, Leposavić G. Micronutrient supplementation supports immune response to seasonal influenza vaccine in mice. in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland. 2024;:1645-1645.
https://hdl.handle.net/21.15107/rcub_intor_983 .

Bufan, Biljana, Arsenović-Ranin, Nevena, Živković, Irena, Blagojević, Veljko, Kotur-Stevuljević, Jelena, Leposavić, Gordana, "Micronutrient supplementation supports immune response to seasonal influenza vaccine in mice" in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland (2024):1645-1645,
https://hdl.handle.net/21.15107/rcub_intor_983 .

TY  - CONF
AU  - Protić-Rosić, Isidora
AU  - Kratzer, Bernhard
AU  - Lopandić, Zorana
AU  - Blagojević, Gordan
AU  - Gavrović-Jankulović, Marija
AU  - Pickl, Winfried
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/982
AB  - Employing effective adjuvants and hypoallergenic isoforms is essential for improving the efficacy and safety of allergenspecific immunotherapy (AIT). Banana lectin (BanLec) exhibits immunomodulatory potential, and mutation of histidine
at position 84 to threonine (BanLecH84T) reduced its mitogenicity. Our study aims to assess the immunomodulatory
potential on antigen-presenting cells (APCs) of recombinant chimeras combining the major birch pollen allergen Bet v
1a and its hypoallergenic isoform Bet v 1l with BanLec and BanLecH84T. After the production in Escherichia coli and
purification, chimeras were co-incubated with peripheral blood mononuclear cells (PBMC) from birch pollen-allergic
patients. Subsequently, secreted cytokines were analyzed. Antigen uptake was evaluated by co-incubating the treated
monocyte-derived dendritic cells (MoDC) or HLA-DR7+ EBV-transformed B cells (EBV-BCL) with T cell receptor
(TCR) transgenic (tg) Jurkat reporter cells. The tg TCR recognizes the immunodominant epitope of the Bet v 1, Bet v
1142-153 in the context of HLA-DR7 (Jurkattg). MoDC were analyzed for the upregulation of activation marker expression
(CD80, CD86, HLA-DR) and secreted cytokines (IL-6, IL-8, IL-10, IL-12p40, IL-23, TNF- α, IFN-γ). Furthermore,
chimera’s uptake by APCs was modulated by incubation with different uptake pathway inhibitors/enhancers during
antigen loading, and co-culture with Jurkattg. We found that the co-incubation of PBMC with Bet v 1a-BanLec prompted
the secretion of the anti-inflammatory cytokine IL-10 and augmented the IFN-γ/IL-4 ratio. Co-incubation of chimeras
with EBV-BCL or MoDC revealed that all chimeras strongly activated the allergen-specific Jurkattg. Additionally,
activation marker expression and secreted cytokines of MoDC co-incubated with the respective chimeras indicated that
they led to the activation of these cells. Supplementation of media with low concentrations of glucose stimulated antigen
uptake and subsequent Jurkattg activation. In contrast, antigen uptake was inhibited by sucrose, suggesting interference
with the macropinocytotic uptake pathway. Our results highlight the potential of chimeras to modulate allergen-specific
immune responses, enhancing anti-inflammatory cytokine secretion and promoting allergen-specific T-cell activation
thereby demonstrating their potential in AIT.
PB  - Wiley
C3  - 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland
T1  - Immunomodulatory impact of the BanLec-Bet v 1 chimera and its variants on antigen-presenting cells
EP  - 1549
SP  - 1549
UR  - https://hdl.handle.net/21.15107/rcub_intor_982
ER  - 

@conference{
author = "Protić-Rosić, Isidora and Kratzer, Bernhard and Lopandić, Zorana and Blagojević, Gordan and Gavrović-Jankulović, Marija and Pickl, Winfried",
year = "2024",
abstract = "Employing effective adjuvants and hypoallergenic isoforms is essential for improving the efficacy and safety of allergenspecific immunotherapy (AIT). Banana lectin (BanLec) exhibits immunomodulatory potential, and mutation of histidine
at position 84 to threonine (BanLecH84T) reduced its mitogenicity. Our study aims to assess the immunomodulatory
potential on antigen-presenting cells (APCs) of recombinant chimeras combining the major birch pollen allergen Bet v
1a and its hypoallergenic isoform Bet v 1l with BanLec and BanLecH84T. After the production in Escherichia coli and
purification, chimeras were co-incubated with peripheral blood mononuclear cells (PBMC) from birch pollen-allergic
patients. Subsequently, secreted cytokines were analyzed. Antigen uptake was evaluated by co-incubating the treated
monocyte-derived dendritic cells (MoDC) or HLA-DR7+ EBV-transformed B cells (EBV-BCL) with T cell receptor
(TCR) transgenic (tg) Jurkat reporter cells. The tg TCR recognizes the immunodominant epitope of the Bet v 1, Bet v
1142-153 in the context of HLA-DR7 (Jurkattg). MoDC were analyzed for the upregulation of activation marker expression
(CD80, CD86, HLA-DR) and secreted cytokines (IL-6, IL-8, IL-10, IL-12p40, IL-23, TNF- α, IFN-γ). Furthermore,
chimera’s uptake by APCs was modulated by incubation with different uptake pathway inhibitors/enhancers during
antigen loading, and co-culture with Jurkattg. We found that the co-incubation of PBMC with Bet v 1a-BanLec prompted
the secretion of the anti-inflammatory cytokine IL-10 and augmented the IFN-γ/IL-4 ratio. Co-incubation of chimeras
with EBV-BCL or MoDC revealed that all chimeras strongly activated the allergen-specific Jurkattg. Additionally,
activation marker expression and secreted cytokines of MoDC co-incubated with the respective chimeras indicated that
they led to the activation of these cells. Supplementation of media with low concentrations of glucose stimulated antigen
uptake and subsequent Jurkattg activation. In contrast, antigen uptake was inhibited by sucrose, suggesting interference
with the macropinocytotic uptake pathway. Our results highlight the potential of chimeras to modulate allergen-specific
immune responses, enhancing anti-inflammatory cytokine secretion and promoting allergen-specific T-cell activation
thereby demonstrating their potential in AIT.",
publisher = "Wiley",
journal = "7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland",
title = "Immunomodulatory impact of the BanLec-Bet v 1 chimera and its variants on antigen-presenting cells",
pages = "1549-1549",
url = "https://hdl.handle.net/21.15107/rcub_intor_982"
}

Protić-Rosić, I., Kratzer, B., Lopandić, Z., Blagojević, G., Gavrović-Jankulović, M.,& Pickl, W.. (2024). Immunomodulatory impact of the BanLec-Bet v 1 chimera and its variants on antigen-presenting cells. in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland
Wiley., 1549-1549.
https://hdl.handle.net/21.15107/rcub_intor_982

Protić-Rosić I, Kratzer B, Lopandić Z, Blagojević G, Gavrović-Jankulović M, Pickl W. Immunomodulatory impact of the BanLec-Bet v 1 chimera and its variants on antigen-presenting cells. in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland. 2024;:1549-1549.
https://hdl.handle.net/21.15107/rcub_intor_982 .

Protić-Rosić, Isidora, Kratzer, Bernhard, Lopandić, Zorana, Blagojević, Gordan, Gavrović-Jankulović, Marija, Pickl, Winfried, "Immunomodulatory impact of the BanLec-Bet v 1 chimera and its variants on antigen-presenting cells" in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland (2024):1549-1549,
https://hdl.handle.net/21.15107/rcub_intor_982 .

TY  - CONF
AU  - Anđelović, Ivana
AU  - Miljković, Radmila
AU  - Ćuruvija, Ivana
AU  - Vasić, Marko
AU  - Petrušić, Marija
AU  - Živković, Irena
AU  - Blagojević, Veljko
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/980
AB  - Trained immunity (TI) has been a hot button issue in current immunology, and we decided to look at the understudied
effect of TI on chemically induced colitis. We used BALB/c male mice, which are more susceptible to TNBS-induced
colitis, and immunized them subcutaneously with BCG (TI-TNBS group) or saline (control-TNBS group). Seven days
later, we induced colitis by intrarectal injection of TNBS in 50% ethanol. We observed the mice over a period of another
week. Most mice in both groups did not survive, but the mice in the control-TNBS group lived significantly longer. We
investigated the mice who survived in an additional study. We immunized the mice with another dose of BCG and paired
them up with adequate controls (mice who didn’t have colitis induced, immunized with BCG or with saline). Prior to the
second immunization we isolated the PBMCs from a blood sample and analyzed them on a flow cytometer. Two days
later mice were euthanized and their bone marrow, peritoneal, and PBMCs were collected and analyzed on a flow
cytometer. The results showed BCG immunization causes an increase in both CD45R+CD19+ B cells and CD4+ T cells in
the circulation, however after colitis induction, the frequency of these cells is reduced in the circulation of the immunized
mice compared to controls. Given that the immunized animals had a shorter lifespan after colitis induction, it is possible
that the infiltration of these lymphocytes from the peripheral circulation could be a causal factor in aggravated
inflammation. In the peritoneal cavity, BCG immunized animals had a greater percentage of F4/80+CD68+ macrophages,
however after colitis, this ratio was reversed, which could indicate a faster depletion of resident macrophages of the
peritoneal cavity, which are known to have a reparative role in visceral tissue damage. Post colitis, a greater percentage
of peritoneal macrophages expressed iNOS, and the BCG vaccination did increase it slightly in the two days after
immunization, compared to the control group. It was shown that TI could aggravate chemically induced colitis in mice,
and our results seem to support this notion, which we will research further.
PB  - Wiley
C3  - 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland
T1  - To BCG or not to BCG – a case against trained immunity?
EP  - 918
SP  - 918
UR  - https://hdl.handle.net/21.15107/rcub_intor_980
ER  - 

@conference{
author = "Anđelović, Ivana and Miljković, Radmila and Ćuruvija, Ivana and Vasić, Marko and Petrušić, Marija and Živković, Irena and Blagojević, Veljko",
year = "2024",
abstract = "Trained immunity (TI) has been a hot button issue in current immunology, and we decided to look at the understudied
effect of TI on chemically induced colitis. We used BALB/c male mice, which are more susceptible to TNBS-induced
colitis, and immunized them subcutaneously with BCG (TI-TNBS group) or saline (control-TNBS group). Seven days
later, we induced colitis by intrarectal injection of TNBS in 50% ethanol. We observed the mice over a period of another
week. Most mice in both groups did not survive, but the mice in the control-TNBS group lived significantly longer. We
investigated the mice who survived in an additional study. We immunized the mice with another dose of BCG and paired
them up with adequate controls (mice who didn’t have colitis induced, immunized with BCG or with saline). Prior to the
second immunization we isolated the PBMCs from a blood sample and analyzed them on a flow cytometer. Two days
later mice were euthanized and their bone marrow, peritoneal, and PBMCs were collected and analyzed on a flow
cytometer. The results showed BCG immunization causes an increase in both CD45R+CD19+ B cells and CD4+ T cells in
the circulation, however after colitis induction, the frequency of these cells is reduced in the circulation of the immunized
mice compared to controls. Given that the immunized animals had a shorter lifespan after colitis induction, it is possible
that the infiltration of these lymphocytes from the peripheral circulation could be a causal factor in aggravated
inflammation. In the peritoneal cavity, BCG immunized animals had a greater percentage of F4/80+CD68+ macrophages,
however after colitis, this ratio was reversed, which could indicate a faster depletion of resident macrophages of the
peritoneal cavity, which are known to have a reparative role in visceral tissue damage. Post colitis, a greater percentage
of peritoneal macrophages expressed iNOS, and the BCG vaccination did increase it slightly in the two days after
immunization, compared to the control group. It was shown that TI could aggravate chemically induced colitis in mice,
and our results seem to support this notion, which we will research further.",
publisher = "Wiley",
journal = "7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland",
title = "To BCG or not to BCG – a case against trained immunity?",
pages = "918-918",
url = "https://hdl.handle.net/21.15107/rcub_intor_980"
}

Anđelović, I., Miljković, R., Ćuruvija, I., Vasić, M., Petrušić, M., Živković, I.,& Blagojević, V.. (2024). To BCG or not to BCG – a case against trained immunity?. in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland
Wiley., 918-918.
https://hdl.handle.net/21.15107/rcub_intor_980

Anđelović I, Miljković R, Ćuruvija I, Vasić M, Petrušić M, Živković I, Blagojević V. To BCG or not to BCG – a case against trained immunity?. in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland. 2024;:918-918.
https://hdl.handle.net/21.15107/rcub_intor_980 .

Anđelović, Ivana, Miljković, Radmila, Ćuruvija, Ivana, Vasić, Marko, Petrušić, Marija, Živković, Irena, Blagojević, Veljko, "To BCG or not to BCG – a case against trained immunity?" in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland (2024):918-918,
https://hdl.handle.net/21.15107/rcub_intor_980 .

TY  - CONF
AU  - Đuretić, Jasmina
AU  - Ćuruvija, Ivana
AU  - Blagojević, Veljko
AU  - Prijić, Ivana
AU  - Bufan, Biljana
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/981
AB  - Purpose: Life expectancy is only slightly affected by multiple sclerosis (MS), and it is a disease that often lasts for several
decades. The proportion of patients with late-onset MS has increased significantly and the mean age at diagnosis has
shifted towards older age. Older age at disease onset is associated with a shorter time to disability. Immune cells, such as
macrophages, microglia and lymphocytes, express N-methyl-D-aspartate receptors (NMDARs). Our aim was to elucidate
the effects of ageing on the role of NMDARs in the pathogenesis of experimental autoimmune encephalomyelitis (EAE).
Methods: Young adult (3-month-old) and aged (24-month-old) female Dark Agouti rats were used in this study.
Memantine, a non-competitive NMDAR antagonist, was administered by oral gavage for 7 consecutive days from the
first day post-immunization (dpi) in the first set of experiments or from the 7th dpi in the second set of experiments.
Mononuclear cells were isolated from the spinal cord or draining lymph nodes and analysed by flow cytometry. Spinal
cord tissue was collected for RT-qPCR.
Results: Administration of an NMDAR antagonist during the induction phase of EAE diminished the proportion of Th1
and Th17 cells and increased the proportion of IL-10+
regulatory T cells in the lymph nodes draining the site of
immunization in aged rats. Memantine increased the proportion of CD163+
and IL-10+
cells within CD11b+
cells in the
draining lymph nodes of aged rats, whereas these effects were absent in young rats. Treatment with memantine from the
7
th dpi resulted in a shift of microglia towards the anti-inflammatory M2 phenotype, characterized by an increase in the
expression of CD163 and a decrease in the expression of MHCII molecules, with an increase in the proportion of IL10+
microglia and the expression level of arginase 1 in the spinal cord of aged rats at the peak of the disease.
Conclusion: NMDARs contribute significantly to the pathogenesis of EAE in aged rats in both the induction and effector
phases. Our results suggest that targeting NMDARs in elderly MS patients may help to tailor treatment to the elderly MS
population.
PB  - Wiley
C3  - 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland
T1  - The beneficial effects of N-methyl-D-aspartate receptor antagonism on experimental autoimmune encephalomyelitis in aged rats
EP  - 1385
SP  - 1385
UR  - https://hdl.handle.net/21.15107/rcub_intor_981
ER  - 

@conference{
author = "Đuretić, Jasmina and Ćuruvija, Ivana and Blagojević, Veljko and Prijić, Ivana and Bufan, Biljana",
year = "2024",
abstract = "Purpose: Life expectancy is only slightly affected by multiple sclerosis (MS), and it is a disease that often lasts for several
decades. The proportion of patients with late-onset MS has increased significantly and the mean age at diagnosis has
shifted towards older age. Older age at disease onset is associated with a shorter time to disability. Immune cells, such as
macrophages, microglia and lymphocytes, express N-methyl-D-aspartate receptors (NMDARs). Our aim was to elucidate
the effects of ageing on the role of NMDARs in the pathogenesis of experimental autoimmune encephalomyelitis (EAE).
Methods: Young adult (3-month-old) and aged (24-month-old) female Dark Agouti rats were used in this study.
Memantine, a non-competitive NMDAR antagonist, was administered by oral gavage for 7 consecutive days from the
first day post-immunization (dpi) in the first set of experiments or from the 7th dpi in the second set of experiments.
Mononuclear cells were isolated from the spinal cord or draining lymph nodes and analysed by flow cytometry. Spinal
cord tissue was collected for RT-qPCR.
Results: Administration of an NMDAR antagonist during the induction phase of EAE diminished the proportion of Th1
and Th17 cells and increased the proportion of IL-10+
regulatory T cells in the lymph nodes draining the site of
immunization in aged rats. Memantine increased the proportion of CD163+
and IL-10+
cells within CD11b+
cells in the
draining lymph nodes of aged rats, whereas these effects were absent in young rats. Treatment with memantine from the
7
th dpi resulted in a shift of microglia towards the anti-inflammatory M2 phenotype, characterized by an increase in the
expression of CD163 and a decrease in the expression of MHCII molecules, with an increase in the proportion of IL10+
microglia and the expression level of arginase 1 in the spinal cord of aged rats at the peak of the disease.
Conclusion: NMDARs contribute significantly to the pathogenesis of EAE in aged rats in both the induction and effector
phases. Our results suggest that targeting NMDARs in elderly MS patients may help to tailor treatment to the elderly MS
population.",
publisher = "Wiley",
journal = "7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland",
title = "The beneficial effects of N-methyl-D-aspartate receptor antagonism on experimental autoimmune encephalomyelitis in aged rats",
pages = "1385-1385",
url = "https://hdl.handle.net/21.15107/rcub_intor_981"
}

Đuretić, J., Ćuruvija, I., Blagojević, V., Prijić, I.,& Bufan, B.. (2024). The beneficial effects of N-methyl-D-aspartate receptor antagonism on experimental autoimmune encephalomyelitis in aged rats. in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland
Wiley., 1385-1385.
https://hdl.handle.net/21.15107/rcub_intor_981

Đuretić J, Ćuruvija I, Blagojević V, Prijić I, Bufan B. The beneficial effects of N-methyl-D-aspartate receptor antagonism on experimental autoimmune encephalomyelitis in aged rats. in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland. 2024;:1385-1385.
https://hdl.handle.net/21.15107/rcub_intor_981 .

Đuretić, Jasmina, Ćuruvija, Ivana, Blagojević, Veljko, Prijić, Ivana, Bufan, Biljana, "The beneficial effects of N-methyl-D-aspartate receptor antagonism on experimental autoimmune encephalomyelitis in aged rats" in 7 th European Conference of Immunology 1-4 September, 2024 Dublin, Ireland (2024):1385-1385,
https://hdl.handle.net/21.15107/rcub_intor_981 .

TY  - CONF
AU  - Lukić, Ivana
AU  - Popović, Mina
AU  - Miljković, Radmila
AU  - Tsibulskaya, Darya
AU  - Panić, Marko
AU  - Dragačević, Luka
AU  - Stojanović, Marijana
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/874
AB  - Limosilactobacillus reuteri demonstrates a significant
role in treating gastrointestinal diseases
through the synthesis of various health-promoting
factors. These include mucus-binding proteins,
reactive oxygen species-scavenging enzymes, antimicrobial
agents (reuterin is capable of inhibiting
the growth of a wide spectrum of microorganisms),
vitamins (folate and vitamin B12), and unique
exopolysaccharides. Different strains of L. reuteri
exhibit strain-specific anti-inflammatory effects,
influencing the expression of immune-related
factors such as IL-10 and TNF-α (PMID: 20798357;
PMID: 22207578). Furthermore, the mitigating
impact of L. reuteri strains on inflammation is confirmed
in vivo and in vitro with the implication of
an interaction between probiotics and immune
cells in the intestinal mucosa (PMID: 22207578).
Our study aimed to investigate the potential anti-
inflammatory effects of daily treatment with autochthonous
probiotic strain L. reuteri B2 (PMID:
33932415) could have an anti-inflammatory effect
on local immune response. In a 14-day experiment
with Intor Swiss: Albino mice (n=10), those treated
with L. reuteri B2 (5x106 CFU/mL, 100 μl) showed a
favorable impact on the gut’s inflammatory environment.
Histological analyses of colon samples
and intraperitoneal macrophages revealed lower
myeloperoxidase (MPO) activity, reduced production
of superoxide ions, IFNγ, IL-6, and TNFα, along
with an enhanced production of IL-10 in L. reuteri
B2 treated mice compared to untreated ones. Notably,
histopathological preparations did not show
significant differences between the groups. The
study suggests that L. reuteri B2 may be valuable
for further evaluation in managing, preventing,
and treating inflammatory bowel diseases. The
presented findings contribute to understanding
the specific anti-inflammatory effects of this strain
on the local immune response, supporting its potential
as a therapeutic agent.
PB  - Serbian Society for Microbiology
C3  - XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april
T1  - The anti-inflammatory effect of Limosilactobacillus Reuteri b2 administration
EP  - 38
SP  - 38
UR  - https://hdl.handle.net/21.15107/rcub_intor_874
ER  - 

@conference{
author = "Lukić, Ivana and Popović, Mina and Miljković, Radmila and Tsibulskaya, Darya and Panić, Marko and Dragačević, Luka and Stojanović, Marijana",
year = "2024",
abstract = "Limosilactobacillus reuteri demonstrates a significant
role in treating gastrointestinal diseases
through the synthesis of various health-promoting
factors. These include mucus-binding proteins,
reactive oxygen species-scavenging enzymes, antimicrobial
agents (reuterin is capable of inhibiting
the growth of a wide spectrum of microorganisms),
vitamins (folate and vitamin B12), and unique
exopolysaccharides. Different strains of L. reuteri
exhibit strain-specific anti-inflammatory effects,
influencing the expression of immune-related
factors such as IL-10 and TNF-α (PMID: 20798357;
PMID: 22207578). Furthermore, the mitigating
impact of L. reuteri strains on inflammation is confirmed
in vivo and in vitro with the implication of
an interaction between probiotics and immune
cells in the intestinal mucosa (PMID: 22207578).
Our study aimed to investigate the potential anti-
inflammatory effects of daily treatment with autochthonous
probiotic strain L. reuteri B2 (PMID:
33932415) could have an anti-inflammatory effect
on local immune response. In a 14-day experiment
with Intor Swiss: Albino mice (n=10), those treated
with L. reuteri B2 (5x106 CFU/mL, 100 μl) showed a
favorable impact on the gut’s inflammatory environment.
Histological analyses of colon samples
and intraperitoneal macrophages revealed lower
myeloperoxidase (MPO) activity, reduced production
of superoxide ions, IFNγ, IL-6, and TNFα, along
with an enhanced production of IL-10 in L. reuteri
B2 treated mice compared to untreated ones. Notably,
histopathological preparations did not show
significant differences between the groups. The
study suggests that L. reuteri B2 may be valuable
for further evaluation in managing, preventing,
and treating inflammatory bowel diseases. The
presented findings contribute to understanding
the specific anti-inflammatory effects of this strain
on the local immune response, supporting its potential
as a therapeutic agent.",
publisher = "Serbian Society for Microbiology",
journal = "XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april",
title = "The anti-inflammatory effect of Limosilactobacillus Reuteri b2 administration",
pages = "38-38",
url = "https://hdl.handle.net/21.15107/rcub_intor_874"
}

Lukić, I., Popović, M., Miljković, R., Tsibulskaya, D., Panić, M., Dragačević, L.,& Stojanović, M.. (2024). The anti-inflammatory effect of Limosilactobacillus Reuteri b2 administration. in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april
Serbian Society for Microbiology., 38-38.
https://hdl.handle.net/21.15107/rcub_intor_874

Lukić I, Popović M, Miljković R, Tsibulskaya D, Panić M, Dragačević L, Stojanović M. The anti-inflammatory effect of Limosilactobacillus Reuteri b2 administration. in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april. 2024;:38-38.
https://hdl.handle.net/21.15107/rcub_intor_874 .

Lukić, Ivana, Popović, Mina, Miljković, Radmila, Tsibulskaya, Darya, Panić, Marko, Dragačević, Luka, Stojanović, Marijana, "The anti-inflammatory effect of Limosilactobacillus Reuteri b2 administration" in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april (2024):38-38,
https://hdl.handle.net/21.15107/rcub_intor_874 .